SW8461B
METHOD STATUS TABLE
SW-846, THIRD EDITION, UPDATES I, II, AND HA
September 1994
• Use this table as a reference guide to identify the
promulgation status of SW-846 methods.
• The methods in this table are listed sequentially by
number.
• This table should not be used as a Table of Contents for
SW-846. Refer to the Table of Contents found in Final
Update II (dated September 1994) for the order in which
the methods appear in SW-846.
-------
SW-846 METHOD STATUS TABLE
September 1994
METH NO.
THIRD EO
DATED
9/86
0010
0020
0030
1010
1020
1110
1310
"
METH NO.
FINAL
UPDATE I
DATED
7/92
~ ™*
~ ~
— •*
— •"
1020A
— w
1310A
1311
~ "*
METH NO.
FINAL
UPDT. II
DATED
9/94
" ""
•w _
"• ""
~ ""
— ~
~ ~
"
1312
METHOD TITLE
Modified Method 5
Sampling Train
Source Assessment
Sampling System
(SASS)
Volatile Organic
Sampling Train
Pensky-Martens
Closed-Cup Method
for Determining
Igm'tability
Setaflash Closed-Cup
Method for
Determining
Ignitability
Corrosivity Toward
Steel
Extraction Procedure
(EP) Toxicity Test
Method and
Structural Integrity
Test
Toxicity
Characteristic
Leaching Procedure
Synthetic
Precipitation
Leaching Procedure
SH-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol II
Chap 10
Vol II
Chap 10
Vol II
Chap 10
Vol 1C
Chap 8
Sec 8.1
Vol 1C
Chap 8
Sec 8.1
Vol 1C
Chap 8
Sec 8.2
Vol 1C
Chap 8
Sec 8.4
Vol 1C
Chap 8
Sec 8.4
Vol 1C
Chap 6
CURRENT
PROMUL-
GATED
METHOD
0010
Rev 0
9/86
0020
Rev 0
9/86
0030
Rev 0
9/86
1010
Rev 0
9/86
1020A
Rev 1
7/92
1110
Rev 0
9/86
1310A
Rev 1
7/92
1311
Rev 0
7/92
1312
Rev 0
9/94
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
KETH NO.
THIRD ED
DATED
9/86
1320
1330
3005
3010
~ ~
3020
3040
3050
METH NO.
FINAL
UPDATE I
DATED
7/92
"
1330A
3005A
3010A
— —
3020A
"
3050A
METH NO.
FINAL
UPDT. II
DATED
9/94
~ —
~ —
3015
"* ~
"
METHOD TITLE
Multiple Extraction
Procedure
Extraction Procedure
for Oily Wastes
Acid Digestion of
Waters for Total
Recoverable or
Dissolved Metals for
Analysis by FLAA or
ICP Spectroscopy
Acid Digestion of
Aqueous Samples and
Extracts for Total
Metals for Analysis
by FLAA or ICP
Spectroscopy
Microwave Assisted
Acid Digestion of
Aqueous Samples and
Extracts
Acid Digestion of
Aqueous Samples and
Extracts for Total
Metals for Analysis
by GFAA Spectroscopy
Dissolution
Procedure for Oils,
Greases, or Waxes
Acid Digestion of
Sediments, Sludges,
and Soils
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol 1C
Chap 6
Vol 1C
Chap 6
Vol IA
Chap 3
Sec 3.2
Vol IA
Chap 3
Sec 3.2
Vol IA
Chap 3
Sec 3.2
Vol IA
Chap 3
Sec 3.2
Vol IA
Chap 3
Sec 3.2
Vol IA
Chap 3
Sec 3.2
CURRENT
PROMUL-
GATED
METHOD
1320
Rev 0
9/86
1330A
Rev 1
7/92
3005A
Rev 1
7/92
3010A
Rev 1
7/92
3015
Rev 0
9/94
3020A
Rev 1
7/92
3040
Rev 0
9/86
3050A
Rev 1
7/92
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
METH NO.
THIRD ED
DATED
9/86
~ ~
3500
3510
3520
3540
~ —
3550
3580
3600
HETH NO.
FINAL
UPDATE I
DATED
7/92
*" "~
3500A
3510A
3520A
3540A
_ -
_ _
3580A
3600A
METH NO.
FINAL
UPDT. II
DATED
9/94
3051
— ~
3510B
3520B
3540B
3541
3550A
~ ~
3600B
METHOD TITLE
Microwave Assisted
Acid Digestion of
Sediments, Sludges,
Soils, and Oils
Organic Extraction
and Sample
Preparation
Separatory Funnel
Liquid-Liquid
Extraction
Continuous Liquid-
Liquid Extraction
Soxhlet Extraction
Automated Soxhlet
Extraction
Ultrasonic Extrac-
tion
Waste Dilution
Cleanup
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IA
Chap 3
Sec 3.2
Vol IB
Chap 4
Sec
4.2.1
Vol IB
Chap 4
Sec
4.2.1
Vol IB
Chap 4
Sec
4.2.1
Vol IB
Chap 4
Sec
4.2.1
Vol IB
Chap 4
Sec
4.2.1
Vol IB
Chap 4
Sec
4.2.1
Vol IB
Chap 4
Sec
4.2.1
Vol IB
Chap 4
Sec
4.2.2
CURRENT
PROMUL-
GATED
METHOD
3051
Rev 0
9/94
3500A
Rev 1
7/92
3510B
Rev 2
9/94
3520B
Rev 2
9/94
3540B
Rev 2
9/94
3541
Rev 0
9/94
3550A
Rev 1
9/94
3580A
Rev 1
7/92
3600B
Rev 2
9/94
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
NETH NO.
THIRD ED
DATED
9/86
3610
3611
3620
3630
3640
3650
3660
"
3810
NETH NO.
FINAL
UPDATE I
DATED
7/92
3610A
3611A
3620A
3630A
~ *
3650A
3660A
• —
"
NETH NO.
FINAL
UPDT. II
DATED
9/94
"
~ ™
~ ~
3630B
3640A
"
~ ~
3665
"
NETHOD TITLE
Alumina Column
Cleanup
Alumina Column
Cleanup and
Separation of
Petroleum Hastes
Florisil Column
Cleanup
Silica Gel Cleanup
Gel -Permeation
Cleanup
Acid-Base Partition
Cleanup
Sulfur Cleanup
Sulfuric
Acid/Permanganate
Cleanup
Headspace
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IB
Chap 4
Sec
4.2.2
Vol IB
Chap 4
Sec
4.2.2
Vol IB
Chap 4
Sec
4.2.2
Vol IB
Chap 4
Sec
4.2.2
Vol IB
Chap 4
Sec
4.2.2
Vol IB
Chap 4
Sec
4.2.2
Vol IB
Chap 4
Sec
4.2.2
Vol IB
Chap 4
Sec
4.2.2
Vol IB
Chap 4
Sec 4.4
CURRENT
PROMUL-
GATED
NETHOD
3610A
Rev 1
7/92
3611A
Rev 1
7/92
3620A
Rev 1
7/92
3630B
Rev 2
9/94
3640A
Rev 1
9/94
3650A
Rev 1
7/92
3660A
Rev 1
7/92
3665
Rev 0
9/94
3810
Rev 0
9/86
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
METH NO.
THIRD ED
DATED
9/86
3820
5030
5040
"
6010
METH NO.
FINAL
UPDATE I
DATED
7/92
— -
5030A
— V
6010A
METH NO.
FINAL
UPDT. II
DATED
9/94
~ ~
4010
(Update
IIA,
dated
8/93)
" ~
5040A
5041
5050
"" ~
METHOD TITLE
Hexadecane
Extraction and
Screening of
Purgeable Organics
Screening for
Pentachlorophenol
by Immunoassay
Purge-and-Trap
Analysis of Sorbent
Cartridges from
Volatile Organic
Sampling Train
(VOST): Gas
Chromatography/Mass
Spectrometry
Technique
Protocol for
Analysis of Sorbent
Cartridges from
Volatile Organic
Sampling Train
(VOST): Wide -bore
Capillary Column
Technique
Bomb Preparation
Method for Solid
Waste
Inductively Coupled
Plasma- Atomic
Emission
Spectroscopy
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IB
Chap 4
Sec 4.4
Vol IB
Chap 4
Sec 4.4
Vol IB
Chap 4
Sec
4.2.1
Vol IB
Chap 4
Sec
4.2.1
Vol IB
Chap 4
Sec
4.2.1
Vol 1C
Chap 5
Vol IA
Chap 3
Sec 3.3
CURRENT
PROMUL-
GATED
METHOD
3820
Rev 0
9/86
4010
Rev 0
8/93
5030A
Rev 1
7/92
5040A
Rev 1
9/94
5041
Rev 0
9/94
5050
Rev 0
9/94
6010A
Rev 1
7/92
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
METH NO.
THIRD ED
DATED
9/86
~ ™*
7000
7020
7040
7041
7060
7061
~ ~ '
7080
METH NO.
FINAL
UPDATE I
DATED
7/92
~ —
7000A
~ "
~ "*
"
"
7061A
™" ~
"
METH NO.
FINAL
UPDT. II
DATED
9/94
6020
_ •
~ *
~ —
"
7060A
™" ~
7062
7080A
METHOD TITLE
Inductively Coupled
Plasma - Mass
Spectrometry
Atomic Absorption
Methods
Aluminum (Atomic
Absorption, Direct
Aspiration)
Antimony (Atomic
Absorption, Direct
Aspiration)
Antimony (Atomic
Absorption, Furnace
Technique)
Arsenic (Atomic
Absorption, Furnace
Technique)
Arsenic (Atomic
Absorption, Gaseous
Hydride)
Antimony and Arsenic
(Atomic Absorption,
Borohydride
Reduction)
Barium (Atomic
Absorption, Direct
Aspiration)
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
CURRENT
PROMUL-
GATED
METHOD
6020
Rev 0
9/94
7000A
Rev 1
7/92
7020
Rev 0
9/86
7040
Rev 0
9/86
7041
Rev 0
9/86
7060A
Rev 1
9/94
7061A
Rev 1
7/92
7062
Rev 0
9/94
7080A
Rev 1
9/94
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
NETH NO.
THIRD ED
DATED
9/86
— -
7090
7091
7130
7131
7140
7190
7191
7195
NETH NO.
FINAL
UPDATE I
DATED
7/92
7081
— —
~ •*
~ ~
~ •"
~ *~
_ _
~ ~
"
NETH NO.
FINAL
UPDT. II
DATED
9/94
_ _
~ *
~ ~*
~ **
7131A
~ ~~
_ _
•" *
"
NETHOD TITLE
Barium (Atomic
Absorption, Furnace
Technique)
Beryllium (Atomic
Absorption, Direct
Aspiration)
Beryllium (Atomic
Absorption, Furnace
Technique)
Cadmium (Atomic
Absorption, Direct
Aspiration)
Cadmium (Atomic
Absorption, Furnace
Technique)
Calcium (Atomic
Absorption, Direct
Aspiration)
Chromium (Atomic
Absorption, Direct
Aspiration)
Chromium (Atomic
Absorption, Furnace
Technique)
Chromium, Hexavalent
(Coprecipitation)
SW-846
VOLUNE/
CHAPTER/
SECTION
LOCATION
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
CURRENT
PROMUL-
GATED
NETHOD
7081
Rev 0
7/92
7090
Rev 0
9/86
7091
Rev 0
9/86
7130
Rev 0
9/86
7131A
Rev 1
9/94
7140
Rev 0
9/86
7190
Rev 0
9/86
7191
Rev 0
9/86
7195
Rev 0
9/86
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
NETH NO.
THIRD ED
DATED
9/86
7196
7197
7198
7200
7201
7210
•"* *~
7380
~ °~
NETH NO.
FINAL
UPDATE I
DATED
7/92
7196A
"
"• ~
"
•" *•
•. m.
7211
™ ~
7381
NETH NO.
FINAL
UPDT. II
DATED
9/94
"
"
— ••
"
•" ~"
* "
"
"
NETHOD TITLE
Chromium, Hexavalent
(Colorimetric)
Chromium, Hexavalent
(Chelation/Extrac-
tion)
Chromium, Hexavalent
(Differential Pulse
Polarography)
Cobalt (Atomic
Absorption, Direct
Aspiration)
Cobalt (Atomic
Absorption, Furnace
Technique)
Copper (Atomic
Absorption, Direct
Aspiration)
Copper (Atomic
Absorption, Furnace
Technique)
Iron (Atomic
Absorption, Direct
Aspiration)
Iron (Atomic
Absorption, Furnace
Technique)
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
CURRENT
PROMUL-
GATED
NETHOD
7196A
Rev 1
7/92
7197
Rev 0
9/86
7198
Rev 0
9/86
7200
Rev 0
9/86
7201
Rev 0
9/86
7210
Rev 0
9/86
7211
Rev 0
7/92
7380
Rev 0
9/86
7381
Rev 0
7/92
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
NETH NO.
THIRD ED
DATED
9/86
7420
7421
— —
7450
7460
"• -*
7470
7471
7480
NETH NO.
FINAL
UPDATE I
DATED
7/92
~ —
~ "•
7430
— —
— ~
7461
"
"™ *
NETH NO.
FINAL
UPDT. II
DATED
9/94
~ ~
w —
_ _
— ~
_ _
™ ~
7470A
7471A
NETHOD TITLE
Lead (Atomic
Absorption, Direct
Aspiration)
Lead (Atomic
Absorption, Furnace
Technique)
Lithium (Atomic
Absorption, Direct
Aspiration)
Magnesium (Atomic
Absorption, Direct
Aspiration)
Manganese (Atomic
Absorption, Direct
Aspiration)
Manganese (Atomic
Absorption, Furnace
Technique)
Mercury in Liquid
Waste (Manual Cold-
Vapor Technique)
Mercury in Solid or
Semi sol id Waste
(Manual Cold-Vapor
Technique)
Molybdenum (Atomic
Absorption, Direct
Aspiration)
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol I A
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
CURRENT
PROMUL-
GATED
NETHOD
7420
Rev 0
9/86
7421
Rev 0
9/86
7430
Rev 0
7/92
7450
Rev 0
9/86
7460
Rev 0
9/86
7461
Rev 0
7/92
7470A
Rev 1
9/94
7471A
Rev 1
9/94
7480
Rev 0
9/86
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
METH NO.
THIRD ED
DATED
9/86
7481
7520
7550
7610
7740
7741
•* —
7760
"" —
METH NO.
FINAL
UPDATE I
DATED
7/92
"
"
~ ' ~
"
_ _
"
"• ~
7760A
7761
METH NO.
FINAL
UPDT. II
DATED
9/94
~ ~
_ •
*• ~
"
~ ~
7741A
7742
~ •*
— —
METHOD TITLE
Molybdenum (Atomic
Absorption, Furnace
Technique)
Nickel (Atomic
Absorption, Direct
Aspiration)
Osmium (Atomic
Absorption, Direct
Aspiration)
Potassium (Atomic
Absorption, Direct
Aspiration)
Selenium (Atomic
Absorption, Furnace
Technique)
Selenium (Atomic
Absorption, Gaseous
Hydride)
Selenium (Atomic
Absorption,
Borohydride
Reduction)
Silver (Atomic
Absorption, Direct
Aspiration)
Silver (Atomic
Absorption, Furnace
Technique)
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
CURRENT
PROMUL-
GATED
METHOD
7481
Rev 0
9/86
7520
Rev 0
9/86
7550
Rev 0
9/86
7610
Rev 0
9/86
7740
Rev 0
9/86
7741A
Rev 1
9/94
7742
Rev 0
9/94
7760A
Rev 1
7/92
7761
Rev 0
7/92
10
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
NETH NO.
THIRD ED
DATED
9/86
7770
~ "•
7840
7841
7870
7910
7911
7950
"
NETH NO.
FINAL
UPDATE I
DATED
7/92
~ **
7780
~ —
*~ "*
"
— ™
~ "
"
7951
NETH NO.
FINAL
UPDT. II
DATED
9/94
*" ~
" ~
<. «
~ ~
"
~ ~
~ ~
"
NETHOD TITLE
Sodium (Atomic
Absorption, Direct
Aspiration)
Strontium (Atomic
Absorption, Direct
Aspiration)
Thallium (Atomic
Absorption, Direct
Aspiration)
Thallium (Atomic
Absorption, Furnace
Technique)
Tin (Atomic
Absorption, Direct
Aspiration)
Vanadium (Atomic
Absorption, Direct
Aspiration)
Vanadium (Atomic
Absorption, Furnace
Technique)
Zinc (Atomic
Absorption, Direct
Aspiration)
Zinc (Atomic
Absorption, Furnace
Technique)
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
Vol IA
Chap 3
Sec 3.3
CURRENT
PROMUL-
GATED
NETHOD
7770
Rev 0
9/86
7780
Rev 0
7/92
7840
Rev 0
9/86
7841
Rev 0
9/86
7870
Rev 0
9/86
7910
Rev 0
9/86
7911
Rev 0
9/86
7950
Rev 0
9/86
7951
Rev 0
7/92
11
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
NETH NO.
THIRD ED
DATED
9/86
8000
8010
8015
8020
8030
"
NETH NO.
FINAL
UPDATE I
DATED
7/92
8000A
8010A
8011
80 ISA
~ •
8021
8030A
— *~
METH NO.
FINAL
UPDT. II
DATED
9/94
"
8010B
~ ~
8020A
8021A
. _
8031
METHOD TITLE
Gas Chromatography
Halogenated Volatile
Organics by Gas
Chromatography
1,2-Dibromoethane
and l,2-Dibromo-3-
chloropropane by
Microextraction and
Gas Chromatography
Nonhalogenated
Volatile Organics by
Gas Chromatography
Aromatic Volatile
Organics by Gas
Chromatography
Halogenated
Volatiles by Gas
Chromatography Using
Photoionization and
Electrolytic
Conductivity
Detectors in Series:
Capillary Column
Technique
Acrolein and
Acrylonitrile by Gas
Chromatography
Acrylonitrile by Gas
Chromatography
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
CURRENT
PROMUL-
GATED
METHOD
8000A
Rev 1
7/92
8010B
Rev 2
9/94
8011
Rev 0
7/92
8015A
Rev 1
7/92
8020A
Rev 1
9/94
8021A
Rev 1
9/94
8030A
Rev 1
7/92
8031
Rev 0
9/94
12
-------
SH-846 METHOD STATUS TABLE (9/94), CONTINUED
HETH NO.
THIRD ED
DATED
9/86
~ ~
8040
8060
~ *"
8080
8090
HETH NO.
FINAL
UPDATE I
DATED
7/92
~ ~
8040A
~ ~
8070
~ ™ '
NETH NO.
FINAL
UPDT. II
DATED
9/94
8032
"
"™ ~
8061
— ~
8080A
8081
"
METHOD TITLE
Acryl amide by Gas
Chromatography
Phenols by Gas
Chromatography
Phthalate Esters
Phthalate Esters by
Capillary Gas
Chromatography with
Electron Capture
Detection (GC/ECD)
Nitrosamines by Gas
Chromatography
Organochlorine Pes-
ticides and
Polychlorinated
Biphenyls by Gas
Chromatography
Organochlorine
Pesticides and PCBs
as Aroclors by Gas
Chromatography:
Capillary Column
Technique
Nitroaromatics and
Cyclic Ketones
SH-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
CURRENT
PROMUL-
GATED
METHOD
8032
Rev 0
9/94
8040A
Rev 1
7/92
8060
Rev 0
9/86
8061
Rev 0
9/94
8070
Rev 0
7/92
8080A
Rev 1
9/94
8081
Rev 0
9/94
8090
Rev 0
9/86
13
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
METH NO.
THIRD ED
DATED
9/86
8100
"" ~
8120
8140
8150
METH NO.
FINAL
UPDATE I
DATED
7/92
"
8110
"* ~
— —
8141
8150A
METH NO.
FINAL
UPDT. II
DATED
9/94
_ *
"
8120A
8121
— ~
8141A
8150B
8151
METHOD TITLE
Polynuclear Aromatic
Hydrocarbons
Haloethers by Gas
Chromatography
Chlorinated
Hydrocarbons by Gas
Chromatography
Chlorinated
Hydrocarbons by Gas
Chromatography:
Capillary Column
Technique
Organophosphorus
Pesticides
Organophosphorus
Compounds by Gas
Chromatography:
Capillary Column
Technique
Chlorinated
Herbicides by Gas
Chromatography
Chlorinated
Herbicides by GC
Using Methylation or
Pentaf 1 uorobenzyl -
ation Derivati-
zation: Capillary
Column Technique
SM-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
Vol IB
Chap 4
Sec
4.3.1
CURRENT
PROMUL-
GATED
METHOD
8100
Rev 0
9/86
8110
Rev 0
7/92
8120A
Rev 1
9/94
8121
Rev 0
9/94
8140
Rev 0
9/86
8141A
Rev 1
9/94
8150B
Rev 2
9/94
8151
Rev 0
9/94
14
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
HETH NO.
THIRD ED
DATED
9/86
8240
8250
8270
8280
METH NO.
FINAL
UPDATE I
DATED
7/92
8240A
8260
8270A
METH NO.
FINAL
UPDT. II
DATED
9/94
8240B
8250A
8260A
8270B
8275
METHOD TITLE
Volatile Organic
Compounds by Gas
Chromatography/Mass
Spectrometry (GC/MS)
Semi volatile Organic
Compounds
by Gas
Chromatography/Mass
Spectrometry (GC/MS)
Volatile Organic
Compounds by Gas
Chromatography/Mass
Spectrometry
(GC/MS): Capillary
Column Technique
Semivolatile Organic
Compounds by Gas
Chromatography/Mass
Spectrometry
(GC/MS): Capillary
Column Technique
Thermal
Chromatography/Mass
Spectrometry (TC/MS)
for Screening
Semivolatile Organic
Compounds
The Analysis of
Polychlorinated
Dibenzo-p-Dioxins
and Polychlorinated
Dibenzofurans
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IB
Chap 4
Sec
4.3.2
Vol IB
Chap 4
Sec
4.3.2
Vol IB
Chap 4
Sec
4.3.2
Vol IB
Chap 4
Sec
4.3.2
Vol IB
Chap 4
Sec 4.4
Vol IB
Chap 4
Sec
4.3.2
CURRENT
PROMUL-
GATED
METHOD
8240B
Rev 2
9/94
8250A
Rev 1
9/94
8260A
Rev 1
9/94
8270B
Rev 2
9/94
8275
Rev 0
9/94
8280
Rev 0
9/86
15
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
METH NO.
THIRD ED
DATED
9/86
8310
"
METH NO.
FINAL
UPDATE I
DATED
7/92
" ~
"
METH NO.
FINAL
UPDT. II
DATED
9/94
8290
— ~
8315
8316
8318
METHOD TITLE
Polychlorinated
Dibenzodioxins
(PCDDs) and
Polychlorinated
Dibenzofurans
(PCDFs) by High-
Resolution Gas
Chromatography/High-
Resolution Mass
Spectrometry
(HRGC/HRMS)
Polynuclear Aromatic
Hydrocarbons
Determination of
Carbonyl Compounds
by High Performance
Liquid
Chromatography
(HPLC)
Aery 1 amide,
Acrylonitrile and
Acrolein by High
Performance Liquid
Chromatography
(HPLC)
N-Methyl carbamates
by High Performance
Liquid Chroma-
tography (HPLC)
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IB
Chap 4
Sec
4.3.2
Vol IB
Chap 4
Sec
4.3.3
Vol IB
Chap 4
Sec
4.3.3
Vol IB
Chap 4
Sec
4.3.3
Vol IB
Chap 4
Sec
4.3.3
CURRENT
PROMUL-
GATED
METHOD
8290
Rev 0
9/94
8310
Rev 0
9/86
8315
Rev 0
9/94
8316
Rev 0
9/94
8318
Rev 0
9/94
16
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
METH NO.
THIRD ED
DATED
9/86
9010
9012
METH NO.
FINAL
UPDATE I
DATED
7/92
9010A
"
METH NO.
FINAL
UPDT. II
DATED
9/94
8321
8330
8331
8410
** ~
"
METHOD TITLE
Solvent Extractable
Non-Volatile
Compounds by High
Performance Liquid
Chromatography/Ther-
mospray/Mass
Spectrometry
(HPLC/TSP/MS) or
Ultraviolet (UV)
Detection
Nitroaromatics and
Nitramines by High
Performance Liquid
Chromatography
(HPLC)
Tetrazene by Reverse
Phase High
Performance Liquid
Chromatography
(HPLC)
Gas Chroma-
tography/Fourier
Transform Infrared
(GC/FT-IR) Spec-
trometry for
Semi volatile
Organics: Capillary
Column
Total and Amenable
Cyanide
(Colorimetric,
Manual)
Total and Amenable
Cyanide
(Colorimetric,
Automated UV)
SM-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol IB
Chap 4
Sec
4.3.3
Vol IB
Chap 4
Sec
4.3.3
Vol IB
Chap 4
Sec
4.3.3
Vol IB
Chap 4
Sec
4.3.4
Vol 1C
Chap 5
Vol 1C
Chap 5
CURRENT
PROMUL-
GATED
METHOD
8321
Rev 0
9/94
8330
Rev 0
9/94
8331
Rev 0
9/94
8410
Rev 0
9/94
9010A
Rev 1
7/92
9012
Rev 0
9/86
17
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
METH NO.
THIRD ED
DATED
9/86
"~ ~
9020
"
9022
9030
"
9035
9036
9038
METH NO.
FINAL
UPDATE I
DATED
7/92
9013
9020A
9021
^ ~
9030A
9031
"
"
METH NO.
FINAL
UPDT. II
DATED
9/94
— ~
90206
~ ~
"
~ ~
*" ~
"
METHOD TITLE
Cyanide Extraction
Procedure for Sol ids
and Oils
Total Organic
Hal ides (TOX)
Purgeable Organic
Hal ides (POX)
Total Organic
Hal ides (TOX) by
Neutron Activation
Analysis
Acid-Soluble and
Acid-Insoluble
Sulfides
Extractable Sulfides
Sulfate
(Colorimetric,
Automated,
Chloranilate)
Sulfate
(Colorimetric,
Automated,
Methyl thymol Blue,
AA II)
Sulfate
(Turbidimetric)
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
CURRENT
PROMUL-
GATED
METHOD
9013
Rev 0
7/92
9020B
Rev 2
9/94
9021
Rev 0
7/92
9022
Rev 0
9/86
9030A
Rev 1
7/92
9031
Rev 0
7/92
9035
Rev 0
9/86
9036
Rev 0
9/86
9038
Rev 0
9/86
18
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
METH NO.
THIRD ED
DATED
9/86
9040
9041
9045
9050
>. ™
9060
9065
9066
9067
METH NO.
FINAL
UPDATE I
DATED
7/92
~ •*
9041A
9045A
*• *•
— —
*• "•
"
NETH NO.
FINAL
UPDT. II
DATED
9/94
9040A
"" ™
9045B
"* *
9056
~" "
— —
"
"
METHOD TITLE
pH Electrometric
Measurement
pH Paper Method
Soil and Waste pH
Specific Conductance
Determination of
Inorganic Anions by
Ion Chromatography
Total Organic Carbon
Phenol ics
(Spectrophotometri c ,
Manual 4-AAP with
Distillation)
Phenol ics
(Colorimetric,
Automated 4-AAP with
Distillation)
Phenol ics
(Spectrophotometri c,
MBTH with
Distillation)
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol 1C
Chap 6
Vol 1C
Chap 6
Vol 1C
Chap 6
Vol 1C
Chap 6
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
CURRENT
PROMUL-
GATED
METHOD
9040A
Rev 1
9/94
9041A
Rev 1
7/92
9045B
Rev 2
9/94
9050
Rev 0
9/86
9056
Rev 0
9/94
9060
Rev 0
9/86
9065
Rev 0
9/86
9066
Rev 0
9/86
9067
Rev 0
9/86
19
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
HETH NO.
THIRD ED
DATED
9/86
9070
9071
9080
9081
HETH NO.
FINAL
UPDATE I
DATED
7/92
"
"
METH NO.
FINAL
UPDT. II
DATED
9/94
9071A
9075
9076
9077
"
"
METHOD TITLE
Total Recoverable
Oil & Grease
(Gravimetric,
Separatory Funnel
Extraction)
Oil and Grease
Extraction Method
for Sludge and
Sediment
Samples
Test Method for
Total Chlorine in
New and Used
Petroleum Products
by X-Ray
Fluorescence
Spectrometry (XRF)
Test Method for
Total Chlorine in
New and Used
Petroleum Products
by Oxidative
Combustion and
Microcoulometry
Test Methods for
Total Chlorine in
New and Used
Petroleum Products
(Field Test Kit
Methods)
Cation-Exchange
Capacity of Soils
(Ammonium Acetate)
Cation-Exchange
Capacity of Soils
(Sodium Acetate)
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 6
Vol 1C
Chap 6
CURRENT
PROMUL-
GATED
METHOD
9070
Rev 0
9/86
9071A
Rev 1
9/94
9075
Rev 0
9/94
9076
Rev 0
9/94
9077
Rev 0
9/94
9080
Rev 0
9/86
9081
Rev 0
9/86
20
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
METH NO.
THIRD ED
DATED
9/86
9090
9095
*~ ~
9100
9131
9132
9200
9250
9251
METH NO.
FINAL
UPDATE I
DATED
7/92
9090A
_ -
~ ~"
"
™* *"
*" ~
METH NO.
FINAL
UPDT. II
DATED
9/94
~ "
~ "
9096
~ ""
— ••
~ "
~ *
"* *"
METHOD TITLE
Compatibility Test
for Wastes and
Membrane Liners
Paint Filter Liquids
Test
Liquid Release Test
(LRT) Procedure
Saturated Hydraulic
Conductivity,
Saturated Leachate
Conductivity, and
Intrinsic
Permeability
Total Col i form:
Multiple Tube
Fermentation
Technique
Total Col i form:
Membrane Filter
Technique
Nitrate
Chloride
(Colorimetric,
Automated
Ferricyanide AAI)
Chloride
(Colorimetric,
Automated
Ferricyanide AAI I)
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol 1C
Chap 6
Vol 1C
Chap 6
Vol 1C
Chap 6
Vol 1C
Chap 6
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 5
CURRENT
PROMUL-
GATED
METHOD
9090A
Rev 1
7/92
9095
Rev 0
9/86
9096
Rev 0
9/94
9100
Rev 0
9/86
9131
Rev 0
9/86
9132
Rev 0
9/86
9200
Rev 0
9/86
9250
Rev 0
9/86
9251
Rev 0
9'S6
21
-------
SW-846 METHOD STATUS TABLE (9/94), CONTINUED
METH NO.
THIRD ED
DATED
9/86
9252
~ *
9310
9315
9320
HCN Test
Method
H2S Test
Method
METH NO.
FINAL
UPDATE I
DATED
7/92
•" ~
*™ *"
— —
— ••
~ —
HCN Test
Method
H2S Test
Method
METH NO.
FINAL
UPDT. II
DATED
9/94
9252A
9253
™* ~
— ~
"~ ~
HCN Test
Method
H2S Test
Method
METHOD TITLE
Chloride
(Titrimetric,
Mercuric Nitrate)
Chloride
(Titrimetric, Silver
Nitrate)
Gross Alpha and
Gross Beta
Alpha- Emitting
Radium Isotopes
Radium-228
Test Method to
Determine Hydrogen
Cyanide Released
from Wastes
Test Method to
Determine Hydrogen
Sulfide Released
from Wastes
SW-846
VOLUME/
CHAPTER/
SECTION
LOCATION
Vol 1C
Chap 5
Vol 1C
Chap 5
Vol 1C
Chap 6
Vol 1C
Chap 6
Vol 1C
Chap 5
Vol 1C
Chap 7
Sec 7.3
Vol 1C
Chap 7
Sec 7.3
CURRENT
PROMUL-
GATED
METHOD
9252A
Rev 1
9/94
9253
Rev 0
9/94
9310
Rev 0
9/86
9315
Rev 0
9/86
9320
Rev 0
9/86
Guidance
Method
Only
Guidance
Method
Only
22
-------
VOLUME ONE,
SECTION B
Revision 0
Date September 1986
-------
For Rale by the Superintendent of Documents, U.S. Government Printing Office Washington, D.C. 20402
-------
DISCLAIMER
Mention of trade names or commercial products does not constitute
endorsement or recommendation for use by the U.S. Environmental Protection
Agency.
SW-846 methods are designed to be used with equipment from any manufacturer
that results In suitable method performance (as assessed by accuracy, precision,
detection limits and matrix compatibility). In several SW-846 methods, equipment
specifications and settings are given for the specific instrument used during
method development, or subsequently approved for use in the method. These
references are made to provide the best possible guidance to laboratories using
this manual. Equipment not specified in the method may be used as long as the
laboratory achieves equivalent or superior method performance. If alternate
equipment is used, the laboratory must follow the manufacturer's instructions for
their particular instrument.
Since many types and sizes of glassware and supplies are commercially
available, and since it is possible to prepare reagents and standards in many
different ways, those specified in these methods may be replaced by any similar
types as long as this substitution does not affect the overall quality of the
analyses.
DISCLAIMER - 1 Revision 0
July 1992
-------
ABSTRACT
Test Methods for Evaluating Solid Waste, Physical/Chemical Methods (SW-846)
provides test procedures and guidance which are recommended for use in conducting
the evaluations and measurements needed to comply with the Resource Conservation
and Recovery Act (RCRA), Public Law 94-580, as amended. These methods are
approved by the U.S. Environmental Protection Agency for obtaining data to
satisfy the requirements of 40 CFR Parts 122 through 270 promulgated under RCRA,
as amended. This manual presents the state-of-the-art in routine analytical
tested adapted for the RCRA program. It contains procedures for field and
laboratory quality control, sampling, determining hazardous constituents in
wastes, determining the hazardous characteristics of wastes (toxicity,
ignitability, reactivity, and corrosivity), and for determining physical
properties of wastes. It also contains guidance on how to select appropriate
methods.
Several of the hazardous waste regulations under Subtitle C of RCRA require
that specific testing methods described in SW-846 be employed for certain
applications. Refer to 40 Code of Federal Regulations (CFR), Parts 260 through
270, for those specific requirements. Any reliable analytical method may be used
to meet other requirements under Subtitle C of RCRA.
ABSTRACT - : Revision 2
September 1994
-------
TABLE OF CONTENTS
VOLUME ONE
SECTION A
DISCLAIMER
ABSTRACT
TABLE OF CONTENTS
METHOD INDEX AND CONVERSION TABLE
PREFACE
ACKNOWLEDGEMENTS
PART I METHODS FOR ANALYTES AND PROPERTIES
CHAPTER ONE -- QUALITY CONTROL
1.0
2.0
3.0
4.0
5.0
6.0
Introduction
QA Project Plan
Field Operations
Laboratory Operations
Definitions
References
CHAPTER TWO -- CHOOSING THE CORRECT PROCEDURE
2.1 Purpose
2.2 Required Information
2.3 Implementing the Guidance
2.4 Characteristics
2.5 Ground Water
2.6 References
CHAPTER THREE -- METALLIC ANALYTES
3.1 Sampling Considerations
3.2 Sample Preparation Methods
Method 3005A: Acid Digestion of Waters for Total Recoverable or
Dissolved Metals for Analysis by Flame Atomic Absorption
(FLAA) or Inductively Coupled Plasma (ICP) Spectroscopy
Method 3010A: Acid Digestion of Aqueous Samples and Extracts for Total
Metals for Analysis by Flame Atomic Absorption (FLAA) or
Inductively Coupled Plasma (ICP) Spectroscopy
Method 3015: Microwave Assisted Acid Digestion of Aqueous Samples and
Extracts
CONTENTS - 1 Revision 2
September 1994
-------
Method 3020A:
Method 3040:
Method 3050A:
Method 3051:
Acid Digestion of Aqueous Samples and Extracts for Total
Metals for Analysis by Graphite Furnace Atomic
Absorption (6FAA) Spectroscopy
Dissolution Procedure for Oils, Greases, or Waxes
Acid Digestion of Sediments, Sludges, and Soils
Microwave Assisted Acid Digestion of Sediments, Sludges,
Soils, and Oils
3.3 Methods for Determination of Metals
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
6010A:
6020:
7000A:
7020:
7040:
7041:
7060A:
7061A:
7062:
7080A:
7081:
7090:
7091:
7130:
7131 A:
7140:
7190:
7191:
7195:
7196A:
7197:
7198:
7200:
7201:
7210:
7211:
7380:
7381:
7420:
7421:
7430:
7450:
7460:
7461:
7470A:
7471A:
Method 7480:
Method 7481:
Method 7520:
Method 7550:
Method 7610:
Method 7740:
Inductively Coupled Plasma-Atomic Emission Spectroscopy
Inductively Coupled Plasma - Mass Spectrometry
Atomic Absorption Methods
Aluminum (AA, Direct Aspiration)
Antimony (AA, Direct Aspiration)
Antimony (AA, Furnace Technique)
Arsenic (AA, Furnace Technique)
Arsenic (AA, Gaseous Hydride)
Antimony and Arsenic (AA, Borohydride Reduction)
Barium (AA, Direct Aspiration)
Barium (AA, Furnace Technique)
Beryllium (AA, Direct Aspiration)
Beryllium (AA, Furnace Technique)
Cadmium (AA, Direct Aspiration)
Cadmium (AA, Furnace Technique)
Calcium (AA, Direct Aspiration)
Chromium (AA, Direct Aspiration)
Chromium (AA, Furnace Technique)
Chromium, Hexavalent (Coprecipitation)
Chromium, Hexavalent (Colorimetric)
Chromium, Hexavalent (Chelation/Extraction)
Chromium, Hexavalent (Differential Pulse Polarography)
Cobalt (AA, Direct Aspiration)
Cobalt (AA, Furnace Technique)
Copper (AA, Direct Aspiration)
Copper (AA, Furnace Technique)
Iron (AA, Direct Aspiration)
Iron (AA, Furnace Technique)
Lead (AA, Direct Aspiration)
Lead (AA, Furnace Technique)
Lithium (AA, Direct Aspiration)
Magnesium (AA, Direct Aspiration)
Manganese (AA, Direct Aspiration)
Manganese (AA, Furnace Technique)
Mercury in Liquid Waste (Manual Cold-Vapor Technique)
Mercury in Solid or Semi sol id Waste (Manual Cold-Vapor
Technique)
Molybdenum (AA, Direct Aspiration)
Molybdenum (AA, Furnace Technique)
Nickel (AA, Direct Aspiration)
Osmium (AA, Direct Aspiration)
Potassium (AA, Direct Aspiration)
Selenium (AA, Furnace Technique)
CONTENTS - 2
Revision 2
September 1994
-------
Method 7741A: Selenium (AA; Gaseous Hydride)
Method 7742: Selenium (AA, Borohydride Reduction)
Method 7760A: Silver (AA, Direct Aspiration)
Method 7761: Silver (AA, Furnace Technique)
Method 7770: Sodium (AA, Direct Aspiration)
Method 7780: Strontium (AA, Direct Aspiration)
Method 7840: Thallium (AA, Direct Aspiration)
Method 7841: Thallium (AA, Furnace Technique)
Method 7870: Tin (AA, Direct Aspiration)
Method 7910: Vanadium (AA, Direct Aspiration)
Method 7911: Vanadium (AA, Furnace Technique)
Method 7950: Zinc (AA, Direct Aspiration)
Method 7951: Zinc (AA, Furnace Technique)
APPENDIX -- COMPANY REFERENCES
NOTE; A suffix of "A" in the method number indicates revision one
(the method has been revised once). A suffix of "B" in the method
number indicates revision two (the method has been revised twice). In
order to properly document the method used for analysis, the entire
method number Including the suffix letter designation (e.g., A or B)
must be Identified by the analyst. A method reference found within
the RCRA regulations and the text of SW-846 methods and chapters
refers to the latest promulgated revision of the method, even though
the method number does not include the appropriate letter suffix.
CONTENTS - 3
Revision
September
-------
VOLUME ONE
SECTION B
DISCLAIMER
ABSTRACT
TABLE OF CONTENTS
METHOD INDEX AND CONVERSION TABLE
PREFACE
ACKNOWLEDGEMENTS
CHAPTER ONE. REPRINTED -- QUALITY CONTROL
1.0 Introduction
2.0 QA Project Plan
3.0 Field Operations
4.0 Laboratory Operations
5.0 Definitions
6.0 References
CHAPTER FOUR -- ORGANIC ANALYTES
4.1 Sampling Considerations
4.2 Sample Preparation Methods
4.2.1 Extractions and Preparations
Method 3500A: Organic Extraction and Sample Preparation
Method 3510B: Separatory Funnel Liquid-Liquid Extraction
Method 3520B: Continuous Liquid-Liquid Extraction
Method 3540B: Soxhlet Extraction
Method 3541: Automated Soxhlet Extraction
Method 3550A: Ultrasonic Extraction
Method 3580A: Waste Dilution
Method 5030A: Purge-and-Trap
Method 5040A: Analysis of Sorbent Cartridges from Volatile Organic
Sampling Train (VOST): Gas Chromatography/Mass
Spectrometry Technique
Method 5041: Protocol for Analysis of Sorbent Cartridges from
Volatile Organic Sampling Train (VOST): Wide-bore
Capillary Column Technique
Method 5100: Determination of the Volatile Organic Concentration of
Waste Samples
Method 5110: Determination of Organic Phase Vapor Pressure in Waste
Samples
4.2.2 Cleanup
Method 3600B: Cleanup
Method 3610A: Alumina Column Cleanup
CONTENTS - 4 Revision 2
September 1994
-------
Method 3611A:
Method
Method
Method
Method
Method
Method
3620A:
3630B:
3640A:
3650A:
3660A:
3665:
Alumina Column
Petroleum Wastes
Florisil Column Cleanup
Silica Gel Cleanup
Gel-Permeation Cleanup
Acid-Base Partition Cleanup
Sulfur Cleanup
Sulfuric Acid/Permanganate Cleanup
Cleanup and Separation of
4.3 Determination of Organic Analytes
4.3.1
Gas Chromatographic Methods
Method 8000A:
Method 8010B:
Method 8011:
Method 8015A:
Method 8020A:
Method 8021A:
Method
Method
Method
Method
Method
Method
8030A:
8031:
8032:
8040A:
8060:
8061:
Method 8070:
Method 8080A:
Method 8081:
Method 8090:
Method 8100:
Method 8110:
Method 8120A:
Method 8121:
Method 8140:
Method 8141A:
Method 8150B:
Method 8151:
by
Gas Chromatography
Halogenated Volatile Organics by Gas Chromatography
1,2-Dibromoethane and l,2-Dibromo-3-chloropropane
Microextraction and Gas Chromatography
Nonhalogenated Volatile Organics by Gas Chromatography
Aromatic Volatile Organics by Gas Chromatography
Halogenated Volatiles by Gas Chromatography Using
Photoionization and Electrolytic Conductivity Detectors
in Series: Capillary Column Technique
Acrolein and Acrylonitrile by Gas Chromatography
Acrylonitrile by Gas Chromatography
Acrylamide by Gas Chromatography
Phenols by Gas Chromatography
Phthalate Esters
Phthalate Esters by Capillary Gas Chromatography with
Electron Capture Detection (GC/ECD)
Nitrosamines by Gas Chromatography
Organochlorine Pesticides and Polychlorinated Biphenyls
by Gas Chromatography
Organochlorine Pesticides and PCBs as Aroclors by Gas
Chromatography: Capillary Column Technique
Nitroaromatics and Cyclic Ketones
Polynuclear Aromatic Hydrocarbons
Haloethers by Gas Chromatography
Chlorinated Hydrocarbons by Gas Chromatography
Chlorinated Hydrocarbons by Gas Chromatography:
Capillary Column Technique
Organophosphorus Pesticides
Organophosphorus Compounds by Gas Chromatography:
Capillary Column Technique
Chlorinated Herbicides by Gas Chromatography
Chlorinated Herbicides by GC Using Methylation or
Pentafluorobenzylation Derivatization: Capillary Column
Technique
CONTENTS - 5
Revision 2
September 1994
-------
4.3.2
Gas Chromatographic/Mass Spectroscopic Methods
Method 8240B:
Method 8250A:
Method 8260A:
Method 8270B:
Method 8280:
Appendix A:
Appendix B:
Method 8290:
Appendix A:
Volatile Organic Compounds by Gas Chromatography/Mass
Spectrometry (GC/MS)
Semivolatile Organic Compounds by Gas
Chromatography/Mass Spectrometry (GC/MS)
Volatile Organic Compounds by Gas Chromatography/Mass
Spectrometry (GC/MS): Capillary Column Technique
Semivolatile Organic Compounds by Gas
Chromatography/Mass Spectrometry (GC/MS): Capillary
Column Technique
The Analysis of Polychlorinated Dibenzo-p-Dioxins and
Polychlorinated Dibenzofurans
Signal-to-Noise Determination Methods
Recommended Safety and Handling Procedures for
PCDDs/PCDFs
Polychlorinated Dibenzodioxins (PCDDs) and
Polychlorinated Dibenzofurans (PCDFs) by High-Resolution
Gas Chromatography/High-Resolution Mass Spectrometry
(HRGC/HRMS)
Procedures for the Collection, Handling,
Analysis, and Reporting of Wipe Tests Performed
within the Laboratory
4.3.3
Method 8310:
Method 8315:
Appendix A:
Method 8316:
Method 8318:
Method 8321:
Method 8330:
Method 8331:
High Performance Liquid Chromatographic Methods
Polynuclear Aromatic Hydrocarbons
Determination of Carbonyl Compounds by High Performance
Liquid Chromatography (HPLC)
Recrystallization of 2,4-Dinitrophenylhydrazine
(DNPH)
Acrylamide, Acrylonitrile and Acrolein by High
Performance Liquid Chromatography (HPLC)
N-Methylcarbamates by High Performance Liquid
Chromatography (HPLC)
Solvent Extractable Non-Volatile Compounds by High
Performance Liquid Chromatography/Thermospray/Mass
Spectrometry (HPLC/TSP/MS) or Ultraviolet (UV) Detection
Nitroaromatics and Nitramines by High Performance Liquid
Chromatography (HPLC)
Tetrazene by Reverse Phase High Performance Liquid
Chromatography (HPLC)
4.3.4
Method 8410:
Fourier Transform Infrared Methods
Gas Chromatography/Fourier Transform Infrared (GC/FT-IR)
Spectrometry for Semivolatile Organics: Capillary
Column
CONTENTS - 6
Revision 2
September 1994
-------
4.4 Miscellaneous Screening Methods
Method 3810: Headspace
Method 3820: Hexadecane Extraction and Screening of Purgeable
Organics
Method 4010: Screening for Pentachlorophenol by Immunoassay
Method 8275: Thermal Chromatography/Mass Spectrometry (TC/MS) for
Screening Semivolatile Organic Compounds
APPENDIX -- COMPANY REFERENCES
NOTE: A suffix of "A" in the method number indicates revision one
(the method has been revised once). A suffix of "B" in the method
number indicates revision two (the method has been revised twice). In
order to properly document the method used for analysis, the entire
method number Including the suffix letter designation (e.g., A or B)
must be Identified by the analyst. A method reference found within
the RCRA regulations and the text of SW-846 methods and chapters
refers to the latest promulgated revision of the method, even though
the method number does not include the appropriate letter suffix.
CGN'ENTS - 7 Revision 2
September 1994
-------
VOLUME ONE
SECTION C
UV)
DISCLAIMER
ABSTRACT
TABLE OF CONTENTS
METHOD INDEX AND CONVERSION TABLE
PREFACE
CHAPTER ONE, REPRINTED -- QUALITY CONTROL
1.0 Introduction
2.0 QA Project Plan
3.0 Field Operations
4.0 Laboratory Operations
5.0 Definitions
6.0 References
CHAPTER FIVE -- MISCELLANEOUS TEST METHODS
Method 5050: Bomb Preparation Method for Solid Waste
Method 9010A: Total and Amenable Cyanide (Colorimetric, Manual)
Method 9012: Total and Amenable Cyanide (Colorimetric, Automated
Method 9013: Cyanide Extraction Procedure for Solids and Oils
Method 9020B: Total Organic Hal ides (TOX)
Method 9021: Purgeable Organic Hal ides (POX)
Method 9022: Total Organic Hal ides (TOX) by Neutron Activation
Analysis
Acid-Soluble and Acid-Insoluble Sulfides
Extractable Sulfides
Sulfate (Colorimetric, Automated,
Sulfate (Colorimetric, Automated,
II)
Method 9038: Sulfate (Turbidimetric)
Method 9056: Determination of Inorganic Anions by Ion Chromatography
Method 9060: Total Organic Carbon
Method 9065: Phenolics (Spectrophotometric,
Distillation)
Phenolics (Colorimetric, Automated
Distillation)
Method 9067: Phenolics (Spectrophotometric, MBTH with Distillation)
Method 9070: Total Recoverable Oil & Grease (Gravimetric, Separatory
Funnel Extraction)
Method 9071A: Oil and Grease Extraction Method for Sludge and Sediment
Samples
Method 9075: Test Method for Total Chlorine in New and Used Petroleum
Products by X-Ray Fluorescence Spectrometry (XRF)
Method 9076: Test Method for Total Chlorine in New and Used Petroleum
Products by Oxidative Combustion and Microcoulometry
Method 9030A:
Method 9031:
Method 9035:
Method 9036:
Chloranilate)
Methyl thymol Blue,
AA
Method 9066:
Manual 4-AAP with
4-AAP with
CONTENTS - 8
Revision 2
September 1994
-------
Method 9077:
Method A:
Method B:
Method C:
Method 9131:
Method 9132:
Method 9200:
Method 9250:
Method 9251:
Method 9252A:
Method 9253:
Method 9320:
CHAPTER SIX -- PROPERTIES
Method 1312:
Method 1320:
Method 1330A:
Method 9040A:
Method 9041A:
Method 9045B:
Method 9050:
Method 9080:
Method 9081:
Method 9090A:
Method 9095:
Method 9096:
Appendix A:
Method 9100:
Method 9310:
Method 9315:
Test Methods for Total Chlorine in New and Used
Petroleum Products (Field Test Kit Methods)
Fixed End Point Test Kit Method
Reverse Titration Quantitative End Point Test Kit
Method
Direct Titration Quantitative End Point Test Kit Method
Total Coliform: Multiple Tube Fermentation Technique
Total Coliform: Membrane Filter Technique
Nitrate
Chloride (Colorimetric, Automated Ferricyanide AAI)
Chloride (Colorimetric, Automated Ferricyanide AAII)
Chloride (Titrimetric, Mercuric Nitrate)
Chloride (Titrimetric, Silver Nitrate)
Radium-228
Synthetic Precipitation Leaching Procedure
Multiple Extraction Procedure
Extraction Procedure for Oily Wastes
pH Electrometric Measurement
pH Paper Method
Soil and Waste pH
Specific Conductance
Cation-Exchange Capacity of Soils (Ammonium Acetate)
Cation-Exchange Capacity of Soils (Sodium Acetate)
Compatibility Test for Wastes and Membrane Liners
Paint Filter Liquids Test
Liquid Release Test (LRT) Procedure
LRT Pre-Test
Saturated Hydraulic Conductivity, Saturated Leachate
Conductivity, and Intrinsic Permeability
Gross Alpha and Gross Beta
Alpha-Emitting Radium Isotopes
PART II CHARACTERISTICS
CHAPTER SEVEN -- INTRODUCTION AND REGULATORY DEFINITIONS
7.1 Ignitability
7.2 Corrosivity
7.3 Reactivity
Test Method to Determine Hydrogen Cyanide Released from Wastes
Test Method to Determine Hydrogen Sulfide Released from Wastes
7.4 Toxicity Characteristic Leaching Procedure
CONTENTS - 9
Revision 2
September 1994
-------
CHAPTER EIGHT -- METHODS FOR DETERMINING CHARACTERISTICS
8.1 Ignitability
Method 1010: Pensky-Martens Closed-Cup Method for Determining
Ignitability
Method 1020A: Setaflash Closed-Cup Method for Determining Ignitability
8.2 Corrosivity
Method 1110: Corrosivity Toward Steel
8.3 Reactivity
8.4 Toxicity
Method 1310A: Extraction Procedure (EP) Toxicity Test Method and
Structural Integrity Test
Method 1311: Toxicity Characteristic Leaching Procedure
APPENDIX -- COMPANY REFERENCES
NOTE: A suffix of "A" in the method number indicates revision one
(the method has been revised once). A suffix of "B" in the method
number indicates revision two (the method has been revised twice). In
order to properly document the method used for analysis, the entire
method number Including the suffix letter designation (e.g., A or B)
must be Identified by the analyst. A method reference found within
the RCRA regulations and the text of SW-846 methods and chapters
refers to the latest promulgated revision of the method, even though
the method number does not include the appropriate letter suffix.
CONTENTS - 10 Revision 2
September 1994
-------
VOLUME TWO
DISCLAIMER
ABSTRACT
TABLE OF CONTENTS
METHOD INDEX AND CONVERSION TABLE
PREFACE
CHAPTER ONE. REPRINTED -- QUALITY CONTROL
1.0 Introduction
2.0 QA Project Plan
3.0 Field Operations
4.0 Laboratory Operations
5.0 Definitions
6.0 References
PART III SAMPLING
CHAPTER NINE -- SAMPLING PLAN
9.1 Design and Development
9.2 Implementation
CHAPTER TEN -- SAMPLING METHODS
Method 0010: Modified Method 5 Sampling Train
Appendix A: Preparation of XAD-2 Sorbent Resin
Appendix B: Total Chromatographable Organic Material Analysis
Method 0020: Source Assessment Sampling System (SASS)
Method 0030: Volatile Organic Sampling Train
PART IV MONITORING
CHAPTER ELEVEN -- GROUND WATER MONITORING
11.1 Background and Objectives
11.2 Relationship to the Regulations and to Other Documents
11.3 Revisions and Additions
11.4 Acceptable Designs and Practices
11.5 Unacceptable Designs and Practices
CHAPTER TWELVE -- LAND TREATMENT MONITORING
12.1 Background
12.2 Treatment Zone
12.3 Regulatory Definition
CONTENTS - 11 Rev is-, or. :
September 1994
-------
12.4 Monitoring and Sampling Strategy
12.5 Analysis
12.6 References and Bibliography
CHAPTER THIRTEEN - INCINERATION
13.1 Introduction
13.2 Regulatory Definition
13.3 Waste Characterization Strategy
13.4 Stack-Gas Effluent Characterization Strategy
13.5 Additional Effluent Characterization Strategy
13.6 Selection of Specific Sampling and Analysis Methods
13.7 References
APPENDIX -- COMPANY REFERENCES
NOTE; A suffix of "A" in the method number indicates revision one
(the method has been revised once). A suffix of "B" in the method
number indicates revision two (the method has been revised twice). In
order to properly document the method used for analysis, the entire
method number including the suffix letter designation (e.g., A or B)
must be Identified by the analyst. A method reference found within
the RCRA regulations and the text of SW-846 methods and chapters
refers to the latest promulgated revision of the method, even though
the method number does not include the appropriate letter suffix.
CONTENTS - 12 Revision 2
September 1994
-------
METHOD INDEX AND CONVERSION TABLE
Method Number,
Third Edition
0010
0020
0030
1010
1020
1110
1310
1320
1330
3005
3010
3020
3040
3050
3500
3510
3520
3540
3550
3580
3600
3610
3611
3620
3630
3640
3650
3660
3810
3820
5030
5040
6010
7000
7020
Chapter Number,
Third Edition
Ten
Ten
Ten
Eight (8.1)
Eight (8.1)
Eight (8.2)
Eight (8.4)
Six
Six
Three
Three
Three
Three
Three
Four (4.2.1)
Four (4.2.1)
Four (4.2.1)
Four (4.2.1)
Four (4.2.1)
Four (4.2.1)
Four (4.2.2)
Four (4.2.2)
Four (4.2.2)
Four (4.2.2)
Four (4.2.2)
Four (4.2.2)
Four (4.2.2)
Four (4.2.2)
Four (4.4)
Four (4.4)
Four (4.2.1)
Four (4.2.1)
Three
Three
Three
Method Number,
Current Revision
Second Edition
0010
0020
0030
1010
1020
1110
1310
1320
1330
3005
3010
3020
3040
3050
None (new method)
3510
3520
3540
3550
None (new method)
None (new method)
None (new method)
3570
None (new method)
None (new method)
None (new method)
None (new method)
None (new method)
5020
None (new method)
5030
3720
6010
7000
7020
Number
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
METHOD INDEX - 1
Revision 0
Date September 1986
-------
METHOD INDEX AND CONVERSION TABLE
(Continued)
Method Number,
Third Edition
Chapter Number,
Third Edition
Method Number,
Second Edition
Current Revision
Number
7040
7041
7060
7061
7080
7090
7091
7130
7131
7140
7190
7191
7195
7196
7197
7198
7200
7201
7210
7380
7420
7421
7450
7460
7470
7471
7480
7481
7520
7550
7610
7740
7741
7760
7770
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
Three
7040
7041
7060
7061
7080
7090
7091
7130
7131
7140
7190
7191
7195
7196
7197
7198
7200
7201
7210
7380
7420
7421
7450
7460
7470
7471
7480
7481
7520
7550
7610
7740
7741
7760
7770
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
METHOD INDEX - 2
Revision 0
Date September 1986
-------
METHOD INDEX AND CONVERSION TABLE
(Continued)
Method Number,
Third Edition
Chapter Number,
Third Edition
Method Number,
Second Edition
Current Revision
Number
7840
7841
7870
7910
7911
7950
8000
8010
8015
8020
8030
8040
8060
8080
8090
8100
8120
8140
8150
8240
8250
8270
8280
8310
9010
9020
9022
9030
9035
9036
9038
9040
9041
9045
9050
Three
Three
Three
Three
Three
Three
Four (4.3.1)
Four (4.3.1)
Four (4.3.1)
Four (4.3.1)
Four (4.3.1)
Four (4.3.1)
Four (4.3.1)
Four (4.3.1)
Four (4.3.1)
Four (4.3.1)
Four (4.3.1)
Four (4.3.1)
Four (4.3.1)
Four (4.3.2)
Four (4.3.2)
Four (4.3.2)
Four (4.3.2)
Four (4.3.3)
Five
Five
Five
Five
Five
Five
Five
Six
Six
Six
Six
7840 0
7841 0
7870 0
7910 0
7911 0
7950 0
None (new method) 0
8010 0
8015 0
8020 0
8030 0
8040 0
8060 0
8080 0
8090 0
8100 0
8120 0
8140 0
8150 0
8240 0
8250 0
8270 0
None (new method) 0
8310 0
9010 0
9020 0
9022 0
9030 0
9035 0
9036 0
9038 0
9040 0
9041 0
9045 0
9050 0
METHOD INDEX - 3
Revision 0
Date September 1986
-------
METHOD INDEX AND CONVERSION TABLE
(Continued)
Method Number,
Third Edition
Chapter Number,
Third Edition
Method Number,
Second Edition
Current Revision
Number
9060 Five
9065 Five
•9066 Five
9067 Five
9070 Five
9071 Five
9080 Six
9081 Six
9090 Six
9095 Six
9100 Six
9131 Five
9132 Five
9200 Five
9250 Five
9251 Five
9252 Five
9310 Six
9315 Six
9320 Five
HCN Test Method Seven
Test Method Seven
9060
9065
9066
9067
9070
9071
9080
9081
9090
9095
9100
9131
9132
9200
9250
9251
9252
9310
9315
9320
HCN Test Method
Test Method
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
METHOD INDEX - 4
Revision 0
Date September 1986
-------
CHAPTER ONE
TABLE OF CONTENTS
Section
1.0
2.0
3.0
INTRODUCTION
QA PROJECT PLAN
2.1 DATA QUALITY OBJECTIVES
2.2 PROJECT OBJECTIVES
2.3 SAMPLE COLLECTION
2.4 ANALYSIS AND TESTING
2.5 QUALITY CONTROL
2.6 PROJECT DOCUMENTATION
2.7 ORGANIZATION PERFORMING FIELD OR LABORATORY
OPERATIONS
2.7.1 Performance Evaluation
2.7.2 Internal Assessment by QA Function
2.7.3 External Assessment
2.7.4 On-Site Evaluation
2.7.4.1 Field Activities
2.7.4.2 Laboratory Activities
2.7.5 QA Reports
FIELD OPERATIONS
3.1 FIELD LOGISTICS
3.2 EQUIPMENT/ INSTRUMENTATION
3.3 OPERATING PROCEDURES
3.3.1 Sample Management
3.3.2 Reagent/Standard Preparation
3.3.3 Decontamination
3.3.4 Sample Collection
3.3.5 Field Measurements
3.3.6 Equipment Calibration And Maintenance . . . .
3.3.7 Corrective Action
3.3.8 Data Reduction and Validation
3.3.9 Reporting
3.3.10 Records Management
3.3.11 Waste Disposal
3.4 FIELD QA AND QC REQUIREMENTS
3.4.1 Control Samples
3.4.2 Acceptance Criteria
3.4.3 Deviations
3.4.4 Corrective Action
3.4.5 Data Handling
3.5 QUALITY ASSURANCE REVIEW
3.6 FIELD RECORDS
Page
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.... 3
.... 4
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.... 5
.... 5
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.... 5
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ONE - i Revision 1
July 1992
-------
TABLE OF CONTENTS
(continued)
Section
4.0
5.0
6.0
LABORATORY OPERATIONS
4.1 FACILITIES
4.2 EQUIPMENT/ INSTRUMENTATION
4.3 OPERATING PROCEDURES
4.3.1 Sample Management
4.3.2 Reagent/Standard Preparation
4.3.3 General Laboratory Techniques
4.3.4 Test Methods
4.3.5 Equipment Calibration and Maintenance . . . .
4.3.6 QC
4.3.7 Corrective Action
4.3.8 Data Reduction and Validation
4.3.9 Reporting
4.3.10 Records Management
4.3.11 Waste Disposal
4.4 LABORATORY QA AND QC PROCEDURES
4.4.1 Method Proficiency
4.4.2 Control Limits
4.4.3 Laboratory Control Procedures
4.4.4 Deviations
4.4.5 Corrective Action
4.4.6 Data Handling
4.5 QUALITY ASSURANCE REVIEW
4.6 LABORATORY RECORDS
DEFINITIONS
REFERENCES
INDEX
Page
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.... 15
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.... 16
.... 16
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ONE - ii Revision 1
July 1992
-------
CHAPTER ONE
QUALITY CONTROL
1.0 INTRODUCTION
It is the goal of the U.S. Environmental Protection Agency's (EPA's)
quality assurance (QA) program to ensure that all data be scientifically valid,
defensible, and of known precision and accuracy. The data should be of
sufficient known quality to withstand scientific and legal challenge relative to
the use for which the data are obtained. The QA program is management's tool for
achieving this goal.
For RCRA analyses, the recommended minimum requirements for a QA program
and the associated quality control (QC) procedures are provided in this chapter.
The data acquired from QC procedures are used to estimate the quality of
analytical data, to determine the need for corrective action in response to
identified deficiencies, and to interpret results after corrective action
procedures are implemented. Method-specific QC procedures are incorporated in
the individual methods since they are not applied universally.
A total program to generate data of acceptable quality should include both
a QA component, which encompasses the management procedures and controls, as well
as an operational day-to-day QC component. This chapter defines fundamental
elements of such a data collection program. Data collection efforts involve:
1. design of a project plan to achieve the data quality objectives
(DQOs);
2. implementation of the project plan; and
3. assessment of the data to determine if the DQOs are met.
The project plan may be a sampling and analysis plan or a waste analysis plan if
it covers the QA/QC goals of the Chapter, or it may be a Quality Assurance
Project Plan as described later in this chapter.
This chapter identifies the minimal QC components that should be used in
the performance of sampling and analyses, including the QC information which
should be documented. Guidance is provided to construct QA programs for field
and laboratory work conducted in support of the RCRA program.
2.0 QA PROJECT PLAN
It is recommended that all projects which generate environment-related data
in support of RCRA have a QA Project Plan (QAPjP) or equivalent. In some
instances, a sampling and analysis plan or a waste analysis plan may be
equivalent if it covers all of the QA/QC goals outlined in this chapter. In
addition, a separate QAPjP need not be prepared for routine analyses or
activities where the procedures to be followed are described in a Standard
ONE - 1 Revision 1
July 1992
-------
Operating Procedures manual or similar document and include the elements of a
QAPjP. These documents should be available and referenced in the documentation
and/or records for the analysis activities. The term "QAPjP" in this chapter
refers to any of these QA/QC documents.
The QAPjP should detail the QA/QC goals and protocols for a specific data
collection activity. The QAPjP sets forth a plan for sampling and analysis
activities that will generate data of a quality commensurate with their intended
use. QAPjP elements should include a description of the project and its
objectives; a statement of the DQOs of the project; identification of those in-
volved in the data collection and their responsibilities and authorities;
reference to (or inclusion of) the specific sample collection and analysis
procedures that will be followed for all aspects of the project; enumeration of
QC procedures to be followed; and descriptions of all project documentation.
Additional elements should be included in the QAPjP if needed to address all
quality related aspects of the data collection project. Elements should be
omitted only when they are inappropriate for the project or when absence of those
elements will not affect the quality of data obtained for the project (see
reference 1).
The role and importance of DQOs and project documentation are discussed
below in Sections 2.1 through 2.6. Management and organization play a critical
role in determining the effectiveness of a QA/QC program and ensuring that all
required procedures are followed. Section 2.7 discusses the elements of an
organization's QA program that have been found to ensure an effective program.
Field operations and laboratory operations (along with applicable QC procedures)
are discussed in Sections 3 and 4, respectively.
2.1 DATA QUALITY OBJECTIVES
Data quality objectives (DQOs) for the data collection activity describe
the overall level of uncertainty that a decision-maker is willing to accept in
results derived from environmental data. This uncertainty is used to specify the
quality of the measurement data required, usually in terms of objectives for
precision, bias, representativeness, comparability and completeness. The DQOs
should be defined prior to the initiation of the field and laboratory work. The
field and laboratory organizations performing the work should be aware of the
DQOs so that their personnel may make informed decisions during the course of the
project to attain those DQOs. More detailed information on DQOs is available
from the U.S. EPA Quality Assurance Management Staff (QAMS) (see references 2 and
4).
2.2 PROJECT OBJECTIVES
A statement of the project objectives and how the objectives are to be
attained should be concisely stated and sufficiently detailed to permit clear
understanding by all parties involved in the data collection effort. This
includes a statement of what problem is to be solved and the information required
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in the process. It also includes appropriate statements of the DQOs (i.e., the
acceptable level of uncertainty in the information).
2.3 SAMPLE COLLECTION
Sampling procedures, locations, equipment, and sample preservation and
handling requirements should be specified in the QAPjP. Further details on
quality assurance procedures for field operations are described in Section 3 of
this chapter. The OSW is developing policies and procedures for sampling in a
planned revision of Chapter Nine of this manual. Specific procedures for
groundwater sampling are provided in Chapter Eleven of this manual.
2.4 ANALYSIS AND TESTING
Analytes and properties of concern, analytical and testing procedures to
be employed, required detection limits, and requirements for precision and bias
should be specified. All applicable regulatory requirements and the project DQOs
should be considered when developing the specifications. Further details on the
procedures for analytical operations are described in Section 4 of this chapter.
2.5 QUALITY CONTROL
The quality assurance program should address both field and laboratory
activities. Quality control procedures should be specified for estimating the
precision and bias of the data. Recommended minimum requirements for QC samples
have been established by EPA and should be met in order to satisfy recommended
minimum criteria for acceptable data quality. Further details on procedures for
field and laboratory operations are described in Sections 3 and 4, respectively,
of this chapter.
2.6 PROJECT DOCUMENTATION
Documents should be prepared and maintained in conjunction with the data
collection effort. Project documentation should be sufficient to allow review
of all aspects of the work being performed. The QAPjP discussed in Sections 3
and 4 is one important document that should be maintained.
The length of storage time for project records should comply with
regulatory requirements, organizational policy, or project requirements,
whichever is more stringent. It is recommended that documentation be stored for
three years from submission of the project final report.
Documentation should be secured in a facility that adequately
addresses/minimizes its deterioration for the length of time that it is to be
retained. A system allowing for the expedient retrieval of information should
exist.
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Access to archived information should be controlled to maintain the
integrity of the data. Procedures should be developed to identify those
individuals with access to the data.
2.7 ORGANIZATION PERFORMING FIELD OR LABORATORY OPERATIONS
Proper design and structure of the organization facilitates effective and
efficient transfer of information and helps to prevent important procedures from
being overlooked.
The organizational structure, functional responsibilities, levels of
authority, job descriptions, and lines of communication for all project
activities should be established and documented. One person may cover more than
one organizational function. Each project participant should have a clear
understanding of his or her duties and responsibilities and the relationship of
those responsibilities to the overall data collection effort.
The management of each organization participating in a project involving
data collection activities should establish that organization's operational and
QA policies. This information should be documented in the QAPjP. The management
should ensure that (1) the appropriate methodologies are followed as documented
in the QAPjPs; (2) personnel clearly understand their duties and
responsibilities; (3) each staff member has access to appropriate project
documents; (4) any deviations from the QAPjP are communicated to the project
management and documented; and (5) communication occurs between the field,
laboratory, and project management, as specified in the QAPjP. In addition, each
organization should ensure that their activities do not increase the risk to
humans or the environment at or about the project location. Certain projects may
require specific policies or a Health and Safety Plan to provide this assurance.
The management of the participating field or laboratory organization should
establish personnel qualifications and training requirements for the project.
Each person participating in the project should have the education, training,
technical knowledge, and experience, or a combination thereof, to enable that
individual to perform assigned functions. Training should be provided for each
staff member as necessary to perform their functions properly. Personnel
qualifications should be documented in terms of education, experience, and
training, and periodically reviewed to ensure adequacy to current
responsibilities.
Each participating field organization or laboratory organization should
have a designated QA function (i.e., a team or individual trained in QA) to
monitor operations to ensure that the equipment, personnel, activities,
procedures, and documentation conform with the QAPjP. To the extent possible,
the QA monitoring function should be entirely separate from, and independent of,
personnel engaged in the work being monitored. The QA function should be
responsible for the QA review.
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2.7.1 Performance Evaluation
Performance evaluation studies are used to measure the performance of the
laboratory on unknown samples. Performance evaluation samples are typically
submitted to the laboratory as blind samples by an independent outside source.
The results are compared to predetermined acceptance limits. Performance
evaluation samples can also be submitted to the laboratory as part of the QA
function during internal assessment of laboratory performance. Records of all
performance evaluation studies should be maintained by the laboratory. Problems
identified through participation in performance evaluation studies should be
immediately investigated and corrected.
2.7.2 Internal Assessment by QA Function
Personnel performing field and laboratory activities are responsible for
continually monitoring individual compliance with the QAPjP. The QA function
should review procedures, results and calculations to determine compliance with
the QAPjP. The results of this internal assessment should be reported to
management with requirements for a plan to correct observed deficiencies.
2.7.3 External Assessment
The field and laboratory activities may be reviewed by personnel external
to the organization. Such an assessment is an extremely valuable method for
identifying overlooked problems. The results of the external assessment should
be submitted to management with requirements for a plan to correct observed
deficiencies.
2.7.4 On-Site Evaluation
On-site evaluations may be conducted as part of both internal and external
assessments. The focus of an on-site evaluation is to evaluate the degree of
conformance of project activities with the applicable QAPjP. On-site evaluations
may include, but are not limited to, a complete review of facilities, staff,
training, instrumentation, procedures, methods, sample collection, analyses, QA
policies and procedures related to the generation of environmental data. Records
of each evaluation should include the date of the evaluation, location, the areas
reviewed, the person performing the evaluation, findings and problems, and
actions recommended and taken to resolve problems. Any problems identified that
are likely to affect data integrity should be brought immediately to the
attention of management.
2.7.4.1 Field Activities
The review of field activities should be conducted by one or more persons
knowledgeable in the activities being reviewed and include evaluating, at a
minimum, the following subjects:
Completeness of Field Reports -- This review determines whether all
requirements for field activities in the QAPjP have been fulfilled, that
complete records exist for each field activity, and that the procedures
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specified in the QAPjP have been implemented. Emphasis on field
documentation will help assure sample integrity and sufficient technical
information to recreate each field event. The results of this
completeness check should be documented, and environmental data affected
by incomplete records should be identified.
Identification of Valid Samples -- This review involves interpretation and
evaluation of the field records to detect problems affecting the repre-
sentativeness of environmental samples. Examples of items that might
indicate potentially invalid samples include improper well development,
improperly screened wells, instability of pH or conductivity, and collec-
tion of volatiles near internal combustion engines. The field records
should be evaluated against the QAPjP and SOPs. The reviewer should docu-
ment the sample validity and identify the environmental data associated
with any poor or incorrect field work.
Correlation of Field Test Data -- This review involves comparing any
available results of field measurements obtained by more than one method.
For example, surface geophysical methods should correlate with direct
methods of site geologic characterization such as lithologic logs
constructed during drilling operations.
Identification of Anomalous Field Test Data -- This review identifies any
anomalous field test data. For example, a water temperature for one well
that is 5 degrees higher than any other well temperature in the same
aquifer should be noted. The reviewer should evaluate the impact of
anomalous field measurement results on the associated environmental data.
Validation of Field Analyses -- This review validates and documents all
data from field analysis that are generated in situ or from a mobile
laboratory as specified in Section 2.7.4.2. The reviewer should document
whether the QC checks meet the acceptance criteria, and whether corrective
actions were taken for any analysis performed when acceptance criteria
were exceeded.
2.7.4.2 Laboratory Activities
The review of laboratory data should be conducted by one or more persons
knowledgeable in laboratory activities and include evaluating, at a minimum, the
following subjects:
Completeness of Laboratory Records -- This review determines whether: (1)
all samples and analyses required by the QAPjP have been processed, (2)
complete records exist for each analysis and the associated QC samples,
and that (3) the procedures specified in the QAPjP have been implemented.
The results of the completeness check should be documented, and
environmental data affected by incomplete records should be identified.
Evaluation of Data with Respect to Detection and Quantitation Limits --
This review compares analytical results to required quantitation limits.
Reviewers should document instances where detection or quantitation limits
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exceed regulatory limits, action levels, or target concentrations
specified in the QAPjP.
Evaluation of Data with Respect to Control Limits -- This review compares
the results of QC and calibration check samples to control criteria.
Corrective action should be implemented for data not within control
limits. The reviewer should check that corrective action reports, and the
results of reanalysis, are available. The review should determine
whether samples associated with out-of-control QC data are identified in
a written record of the data review, and whether an assessment of the
utility of such analytical results is recorded.
Review of Holding Time Data -- This review compares sample holding times
to those required by the QAPjP, and notes all deviations.
Review of Performance Evaluation (PE) Results -- PE study results can be
helpful in evaluating the impact of out-of-control conditions. This review
documents any recurring trends or problems evident in PE studies and
evaluates their effect on environmental data.
Correlation of Laboratory Data -- This review determines whether the
results of data obtained from related laboratory tests, e.g., Purgeable
Organic Hal ides (POX) and Volatile Organics, are documented, and whether
the significance of any differences is discussed in the reports.
2.7.5 QA Reports
There should be periodic reporting of pertinent QA/QC information to the
project management to allow assessment of the overall effectiveness of the QA
program. There are three major types of QA reports to project management:
Periodic Report on Key QA Activities -- Provides summary of key QA activi-
ties during the period, stressing measures that are being taken to improve
data quality; describes significant quality problems observed and
corrective actions taken; reports information regarding any changes in
certification/accreditation status; describes involvement in resolution of
quality issues with clients or agencies; reports any QA organizational
changes; and provides notice of the distribution of revised documents
controlled by the QA organization (i.e., procedures).
Report on Measurement Quality Indicators -- Includes the assessment of QC
data gathered over the period, the frequency of analyses repeated due to
unacceptable QC performance, and, if possible, the reason for the unac-
ceptable performance and corrective action taken.
Reports on QA Assessments -- Includes the results of the assessments and
the plan for correcting identified deficiencies; submitted immediately
following any internal or external on-site evaluation or upon receipt of
the results of any performance evaluation studies.
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3.0 FIELD OPERATIONS
The field operations should be conducted in such a way as to provide
reliable information that meets the DQOs. To achieve this, certain minimal
policies and procedures should be implemented. The OSW is considering revisions
of Chapter Nine and Eleven of this manual. Supplemental information and guidance
is available in the RCRA Ground-Water Monitoring Technical Enforcement Guidance
Document (TEGD) (Reference 3). The project documentation should contain the
information specified below.
3.1 FIELD LOGISTICS
The QAPjP should describe the type(s) of field operations to be performed
and the appropriate area(s) in which to perform the work. The QAPjP should
address ventilation, protection from extreme weather and temperatures, access to
stable power, and provision for water and gases of required purity.
Whenever practical, the sampling site facilities should be examined prior
to the start of work to ensure that all required items are available. The actual
area of sampling should be examined to ensure that trucks, drilling equipment,
and personnel have adequate access to the site.
The determination as to whether sample shipping is necessary should be made
during planning for the project. This need is established by evaluating the
analyses to be performed, sample holding times, and location of the site and the
laboratory. Shipping or transporting of samples to a laboratory should be done
within a timeframe such that recommended holding times are met.
Samples should be packaged, labelled, preserved (e.g., preservative added,
iced, etc.), and documented in an area which is free of contamination and
provides for secure storage. The level of custody and whether sample storage is
needed should be addressed in the QAPjP.
Storage areas for solvents, reagents, standards, and reference materials
should be adequate to preserve their identity, concentration, purity, and
stability prior to use.
Decontamination of sampling equipment may be performed at the location
where sampling occurs, prior to going to the sampling site, or in designated
areas near the sampling site. Project documentation should specify where and how
this work is accomplished. If decontamination is to be done at the site, water
and solvents of appropriate purity should be available. The method of
accomplishing decontamination, including the required materials, solvents, and
water purity should be specified.
During the sampling process and during on-site or in situ analyses, waste
materials are sometimes generated. The method for storage and disposal of these
waste materials that complies with applicable local, state and Federal
regulations should be specified. Adequate facilities should be provided for the
collection and storage of all wastes, and these facilities should be operated so
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as to minimize environmental contamination. Waste storage and disposal
facilities should comply with applicable federal, state, and local regulations.
The location of long-term and short-term storage for field records, and the
measures to ensure the integrity of the data should be specified.
3.2 EQUIPMENT/INSTRUMENTATION
The equipment, instrumentation, and supplies at the sampling site should
be specified and should be appropriate to accomplish the activities planned. The
equipment and instrumentation should meet the requirements of specifications,
methods, and procedures as specified in the QAPjP.
3.3 OPERATING PROCEDURES
The QAPjP should describe or make reference to all field activities that
may affect data quality. For routinely performed activities, standard operating
procedures (SOPs) are often prepared to ensure consistency and to save time and
effort in preparing QAPjPs. Any deviation from an established procedure during
a data collection activity should be documented. The procedures should be
available for the indicated activities, and should include, at a minimum, the
information described below.
3.3.1 Sample Management
The numbering and labeling system, chain-of-custody procedures, and how the
samples are to be tracked from collection to shipment or receipt by the
laboratory should be specified. Sample management procedures should also specify
the holding times, volumes of sample required by the laboratory, required
preservatives, and shipping requirements.
3.3.2 Reagent/Standard Preparation
The procedures describing how to prepare standards and reagents should be
specified. Information concerning specific grades of materials used in reagent
and standard preparation, appropriate glassware and containers for preparation
and storage, and labeling and record keeping for stocks and dilutions should be
included.
3.3.3 Decontamination
The procedures describing decontamination of field equipment before and
during the sample collection process should be specified. These procedures
should include cleaning materials used, the order of washing and rinsing with the
cleaning materials, requirements for protecting or covering cleaned equipment,
and procedures for disposing of cleaning materials.
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3.3.4 Sample Collection
The procedures describing how the sampling operations are actually
performed in the field should be specified. A simple reference to standard
methods is not sufficient, unless a procedure is performed exactly as described
in the published method. Methods from source documents published by the EPA,
American Society for Testing and Materials, U.S. Department of the Interior,
National Water Well Association, American Petroleum Institute, or other
recognized organizations with appropriate expertise should be used, if possible.
The procedures for sample collection should include at least the following:
• Applicability of the procedure,
Equipment required,
• Detailed description of procedures to be followed in collecting the
samples,
• Common problems encountered and corrective actions to be followed, and
• Precautions to be taken.
3.3.5 Field Measurements
The procedures describing all methods used in the field to determine a
chemical or physical parameter should be described in detail. The procedures
should address criteria from Section 4, as appropriate.
3.3.6 Equipment Calibration And Maintenance
The procedures describing how to ensure that field equipment and
instrumentation are in working order should be specified. These describe
calibration procedures and schedules, maintenance procedures and schedules,
maintenance logs, and service arrangements for equipment. Calibration and
maintenance of field equipment and instrumentation should be in accordance with
manufacturers' specifications or applicable test specifications and should be
documented.
3.3.7 Corrective Action
The procedures describing how to identify and correct deficiencies in the
sample collection process should be specified. These should include specific
steps to take in correcting deficiencies such as performing additional
decontamination of equipment, resampling, or additional training of field
personnel. The procedures should specify that each corrective action should be
documented with a description of the deficiency and the corrective action taken,
and should include the person(s) responsible for implementing the corrective
action.
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3.3.8 Data Reduction and Validation
The procedures describing how to compute results from field measurements
and to review and validate these data should be specified. They should include
all formulas used to calculate results and procedures used to independently
verify that field measurement results are correct.
3.3.9 Reporting
The procedures describing the process for reporting the results of field
activities should be specified.
3.3.10 Records Management
The procedures describing the means for generating, controlling, and
archiving project-specific records and field operations records should be
specified. These procedures should detail record generation and control and the
requirements for record retention, including type, time, security, and retrieval
and disposal authorities.
Project-specific records relate to field work performed for a project.
These records may include correspondence, chain-of-custody records, field
notes, all reports issued as a result of the work, and procedures used.
Field operations records document overall field operations and may include
equipment performance and maintenance logs, personnel files, general field
procedures, and corrective action reports.
3.3.11 Waste Disposal
The procedures describing the methods for disposal of waste materials
resulting from field operations should be specified.
3.4 FIELD QA AND QC REQUIREMENTS
The QAPjP should describe how the following elements of the field QC
program will be implemented.
3.4.1 Control Samples
Control samples are QC samples that are introduced into a process to
monitor the performance of the system. Control samples, which may include blanks
(e.g., trip, equipment, and laboratory), duplicates, spikes, analytical
standards, and reference materials, can be used in different phases of the data
collection process beginning with sampling and continuing through transportation,
storage, and analysis.
Each day of sampling, at least one field duplicate and one equipment
rinsate should be collected for each matrix sampled. If this frequency is not
appropriate for the sampling equipment and method, then the appropriate changes
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should be cTearly identified in the QAPjP. When samples are collected for
volatile organic analysis, a trip blank is also recommended for each day that
samples are collected. In addition, for each sampling batch (20 samples of one
matrix type), enough volume should be collected for at least one sample so as to
allow the laboratory to prepare one matrix spike and either one matrix duplicate
or one matrix spike duplicate for each analytical method employed. This means
that the following control samples are recommended:
•Field duplicate (one per day per matrix type)
•Equipment rinsate (one per day per matrix type)
•Trip blank (one per day, volatile organics only)
•Matrix spike (one per batch [20 samples of each matrix type])
•Matrix duplicate or matrix spike duplicate (one per batch)
Additional control samples may be necessary in order to assure data quality to
meet the project-specific DQOs. '
3.4.2 Acceptance Criteria
Procedures should be in place for establishing acceptance criteria for
field activities described in the QAPjP. Acceptance criteria may be qualitative
or quantitative. Field events or data that fall outside of established
acceptance criteria may indicate a problem with the sampling process that should
be investigated.
3.4.3 Deviations
All deviations from plan should be documented as to the extent of, and
reason for, the deviation. Any activity not performed in accordance with
procedures or QAPjPs is considered a deviation from plan. Deviations from plan
may or may not affect data quality.
3.4.4 Corrective Action
Errors, deficiencies, deviations, certain field events, or data that fall
outside established acceptance criteria should be investigated. In some in-
stances, corrective action may be needed to resolve the problem and restore
proper functioning to the system. The investigation of the problem and any
subsequent corrective action taken should be documented.
3.4.5 Data Handling
All field measurement data should be reduced according to protocols
described or referenced in the QAPjP. Computer programs used for data reduction
should be validated before use and verified on a regular basis. All information
used in the calculations should be recorded to enable reconstruction of the final
result at a later date.
Data should be reported in accordance with the requirements of the end-user
as described in the QAPjP.
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3.5 QUALITY ASSURANCE REVIEW
The QA Review consists of internal and external assessments to ensure that
QA/QC procedures are in use and to ensure that field staff conform to these
procedures. QA review should be conducted as deemed appropriate and necessary.
3.6 FIELD RECORDS
Records provide the direct evidence and support for the necessary technical
interpretations, judgments, and discussions concerning project activities. These
records, particularly those that are anticipated to be used as evidentiary data,
should directly support current or ongoing technical studies and activities and
should provide the historical evidence needed for later reviews and analyses.
Records should be legible, identifiable, and retrievable and protected against
damage, deterioration, or loss. The discussion in this section (3.6) outlines
recommended procedures for record keeping. Organizations which conduct field
sampling should develop appropriate record keeping procedures which satisfy
relevant technical and legal requirements.
Field records generally consist of bound field notebooks with prenumbered
pages, sample collection forms, personnel qualification and training forms,
sample location maps, equipment maintenance and calibration forms, chain-of-
custody forms, sample analysis request forms, and field change request forms.
All records should be written in indelible ink.
Procedures for reviewing, approving, and revising field records should be
clearly defined, with the lines of authority included. It is recommended that
all documentation errors should be corrected by drawing a single line through the
error so it remains legible and should be initialed by the responsible
individual, along with the date of change. The correction should be written
adjacent to the error.
Records should include (but are not limited to) the following:
Calibration Records & Traceability of Standards/Reagents -- Calibration is
a reproducible reference point to which all sample measurements can be
correlated. A sound calibration program should include provisions for
documentation of frequency, conditions, standards, and records reflecting
the calibration history of a measurement system. The accuracy of the
calibration standards is important because all data will be in reference
to the standards used. A program for verifying and documenting the
accuracy of all working standards against primary grade standards should
be routinely followed.
Sample Collection -- To ensure maximum utility of the sampling effort and
resulting data, documentation of the sampling protocol, as performed in
the field, is essential. It is recommended that sample collection records
contain, at a minimum, the names of persons conducting the activity,
sample number, sample location, equipment used, climatic conditions,
documentation of adherence to protocol, and unusual observations. The
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actual sample collection record is usually one of the following: a bound
field notebook with prenumbered pages, a pre-printed form, or digitized
information on a computer tape or disc.
Chain-of-Custody Records -- The chain-of-custody involving the possession
of samples from the time they are obtained until they are disposed or
shipped off-site should be documented as specified in the QAPjP and should
include the following information: (1) the project name; (2) signatures
of samplers; (3) the sample number, date and time of collection, and grab
or composite sample designation; (4) signatures of individuals involved in
sample transfer; and (5) if applicable, the air bill or other shipping
number.
Maps and Drawings -- Project planning documents and reports often contain
maps. The maps are used to document the location of sample collection
points and monitoring wells and as a means of presenting environmental
data. Information used to prepare maps and drawings is normally obtained
through field surveys, property surveys, surveys of monitoring wells,
aerial photography or photogrammetric mapping. The final, approved maps
and/or drawings should have a revision number and date and should be sub-
ject to the same controls as other project records.
QC Samples -- Documentation for generation of QC samples, such as trip and
equipment rinsate blanks, duplicate samples, and any field spikes should
be maintained.
Deviations -- All deviations from procedural documents and the QAPjP
should be recorded in the site logbook.
Reports -- A copy of any report issued and any supporting documentation
should be retained.
4.0 LABORATORY OPERATIONS
The laboratory should conduct its operations in such a way as to provide
reliable information. To achieve this, certain minimal policies and procedures
should be implemented.
4.1 FACILITIES
The QAPjP should address all facility-related issues that may impact
project data quality. Each laboratory should be of suitable size and
construction to facilitate the proper conduct of the analyses. Adequate bench
space or working area per analyst should be provided. The space requirement per
analyst depends on the equipment or apparatus that is being utilized, the number
of samples that the analyst is expected to handle at any one time, and the number
of operations that are to be performed concurrently by a single analyst. Other
issues to be considered include, but are not limited to, ventilation, lighting,
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control of dust and drafts, protection from extreme temperatures, and access to
a source of stable power.
Laboratories should be designed so that there is adequate separation of
functions to ensure that no laboratory activity has an adverse effect on the
analyses. The laboratory may require specialized facilities such as a perchloric
acid hood or glovebox.
Separate space for laboratory operations and appropriate ancillary support
should be provided, as needed, for the performance of routine and specialized
procedures.
As necessary to ensure secure storage and prevent contamination or
misidentification, there should be adequate facilities for receipt and storage
of samples. The level of custody required and any special requirements for
storage such as refrigeration should be described in planning documents.
Storage areas for reagents, solvents, standards, and reference materials
should be adequate to preserve their identity, concentration, purity, and
stability.
Adequate facilities should be provided for the collection and storage of
all wastes, and these facilities should be operated so as to minimize environ-
mental contamination. Waste storage and disposal facilities should comply with
applicable federal, state, and local regulations.
The location of long-term and short-term storage of laboratory records and
the measures to ensure the integrity of the data should be specified.
4.2 EQUIPMENT/INSTRUMENTATION
Equipment and instrumentation should meet the requirements and specifica-
tions of the specific test methods and other procedures as specified in the
QAPjP. The laboratory should maintain an equipment/instrument description list
that includes the manufacturer, model number, year of purchase, accessories, and
any modifications, updates, or upgrades that have been made.
4.3 OPERATING PROCEDURES
The QAPjP should describe or make reference to all laboratory activities
that may affect data quality. For routinely performed activities, SOPs are often
prepared to ensure consistency and to save time and effort in preparing QAPjPs.
Any deviation from an established procedure during a data collection activity
should be documented. It is recommended that procedures be available for the
indicated activities, and include, at a minimum, the information described
below.
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4.3.1 Sample Management
The procedures describing the receipt, handling, scheduling, and storage
of samples should be specified.
Sample Receipt and Handling -- These procedures describe the precautions
to be used in opening sample shipment containers and how to verify that
chain-of-custody has been maintained, examine samples for damage, check
for proper preservatives and temperature, and log samples into the
laboratory sample streams.
Sample Scheduling -- These procedures describe the sample scheduling in
the laboratory and includes procedures used to ensure that holding time
requirements are met.
Sample Storage -- These procedures describe the storage conditions for all
samples, verification and documentation of daily storage temperature, and
how to ensure that custody of the samples is maintained while in the
laboratory.
4.3.2 Reagent/Standard Preparation
The procedures describing how to prepare standards and reagents should be
specified. Information concerning specific grades of materials used in reagent
and standard preparation, appropriate glassware and containers for preparation
and storage, and labeling and recordkeeping for stocks and dilutions should be
included.
4.3.3 General Laboratory Technigues
The procedures describing all essentials of laboratory operations that are
not addressed elsewhere should be specified. These techniques should include,
but are not limited to, glassware cleaning procedures, operation of analytical
balances, pipetting techniques, and use of volumetric glassware.
4.3.4 Test Methods
Procedures for test methods describing how the analyses are actually
performed in the laboratory should be specified. A simple reference to standard
methods is not sufficient, unless the analysis is performed exactly as described
in the published method. Whenever methods from SW-846 are not appropriate,
recognized methods from source documents published by the EPA, American Public
Health Association (APHA), American Society for Testing and Materials (ASTM), the
National Institute for Occupational Safety and Health (NIOSH), or other
recognized organizations with appropriate expertise should be used, if possible.
The documentation of the actual laboratory procedures for analytical methods
should include the following:
Sample Preparation and Analysis Procedures -- These include applicable
holding time, extraction, digestion, or preparation steps as appropriate
to the method; procedures for determining the appropriate dilution to
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analyze; and any other information required to perform the analysis
accurately and consistently.
Instrument Standardization -- This includes concentration(s) and frequency
of analysis of calibration standards, linear range of the method, and
calibration acceptance criteria.
Sample Data -- This includes recording requirements and documentation in-
cluding sample identification number, analyst, data verification, date of
analysis and verification, and computational method(s).
Precision and Bias -- This includes all analytes for which the method is
applicable and the conditions for use of this information.
Detection and Reporting Limits -- This includes all analytes in the
method.
Test-Specific QC -- This describes QC activities applicable to the
specific test and references any applicable QC procedures.
4.3.5 Equipment Calibration and Maintenance
The procedures describing how to ensure that laboratory equipment and
instrumentation are in working order should be specified. These procedures
include calibration procedures and schedules, maintenance procedures and
schedules, maintenance logs, service arrangements for all equipment, and spare
parts available in-house. Calibration and maintenance of laboratory equipment
and instrumentation should be in accordance with manufacturers' specifications
or applicable test specifications and should be documented.
4.3.6 QC
The type, purpose, and frequency of QC samples to be analyzed in the
laboratory and the acceptance criteria should be specified. Information should
include the applicability of the QC sample to the analytical process, the
statistical treatment of the data, and the responsibility of laboratory staff and
management in generating and using the data. Further details on development of
project-specific QC protocols are described in Section 4.4.
4.3.7 Corrective Action
The procedures describing how to identify and correct deficiencies in the
analytical process should be specified. These should include specific steps to
take in correcting the deficiencies such as preparation of new standards and
reagents, recalibration and restandardization of equipment, reanalysis of
samples, or additional training of laboratory personnel in methods and
procedures. The procedures should specify that each corrective action should be
documented with a description of the deficiency and the corrective action taken,
and should include the person(s) responsible for implementing the corrective
action.
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4.3.8 Data Reduction and Validation
The procedures describing how to review and validate the data should be
specified. They should include procedures for computing and interpreting the
results from QC samples, and independent procedures to verify that the analytical
results are reported correctly. In addition, routine procedures used to monitor
precision and bias, including evaluations of reagent, equipment rinsate, and trip
blanks, calibration standards, control samples, duplicate and matrix spike
samples, and surrogate recovery, should be detailed in the procedures. More
detailed validation procedures should be performed when required in the contract
or QAPjP.
4.3.9 Reporting
The procedures describing the process for reporting the analytical results
should be specified.
4.3.10 Records Management
The procedures describing the means for generating, controlling, and
archiving laboratory records should be specified. The procedures should detail
record generation and control, and the requirements for record retention, includ-
ing type, time, security, and retrieval and disposal authorities.
Project-specific records may include correspondence, chain-of-custody
records, request for analysis, calibration data records, raw and finished
analytical and QC data, data reports, and procedures used.
Laboratory operations records may include laboratory notebooks, instrument
performance logs and maintenance logs in bound notebooks with prenumbered
pages; laboratory benchsheets; software documentation; control charts;
reference material certification; personnel files; laboratory procedures;
and corrective action reports.
4.3.11 Waste Disposal
The procedures describing the methods for disposal of chemicals including
standard and reagent solutions, process waste, and samples should be specified.
4.4 LABORATORY QA AND QC PROCEDURES
The QAPjP should describe how the following required elements of the
laboratory QC program are to be implemented.
4.4.1 Method Proficiency
Procedures should be in place for demonstrating proficiency with each
analytical method routinely used in the laboratory. These should include
procedures for demonstrating the precision and bias of the method as performed
by the laboratory and procedures for determining the method detection limit
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(MDL). All terminology, procedures and frequency of determinations associated
with the laboratory's establishment of the MDL and the reporting limit should be
well-defined and well-documented. Documented precision, bias, and MDL
information should be maintained for all methods performed in the laboratory.
4.4.2 Control Limits
Procedures should be in place for establishing and updating control limits
for analysis. Control limits should be established to evaluate laboratory
precision and bias based on the analysis of control samples. Typically, control
limits for bias are based on the historical mean recovery plus or minus three
standard deviation units, and control limits for precision range from zero (no
difference between duplicate control samples) to the historical mean relative
percent difference plus three standard deviation units. Procedures should be in
place for monitoring historical performance and should include graphical (control
charts) and/or tabular presentations of the data.
4.4.3 Laboratory Control Procedures
Procedures should be in place for demonstrating that the laboratory is in
control during each data collection activity. Analytical data generated with
laboratory control samples that fall within prescribed limits are judged to be
generated while the laboratory was in control. Data generated with laboratory
control samples that fall outside the established control limits are judged to
be generated during an "out-of-control" situation. These data are considered
suspect and should be repeated or reported with qualifiers.
Laboratory Control Samples -- Laboratory control samples should be
analyzed for each analytical method when appropriate for the method. A
laboratory control sample consists of either a control matrix spiked with
analytes representative of the target analytes or a certified reference
material.
Laboratory control sample(s) should be analyzed with each batch of samples
processed to verify that the precision and bias of the analytical process
are within control limits. The results of the laboratory control
sample(s) are compared to control limits established for both precision
and bias to determine usability of the data.
Method Blank -- When appropriate for the method, a method blank should be
analyzed with each batch of samples processed to assess contamination
levels in the laboratory. Guidelines should be in place for accepting or
rejecting data based on the level of contamination in the blank.
Procedures should be in place for documenting the effect of the matrix on
method performance. When appropriate for the method, there should be at least
one matrix spike and either one matrix duplicate or one matrix spike duplicate
per analytical batch. Additional control samples may be necessary to assure data
quality to meet the project-specific DQOs.
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Matrix-Specific Bias -- Procedures should be in place for determining the
bias of the method due to the matrix. These procedures should include
preparation and analysis of matrix spikes, selection and use of surrogates
for organic methods, and the method of standard additions for metal and
inorganic methods. When the concentration of the analyte in the sample is
greater than 0.1%, no spike is necessary.
Matrix-Specific Precision -- Procedures should be in place for determining
the precision of the method for a specific matrix. These procedures
should include analysis of matrix duplicates and/or matrix spike
duplicates. The frequency of use of these techniques should be based on
the DQO for the data collection activity.
Matrix-Specific Detection Limit -- Procedures should be in place for
determining the MDL for a specific matrix type (e.g., wastewater treatment
sludge, contaminated soil, etc).
4.4.4 Deviations
Any activity not performed in accordance with laboratory procedures or
QAPjPs is considered a deviation from plan. All deviations from plan should be
documented as to the extent of, and reason for, the deviation.
4.4.5 Corrective Action
Errors, deficiencies, deviations, or laboratory events or data that fall
outside of established acceptance criteria should be investigated. In some
instances, corrective action may be needed to resolve the problem and restore
proper functioning to the analytical system. The investigation of the problem
and any subsequent corrective action taken should be documented.
4.4.6 Data Handling
Data resulting from the analyses of samples should be reduced according to
protocols described in the laboratory procedures. Computer programs used for
data reduction should be validated before use and verified on a regular basis.
All information used in the calculations (e.g., raw data, calibration files,
tuning records, results of standard additions, interference check results, and
blank- or background-correction protocols) should be recorded in order to enable
reconstruction of the final result at a later date. Information on the
preparation of the sample (e.g., weight or volume of sample used, percent dry
weight for solids, extract volume, dilution factor used) should also be
maintained in order to enable reconstruction of the final result at a later date.
All data should be reviewed by a second analyst or supervisor according to
laboratory procedures to ensure that calculations are correct and to detect
transcription errors. Spot checks should be performed on computer calculations
to verify program validity. Errors detected in the review process should be
referred to the analyst(s) for corrective action. Data should be reported in
accordance with the requirements of the end-user. It is recommended that the
supporting documentation include at a minimum:
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Laboratory name and address.
Sample information (including unique sample identification, sample
collection date and time, date of sample receipt, and date(s) of sample
preparation and analysis).
Analytical results reported with an appropriate number of significant
figures.
Detection limits that reflect dilutions, interferences, or correction for
equivalent dry weight.
Method reference.
Appropriate QC results (correlation with sample batch should be traceable
and documented).
Data qualifiers with appropriate references and narrative on the quality
of the results.
4.5 QUALITY ASSURANCE REVIEW
The QA review consists of internal and external assessments to ensure that
QA/QC procedures are in use and to ensure that laboratory staff conform to these
procedures. QA review should be conducted as deemed appropriate and necessary.
4.6 LABORATORY RECORDS
Records provide the direct evidence and support for the necessary technical
interpretations, judgements, and discussions concerning project activities.
These records, particularly those that are anticipated to be used as evidentiary
data, should directly support technical studies and activities, and provide the
historical evidence needed for later reviews and analyses. Records should be
legible, identifiable, and retrievable, and protected against damage,
deterioration, or loss. The discussion in this section (4.6) outlines
recommended procedures for record keeping. Organizations which conduct field
sampling should develop appropriate record keeping procedures which satisfy
relevant technical and legal requirements.
Laboratory records generally consist of bound notebooks with prenumbered
pages, personnel qualification and training forms, equipment maintenance and
calibration forms, chain-of-custody forms, sample analysis request forms, and
analytical change request forms. All records should be written in indelible ink.
Procedures for reviewing, approving, and revising laboratory records should
be clearly defined, with the lines of authority included. Any documentation
errors should be corrected by drawing a single line through the error so that it
remains legible and should be initialed by the responsible individual, along with
the date of change. The correction is written adjacent to the error.
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Strip-chart recorder printouts should be signed by the person who performed
the instrumental analysis. If corrections need to be made in computerized data,
a system parallel to the corrections for handwritten data should be in place.
Records of sample management should be available to permit the re-creation
of an analytical event for review in the case of an audit or investigation of a
dubious result.
Laboratory records should include, at least, the following:
Operating Procedures -- Procedures should be available to those performing
the task outlined. Any revisions to laboratory procedures should be
written, dated, and distributed to all affected individuals to ensure
implementation of changes. Areas covered by operating procedures are
given in Sections 3.3 and 4.3.
Quality Assurance Plans -- The QAPjP should be on file.
Equipment Maintenance Documentation -- A history of the maintenance record
of each system serves as an indication of the adequacy of maintenance
schedules and parts inventory. As appropriate, the maintenance guidelines
of the equipment manufacturer should be followed. When maintenance is
necessary, it should be documented in either standard forms or in
logbooks. Maintenance procedures should be clearly defined and written
for each measurement system and required support equipment.
Proficiency -- Proficiency information on all compounds reported should be
maintained and should include (1) precision; (2) bias; (3) method detec-
tion limits; (4) spike recovery, where applicable; (5) surrogate recovery,
where applicable; (6) checks on reagent purity, where applicable; and
(7) checks on glassware cleanliness, where applicable.
Calibration Records & Traceability of Standards/Reagents -- Calibration is
a reproducible reference point to which all sample measurements can be
correlated. A sound calibration program should include provisions for
documenting frequency, conditions, standards, and records reflecting the
calibration history of a measurement system. The accuracy of the
calibration standards is important because all data will be in reference
to the standards used. A program for verifying and documenting the
accuracy and traceability of all working standards against appropriate
primary grade standards or the highest quality standards available should
be routinely followed.
Sample Management --All required records pertaining to sample management
should be maintained and updated regularly. These include chain-of-
custody forms, sample receipt forms, and sample disposition records.
Original Data -- The raw data and calculated results for all samples
should be maintained in laboratory notebooks, logs, benchsheets, files or
other sample tracking or data entry forms. Instrumental output should be
stored in a computer file or a hardcopy report.
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QC Data -- The raw data and calculated results for all QC and field
samples and standards should be maintained in the manner described in the
preceding paragraph. Documentation should allow correlation of sample
results with associated QC data. Documentation should also include the
source and lot numbers of standards for traceability. QC samples include,
but are not limited to, control samples, method blanks, matrix spikes, and
matrix spike duplicates.
Correspondence -- Project correspondence can provide evidence supporting
technical interpretations. Correspondence pertinent to the project should
be kept and placed in the project files.
Deviations -- All deviations from procedural and planning documents should
be recorded in laboratory notebooks. Deviations from QAPjPs should be
reviewed and approved by the authorized personnel who performed the
original technical review or by their designees.
Final Report -- A copy of any report issued and any supporting documenta-
tion should be retained.
5.0 DEFINITIONS
The following terms are defined for use in this document:
ACCURACY The closeness of agreement between an observed value and
an accepted reference value. When applied to a set of
observed values, accuracy will be a combination of a
random component and of a common systematic error (or
bias) component.
BATCH: A group of samples which behave similarly with respect to
the sampling or the testing procedures being employed and
which are processed as a unit (see Section 3.4.1 for field
samples and Section 4.4.3 for laboratory samples). For QC
purposes, if the number of samples in a group is greater
than 20, then each group of 20 samples or less will all be
handled as a separate batch.
BIAS: The deviation due to matrix effects of the measured value
(x. - xu) from a known spiked amount. Bias can be assessed
by comparing a measured value to an accepted reference
value in a sample of known concentration or by determining
the recovery of a known amount of contaminant spiked into
a sample (matrix spike). Thus, the bias (B) due to matrix
effects based on a matrix spike is calculated as:
B - (x. - xu ) - K
where:
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BLANK:
CONTROL SAMPLE:
DATA QUALITY
OBJECTIVES (DQOs):
DATA VALIDATION:
DUPLICATE:
EQUIPMENT BLANK:
EQUIPMENT RINSATE:
ESTIMATED
QUANTITATION
LIMIT (EQL):
xt = measured value for spiked sample,
xu = measured value for unspiked sample, and
K = known value of the spike in the sample.
Using the following equation yields the percent recovery
%R = 100 (x. - xu)/ K
see Equipment Rinsate, Method Blank, Trip Blank.
A QC sample introduced into a process to monitor the
performance of the system.
A statement of the overall level of uncertainty that a
decision-maker is willing to accept in results derived
from environmental data (see reference 2, EPA/QAMS, July
16, 1986). This is qualitatively distinct from quality
measurements such as precision, bias, and detection limit.
The process of evaluating the available data against the
project DQOs to make sure that the objectives are met.
Data validation may be very rigorous, or cursory,
depending on project DQOs. The available data reviewed
will include analytical results, field QC data and lab QC
data, and may also include field records.
see Matrix Duplicate,
Duplicate.
see Equipment Rinsate.
Field Duplicate, Matrix Spike
A sample of analyte-free media which has been used to
rinse the sampling equipment. It is collected after
completion of decontamination and prior to sampling. This
blank is useful in documenting adequate decontamination of
sampling equipment.
The lowest concentration that can be reliably achieved
within specified limits of precision and accuracy during
routine laboratory operating conditions. The EQL is
generally 5 to 10 times the MDL. However, it may be
nominally chosen within these guidelines to simplify data
reporting. For many analytes the EQL analyte
concentration is selected as the lowest non-zero standard
in the calibration curve. Sample EQLs are highly matrix-
dependent. The EQLs in SW-846 are provided for guidance
and may not always be achievable.
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FIELD DUPLICATES:
LABORATORY CONTROL
SAMPLE:
MATRIX:
MATRIX DUPLICATE:
MATRIX SPIKE:
MATRIX SPIKE
DUPLICATES:
METHOD BLANK:
METHOD DETECTION
LIMIT (MDL):
Independent samples which are collected as close as
possible to the same point In space and time. They are
two separate samples taken from the same source, stored in
separate containers, and analyzed independently. These
duplicates are useful in documenting the precision of the^
sampling process.
A known matrix spiked with compound(s) representative of
the target analytes. This is used to document laboratory
performance.
The component or substrate (e.g., surface water, drinking
water) which contains the analyte of interest.
An intralaboratory split sample which is used to document
the precision of a method in a given sample matrix.
An aliquot of sample spiked with a known concentration of
target analyte(s). The spiking occurs prior to sample
preparation and analysis. A matrix spike is used to
document the bias of a method in a given sample matrix.
Intralaboratory split samples spiked with identical
concentrations of target analyte(s). The spiking occurs
prior to sample preparation and analysis. They are used
to document the precision and bias of a method in a given
sample matrix.
An analyte-free matrix to which all reagents are added in
the same volumes or proportions as used in sample
processing. The method blank should be carried through
the complete sample preparation and analytical procedure.
The method blank is used to document contamination
resulting from the analytical process.
For a method blank to be acceptable for use with the
accompanying samples, the concentration in the blank of
any analyte of concern should not be higher than the
highest of either:
(l)The method detection limit, or
(2)Five percent of the regulatory limit for that analyte,
or
(3)Five
sample.
percent of the measured concentration in the
The minimum concentration of a substance that can be
measured and reported with 99% confidence that the analyte
concentration is greater than zero and is determined from
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analysis of a sample In a given matrix type containing
the analyte.
For operational purposes, when it is necessary to
determine the MDL in the matrix, the MDL should be
determined by multiplying the appropriate one-sided 99% t-
statistic by the standard deviation obtained from a
minimum of three analyses of a matrix spike containing the
analyte of interest at a concentration three to five times
the estimated MDL, where the t-statistic is obtained from
standard references or the table below.
No. of samples; t-statistic
3 6.96
4 4.54
5 3.75
6 3.36
7 3.14
8 3.00
9 2.90
10 2.82
Estimate the MDL as follows:
Obtain the concentration value that corresponds to:
a) an instrument signal/noise ratio within the range of
2.5 to 5.0, or
b) the region of the standard curve where there is a
significant change in sensitivity (i.e., a break in the
slope of the standard curve).
Determine the variance (S2) for each analyte as follows:
-"^2
where Xj * the ith measurement of the variable x
and x = the average value of x;
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ORGANIC-FREE
REAGENT WATER:
PRECISION:
Determine the standard deviation (s) for each analyte as
follows:
2\1/2
s = (S2)
Determine the MDL for each analyte as follows:
MDL
"
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PROJECT:
QUALITY ASSURANCE
PROJECT PLAN
(QAPJP):
RCRA:
REAGENT BLANK:
REAGENT GRADE:
REAGENT WATER:
REFERENCE MATERIAL:
SPLIT SAMPLES:
STANDARD ADDITION:
STANDARD CURVE:
where:
x = the arithmetic mean of the Xj measurements, and S «
variance; and the relative percent difference (RPD) when
only two samples are available.
RPD = 100 [(x, -
+ x2)/2}].
Single or multiple data collection activities that are
related through the same planning sequence.
An orderly assemblage of detailed procedures designed to
produce data of sufficient quality to meet the data
quality objectives for a specific data collection
activity.
The Resource Conservation and Recovery Act.
See Method Blank.
Analytical reagent (AR) grade, ACS reagent grade, and
reagent grade are synonymous terms for reagents which
conform to the current specifications of the Committee on
Analytical Reagents of the American Chemical Society.
Water that has been generated by any method which would
achieve the performance specifications for ASTM Type II
water. For organic analyses, see the definition of
organic-free reagent water.
A material containing known quantities of target analytes
in solution or in a homogeneous matrix. It is used to
document the bias of the analytical process.
Aliquots of sample taken from the same container and
analyzed independently. In cases where aliquots of
samples are impossible to obtain, field duplicate samples
should be taken for the matrix duplicate analysis. These
are usually taken after mixing or compositing and are used
to document intra- or interlaboratory precision.
The practice of adding a known amount of an analyte to a
sample immediately prior to analysis. It is typically
used to evaluate interferences.
A plot of concentrations of known analyte standards versus
the instrument response to the analyte. Calibration
standards are prepared by successively diluting a standard
solution to produce working standards which cover the
working range of the instrument. Standards should be
prepared at the frequency specified in the appropriate
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section. The calibration standards should be prepared
using the same type of acid or solvent and at the same
concentration as will result in the samples following
sample preparation. This is applicable to organic and
inorganic chemical analyses.
SURROGATE: An organic compound which is similar to the target
analyte(s) in chemical composition and behavior in the
analytical process, but which is not normally found in
environmental samples.
TRIP BLANK: A sample of analyte-free media taken from the laboratory
to the sampling site and returned to the laboratory
unopened. A trip blank is used to document contamination
attributable to shipping and field handling procedures.
This type of blank is useful in documenting contamination
of volatile organics samples.
6.0 REFERENCES
1. Interim Guidelines and Specifications for Preparing Quality Assurance
Project Plans, QAMS-005/80, December 29, 1980, Office of Monitoring Systems
and Quality Assurance, ORD, U.S. EPA, Washington, DC 20460.
2. Development of Data Quality Objectives, Description of Stages I and II, July
16, 1986, Quality Assurance Management Staff, ORD, U.S. EPA, Washington, DC
20460.
3. RCRA Ground-Water Monitoring Technical Enforcement Guidance Document,
September, 1986, Office of Waste Programs Enforcement. OSWER, U.S. EPA,
Washington, DC, 20460.
4. DQO Training Software, Version 6.5, December, 1988, Quality Assurance
Management Staff, ORD, U.S. EPA, Washington, DC 20460.
5. Preparing Perfect Project Plans, EPA/600/9-89/087, October 1989, Risk
Reduction Engineering Laboratory (Guy Simes), Cincinnati OH.
6. ASTM Method D 1129-77, Specification for Reagent Water. 1991 Annual Book
of ASTM Standards. Volume 11.01 Water and Environmental Technology.
7. Generation of Environmental Data Related to Waste Management Activities
(Draft). February 1989. ASTM.
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INDEX
Accuracy 1, 13, 22, 23", 24
Batch 12, 19, 21, 23"
Bias 2, 3, 17-20, 22, 23"-25, 28
Blank 11, 12, 14, 18-20, 23", 24, 25, 28, 29
Equipment Rinsate 11, 12, 14, 18, 24*
Method Blank 19, 24, 25", 28
Reagent Blank 28"
Trip Blank 12, 18, 24, 29*
Chain-of-Custody 9, 11, 13, 14, 18, 21, 22
Control Chart 18, 19
Control Sample 11, 12, 18, 19, 23, 24"
Data Quality Objectives (DQO) 1-3, 8, 12, 19, 20, 24", 28
Decision-maker 2, 24
Duplicate 11, 12, 14, 18-20, 23, 24", 25, 27, 28
Field Duplicate 11, 12, 24, 25", 28
Matrix Duplicate 12, 19, 20, 24, 25", 28
Matrix Spike Duplicate 12, 19, 20, 23, 24, 25"
Equipment Blank 11, 24*
Equipment Rinsate 11, 12, 14, 18, 24"
Estimated Quantitation Limit (EQL) 24"
Field Duplicate 12, 24, 25", 28
Laboratory Control Sample 19, 25"
Matrix 11, 12, 18-20, 23-25", 26-28
Matrix Duplicate 12, 19, 20, 24, 25", 28
Matrix Spike 12, 18-20, 23, 25", 26, 27
Matrix Spike Duplicate 12, 19, 20, 23, 24, 25"
Method Blank 19, 24, 25", 28
Method Detection Limit (MDL) 18-20, 22, 24, 25*-27
Organic-Free Reagent Water 27", 28
Precision 1-3, 17-20, 22, 24, 25, 27", 28
Project 1-5, 7, 8, 11-14, 17-19, 21, 23, 24, 28"
Quality Assurance Project Plan (QAPjP) 1-9, 11, 12, 14, 15, 18, 20, 22, 23, 28"
RCRA 1, 8, 28"
Reagent Blank 28"
Reagent Grade 28"
Reagent Water 27, 28*
Reference Material 8, 11, 15, 18, 19, 28*
Split Samples 25, 28"
Standard Addition 20, 28"
Standard Curve 26, 28"
Surrogate 18, 20, 22, 29"
Trip Blank 12, 18, 24, 29"
Definition of term.
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PREFACE AND OVERVIEW
PURPOSE OF THE MANUAL
Test Methods for Evaluating Solid Waste (SW-846) is intended to provide a
unified, up-to-date source of information on sampling and analysis related to
compliance with RCRA regulations. It brings together into one reference all
sampling and testing methodology approved by the Office of Solid Waste for use
in implementing the RCRA regulatory program. The manual provides methodology
for collecting and testing representative samples of waste and other materials
to be monitored. Aspects of sampling and testing covered in SW-846 include
quality control, sampling plan development and implementation, analysis of
inorganic and organic constituents, the estimation of intrinsic physical
properties, and the appraisal of waste characteristics.
The procedures described in this manual are meant to be comprehensive and
detailed, coupled with the realization that the problems encountered in
sampling and analytical situations require a certain amount of flexibility.
The solutions to these problems will depend, in part, on the skill, training,
and experience of the analyst. For some situations, it is possible to use
this manual in rote fashion. In other situations, it will require a
combination of technical abilities, using the manual as guidance rather than
in a step-by-step, word-by-word fashion. Although this puts an extra burden
on the user, it is unavoidable because of the variety of sampling and
analytical conditions found with hazardous wastes.
ORGANIZATION AND FORMAT
This manual is divided into two volumes. Volume I focuses on laboratory
activities and is divided for convenience into three sections. Volume IA
deals with quality control, selection of appropriate test methods, and
analytical methods for metallic species. Volume IB consists of methods for
organic analytes. Volume 1C includes a variety of test methods for
miscellaneous analytes and properties for use in evaluating the waste
characteristics. Volume II deals with sample acquisition and includes quality
control, sampling plan design and implementation, and field sampling methods.
Included for the convenience of sampling personnel are discusssions of the
ground water, land treatment, and incineration monitoring regulations.
Volume I begins with an overview of the quality control precedures to be
imposed upon the sampling and analytical methods. The quality control chapter
(Chapter One) and the methods chapters are interdependent. The analytical
procedures cannot be used without a thorough understanding of the quality
control requirements and the means to implement them. This understanding can
be achieved only be reviewing Chapter One and the analytical methods together.
It is expected that individual laboratories, using SW-846 as the reference
PREFACE - 1
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source, will select appropriate methods and develop a standard operating
procedure (SOP) to be followed by the laboratory. The SOP should incorporate
the pertinent information from this manual adopted to the specific needs and
circumstances of the individual laboratory as well as to the materials to be
evaluated.
The method selection chapter (Chapter Two) presents a comprehensive
discussion of the application of these methods to various matrices in the
determination of groups of analytes or specific analytes. It aids the chemist
in constructing the correct analytical method from the array of procedures
which may cover the matrix/analyte/concentration combination of interests.
The section discusses the objective of the testing program and its
relationship to the choice of an analytical method. Flow charts are presented
along with tables to guide in the selection of the correct analytical
procedures to form the appropriate method.
The analytical methods are separated into distinct procedures describing
specific, independent analytical operations. These include extraction,
digestion, cleanup, and determination. This format allows linking of the
various steps in the analysis according to: the type of sample (e.g., water,
soil, sludge, still bottom); analytes(s) of interest; needed sensitivity; and
available analytical instrumentation. The chapters describing Miscellaneous
Test Methods and Properties, however, give complete methods which are not
amenable to such segmentation to form discrete procedures.
The introductory material at the beginning of each section containing
analytical procedures presents information on sample handling and
preservation, safety, and sample preparation.
Part II of Volume I (Chapters Seven and Eight) describes the
characteristics of a waste. Sections following the regulatory descriptions
contain the methods used to determine if the waste is hazardous because it
exhibits a particular characteristic.
Volume II gives background information on statistical and nonstatistical
aspects of sampling. It also presents practical sampling techniques
appropriate for situations presenting a variety of physical conditions.
A discussion of the regulatory requirements with respect to several
monitoring categories is also given in this volume. These include ground
water monitoring, land treatment, and incineration. The purpose of this
guidance is to orient the user to the objective of the analysis, and to assist
in developing data quality objectives, sampling plans, and laboratory SOP's.
Significant interferences, or other problems, may be encountered with
certain samples. In these situations, the analyst 1s advised to contact the
Chief, Methods Section (WH-562B) Technical Assessment Branch, Office of Solid
Waste, US EPA, Washington, DC 20460 (202-382-4761) for assistance. The
manual is intended to serve all those with a need to evaluate solid waste.
Your comments, corrections, suggestions, and questions concerning any material
contained in, or omitted from, this manual will be gratefully appreciated.
Please direct your comments to the above address.
PREFACE - 2
Revision 0
Date September 1986
-------
CHAPTER FOUR
ORGANIC ANALYTES
4.1 SAMPLING CONSIDERATIONS
4.1.1 Introduction
Following the initial and critical step of designing a sampling plan
(Chapter Nine) is the implementation of that plan such that a representative
sample of the solid waste is collected. Once the sample has been collected it
must be stored and preserved to maintain the chemical and physical properties
that it possessed at the time of collection. The sample type, type of containers
and their preparation, possible forms of contamination, and preservation methods
are all items which must be thoroughly examined in order to maintain the
integrity of the samples. This section highlights considerations which must be
addressed in order to maintain a sample's integrity and representativeness. This
section is, however, applicable only to trace analyses.
Quality Control (QC) requirements need not be met for all compounds
presented in the Table of Analytes for the method in use, rather, they must be
met for all compounds reported. A report of non-detect is considered a
quantitative report, and must meet all applicable QC requirements for that
compound and the method used.
4.1.2 Sample Handling and Preservation
This section deals separately with volatile and semivolatile organics.
Refer to Chapter Two and Table 4-1 of this section for sample containers, sample
preservation, and sample holding time information.
Volatile Organics
Standard 40 ml glass screw-cap VOA vials with Teflon lined silicone septa
may be used for both liquid and solid matrices. The vials and septa should be
washed with soap and water and rinsed with distilled deionized water. After
thoroughly cleaning the vials and septa, they should be placed in an oven and
dried at 100°C for approximately one hour.
NOTE: Do not heat the septa for extended periods of time (i.e., more than one
hour, because the silicone begins to slowly degrade at 105°C).
When collecting the samples, liquids and solids should be introduced into
the vials gently to reduce agitation which might drive off volatile compounds.
In general, liquid samples should be poured into the vial without introducing any
air bubbles within the vial as it is being filled. Should bubbling occur as a
result of violent pouring, the sample must be poured out and the vial refilled.
The vials should be completely filled at the time of sampling, so that when the
septum cap is fitted and sealed, and the vial inverted, no headspace is visible.
The sample should be hermetically sealed in the vial at the time of sampling, and
must not be opened prior to analysis to preserve their integrity.
FOUR - 1 Revision 2
September 1994
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due to differing solubility and diffusion properties of gases in
LIQUID matrices at different temperatures, it is possible for the
sample to generate some headspace during storage. This headspace
will appear in the form of micro bubbles, and should not invalidate
a sample for volatiles analysis.
The presence of a macro bubble in a sample vial generally indicates
either improper sampling technique or a source of gas evolution
within the sample. The latter case is usually accompanied by a
buildup of pressure within the vial, (e.g. carbonate-containing
samples preserved with acid). Studies conducted by the USEPA
(EMSL-Ci, unpublished data) indicate that "pea-sized" bubbles (i.e.,
bubbles not exceeding 1/4 inch or 6 mm in diameter) did not
adversely affect volatiles data. These bubbles were generally
encountered in wastewater samples, which are more susceptible to
variations in gas solubility than are groundwater samples.
At the time of analysis, the aliquot to be analyzed should be taken from the
vial with a gas-tight syringe inserted directly through the septum of the vial.
Only one analytical sample can be taken from each vial. If these guidelines are
not followed, the validity of the data generated from the samples is suspect.
VGA vials for samples with solid or semi-solid matrices (e.g., sludges)
should be completely filled as best as possible. The vials should be tapped
slightly as they are filled to try and eliminate as much free air space as
possible. Two vials should also be filled per sample location.
At least two VGA vials should be filled and labeled immediately at the
point at which the sample is collected. They should NOT be filled near a running
motor or any type of exhaust system because discharged fumes and vapors may
contaminate the samples. The two vials from each sampling location should then
be sealed in separate plastic bags to prevent cross-contamination between
samples, particularly if the sampled waste is suspected of containing high levels
of volatile organics. (Activated carbon may also be included in the bags to
prevent cross-contamination from highly contaminated samples). VOA samples may
also be contaminated by diffusion of volatile organics through the septum during
shipment and storage. To monitor possible contamination, a trip blank prepared
from organic-free reagent water (as defined in Chapter One) should be carried
throughout the sampling, storage, and shipping process.
Semivolatile Organics (including Pesticides, PCBs and Herbicides.)
Containers used to collect samples for the determination of semivolatile
organic compounds should be soap and water washed followed by methanol (or
isopropanol) rinsing (see Sec. 4.1.4 for specific instructions on glassware
cleaning). The sample containers should be of glass or Teflon, and have screw-
caps with Teflon lined septa. In situations where Teflon is not available,
solvent-rinsed aluminum foil may be used as a liner. However, acidic or basic
samples may react with the aluminum foil, causing eventual contamination of the
sample. Plastic containers or lids may NOT be used for the storage of samples
due to the possibility of sample contamination from the phthalate esters and
other hydrocarbons within the plastic. Sample containers should be filled with
care so as to prevent any portion of the collected sample coming in contact with
FOUR - 2 Revision 2
September 1994
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the sampler's gloves, thus causing contamination. Samples should not be
collected or stored in the presence of exhaust fumes. If the sample comes in
contact with the sampler (e.g. if an automatic sampler is used), run organic-free
reagent water through the sampler and use as a field blank.
4.1.3 Safety
Safety should always be the primary consideration in the collection of
samples. A thorough understanding of the waste production process, as well as
all of the potential hazards making up the waste, should be investigated whenever
possible. The site should be visually evaluated just prior to sampling to
determine additional safety measures. Minimum protection of gloves and safety
glasses should be worn to prevent sample contact with the skin and eyes. A
respirator should be worn even when working outdoors if organic vapors are
present. More hazardous sampling missions may require the use of supplied air
and special clothing.
4.1.4 Cleaning of Glassware
In the analysis of samples containing components in the parts per billion
range, the preparation of scrupulously clean glassware is necessary. Failure to
do so can lead to a myriad of problems in the interpretation of the final
chromatograms due to the presence of extraneous peaks resulting from
contamination. Particular care must be taken with glassware such as Soxhlet
extractors, Kuderna-Danish evaporative concentrators, sampling-train components,
or any other glassware coming in contact with an extract that will be evaporated
to a smaller volume. The process of concentrating the compounds of interest in
this operation may similarly concentrate the contaminating substance(s), which
may seriously distort the results.
The basic cleaning steps are:
1. Removal of surface residuals immediately after use;
2. Hot soak to loosen and float most particulate material;
3. Hot water rinse to flush away floated particulates;
4. Soak with an oxidizing agent to destroy traces of organic compounds;
5. Hot water rinse to flush away materials loosened by the deep penetrant
soak;
6. Distilled water rinse to remove metallic deposits from the tap water;
7. Alcohol, e.g., isopropanol or methanol, rinse to flush off any final
traces of organic materials and remove the water; and
8. Flushing the item immediately before use with some of the same solvent
that will be used in the analysis.
FOUR - 3 Revision 2
September 1994
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Each of these eight fundamental steps are discussed here in the order in
which they appeared on the preceeding page.
1. As soon possible after glassware (i.e., beakers, pipets, flasks, or
bottles) has come in contact with sample or standards, the glassware
should be flushed with alcohol before it is placed in the hot
detergent soak. If this is not done, the soak bath may serve to
contaminate all other glassware placed therein.
2. The hot soak consists of a bath of a suitable detergent in water of
50°C or higher. The detergent, powder or liquid, should be entirely
synthetic and not a fatty acid base. There are very few areas of the
country where the water hardness is sufficiently low to avoid the
formation of some hard-water scum resulting from the reaction between
calcium and magnesium salts with a fatty acid soap. This hard-water
scum or curd would have an affinity particularly for many chlorinated
compounds and, being almost wholly water-insoluble, would deposit on
all glassware in the bath in a thin film.
There are many suitable detergents on the wholesale and retail market.
Most of the common liquid dishwashing detergents sold at retail are
satisfactory but are more expensive than other comparable products
sold industrially. Alconox, in powder or tablet form, is manufactured
by Alconox, Inc., New York, and is marketed by a number of laboratory
supply firms. Sparkleen, another powdered product, is distributed by
Fisher Scientific Company.
3. No comments required.
4. The most common and highly effective oxidizing agent for removal of
traces of organic compounds is the traditional chromic acid solution
made up of concentrated sulfuric acid and potassium or sodium
dichromate. For maximum efficiency, the soak solution should be hot
(40-50°C). Safety precautions must be rigidly observed in the
handling of this solution. Prescribed safety gear should include
safety goggles, rubber gloves, and apron. The bench area where this
operation is conducted should be covered with fluorocarbon sheeting
because spattering will disintegrate any unprotected surfaces.
The potential hazards of using chromic-sulfuric acid mixture are great
and have been well publicized. There are now commercially available
substitutes that possess the advantage of safety in handling. These
are biodegradable concentrates with a claimed cleaning strength equal
to the chromic acid solution. They are alkaline, equivalent to ca.
0.1 N NaOH upon dilution, and are claimed to remove dried blood,
silicone greases, distillation residues, insoluble organic residues,
etc. They are further claimed to remove radioactive traces and will
not attack glass or exert a corrosive effect on skin or clothing. One
such product is "Chem Solv 2157," manufactured by Mallinckrodt and
available through laboratory supply firms. Another comparable product
is "Detex," a product of Borer-Chemie, Solothurn, Switzerland.
FOUR - 4 Revision 2
September 1994
-------
5, 6, and 7. No comments required.
8. There is always a possibility that between the time of washing and the
next use, the glassware could pick up some contamination from either
the air or direct contact. To ensure against this, it is good
practice to flush the item immediately before use with some of the
same solvent that will be used in the analysis.
The drying and storage of the cleaned glassware is of critical importance
to prevent the beneficial effects of the scrupulous cleaning from being
nullified. Pegboard drying is not recommended. It is recommended that
laboratory glassware and equipment be dried at 100°C. Under no circumstances
should such small items be left in the open without protective covering. The
dust cloud raised by the daily sweeping of the laboratory floor can most
effectively recontaminate the clean glassware.
As an alternate to solvent rinsing, the glassware can be heated to a
minimum of 300°C to vaporize any organics. Do not use this high temperature
treatment on volumetric glassware, glassware with ground glass joints, or
sintered glassware.
4.1.5 High Concentration Samples
Cross contamination of trace concentration samples may occur when
prepared in the same laboratory with high concentration samples. Ideally,
if both type samples are being handled, a laboratory and glassware
dedicated solely to the preparation of high concentration samples would be
available for this purpose. If this is not feasible, as a minimum when
preparing high concentration samples, disposable glassware should be used
or, at least, glassware dedicated entirely to the high concentration
samples. Avoid cleaning glassware used for both trace and high
concentration samples in the same area.
FOUR - 5 Revision 2
September 1994
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4.2 SAMPLE PREPARATION METHODS
4.2.1 EXTRACTIONS AND PREPARATIONS
The following methods are included in this section:
Method 3500A: Organic Extraction and Sample Preparation
Method 3510B: Separatory Funnel Liquid-Liquid Extraction
Method 3520B: Continuous Liquid-Liquid Extraction
Method 3540B: Soxhlet Extraction
Method 3541: Automated Soxhlet Extraction
Method 3550A: Ultrasonic Extraction
Method 3580A: Waste Dilution
Method 5030A: Purge-and-Trap
Method 5040A: Analysis of Sorbent Cartridges from Volatile
Organic Sampling Train (VOST): Gas
Chromatography/Mass Spectrometry Technique
Method 5041: Protocol for Analysis of Sorbent Cartridges from
Volatile Organic Sampling Train (VOST): Wide-
bore Capillary Column Technique
FOUR - 8 Revision 2
September 1994
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METHOD 3500A
ORGANIC EXTRACTION AND SAMPLE PREPARATION
1.0 SCOPE AND APPLICATION
1.1 The 3500 Methods are procedures for quantitatively extracting
nonvolatile and semivolatile organic compounds from various sample matrices.
Cleanup and/or analysis of the resultant extracts are described in Chapter Two,
Sections 2.3.2 and 2.3.1, respectively.
1.2 Method 3580 describes a solvent dilution technique that may be used
on non-aqueous nonvolatile and semivolatile organic samples prior to cleanup
and/or analysis.
1.3 The 5000 Methods are procedures for preparing samples containing
volatile organic compounds for quantitative analysis.
1.4 Refer to the specific method of interest for further details.
2.0 SUMMARY OF METHOD
2.1 3500 Methods: A sample of a known volume or weight is solvent
extracted. The resultant extract is dried and then concentrated in a Kuderna-
Danish apparatus (if necessary). Other concentration devices or techniques may
be used in place of the Kuderna-Danish concentrator if the quality control
requirements of the determinative methods are met (Method 8000, Section 8.0).
2.2 5000 Methods: Refer to the specific method of interest.
3.0 INTERFERENCES
3.1 Samples requiring analysis for volatile organic compounds, can be
contaminated by diffusion of volatile organics (particularly chlorofluoro-carbons
and methylene chloride) through the sample container septum during shipment and
storage. A field blank prepared from organic-free reagent water and carried
through sampling and subsequent storage and handling can serve as a check on such
contamination.
3.2 Solvents, reagents, glassware, and other sample processing hardware
may yield artifacts and/or interferences to sample analysis. All these materials
must be demonstrated to be free from interferences under the conditions of the
analysis by analyzing method blanks. Specific selection of reagents and
purification of solvents by distillation in all-glass systems may be required.
Refer to Chapter One for specific guidance on quality control procedures.
3.3 Interferences coextracted from the samples will vary considerably
from source to source. If analysis of an extracted sample is prevented due to
interferences, further cleanup of the sample extract may be necessary. Refer to
Method 3600 for guidance on cleanup procedures.
3500A - 1 Revision 1
July 1992
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3.4 Phthalate esters contaminate many types of products commonly found
in the laboratory. Plastics, in particular, must be avoided because phthalates
are commonly used as plasticizers and are easily extracted from plastic
materials. Serious phthalate contamination may result at any time if consistent
quality control is not practiced.
3.5 Glassware contamination resulting in analyte degradation: Soap
residue on glassware may cause degradation of certain analytes. Specifically,
aldrin, heptachlor, and most organophosphorus pesticides will degrade in this
situation. This problem is especially pronounced with glassware that may be
difficult to rinse (e.g., 500 ml K-D flask). These items should be hand-rinsed
very carefully to avoid this problem.
4.0 APPARATUS AND MATERIALS
4.1 Refer to the specific method of interest for a description of the
apparatus and materials needed.
5.0 REAGENTS
5.1 Refer to the specific method of interest for a description of the
solvents needed.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water as defined in Chapter One.
5.3 Stock standards: Stock solutions may be prepared from pure standard
materials or purchased as certified solutions.
5.3.1 Purgeable stock standards: Prepare stock standards in
methanol using assayed liquids or gases, as appropriate. Because of the
toxicity of some of the organohalides, primary dilutions of these
materials should be prepared in a hood.
5.3.1.1 Place about 9.8 ml of methanol in a 10-mL tared
ground-glass-stoppered volumetric flask. Allow the flask to stand,
unstoppered, for about 10 min or until all alcohol-wetted surfaces
have dried. Weigh the flask to the nearest 0.0001 g.
5.3.1.2 Using a 100-/itL syringe, immediately add two or
more drops of assayed reference material to the flask, then reweigh.
The liquid must fall directly into the alcohol without contacting
the neck of the flask.
5.3.1.3 Reweigh, dilute to volume, stopper, then mix by
inverting the flask several times. Calculate the concentration in
milligrams per liter (mg/L) from the net gain in weight. When
compound purity is assayed to be 96% or greater, the weight may be
used without correction to calculate the concentration of the stock
standard. Commercially prepared stock standards may be used at any
concentration if they are certified by the manufacturer or by an
independent source.
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5.3.1.4 Transfer the stock standard solution into a
Teflon-sealed screw-cap bottle. Store, with minimal headspace, at -
10°C to -20°C and protect from light.
5.3.1.5 All standards must be replaced after 1 month, or
sooner if comparison with check standards indicates a problem.
5.3.2 Semivolatile stock standards: Base/neutral and acid stock
standards are prepared in methanol. Organochlorine pesticide standards
are prepared in acetone.
5.3.2.1 Stock standard solutions should be stored in
Teflon-sealed containers at 4°C. The solutions should be checked
frequently for stability. These solutions must be replaced after
six months, or sooner if comparison with quality control check
samples indicate a problem.
5.4 Surrogate standards: A surrogate standard (i.e., a chemically inert
compound not expected to occur in an environmental sample) should be added to
each sample, blank, and matrix spike sample just prior to extraction or
processing. The recovery of the surrogate standard is used to monitor for
unusual matrix effects, gross sample processing errors, etc. Surrogate recovery
is evaluated for acceptance by determining whether the measured concentration
falls within the acceptance limits. Recommended surrogates for different analyte
groups follow. However, these compounds, or others that better correspond to the
analyte group, may be used for other analyte groups as well. Normally three or
more standards are added for each analyte group.
5.4.1 Base/neutral and acid surrogate spiking solutions: The
following are recommended surrogate standards.
Base/neutral Acid
2-Fluorobiphenyl 2-Fluorophenol
Ni trobenzene-d5 2,4,6-Tri bromophenol
Terphenyl-du Phenol-d6
5.4.1.1 Prepare a surrogate standard spiking solution in
methanol that contains the base/neutral compounds at a concentration
of 100 mg/L, and the acid compounds at 200 mg/L for water and
sediment/soil samples (low- and medium-level). For waste samples,
the concentration should be 500 mg/L for base/neutrals and 1000 mg/L
for acids.
5.4.2 Organochlorine pesticide/PCB surrogate spiking solution: The
following are recommended surrogate standards for Organochlorine
pesticides/PCBs.
Orqanochlorine pesticides/PCBs
Dibutylchlorendate (DBC) (if available)
2,4,5,6-Tetrachloro-meta-xylene (TCMX)
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5.4.2.1 Prepare a surrogate standard spiking solution at
a concentration of 1 mg/L in acetone for water and sediment/soil
samples. For waste samples, the concentration should be 5 mg/L.
5.4.3 Purgeable surrogate spiking solution: The following are
recommended surrogate standards for volatile organics.
Purqeable organics
p-Bromof1uorobenzene
l,2-Dichloroethane-d4
Toluene-d8
5.4.3.1 Prepare a surrogate spiking solution (as described
in Section 5.3.1 or through secondary dilution of the stock
standard) in methanol containing the surrogate standards at a
concentration of 25 mg/L.
5.5 Matrix spike standards: Select five or more analytes from each
analyte group for use in a spiking solution. The following are recommended
matrix spike standard mixtures for a few analyte groups. These compounds, or
others that better correspond to the analyte group, may be used for other analyte
groups as well.
5.5.1 Base/neutral and acid matrix spiking solution: Prepare a
spiking solution in methanol that contains each of the following
base/neutral compounds at 100 mg/L and the acid compounds at 200 mg/L for
water and sediment/soil samples. The concentration of these compounds
should be five times higher for waste samples.
Base/neutrals Acids
1,2,4-Tri chlorobenzene Pentachlorophenol
Acenaphthene Phenol
2,4-Dinitrotoluene 2-Chlorophenol
Pyrene 4-Chloro-3-methylphenol
N-Nitroso-di-n-propylamine 4-Nitrophenol
1,4-Dichlorobenzene
5.5.2 Organochlorine pesticide matrix spiking solution: Prepare a
spiking solution in acetone or methanol that contains the following
pesticides in the concentrations specified for water and sediment/soil.
The concentration should be five times higher for waste samples.
Pesticide Concentration (mq/L)
Lindane 0.2
Heptachlor 0.2
Aldrin 0.2
Dieldrin 0.5
Endrin 0.5
4,4'-DDT 0.5
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5.5.3 Purgeable matrix spiking solution: Prepare a spiking solution
in methanol that contains the following compounds at a concentration of 25
mg/L.
Purgeable organics
1,1-Dichloroethene
Trichloroethene
Chlorobenzene
Toluene
Benzene
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to the Organic Analyte Chapter,
Section 4.1.
7.0 PROCEDURE
7.1 Semivolatile organic sample extraction: Water, soil/sediment,
sludge, and waste samples requiring analysis for base/neutral and acid
extractables and/or organochlorine pesticides must undergo solvent extraction
prior to analysis. This manual contains four methods that may be used for this
purpose: Method 3510; Method 3520; Method 3540; and Method 3550. The method
that should be used on a particular sample, is highly dependent upon the physical
characteristics of that sample. Therefore, review these four methods prior to
choosing one in particular. Appropriate surrogate standards and, if necessary,
matrix spiking solutions are added to the sample prior to extraction for all four
methods.
7.1.1 Method 3510: Applicable to the extraction and concentration
of water-insoluble and slightly water-soluble organics from aqueous
samples. A measured volume of sample is solvent extracted using a
separatory funnel. The extract is dried, concentrated and, if necessary,
exchanged into a solvent compatible with further analysis. Method 3520
should be used if an emulsion forms between the solvent-sample phases,
which can not be broken up by mechanical techniques.
7.1.2 Method 3520: Applicable to the extraction and concentration
of water-insoluble and slightly water-soluble organics from aqueous
samples. A measured volume of sample is extracted with an organic solvent
in a continuous liquid-liquid extractor. The solvent must have a density
greater than that of the sample. The extract is dried, concentrated and,
if necessary, exchanged into a solvent compatible with further analysis.
The limitations of Method 3510 concerning solvent-sample phase separation
do not interfere with this procedure.
7.1.3 Method 3540: This is a procedure for extracting nonvolatile
and semivolatile organic compounds from solids such as soils, sludges, and
wastes. A solid sample is mixed with anhydrous sodium sulfate, placed
into an extraction thimble or between two plugs of glass wool, and
extracted using an appropriate solvent in a Soxhlet extractor. The
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extract is dried, concentrated and, if necessary, exchanged into a solvent
compatible with further analysis.
7.1.4 Method 3550: This method is applicable to the extraction of
nonvolatile and semivolatile organic compounds from solids such as soils,
sludges, and wastes using the technique of ultrasonic extraction. Two
procedures are detailed depending upon the expected concentration of
organics in the sample; a low concentration and a high concentration
method. In both, a known weight of sample is mixed with anhydrous sodium
sulfate and solvent extracted using ultrasonic extraction. The extract is
dried, concentrated and, if necessary, exchanged into a solvent compatible
with further analysis.
7.1.5 Method 3580: This method describes the technique of solvent
dilution of non-aqueous waste samples. It is designed for wastes that may
contain organic chemicals at a level greater than 20,000 mg/kg and that
are soluble in the dilution solvent. When using this method, the analyst
must use caution in determining the correct concentration of spike and
surrogate solution to avoid diluting out these compounds when diluting the
sample. The loss of surrogate and spike data should only occur in samples
containing a high concentration of analytes which is unknown at the time
of extraction or where sample interferences could not be eliminated
following the best attempts at extract cleanup .by the laboratory.
7.2 Volatile organic sample preparation: There are three methods for
volatile sample preparation: Method 5030; Method 5040; and direct injection.
Method 5030 is the most widely applicable procedure for analysis of volatile
organics, while the direct injection technique may have limited applicability to
aqueous matrices.
7.2.1 Method 5030: This method describes the technique of purge-
and-trap for the introduction of purgeable organics into a gas
chromatograph. This procedure is applicable for use with aqueous samples
directly and to solids, wastes, soils/sediments, and water-miscible
liquids following appropriate preparation. An inert gas is bubbled
through the sample, which will efficiently transfer the purgeable organics
from the aqueous phase to the vapor phase. The vapor phase is swept
through a sorbent trap where the purgeables are trapped. After purging is
completed, the trap is heated and backflushed with the inert gas to desorb
the purgeables onto a gas chromatographic column. Prior to application of
the purge-and-trap procedure, all samples (including blanks, spikes, and
duplicates) should be spiked with surrogate standards and, if required,
with matrix spiking compounds.
7.2.2 Method 5040: This method is applicable to the investigation
of sorbent cartridges from volatile organic sampling train (VOST).
7.3 Sample analysis: Following preparation of a sample by one of the
methods described above, the sample is ready for further analysis. For samples
requiring volatile organic analysis, application of one of the methods described
above is followed directly by gas chromatographic analysis (Methods 8010, 8011,
8015, 8020, 8021, 8030, 8240 and 8260). Samples prepared for semivolatile
analysis may, if necessary, undergo cleanup (See Method 3600) prior to
application of a specific determinative method.
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8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific guidance on quality control
procedures.
8.2 Before processing any samples, the analyst should demonstrate through
the analysis of a reagent water blank that all glassware and reagents are
interference free. Each time a set of samples is processed, a method blank(s)
should be processed as a safeguard against chronic laboratory contamination. The
blank samples should be carried through all stages of the sample preparation and
measurement.
8.3 Surrogate standards should be added to all samples when specified in
the appropriate determinative method in Chapter Four, Section 4.3
8.4 A reagent blank, a matrix spike, and a duplicate or matrix spike
duplicate must be performed for each analytical batch (up to a maximum of 20
samples) analyzed.
8.5 For GC or GC/MS analysis, the analytical system performance must be
verified by analyzing quality control (QC) check samples. Method 8000, Section
8.0 discusses in detail the process of verification; however, preparation of the
QC check sample concentrate is dependent upon the method being evaluated.
8.5.1 Volatile organic QC check samples: QC check sample
concentrates containing each analyte of interest are spiked into reagent
water (defined as the QC check sample) and analyzed by purge-and-trap
(Method 5030). The concentration of each analyte in the QC check sample
is 20 ng/L. The evaluation of system performance is discussed in detail
in Method 8000, beginning with Paragraph 8.6
8.5.2 Semi volatile organic QC check samples: To evaluate the
performance of the analytical method, the QC check samples must be handled
in exactly the same manner as actual samples. Therefore, 1.0 ml of the QC
check sample concentrate is spiked into each of four 1-L aliquots of
reagent water (now called the QC check sample), extracted, and then
analyzed by GC. The variety of semivolatile analytes which may be
analyzed by GC is such that the concentration of the QC check sample
concentrate is different for the different analytical techniques presented
in the manual. Method 8000 discusses in detail the procedure of verifying
the detection system once the QC check sample has been prepared. The
concentrations of the QC check sample concentrate for the various methods
are as follows:
8.5.2.1 Method 8040 - Phenols: The QC check sample
concentrate should contain each analyte at a concentration of 100
mg/L in 2-propanol.
8.5.2.2 Method 8060 - Phthalate esters: The QC check
sample concentrate should contain the following analytes at the
following concentrations in acetone: butyl benzyl phthalate, 10
mg/L; bis(2-ethylhexyl) phthalate, 50 mg/L; di-n-octylphthalate, 50
mg/L; and any other phthalate at 25 mg/L.
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8.5.2.3 Method 8070 - Nitrosamines: The QC check sample
concentrate should contain each analyte at 20 mg/L in methanol or
some other water miscible solvent.
8.5.2.4 Method 8080 - Orqanochlorine pesticides and PCBs;
The QC check sample concentrate should contain each single-component
analyte at the following concentrations in acetone or some other
water miscible solvent: 4,4'-DDD, 10 mg/L; 4,4'-DDT, 10 mg/L;
endosulfan II, 10 mg/L; endosulfan sulfate, 10 mg/L; endrin, 10
mg/L; and any other single-component pesticide at 2 mg/L . If the
method is only to be used to analyze PCBs, chlordane, or toxaphene,
the QC check sample concentrate should contain the most
representative multicomponent parameter at a concentration of 50
mg/L in acetone.
8.5.2.5 Method 8090 - Nitroaromatics and Cyclic Ketones:
The QC check sample concentrate should contain each analyte at the
following concentrations in acetone: each dinitrotoluene at 20
mg/L; and isophorone and nitrobenzene at 100 mg/L.
8.5.2.6 Method 8100 - Polynuclear aromatic hydrocarbons;
The QC check sample concentrate should contain each analyte at the
following concentrations in acetonitrile: naphthalene, 100 mg/L;
acenaphthylene, 100 mg/L; acenaphthene, 100 mg/L; fluorene, 100
mg/L; phenanthrene, 100 mg/L; anthracene, 100 mg/L;
benzo(k)fluoranthene, 5 mg/L; and any other PAH at 10 mg/L .
8.3.2.7 Method 8110 - Haloethers; The QC check sample
concentrate should contain each analyte at a concentration of
20 mg/L in methanol or some other water miscible solvent.
8.5.2.8 Method 8120 - Chlorinated hydrocarbons: The QC
check sample concentrate should contain each analyte at the
following concentrations in acetone: hexachloro-substituted
hydrocarbons, 10 mg/L; and any other chlorinated hydrocarbon, 100
mg/L.
8.3.2.9 Method 8140/8141 - Organophosphorus compounds:
The QC check sample concentrate should contain each analyte in
acetone at a concentration 1,000 times more concentrated than the
selected spike concentration.
8.3.2.10 Method 8150 - Chlorinated herbicides: The QC
check sample concentrate should contain each analyte in acetone at
a concentration 1,000 times more concentrated than the selected
spike concentration.
8.3.2.11 Method 8250/8270 - Semi volatile organics: The QC
check sample concentrate should contain each analyte in acetone at
a concentration of 100 mg/L.
8.3.2.12 Method 8310 - Polynuclear aromatic hydrocarbons:
The QC check sample concentrate should contain each analyte at the
following concentrations in acetonitrile: naphthalene, 100 mg/L;
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acenaphthylene, 100 mg/L; acenaphthene, 100 mg/L; fluorene,
100 mg/L; phenanthrene, 100 mg/L; anthracene, 100 mg/L;
benzo(k)fluoranthene, 5 mg/L; and any other PAH at 10 mg/L.
9.0 METHOD PERFORMANCE
9.1 The recovery of surrogate standards is used to monitor unusual matrix
effects, sample processing problems, etc. The recovery of matrix spiking
compounds indicates the presence or absence of unusual matrix effects.
9.2 The performance of this method will be dictated by the overall
performance of the sample preparation in combination with the analytical
determinative method.
10.0 REFERENCES
10.1 None required.
3500A - 9 Revision 1
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METHOD 3500A
ORGANIC EXTRACTION AND SAMPLE PREPARATION
7 1 1 Method
3510
Non- aqueous
Semi volatile
Organics
No
713 Method
3510
C1eanup,
Determina 11v
Method
Yes
) >
7 1 5 Method
3580
7 1 4 Method
3550
\,
So rbent
Ca r tridges
from VOST
722 Method
5030
Direct
Injection
3500A - 10
Revision 1
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METHOD 3510B
SEPARATORY FUNNEL LIQUID-LIQUID EXTRACTION
1.0 SCOPE AND APPLICATION
1.1 This method describes a procedure for isolating organic compounds
is samples. The method also describes concentration techniques
r preparing the extract for the appropriate determinative methods
n Co*- A 7 f\-f f""h !in4"a v» Cnnv*
_,_ ..--_ — __ _____ ____ _. r
from aqueous samples. The method al<
suitable for preparing the extract for
described in Sec. 4.3 of Chapter Four.
1.2 This method is applicable to the isolation and concentration of
water-insoluble and slightly water-soluble organics in preparation for a variety
of chromatographic procedures.
2.0 SUMMARY OF METHOD
2.1 A measured volume of sample, usually 1 liter, at a specified pH (see
Table 1), is serially extracted with methylene chloride using a separatory
funnel. The extract is dried, concentrated (if necessary), and, as necessary,
exchanged into a solvent compatible with the cleanup or determinative method to
be used (see Table 1 for appropriate exchange solvents).
3.0 INTERFERENCES
3.1 Refer to Method 3500.
3.2 Under basic extraction conditions required to separate analytes for
the packed columns of Method 8250, the decomposition of some analytes has been
demonstrated. Organochlorine pesticides may dechlorinate, phthalate esters may
exchange, and phenols may react to form tannates. These reactions increase with
increasing pH, and are decreased by the shorter reaction times available in
Method 3510. Methods 3520/8270, 3510/8270, and 3510/8250, respectively, are
preferred over Method 3520/8250 for the analysis of these classes of compounds.
4.0 APPARATUS AND MATERIALS
4.1 Separatory funnel - 2 liter, with Teflon stopcock.
4.2 Drying column - 20 mm ID Pyrex chromatographic column with Pyrex
glass wool at bottom and a Teflon stopcock.
NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without
frits may be purchased. Use a small pad of Pyrex glass wool to
retain the adsorbent. Prewash the glass wool pad with 50 mL of
acetone followed by 50 mL of elution solvent prior to packing the
column with adsorbent.
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4.3 Kuderna-Danish (K-D) apparatus.
4.3.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025 or
equivalent). A ground-glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.5 Water bath - Heated, with concentric ring cover, capable of
temperature control (±5°C). The bath should be used in a hood.
4.6 Vials - 2 ml, glass with Teflon lined screw-caps or crimp tops.
4.7 pH indicator paper - pH range including the desired extraction pH.
4.8 Erlenmeyer flask - 250 ml.
4.9 Syringe - 5 ml.
4.10 Graduated cylinder - 1 liter.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination. Reagents should be stored
in glass to prevent the leaching of contaminants from plastic containers.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium hydroxide solution (ION), NaOH. Dissolve 40 g NaOH in
organic-free reagent water and dilute to 100 ml.
5.4 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
3510B - 2 Revision 2
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methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.5 Sulfuric acid solution (1:1 v/v), H2S04. Slowly add 50 ml of H2S04
(sp. gr. 1.84) to 50 ml of organic-free reagent water.
5.6 Extraction/exchange solvents
5.6.1 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.6.2 Hexane, C6H14 - Pesticide quality or equivalent.
5.6.3 2-Propanol, CH3CH(OH)CH3 - Pesticide quality or equivalent.
5.6.4 Cyclohexane, C6H12 - Pesticide quality or equivalent.
5.6.5 Acetonitrile, CH3CN - Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Using a 1 liter graduated cylinder, measure 1 liter (nominal) of
sample and transfer it quantitatively to the separatory funnel. If high
concentrations are anticipated, a smaller volume may be used and then diluted
with organic-free reagent water to 1 liter. Add 1.0 mL of the surrogate
standards to all samples, spikes, and blanks (see Method 3500 and the
determinative method to be used, for details on the surrogate standard solution
and the matrix spike solution). For the sample in each analytical batch selected
for spiking, add 1.0 mL of the matrix spiking standard. For base/neutral-acid
analysis, the amount added of the surrogates and matrix spiking compounds should
result in a final concentration of 100 ng/juL of each base/neutral analyte and
200 ng//uL of each acid analyte in the extract to be analyzed (assuming a 1 /nL
injection). If Method 3640, Gel-Permeation Cleanup, is to be used, add twice the
volume of surrogates and matrix spiking compounds since half the extract is lost
due to loading of the GPC column.
7.2 Check the pH of the sample with wide-range pH paper and, if
necessary, adjust the pH to that indicated in Table 1 for the specific
determinative method that will be used to analyze the extract.
7.3 Add 60 mL of methylene chloride to the separatory funnel.
7.4 Seal and shake the separatory funnel vigorously for 1-2 minutes with
periodic venting to release excess pressure.
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NOTE: Methylene chloride creates excessive pressure very rapidly;
therefore, initial venting should be done immediately after the
separatory funnel has been sealed and shaken once. Venting of the
separatory funnel should be into a hood to avoid needless exposure
of the analyst to solvent vapors.
7.5 Allow the organic layer to separate from the water phase for a
minimum of 10 minutes. If the emulsion interface between layers is more than
one-third the size of the solvent layer, the analyst must employ mechanical
techniques to complete the phase separation. The optimum technique depends upon
the sample and may include stirring, filtration of the emulsion through glass
wool, centrifugation, or other physical methods. Collect the solvent extract in
an Erlenmeyer flask. If the emulsion cannot be broken (recovery of < 80% of the
methylene chloride, corrected for the water solubility of methylene chloride),
transfer the sample, solvent, and emulsion into the extraction chamber of a
continuous extractor and proceed as described in Method 3520, Continuous Liquid-
Liquid Extraction.
7.6 Repeat the extraction two more times using fresh portions of solvent
(Sees. 7.3 through 7.5). Combine the three solvent extracts.
7.7 If further pH adjustment and extraction is required, adjust the pH
of the aqueous phase to the desired pH indicated in Table 1. Serially extract
three times with 60 mL of methylene chloride, as outlined in Sees. 7.3
through 7.5. Collect and combine the extracts and label the combined extract
appropriately.
7.8 If performing GC/MS analysis (Method 8270), the acid/neutral and base
extracts may be combined prior to concentration. However, in some situations,
separate concentration and analysis of the acid/neutral and base extracts may be
preferable (e.g. if for regulatory purposes the presence or absence of specific
acid/neutral or base compounds at low concentrations must be determined, separate
extract analyses may be warranted).
7.9 Perform the concentration (if necessary) using the Kuderna-Danish
(K-D) Technique (Sees. 7.10.1 through 7.10.4).
7.10 K-D Technique
7.10.1 Assemble a Kuderna-Danish (K-D) concentrator by
attaching a 10 mL concentrator tube to a 500 mL evaporation flask. Dry
the extract by passing it through a drying column containing about 10 cm
of anhydrous sodium sulfate. Collect the dried extract in a K-D
concentrator. Rinse the Erlenmeyer flask, which contained the solvent
extract, with 20-30 mL of methylene chloride and add it to the column to
complete the quantitative transfer.
7.10.2 Add one or two clean boiling chips to the flask and
attach a three ball Snyder column. Prewet the Snyder column by adding
about 1 mL of methylene chloride to the top of the column. Place the K-D
apparatus on a hot water bath (15-20°C above the boiling point of the
solvent) so that the concentrator tube is partially immersed in the hot
water and the entire lower rounded surface of the flask is bathed with hot
3510B - 4 Revision 2
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vapor. Adjust the vertical position of the apparatus and the water
temperature as required to complete the concentration in 10-20 minutes.
At the proper rate of distillation the balls of the column will actively
chatter, but the chambers will not flood. When the apparent volume of
liquid reaches 1 ml, remove the K-D apparatus from the water bath and
allow it to drain and cool for at least 10 minutes.
7.10.3 If a solvent exchange is required (as indicated in Table
1), momentarily remove the Snyder column, add 50 ml of the exchange
solvent, a new boiling chip, and reattach the Snyder column. Concentrate
the extract, as described in Sec. 7.11, raising the temperature of the
water bath, if necessary, to maintain proper distillation.
7.10.4 Remove the Snyder column and rinse the flask and its
lower joints into the concentrator tube with 1-2 ml of methylene chloride
or exchange solvent. If sulfur crystals are a problem, proceed to
Method 3660 for cleanup. The extract may be further concentrated by using
the technique outlined in Sec. 7.11 or adjusted to 10.0 ml with the
solvent last used.
7.11 If further concentration is indicated in Table 1, either the micro-
Snyder column technique (7.11.1) or nitrogen blowdown technique (7.11.2) is used
to adjust the extract to the final volume required.
7.11.1 Micro-Snyder Column Technique
7.11.1.1 If further concentration is indicated in Table 1,
add another clean boiling chip to the concentrator tube and attach
a two ball micro-Snyder column. Prewet the column by adding 0.5 ml
of methylene chloride or exchange solvent to the top of the column.
Place the K-D apparatus in a hot water bath so that the concentrator
tube is partially immersed in the hot water. Adjust the vertical
position of the apparatus and the water temperature, as required, to
complete the concentration in 5-10 minutes. At the proper rate of
distillation the balls of the column will actively chatter, but the
chambers will not flood. When the apparent volume of liquid reaches
0.5 ml, remove the K-D apparatus from the water bath and allow it to
drain and cool for at least 10 minutes. Remove the Snyder column
and rinse the flask and its lower joints into the concentrator tube
with 0.2 ml of extraction solvent. Adjust the final volume to 1.0-
2.0 ml, as indicated in Table 1, with solvent.
7.11.2 Nitrogen Blowdown Technique
7.11.2.1 Place the concentrator tube in a warm bath (35°C)
and evaporate the solvent volume to 0.5 ml using a gentle stream of
clean, dry nitrogen (filtered through a column of activated carbon).
CAUTION: New plastic tubing must not be used between the
carbon trap and the sample, since it may
introduce interferences.
3510B - 5 Revision 2
September 1994
-------
7.11.2.2 The internal wall of the tube must be rinsed down
several times with methylene chloride or appropriate solvent during
the operation. During evaporation, the tube solvent level must be
positioned to avoid water condensation. Under normal procedures,
the extract must not be allowed to become dry.
CAUTION: When the volume of solvent is reduced below 1 ml,
semivolatile analytes may be lost.
7.12 The extract may now be analyzed for the target analytes using the
appropriate determinative technique(s) (see Sec. 4.3 of this Chapter). If
analysis of the extract will not be performed immediately, stopper the
concentrator tube and store refrigerated. If the extract will be stored longer
than 2 days it should be transferred to a vial with a Teflon lined screw-cap or
crimp top, and labeled appropriately.
8.0 QUALITY CONTROL
8.1 Any reagent blanks or matrix spike samples should be subjected to
exactly the same analytical procedures as those used on actual samples.
8.2 Refer to Chapter One for specific quality control procedures and
Method 3500 for extraction and sample preparation procedures.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
3510B - 6 Revision 2
September 1994
-------
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METHOD 3510B
SEPARATORY FUNNEL LIQUID-LIQUID EXTRACTION
j Start J
IT
7.1 Add surrogate
standards to all
samples, spikes,
and blanks.
7.7 Collect
and combine
extracts and label
7.8
GC/MS
analysis (Metho
8270) being
performed?
7.2 Check
and adjust pH.
7.8 Combine
base/neutral
extracts prior
to concentration
7.3 - 7.6
Extract 3
times.
7.9 - 7.11
Concentrate
extract.
7.7
Further
extractions
required?
7.12
Ready for
analysis.
3510B - 8
Revision 2
September 1994
-------
METHOD 3520B
CONTINUOUS LIQUID-LIQUID EXTRACTION
1.0 SCOPE AND APPLICATION
1.1 This method describes a procedure for isolating organic compounds
from aqueous samples. The method also describes concentration techniques
suitable for preparing the extract for the appropriate determinative steps
described in Sec. 4.3 of Chapter Four.
1.2 This method is applicable to the isolation and concentration of
water-insoluble and slightly soluble organics in preparation for a variety of
chromatographic procedures.
1.3 Method 3520 is designed for extraction solvents with greater density
than the sample. Continuous extraction devices are available for extraction
solvents that are less dense than the sample. The analyst must demonstrate the
effectiveness of any such automatic extraction device before employing it in
sample extraction.
2.0 SUMMARY OF METHOD
2.1 A measured volume of sample, usually 1 liter, is placed into a
continuous liquid-liquid extractor, adjusted, if necessary, to a specific pH (see
Table 1), and extracted with organic solvent for 18-24 hours. The extract is
dried, concentrated (if necessary), and, as necessary, exchanged into a solvent
compatible with the cleanup or determinative method being employed (see Table 1
for appropriate exchange solvents).
3.0 INTERFERENCES
3.1 Refer to Method 3500.
3.2 Under basic extraction conditions required to separate analytes for
the packed columns of Method 8250, the decomposition of some analytes has been
demonstrated. Organochlorine pesticides may dechlorinate, phthalate esters may
exchange, and phenols may react to form tannates. These reactions increase with
increasing pH, and are decreased by the shorter reaction times available in
Method 3510. Methods 3520/8270, 3510/8270, and 3510/8250, respectively, are
preferred over Method 3520/8250 for the analysis of these classes of compounds.
4.0 APPARATUS AND MATERIALS
4.1 Continuous liquid-liquid extractor - Equipped with Teflon or glass
connecting joints and stopcocks requiring no lubrication (Kontes 584200-0000,
584500-0000, 583250-0000, or equivalent).
4.2 Drying column - 20 mm ID Pyrex chromatographic column with Pyrex
glass wool at bottom and a Teflon stopcock.
3520B - 1 Revision 2
September 1994
-------
NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without
frits may be purchased. Use a small pad of Pyrex glass wool to
retain the adsorbent. Prewash the glass wool pad with 50 ml of
acetone followed by 50 ml of elution solvent prior to packing the
column with adsorbent.
4.3 Kuderna-Danish (K-D) apparatus
4.3.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.5 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 5°C). The bath should be used in a hood.
4.6 Vials - 2 ml, glass with Teflon lined screw-caps or crimp tops.
4.7 pH indicator paper - pH range including the desired extraction pH.
4.8 Heating mantle - Rheostat controlled.
4.9 Syringe - 5 ml.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination. Reagents should be stored
in glass to prevent the leaching of contaminants from plastic containers.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
3520B - 2 Revision 2
September 1994
-------
5.3 Sodium hydroxide solution (ION), NaOH. Dissolve 40 g NaOH in
organic-free reagent water and dilute to 100 mL.
5.4 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.5 Sulfuric acid solution (1:1 v/v), H2S04. Slowly add 50 ml of H2S04
(sp. gr. 1.84) to 50 ml of organic-free reagent water.
5.6 Extraction/exchange solvents
5.6.1 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.6.2 Hexane, C6H14 - Pesticide quality or equivalent.
5.6.3 2-Propanol, (CH3)2CHOH - Pesticide quality or equivalent.
5.6.4 Cyclohexane, C6H12 - Pesticide quality or equivalent.
5.6.5 Acetonitrile, CH3CN - Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Using a 1 liter graduated cylinder, measure out 1 liter (nominal) of
sample and transfer it quantitatively to the continuous extractor. If high
concentrations are anticipated, a smaller volume may be used and then diluted
with organic-free reagent water to 1 liter. Check the pH of the sample with wide-
range pH paper and adjust the pH, if necessary, to the pH indicated in Table 1
using 1:1 (V/V) sulfuric acid or 10 N sodium hydroxide. Pipet 1.0 mL of the
surrogate standard spiking solution into each sample into the extractor and mix
well. (See Method 3500 and the determinative method to be used, for details on
the surrogate standard solution and the matrix spike solution.) For the sample
in each analytical batch selected for spiking, add 1.0 mL of the matrix spiking
standard. For base/neutral-acid analysis, the amount of the surrogates and
matrix spiking compounds added to the sample should result in a final
concentration of 100 ng//iL of each base/neutral analyte and 200 ng/juL of each
acid analyte in the extract to be analyzed (assuming a 1 /nL injection). If
Method 3640, Gel-Permeation Cleanup, is to be used, add twice the volume of
surrogates and matrix spiking compounds since half the extract is lost due to
loading of the GPC column.
7.2 Add 300-500 mL of methylene chloride to the distilling flask. Add
several boiling chips to the flask.
3520B - 3 Revision 2
September 1994
-------
7.3 Add sufficient water to the extractor to ensure proper operation and
extract for 18-24 hours.
7.4 Allow to cool; then detach the boiling flask. If extraction at a
secondary pH is not required (see Table 1), the extract is dried and concentrated
using one of the techniques referred to in Sec. 7.7.
7.5 Carefully, while stirring, adjust the pH of the aqueous phase to the
second pH indicated in Table 1. Attach a clean distilling flask containing
500 ml of methylene chloride to the continuous extractor. Extract for 18-24
hours, allow to cool, and detach the distilling flask.
7.6 If performing GC/MS analysis (Method 8270), the acid/neutral and base
extracts may be combined prior to concentration. However, in some situations,
separate concentration and analysis of the acid/neutral and base extracts may be
preferable (e.g. if for regulatory purposes the presence or absence of specific
acid/neutral and base compounds at low concentrations must be determined,
separate extract analyses may be warranted).
7.7 Perform concentration (if necessary) using the Kuderna-Danish (K-D)
Technique (Sees. 7.8.1 through 7.8.4).
7.8 K-D Technique
7.8.1 Assemble a Kuderna-Danish (K-D) concentrator by attaching a 10
ml concentrator tube to a 500 ml evaporation flask. Dry the extract by
passing it through a drying column containing about 10 cm of anhydrous
sodium sulfate. Collect the dried extract in a K-D concentrator. Rinse
the flask which contained the solvent extract with 20-30 mL of methylene
chloride and add it to the column to complete the quantitative transfer.
7.8.2 Add one or two clean boiling chips to the flask and attach a
three ball Snyder column. Prewet the Snyder column by adding about 1 ml
of methylene chloride to the top of the column. Place the K-D apparatus
on a hot water bath (15-20°C above the boiling point of the solvent) so
that the concentrator tube is partially immersed in the hot water and the
entire lower rounded surface of the flask is bathed with hot vapor.
Adjust the vertical position of the apparatus and the water temperature,
as required, to complete the concentration in 10-20 minutes. At the
proper rate of distillation the balls of the column will actively chatter,
but the chambers will not flood. When the apparent volume of liquid
reaches 1 ml, remove the K-D apparatus from the water bath and allow it to
drain and cool for at least 10 minutes. Remove the Snyder column and
rinse the flask and its lower joints into the concentrator tube with 1-2
ml of extraction solvent.
7.8.3 If a solvent exchange is required (as indicated in Table 1),
momentarily remove the Snyder column, add 50 ml of the exchange solvent,
a new boiling chip, and reattach the Snyder column. Concentrate the
extract, as described in Sec. 7.9, raising the temperature of the water
bath, if necessary, to maintain proper distillation.
3520B - 4 Revision 2
September 1994
-------
7.8.4 Remove the Snyder column and rinse the flask and its lower
joints into the concentrator tube with 1-2 mL of methylene chloride or
exchange solvent. If sulfur crystals are a problem, proceed to Method
3660 for cleanup. The extract may be further concentrated by using the
techniques outlined in Sec. 7.9 or adjusted to 10.0 ml with the solvent
last used.
7.9 If further concentration is indicated in Table 1, either the micro-
Snyder column technique (7.9.1) or nitrogen blowdown technique (7.9.2) is used
to adjust the extract to the final volume required.
7.9.1 Micro-Snyder Column Technique
7.9.1.1 Add another one or two clean boiling chips to the
concentrator tube and attach a two ball micro-Snyder column. Prewet
the column by adding 0.5 ml of methylene chloride or exchange
solvent to the top of the column. Place the K-D apparatus in a hot
water bath so that the concentrator tube is partially immersed in
the hot water. Adjust the vertical position of the apparatus and
the water temperature, as required, to complete the concentration in
5-10 minutes. At the proper rate of distillation the balls of the
column will actively chatter, but the chambers will not flood. When
the apparent volume of liquid reaches 0.5 ml, remove the K-D
apparatus from the water bath and allow it to drain and cool for at
least 10 minutes. Remove the Snyder column, rinse the flask and its
lower joints into the concentrator tube with 0.2 mL of methylene
chloride or exchange solvent, and adjust the final volume to 1.0 to
2.0 ml, as indicated in Table 1, with solvent.
7.9.2 Nitrogen Blowdown Technique
7.9.2.1 Place the concentrator tube in a warm bath (35°C)
and evaporate the solvent volume to 0.5 ml using a gentle stream of
clean, dry nitrogen (filtered through a column of activated carbon).
CAUTION: New plastic tubing must not be used between the
carbon trap and the sample, since it may
introduce interferences.
7.9.2.2 The internal wall of the tube must be rinsed down
several times with methylene chloride or appropriate solvent during
the operation. During evaporation, the tube solvent level must be
positioned to avoid water condensation. Under normal procedures,
the extract must not be allowed to become dry.
CAUTION: When the volume of solvent is reduced below 1 ml,
semivolatile analytes may be lost.
7.10 The extract may now be analyzed for the target analytes using the
appropriate determinative technique(s) (see Sec. 4.3 of this Chapter). If
analysis of the extract will not be performed immediately, stopper the
concentrator tube and store refrigerated. If the extract will be stored longer
3520B - 5 Revision 2
September 1994
-------
than 2 days it should be transferred to a vial with a Teflon lined screw-cap or
crimp top, and labeled appropriately.
8.0 QUALITY CONTROL
8.1 Any reagent blanks, matrix spike, or replicate samples should be
subjected to exactly the same analytical procedures as those used on actual
samples.
8.2 Refer to Chapter One for specific quality control procedures and
Method 3500 for extraction and sample-preparation procedures.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
3520B - 6 Revision 2
September 1994
-------
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METHOD 3520B
CONTINUOUS LIQUID-LIQUID EXTRACTION
7.1 Add appropriate
surrogate and
matrix spiking
solutions.
7.7 - 7.8
Concentrate extract
7.2 Add methylene
chloride to
distilling flask.
7.8.3 Is
solvent
exchange
required?
7.8.3 Add
exchange solvent;
concentration extract
7.3 Add reagent
water to extractor
extract for 18-24
hours.
7.9 Further
concentrate extract
if necessary;
adjust final volume.
7.5 Adjust pH of
aqueous phase;
extract for 18-24
hours with clean
flask.
7.10 Analyze using
organic techniques.
7.6
GC/MS
malysis
(Method 8270)
performed?
8000
Series
Methods
7.6 Combine acid
and base/neutral
extracts prior to
concentration.
3520B - 8
Revision 2
September 1994
-------
METHOD 3540B
SOXHLET EXTRACTION
1.0 SCOPE AND APPLICATION
1.1 Method 3540 is a procedure for extracting nonvolatile and semi-
volatile organic compounds from solids such as soils, sludges, and wastes. The
Soxhlet extraction process ensures intimate contact of the sample matrix with the
extraction solvent.
1.2 This method is applicable to the isolation and concentration of water
insoluble and slightly water soluble organics in preparation for a variety of
chromatographic procedures.
2.0 SUMMARY OF METHOD
2.1 The solid sample is mixed with anhydrous sodium sulfate, placed in
an extraction thimble or between two plugs of glass wool, and extracted using an
appropriate solvent in a Soxhlet extractor. The extract is then dried,
concentrated (if necessary), and, as necessary, exchanged into a solvent
compatible with the cleanup or determinative step being employed.
3.0 INTERFERENCES
3.1 Refer to Method 3500.
4.0 APPARATUS AND MATERIALS
4.1 Soxhlet extractor - 40 mm ID, with 500 mL round bottom flask.
4.2 Drying column - 20 mm ID Pyrex chromatographic column with Pyrex
glass wool at bottom.
NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without
frits may be purchased. Use a small pad of Pyrex glass wool to
retain the adsorbent. Prewash the glass wool pad with 50 ml of
acetone followed by 50 mL of elution solvent prior to packing the
column with adsorbent.
4.3 Kuderna-Danish (K-D) apparatus
4.3.1 Concentrator tube - 10 mL, graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
3540B - 1 Revision 2
September 1994
-------
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.5 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 5°C). The bath should be used in a hood.
4.6 Vials - Glass, 2 ml capacity, with Teflon lined screw or crimp top.
4.7 Glass or paper thimble or glass wool - Contaminant free.
4.8 Heating mantle - Rheostat controlled.
4.9 Disposable glass pasteur pipet and bulb.
4.10 Apparatus for determining percent dry weight.
4.10.1 Oven - Drying.
4.10.2 Desiccator.
4.10.3 Crucibles - Porcelain or disposable aluminum.
4.11 Apparatus for grinding
4.12 Analytical balance - 0.0001 g.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
3540B - 2 Revision 2
September 1994
-------
5.4 Extraction solvents
5.4.1 Soil/sediment and aqueous sludge samples shall be extracted
using either of the following solvent systems:
5.4.1.1 Acetone/Hexane (1:1) (v/v), CH3COCH3/C6H14.
Pesticide quality or equivalent.
NOTE: This solvent system has lower disposal cost and lower
toxicity.
5.4.1.2 Methylene chloride/Acetone (1:1 v/v),
CH2C12/CH3COCH3. Pesticide quality or equivalent.
5.4.2 Other samples shall be extracted using the following:
5.4.2.1 Methylene chloride, CH2C12. Pesticide quality or
equivalent.
5.4.2.2 Toluene/Methanol (10:1) (v/v), C6H5CH3/CH3OH.
Pesticide quality or equivalent.
5.5 Exchange solvents
5.5.1 Hexane, C6H14. Pesticide quality or equivalent.
5.5.2 2-Propanol, (CH3)2CHOH. Pesticide quality or equivalent.
5.5.3 Cyclohexane, C6H12. Pesticide quality or equivalent.
5.5.4 Acetonitrile, CH3CN. Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analysis, Sec.
4.1.
7.0 PROCEDURE
7.1 Sample Handling
7.1.1 Sediment/soil samples - Decant and discard any water layer on
a sediment sample. Mix sample thoroughly, especially composited samples.
Discard any foreign objects such as sticks, leaves, and rocks.
7.1.2 Waste samples - Samples consisting of multiphases must be
prepared by the phase separation method in Chapter Two before extraction.
This procedure is for solids only.
7.1.3 Dry waste samples amenable to grinding - Grind or otherwise
subdivide the waste so that it either passes through a 1 mm sieve or can
3540B - 3 Revision 2
September 1994
-------
be extruded through a 1 mm hole. Introduce sufficient sample into the
grinding apparatus to yield at least 10 g after grinding.
7.1.4 Gummy, fibrous, or oily materials not amenable to grinding
should be cut, shredded, or otherwise broken up to allow mixing, and
maximum exposure of the sample surfaces for extraction. The professional
judgment of the analyst is required for handling these difficult matrices.
7.2 Determination of sample % dry weight - In certain cases, sample
results are desired based on dry weight basis. When such data are desired, a
portion of sample for this determination should be weighed out at the same time
as the portion used for analytical determination.
WARNING: The drying oven should be contained in a hood or vented.
Significant laboratory contamination may result from a heavily
contaminated hazardous waste sample.
However, samples known or suspected to contain significant concentrations
of toxic, flammable, or explosive constituents should not be oven dried because
of concerns for personal safety. Laboratory discretion is advised. It may be
prudent to delay oven drying of the weighed-out portion until other analytical
results are available.
7.2.1 Immediately after weighing the sample for extraction, weigh 5-
10 g of the sample into a tared crucible. Determine the % dry weight of
the sample by drying overnight at 105°C. Allow to cool in a desiccator
before weighing:
% dry weight = q of dry sample x 100
g of sample
7.3 Blend 10 g of the solid sample with 10 g of anhydrous sodium sulfate
and place in an extraction thimble. The extraction thimble must drain freely for
the duration of the extraction period. A glass wool plug above and below the
sample in the Soxhlet extractor is an acceptable alternative for the thimble.
Add 1.0 ml of the surrogate standard spiking solution onto the sample (see Method
3500 for details on the surrogate standard and matrix spiking solutions). For
the sample in each analytical batch selected for spiking, add 1.0 ml of the
matrix spiking standard. For base/neutral-acid analysis, the amount added of the
surrogates and matrix spiking compounds should result in a final concentration
of 100 ng/juL of each base/neutral analyte and 200 ng//iL of each acid analyte in
the extract to be analyzed (assuming a 1 /xL injection). If Method 3640, Gel
Permeation Chromatography Cleanup, is to be used, add twice the volume of
surrogates and matrix spiking compounds since half the extract is lost due to
loading of the GPC column.
7.4 Place approximately 300 mL of the extraction solvent (Sec. 5.4) into
a 500 ml round bottom flask containing one or two clean boiling chips. Attach
the flask to the extractor and extract the sample for 16-24 hours at 4-6
cycles/hr.
7.5 Allow the extract to cool after the extraction is complete.
3540B - 4 Revision 2
September 1994
-------
7.6 Assemble a Kuderna-Danish (K-D) concentrator (if necessary) by
attaching a 10 ml concentrator tube to a 500 ml evaporation flask.
7.7 Dry the extract by passing it through a drying column containing
about 10 cm of anhydrous sodium sulfate. Collect the dried extract in a K-D
concentrator. Wash the extractor flask and sodium sulfate column with 100 to 125
ml of extraction solvent to complete the quantitative transfer.
7.8 Add one or two clean boiling chips to the flask and attach a three
ball Snyder column. Prewet the Snyder column by adding about 1 mL of methylene
chloride to the top of the column. Place the K-D apparatus on a hot water bath
(15-20°C above the boiling point of the solvent) so that the concentrator tube
is partially immersed in the hot water and the entire lower rounded surface of
the flask is bathed with hot vapor. Adjust the vertical position of the
apparatus and the water temperature, as required, to complete the concentration
in 10-20 minutes. At the proper rate of distillation the balls of the column
will actively chatter, but the chambers will not flood. When the apparent volume
of liquid reaches 1-2 mL, remove the K-D apparatus from the water bath and allow
it to drain and cool for at least 10 minutes.
7.9 If a solvent exchange is required (as indicated in Table 1),
momentarily remove the Snyder column, add approximately 50 ml of the exchange
solvent and a new boiling chip, and reattach the Snyder column. Concentrate the
extract as described in Sec. 7.8, raising the temperature of the water bath, if
necessary, to maintain proper distillation. When the apparent volume again
reaches 1-2 mL, remove the K-D apparatus from the water batch and allow it to
drain and cool for at least 10 minutes.
7.10 Remove the Snyder column and rinse the flask and its lower joints
into the concentrator tube with 1-2 mL of methylene chloride or exchange solvent.
If sulfur crystals are a problem, proceed to Method 3660 for cleanup. The
extract may be further concentrated by using the techniques described in Sec.
7.11 or adjusted to 10.0 mL with the solvent last used.
7.11 If further concentration is indicated in Table 1, either micro Snyder
column technique (Sec. 7.11.1) or nitrogen blowdown technique (Sec. 7.11.2) is
used to adjust the extract to the final volume required.
7.11.1 Micro Snyder Column Technique
7.11.1.1 Add another one or two clean boiling chips to the
concentrator tube and attach a two ball micro Snyder column. Prewet
the column by adding about 0.5 mL of methylene chloride or exchange
solvent to the top of the column. Place the K-D apparatus in a hot
water bath so that the concentrator tube is partially immersed in
the hot water. Adjust the vertical position of the apparatus and
the water temperature, as required, to complete the concentration in
5-10 minutes. At the proper rate of distillation the balls of the
column will actively chatter, but the chambers will not flood. When
the apparent volume of liquid reaches 0.5 mL, remove the K-D
apparatus from the water bath and allow it to drain and cool for at
least 10 minutes. Remove the Snyder column and rinse the flask and
its lower joints with about 0.2 mL of solvent and add to the
3540B - 5 Revision 2
September 1994
-------
concentrator tube. Adjust the final volume to 1.0-2.0 ml, as
indicated in Table 1, with solvent.
7.11.2 Nitrogen Slowdown Technique
7.11.2.1 Place the concentrator tube in a warm water bath
(approximately 35°C) and evaporate the solvent volume to the
required level using a gentle stream of clean, dry nitrogen
(filtered through a column of activated carbon).
CAUTION: Do not use plasticized tubing between the carbon
trap and the sample.
7.11.2.2 The internal wall of the tube must be rinsed down
several times with the appropriate solvent during the operation.
During evaporation, the solvent level in the tube must be positioned
to prevent water from condensing into the sample (i.e., the solvent
level should be below the level of the water bath). Under normal
operating conditions, the extract should not be allowed to become
dry.
CAUTION: When the volume of solvent is reduced below 1 ml,
semi volatile analytes may be lost.
7.12 The extracts obtained may now be analyzed for the target analytes
using the appropriate organic technique(s) (see Sec. 4.3 of this Chapter). If
analysis of the extract will not be performed immediately, stopper the
concentrator tube and store in a refrigerator. If the extract will be stored
longer than 2 days, it should be transferred to a vial with a Teflon lined screw
cap or crimp top, and labeled appropriately.
8.0 QUALITY CONTROL
8.1 Any reagent blanks or matrix spike samples should be subjected to
exactly the same analytical procedures as those used on actual samples.
8.2 Refer to Chapter One for specific quality control procedures and
Method 3500 for extraction and sample preparation procedures.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
3540B - 6 Revision 2
September 1994
-------
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METHOD 3540B
SOXHLET EXTRACTION
'
7.1
Use appropriate
sample handling
technique
'
7.2
Determine sample %
dry weight
\
t
7.3
Add appropriate
surrogate and matrix
spiking standards
i
7.4
Add extraction
solvent to flask:
extract for 16-24
hours
i
75
Cool extract
7.6
Assemble K-D
concentrator
7.7
Dry and collect
extract in K-D
concentrator
7.8
Concentrate using
Snyder column
and K-D apparatus
7.9
Is solvent
exchange required?
7.12
Analyze using
organic techniques
Proceed
to Method
3660 for
cleanup
8000
Series
Methods
7.9
Add exchange
solvent,
reconcentrate extract
3540B - 8
Revision 2
September 1994
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METHOD 3541
AUTOMATED SOXHLET EXTRACTION
1.0 SCOPE AND APPLICATION
1.1 Method 3541 describes the extraction of organic analytes from soil,
sediment, sludges, and waste solids. The method uses a commercially available,
unique, three stage extraction system to achieve analyte recovery comparable to
Method 3540, but in a much shorter time. There are two differences between this
extraction method and Method 3540. In the initial extraction stage of Method
3541, the sample-loaded extraction thimble is immersed into the boiling solvent.
This ensures very rapid intimate contact between the specimen and solvent and
rapid extraction of the organic analytes. In the second stage the thimble is
elevated above the solvent, and is rinse-extracted as in Method 3540. In the
third stage, the solvent is evaporated, as would occur in the Kuderna-Danish
(K-D) concentration step in Method 3540. The concentrated extract is then ready
for cleanup (Method 3600) followed by measurement of the organic analytes.
1.2 The method is applicable to the extraction and concentration of water
insoluble or slightly water soluble polychlorinated biphenyls (PCBs) in
preparation for gas chromatographic determination using either Method 8080 or
8081. This method is applicable to soils, clays, solid wastes and sediments
containing from 1 to 50 jug of PCBs (measured as Arochlors) per gram of sample.
It has been statistically evaluated at 5 and 50 /ng/g of Arochlors 1254 and 1260,
and found to be equivalent to Method 3540 (Soxhlet Extraction). Higher
concentrations of PCBs are measured following volumetric dilution with hexane.
1.3 The method is also applicable the extraction and concentration of
semivolatile organics in preparation for GC/MS analysis by Method 8270 or by
analysis using specific GC or HPLC methods.
2.0 SUMMARY OF METHOD
2.1 PCBs: Moist solid samples (e.g., soil/sediment samples) may be air-
dried and ground prior to extraction or chemically dried with anhydrous sodium
sulfate. The prepared sample is extracted using 1:1 (v/v) acetone:hexane in the
automated Soxhlet following the same procedure as outlined for semivolatile
organics in Sec. 2.1. The extract is then concentrated and exchanged into pure
hexane prior to final gas chromatographic PCB measurement.
2.2 Other semivolatile organics: A 10-g solid sample (the sample is pre-
mixed with anhydrous sodium sulfate for certain matrices) is placed in an
extraction thimble and usually extracted with 50 ml of 1:1 (v/v) acetone/hexane
for 60 minutes in the boiling extraction solvent. The thimble with sample is
then raised into the rinse position and extracted for an additional 60 minutes.
Following the extraction steps, the extraction solvent is concentrated to 1 to
2 ml.
3541 - 1 Revision 0
September 1994
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3.0 INTERFERENCES
3.1 Refer to Method 3500.
3.2 The extraction thimble and the o-rings used to seal the extraction
cup are both a source of interference. Both should be checked by including a
method blank and following the extraction procedure as written. Solvent rinsing
or extraction, prior to use, may be necessary to eliminate or reduce
interferences. Viton seals contributed least to the interference problem,
however, even they contributed some interference peaks when the extraction
solvent was analyzed by the electron capture detector. Use of butyl or EPDM
rings are not recommended since they were found to contribute significant
background when the extraction solvent was 1:1 v/v hexane/acetone or 1:1 v/v
methylene chloride/acetone.
4.0 APPARATUS AND MATERIALS
4.1 Automated Soxhlet Extraction System - with temperature-controlled oil
bath (Soxtec, or equivalent). Tecator bath oil (catalog number 1000-1886) should
be used with the Soxtec. Silicone oil must not be used because it destroys the
rubber parts. See Figure 1. The apparatus is used in a hood.
4.2 Accessories and consumables for the automated Soxhlet system. (The
catalog numbers are Fisher Scientific based on the use of the Soxtec HT-6,
however, other sources that are equivalent are acceptable.)
4.2.1 Cellulose extraction thimbles - 26 mm ID x 60 mm
contamination free, catalog number 1522-0034, or equivalent.
4.2.2 Glass extraction cups (80 ml) - (set of six required for the
HT-6), catalog number 1000-1820.
4.2.3 Thimble adapters - (set of six required for the HT-6),
catalog number 1000-1466.
4.2.4 Viton seals - catalog number 1000-2516.
4.3 Syringes - 100 and 1000 juL and 5 mL.
4.4 Apparatus for Determining Percent Dry Weight
4.4.1 Drying Oven.
4.4.2 Desiccator.
4.4.3 Crucibles, porcelain.
4.4.4 Balance, analytical.
4.5 Apparatus for grinding - Fisher Cyclotec, Fisher Scientific catalog
number 1093, or equivalent.
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4.6 Spatula
4.7 Graduated cylinder - 100 ml.
4.8 Aluminum weighing dish - VWR Scientific catalog number 25433-008 or
equivalent.
4.9 Graduated, conical-bottom glass tubes - 15 ml, Kimble catalog number
45166 or equivalent, or 10 ml KD concentrator tube.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. A method blank must be analyzed, demonstrating that there
is no interference from the sodium sulfate.
5.4 Extraction solvents:
5.4.1 Organochlorine pesticides/PCB extraction:
5.4.1.1 Acetone/hexane (1:1 v/v), CH3COCH3/C6H14.
Pesticide quality or equivalent.
5.4.2 Semivolatile organics extraction:
5.4.2.1 Acetone/hexane (1:1 v/v), CH3COCH3/C6H14.
Pesticide quality or equivalent.
5.4.2.2 Acetone/methylene chloride (1:1 v/v),
CH3COCH3/CH2C12. Pesticide quality or equivalent.
5.5 Hexane, C6H14. Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
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7.0 PROCEDURE
7.1 Sample handling
7.1.1 Sediment/soil samples - Decant and discard any water layer
on a sediment sample. Mix sample thoroughly, especially composited
samples. Discard any foreign objects such as sticks, leaves, and rocks.
7.1.1.1 PCBs or high-boiling organochlorine pesticides -
Air-dry the sample at room temperature for 48 hours in a glass tray
or on hexane-cleaned aluminum foil, or dry the sample by mixing with
anhydrous sodium sulfate until a free-flowing powder is obtained
(see Sec. 7.2).
NOTE: Dry, finely ground soil/sediment allows the best
extraction efficiency for non-volatile, non-polar
organics, e.g., PCBs, 4,4'-DDT, etc. Air-drying
is not appropriate for the analysis of the more
volatile organochlorine pesticides (e.g. the
BHCs) or the more volatile of the semivolatile
organics because of losses during the drying
process.
7.1.2 Dried sediment/soil and dry waste samples amenable to
grinding - Grind or otherwise subdivide the waste so that it either passes
through a 1 mm sieve or can be extruded through a 1 mm hole. Introduce
sufficient sample into the grinding apparatus to yield at least 20 g after
grinding. Disassemble grinder between samples, according to
manufacturer's instructions, and clean with soap and water, followed by
acetone and hexane rinses.
NOTE: The same warning on loss of volatile analytes applies to the
grinding process. Grinding should only be performed when
analyzing for non-volatile organics.
7.1.3 Gummy, fibrous, or oily materials not amenable to grinding
should be cut, shredded, or otherwise broken up to allow mixing, and
maximum exposure of the sample surfaces for extraction. If grinding of
these materials is preferred, the addition and mixing of anhydrous sodium
sulfate with the sample (1:1) may improve grinding efficiency. The
professional judgment of the analyst is required for handling such
difficult matrices.
7.1.4 Multiple phase waste samples - Samples consisting of multiple
phases must be prepared by the phase separation method in Chapter Two
before extraction. This procedure is for solids only.
7.2 For sediment/soil (especially gummy clay) that is moist and cannot
be air-dried because of loss of volatile analytes - Mix 5 g of sample with 5 g
of anhydrous sodium sulfate in a small beaker using a spatula. Use this approach
for any solid sample that requires dispersion of the sample particles to ensure
greater solvent contact throughout the sample mass.
3541 - 4 Revision 0
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7.3 Determination of sample percent dry weight - In certain cases, sample
results are desired based on dry weight basis. When such data are desired, a
portion of sample for this determination should be weighed out at the same time
as the portion used for analytical determination.
WARNING: The drying oven should be contained in a hood or vented.
Significant laboratory contamination may result from the
drying of a heavily contaminated hazardous waste sample.
7.3.1 Immediately after weighing the sample for extraction, weigh
5-10 g of the sample into a tared crucible. Determine the % dry weight of
the sample by drying overnight at 105°C. Allow to cool in a desiccator
before weighing:
% dry weight = g of dry sample x 100
g of sample
7.4 Check the heating oil level in the automated Soxhlet unit and add oil
if needed. See service manual for details. Set the temperature on the service
unit at 140°C when using hexane-acetone (1:1, v/v) as the extraction solvent.
7.5 Press the "MAINS" button; observe that the switch lamp is now "ON".
7.6 Open the cold water tap for the reflux condensers. Adjust the flow
to 2 L/min to prevent solvent loss through the condensers.
7.7 Weigh 10 g of sample into extraction thimbles. For samples mixed
with anhydrous sodium sulfate, transfer the entire contents of the beaker (Sec.
7.2) to the thimble. Add surrogate spikes to each sample and the matrix
spike/matrix spike duplicate to the selected sample.
NOTE: When surrogate spikes and/or matrix spikes contain relatively
volatile compounds (e.g., trichlorobenzenes, BHCs, etc.), steps 7.8,
7.9, and 7.10 must be performed quickly to avoid evaporation losses
of these compounds. As the spike is added to the sample in each
thimble, the thimble should immediately be transferred to the
condenser and lowered into the extraction solvent.
7.8 Immediately transfer the thimbles containing the weighed samples into
the condensers. Raise the knob to the "BOILING" position. The magnet will now
fasten to the thimble. Lower the knob to the "RINSING" position. The thimble
will now hang just below the condenser valve.
7.9 Insert the extraction cups containing boiling chips, and load each
with 50 mL of extraction solvent (normally 1:1 (v/v) hexane:acetone, see Sec.
5.4). Using the cup holder, lower the locking handle, ensuring that the safety
catch engages. The cups are now clamped into position. (The seals must be pre-
rinsed or pre-extracted with extraction solvent prior to initial use.)
7.10 Move the extraction knobs to the "BOILING" position. The thimbles
are now immersed in solvent. Set the timer for 60 minutes. The condenser valves
must be in the "OPEN" position. Extract for the preset time.
3541 - 5 Revision 0
September 1994
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7.11 Move the extraction knobs to the "RINSING" position. The thimbles
will now hang above the solvent surface. Set timer for 60 minutes. Condenser
valves are still open. Extract for the preset time.
7.12 After rinse time has elapsed, close the condenser valves by turning
each a quarter-turn, clockwise.
7.13 When all but 2 to 5 ml of solvent have been collected, open the
system and remove the cups.
7.14 Transfer the contents of the cups to 15 ml graduated, conical-bottom
glass tubes. Rinse the cups using hexane (methylene chloride if 1:1 methylene
chloride-acetone was used for extraction and analysis is by GC/MS) and add the
rinsates to the glass tubes. Concentrate the extracts to 1 to 10 ml. The final
volume is dependent on the determinative method and the quantitation limit
required. Transfer a portion to a GC vial and store at 4°C until analyses are
performed.
NOTE: The recovery solvent volume can be adjusted by adding
solvent at the top of the condensers. For more details
concerning use of the extractor, see the operating manual
for the automated extraction system.
7.15 Shutdown
7.15.1 Turn "OFF" main switch.
7.15.2 Turn "OFF" cold water tap.
7.15.3 Ensure that all condensers are free of solvent. Empty
the solvent that is recovered in the evaporation step into an appropriate
storage container.
7.16 The extract is now ready for cleanup or analysis, depending on the
extent of interfering co-extractives. See Method 3600 for guidance on cleanup
methods and Method 8000 for guidance on determinative methods. Certain cleanup
and/or determinative methods may require a solvent exchange prior to cleanup
and/or determination.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for general quality control procedures and to
Method 3500 for specific extraction and sample preparation QC procedures.
8.2 Before processing any samples, the analyst should demonstrate through
the analysis of an organic-free solid matrix (e.g., reagent sand) method blank
that all glassware and reagents are interference-free. Each time a set of
samples is extracted, or when there is a change in reagents, a method blank
should be processed as a safeguard against chronic laboratory contamination. The
blank samples should be carried through all stages of the sample preparation and
measurement. This is especially important because of the possibility of
interferences being extracted from the extraction cup seal.
3541 - 6 Revision 0
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8.3 Standard quality assurance practices should be used with this method.
Field duplicates should be collected to validate the precision of the sampling
technique. Each analysis batch of 20 or less samples must contain: a method
blank, either a matrix spike/matrix spike duplicate or a matrix spike and
duplicate sample analysis, and a laboratory control sample, unless the
determinative method provides other guidance. Also, routinely check the
integrity of the instrument seals.
8.4 Surrogate standards must be added to all samples when specified in
the appropriate determinative method.
9.0 METHOD PERFORMANCE
9.1 Multi-laboratory accuracy and precision data were obtained for PCBs
in soil. Eight laboratories spiked Arochlors 1254 and 1260 into three portions
of 10 g of Fuller's Earth on three non-consecutive days followed by immediate
extraction using Method 3541. Six of the laboratories spiked each Arochlor at
5 and 50 mg/kg and two laboratories spiked each Arochlor at 50 and 500 mg/kg.
All extracts were analyzed by Oak Ridge National Laboratory, Oak Ridge, TN, using
Method 8081. These data are listed in a table found in Method 8081, and were
taken from Reference 1.
9.2 Single-laboratory accuracy data were obtained for chlorinated
hydrocarbons, nitroaromatics, haloethers, and organochlorine pesticides in a clay
soil. The spiking concentrations ranged from 500 to 5000 jixg/kg, depending on
the sensitivity of the analyte to the electron capture detector. The spiking
solution was mixed into the soil during addition and then immediately transferred
to the extraction device and immersed in the extraction solvent. The data
represents a single determination. Analysis was by capillary column gas
chromatography/electron capture detector following Methods 8081 for the
organochlorine pesticides, 8091 for the nitroaromatics, 8111 for the
hydrocarbons, and 8121 for the chlorinated hydrocarbons. These data are listed
in a table located in their respective methods and were taken from Reference 2.
9.3 Single-laboratory accuracy and precision data were obtained for
semivolatile organics in soil by spiking at a concentration of 6 mg/kg for each
compound. The spiking solution was mixed into the soil during addition and then
allowed to equilibrate for approximately 1 hr prior to extraction. Three
determinations were performed and each extract was analyzed by gas
chromatography/mass spectrometry following Method 8270. The low recovery of the
more volatile compounds is probably due to volatilization losses during
equilibration. These data are listed in a Table located in Method 8270 and were
taken from Reference 2.
10.0 REFERENCES
1. Stewart, J. "Intra-Laboratory Recovery Data for the PCB Extraction
Procedure"; Oak Ridge National Laboratory, Oak Ridge, TN, 37831-6138;
October 1989.
3541 - 7 Revision 0
September 1994
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2. Lopez-Avila, V. (Beckert, W., Project Officer), "Development of a Soxtec
Extraction Procedure for Extracting Organic Compounds from Soils and
Sediments", EPA 600/X-91/140, US EPA, Environmental Monitoring Systems
Laboratory-Las Vegas, October 1991.
3541 - 8 Revision 0
September 1994
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Figure 1
Automated Soxhlet Extraction System
Condenser
Thimble
Glass Wool Plug
Sample
Aluminum beaker (cup)
Hot plate
3541 - 9
Revision 0
September 1994
-------
METHOD 3541
AUTOMATED SOXHLET EXTRACTION
i
7.1
Use appropriate
sample handling
technique.
7.2
Add anhydrous
Na2SO4if
necessary
7.3
Determine percent
dry weight.
7.4
Check oil
level in
Soxhlet unit.
7.5
Press "Mains"
button.
I
7.6
Open Cold water
tap. Adjust flow.
7.7
Weigh sample into
extraction thimbles.
Add surrogate
spike.
7.8
Transfer samples
into condensers.
Adjust position of
magnet and thimble.
1
7.9
Insert extraction
cups and toad
with solvent.
I
7.10
Move extraction
knobs to
"Boiling" for
60 mins.
©
3541 - 10
7.11
Move extraction
knobs to
"Rinsing" for
60 mins.
7.12
Close
condenser valves.
7.13
Remove cups.
I
7.14
Transfer contents
to collection
vials, dilute or
concentrate to
volume.
I
7.15
Shutdown
Revision 0
September 1994
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METHOD 3550A
ULTRASONIC EXTRACTION
See DISCLAIMER-1. See manufacturer's specifications for operational settings.
1.0 SCOPE AND APPLICATION
1.1 Method 3550 is a procedure for extracting nonvolatile and semi-
volatile organic compounds from solids such as soils, sludges, and wastes. The
ultrasonic process ensures intimate contact of the sample matrix with the
extraction solvent.
1.2 The method is divided into two sections, based on the expected
concentration of organics in the sample. The low concentration method
(individual organic components of < 20 mg/kg) uses a larger sample size and a
more rigorous extraction procedure (lower concentrations are more difficult to
extract). The medium/high concentration method (individual organic components
of > 20 mg/kg) is much simpler and therefore faster.
1.3 It is highly recommended that the extracts be cleaned up prior to
analysis. See Chapter Four (Cleanup), Sec. 4.2.2, for applicable methods.
2.0 SUMMARY OF METHOD
2.1 Low concentration method - A 30 g sample is mixed with anhydrous
sodium sulfate to form a free-flowing powder. This is solvent extracted three
times using ultrasonic extraction. The extract is separated from the sample by
vacuum filtration or centrifugation. The extract is ready for cleanup and/or
analysis following concentration.
2.2 Medium/high concentration method - A 2 g sample is mixed with
anhydrous sodium sulfate to form a free-flowing powder. This is solvent
extracted once using ultrasonic extraction. A portion of the extract is removed
for cleanup and/or analysis.
3.0 INTERFERENCES
3.1 Refer to Method 3500.
4.0 APPARATUS AND MATERIALS
4.1 Apparatus for grinding dry waste samples.
4.2 Ultrasonic preparation - A horn type device equipped with a titanium
tip, or a device that will give equivalent performance, shall be used.
3550A - 1 Revision 1
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4.2.1 Ultrasonic Disrupter - The disrupter must have a minimum power
wattage of 300 watts, with pulsing capability. A device designed to
reduce the cavitation sound is recommended. Follow the manufacturers
instructions for preparing the disrupter for extraction of samples with
low and medium/high concentration.
Use a 3/4" horn for the low concentration method and a 1/8" tapered
microtip attached to a 1/2" horn for the medium/high concentration method.
4.3 Sonabox - Recommended with above disrupters for decreasing cavitation
sound (Heat Systems - Ultrasonics, Inc., Model 432B or equivalent).
4.4 Apparatus for determining percent dry weight.
4.4.1 Oven - Drying.
4.4.2 Desiccator.
4.4.3 Crucibles - Porcelain or disposable aluminum.
4.5 Pasteur glass pipets - 1 ml, disposable.
4.6 Beakers - 400 ml.
4.7 Vacuum or pressure filtration apparatus.
4.7.1 Buchner funnel.
4.7.2 Filter paper - Whatman No. 41 or equivalent.
4.8 Kuderna-Danish (K-D) apparatus.
4.8.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.8.2 Evaporation flask - 500 mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.8.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.8.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.8.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.9 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
3550A - 2 Revision 1
September 1994
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4.10 Water bath - Heated, with concentric ring cover, capable of
temperature control (+ 5°C). The batch should be used in a hood.
4.11 Balance - Top loading, capable of accurately weighing to the nearest
0.01 g.
4.12 Vials - 2 ml, for GC autosampler, with Teflon lined screw caps or
crimp tops.
4.13 Glass scintillation vials - 20 ml, with Teflon lined screw caps.
4.14 Spatula - Stainless steel or Teflon.
4.15 Drying column - 20 mm ID Pyrex chromatographic column with Pyrex
glass wool at bottom.
NOTE:
Fritted glass discs are difficult to decontaminate after
highly contaminated extracts have been passed through.
Columns without frits may be purchased. Use a small pad of
Pyrex glass wool to retain the adsorbent. Prewash the glass
wool pad with 50 ml of acetone followed by 50 ml of elution
solvent prior to packing the column with adsorbent.
4.16 Syringe - 5 ml.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise specified, it is intended that all inorganic reagents shall conform to
the specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may be
used, provided it is first ascertained that the reagent is of sufficiently high
purity to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.4 Extraction solvents.
5.4.1 Low concentration soil/sediment and aqueous sludge samples
shall be extracted using a solvent system that gives optimum, reproducible
recovery for the matrix/analyte combination to be measured. Suitable
solvent choices are given in Table 1.
3550A - 3
Revision 1
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5.4.2 Methylene chloride:Acetone, CH2C12:CH3COCH3 (1:1, v:v).
Pesticide quality or equivalent.
5.4.3 Methylene chloride, CH2C12. Pesticide quality or equivalent.
5.4.4 Hexane, C6H14. Pesticide quality or equivalent.
5.5 Exchange solvents.
5.5.1 Hexane, C6H14. Pesticide quality or equivalent.
5.5.2 2-Propanol, (CH3)2CHOH. Pesticide quality or equivalent.
5.5.3 Cyclohexane, C6H12. Pesticide quality or equivalent.
5.5.4 Acetonitrile, CH3CN. Pesticide quality or equivalent.
5.5.5 Methanol, CH3OH. Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes, Sec.
4.1.
7.0 PROCEDURE
7.1 Sample handling
7.1.1 Sediment/soil samples - Decant and discard any water layer on
a sediment sample. Mix sample thoroughly, especially composited samples.
Discard any foreign objects such as sticks, leaves, and rocks.
7.1.1.2 Determine the dry weight of the sample (Sec. 7.2)
remaining after decanting. Measurement of soil pH may be required.
7.1.2 Waste samples - Samples consisting of multiphases must be
prepared by the phase separation method in Chapter Two before extraction.
This procedure is for solids only.
7.1.3 Dry waste samples amenable to grinding - Grind or otherwise
subdivide the waste so that it either passes through a 1 mm sieve or can
be extruded through a 1 mm hole. Introduce sufficient sample into the
grinder to yield at least 100 g after grinding.
7.1.4 Gummy, fibrous or oily materials not amenable to grinding
should be cut, shredded, or otherwise broken up to allow mixing, and
maximum exposure of the sample surfaces for extraction. The professional
judgment of the analyst is required for handling of these difficult
matrices.
3550A - 4 Revision 1
September 1994
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7.2 Determination of percent dry weight - In certain cases, sample
results are desired based on a dry weight basis. When such data are desired, or
required, a portion of sample for this determination should be weighed out at the
same time as the portion used for analytical determination.
WARNING: The drying oven should be contained in a hood or vented.
Significant laboratory contamination may result from drying a
heavily contaminated hazardous waste sample.
However, samples known or suspected to contain significant concentrations
of toxic, flammable, or explosive constituents should not be overdried because
of concerns for personal safety. Laboratory discretion is advised. It may be
prudent to delay overdrying of the weighed-out portion until other analytical
results are available.
7.2.1 Immediately after weighing the sample for extraction, weigh 5-
10 g of the sample into a tared crucible. Determine the % dry weight of
the sample by drying overnight at 105°C. Allow to cool in a desiccator
before weighing:
% dry weight = q of dry sample x 100
g of sample
7.3 Extraction method for samples expected to contain low concentrations
of organics and pesticides (< 20 mg/kg):
7.3.1 The following step should be performed rapidly to avoid loss
of the more volatile extractables. Weigh approximately 30 g of sample
into a 400 ml beaker. Record the weigh to the nearest 0.1 g. Nonporous
or wet samples (gummy or clay type) that do not have a free-flowing sandy
texture must be mixed with 60 g of anhydrous sodium sulfate, using a
spatula. If required, more sodium sulfate may be added. After addition
of sodium sulfate, the sample should be free flowing. Add 1 ml of
surrogate standards to all samples, spikes, standards, and blanks (see
Method 3500 for details on the surrogate standard solution and the matrix
spike solution). For the sample in each analytical batch selected for
spiking, add 1.0 ml of the matrix spiking standard. For base/neutral-acid
analysis, the amount added of the surrogates and matrix spiking compounds
should result in a final concentration of 100 ng/ni of each base/neutral
analyte and 200 ng//LtL of each acid analyte in the extract to be analyzed
(assuming a 1 juL injection). If Method 3640, Gel-Permeation Cleanup, is
to be used, add twice the volume of surrogates and matrix spiking
compounds since half of the extract is lost due to loading of the GPC
column. Immediately add 100 ml of 1:1 methylene chloride:acetone.
7.3.2 Place the bottom surface of the tip of the #207 3/4 in.
disrupter horn about 1/2 in. below the surface of the solvent, but above
the sediment layer.
7.3.3 Extract ultrasonically for 3 minutes, with output control knob
set at 10 (full power) and with mode switch on Pulse (pulsing energy
3550A - 5 Revision 1
September 1994
-------
rather than continuous energy) and percent-duty cycle knob set at 50%
(energy on 50% of time and off 50% of time). Do not use microtip probe.
7.3.4 Decant the extract and filter it through Whatman No. 41 filter
paper (or equivalent) in a Buchner funnel that is attached to a clean 500
ml filtration flask. Alternatively, decant the extract into a centrifuge
bottle and centrifuge at low speed to remove particles.
7.3.5 Repeat the extraction two or more times with two additional
100 ml portions of solvent. Decant off the solvent after each ultrasonic
extraction. On the final ultrasonic extraction, pour the entire sample
into the Buchner funnel and rinse with extraction solvent. Apply a vacuum
to the filtration flask, and collect the solvent extract. Continue
filtration until all visible solvent is removed from the funnel, but do
not attempt to completely dry the sample, as the continued application of
a vacuum may result in the loss of some analytes. Alternatively, if
centrifugation is used in Sec. 7.3.4, transfer the entire sample to the
centrifuge bottle. Centrifuge at low speed, and then decant the solvent
from the bottle.
7.3.6 Assemble a Kuderna-Danish (K-D) concentrator (if necessary) by
attaching a 10 ml concentrator tube to a 500 mL evaporator flask.
Transfer filtered extract to a 500 ml evaporator flask and proceed to the
next section.
7.3.7 Add one to two clean boiling chips to the evaporation flask,
and attach a three ball Snyder column. Prewet the Snyder column by adding
about 1 ml methylene chloride to the top. Place the K-D apparatus on a
hot water bath (80-90 °C) so that the concentrator tube is partially
immersed in the hot water and the entire lower rounded surface of the
flask is bathed with hot vapor. Adjust the vertical position of the
apparatus and the water temperature, as required, to complete the
concentration in 10-15 min. At the proper rate of distillation the balls
of the column will actively chatter, but the chambers will not flood with
condensed solvent. When the apparent volume of liquid reaches 1 ml,
remove the K-D apparatus and allow it to drain and cool for at least 10
min.
7.3.8 If a solvent exchange is required (as indicated in Table 1),
momentarily remove the Snyder column, add 50 ml of the exchange solvent
and a new boiling chip, and re-attach the Snyder column. Concentrate the
extract as described in Sec. 7.3.10, raising the temperature of the water
bath, if necessary, to maintain proper distillation. When the apparent
volume again reaches 1-2 ml, remove the K-D apparatus and allow it to
drain and cool for at least 10 minutes.
7.3.9 Remove the Snyder column and rinse the flask and its lower
joints into the concentrator tube with 1-2 ml of methylene chloride or
exchange solvent. If sulfur crystals are a problem, proceed to Method
3660 for cleanup. The extract may be further concentrated by using the
technique outlined in Sec. 7.3.10 or adjusted to 10.0 ml with the solvent
last used.
3550A - 6 Revision 1
September 1994
-------
7.3.10 If further concentration is indicated in Table 1, either
micro Snyder column technique (Sec. 7.3.10.1) or nitrogen blow down
technique (Sec. 7.3.10.2) is used to adjust the extract to the final
volume required.
7.3.10.1 Micro Snyder Column Technique
7.3.10.1.1 Add a clean boiling chip and attach a
two ball micro Snyder column to the concentrator tube. Prewet
the column by adding approximately 0.5 mL of methylene
chloride or exchange solvent through the top. Place the
apparatus in the hot water bath. Adjust the vertical position
and the water temperature, as required, to complete the
concentration in 5-10 minutes. At the proper rate of
distillation the balls of the column will actively chatter,
but the chambers will not flood. When the liquid reaches an
apparent volume of approximately 0.5 ml, remove the apparatus
from the water bath and allow to drain and cool for at least
10 minutes. Remove the micro Snyder column and rinse its
lower joint with approximately 0.2 ml of appropriate solvent
and add to the concentrator tube. Adjust the final volume to
the volume required for cleanup or for the determinative
method (see Table 1).
7.3.10.2 Nitrogen Slowdown Technique
7.3.10.2.1 Place the concentrator tube in a warm
water bath (approximately 35 °C) and evaporate the solvent
volume to the required level using a gentle stream of clean,
dry nitrogen (filtered through a column of activated carbon).
CAUTION: Do not use plasticized tubing between the
carbon trap and the sample.
7.3.10.2.2 The internal wall of the tube must be
rinsed down several times with the appropriate solvent during
the operation. During evaporation, the solvent level in the
tube must be positioned to prevent water from condensing into
the sample (i.e., the solvent level should be below the level
of the water bath). Under normal operating conditions, the
extract should not be allowed to become dry.
CAUTION: When the volume of solvent is reduced below
1 ml, semivolatile analytes may be lost.
7.4 If analysis of the extract will not be performed immediately, stopper
the concentrator tube and store refrigerated. If the extract will be stored
longer than 2 days, it should be transferred to a vial with a Teflon lined cap
and labeled appropriately.
3550A - 7 Revision 1
September 1994
-------
7.5 Extraction method for samples expected to contain high concentrations
of organics (> 20 mg/kg):
7.5.1 Transfer approximately 2 g (record weight to the nearest 0.1
g) of sample to a 20 ml vial. Wipe the mouth of the vial with a tissue to
remove any sample material. Record the exact weight of sample taken. Cap
the vial before proceeding with the next sample to avoid any cross
contamination.
7.5.2 Add 2 g of anhydrous sodium sulfate to sample in the 20 ml
vial and mix well.
7.5.3 Surrogate standards are added to all samples, spikes, and
blanks (see Method 3500 for details on the surrogate standard solution and
on the matrix spike solution). Add 1.0 ml of surrogate spiking solution
to sample mixture. For the sample in each analytical batch selected for
spiking, add 1.0 ml of the matrix spiking standard. For base/neutral-acid
analysis, the amount added of the surrogates and matrix spiking compounds
should result in a final concentration of 100 ng//xL of each base/neutral
analyte and 200 ng/^L of each acid analyte in the extract to be analyzed
(assuming a 1 fj.1 injection). If Method 3640, Gel-Permeation Cleanup, is
to be used, add twice the volume of surrogates and matrix spiking
compounds since half the extract is lost due to loading of the GPC column.
7.5.4 Immediately add whatever volume of solvent is necessary to
bring the final volume to 10.0 ml considering the added volume of
surrogates and matrix spikes. Disrupt the sample with the 1/8 in. tapered
microtip ultrasonic probe for 2 minutes at output control setting 5 and
with mode switch on pulse and percent duty cycle at 50%. Extraction
solvents are:
1. For nonpolar compounds (i.e., organochlorine pesticides and
PCBs), use hexane or appropriate solvent.
2. For extractable priority pollutants, use methylene chloride.
7.5.5 Loosely pack disposable Pasteur pipets with 2 to 3 cm Pyrex
glass wool plugs. Filter the extract through the glass wool and collect
5.0 ml in a concentrator tube if further concentration is required.
Follow Sec. 7.3.10 for details on concentration. Normally, the 5.0 mL
extract is concentrated to approximately 1.0 ml or less.
7.5.6 The extract is ready for cleanup or analysis, depending on the
extent of interfering co-extractives.
8.0 QUALITY CONTROL
8.1 Any reagent blanks or matrix spike samples should be subjected to
exactly the same analytical procedures as those used on actual samples.
3550A - 8 Revision 1
September 1994
-------
8.2 Refer to Chapter One for specific quality control procedures and
Method 3500 for extraction and sample preparation procedures.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative method for performance data.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
2. U.S. EPA, Inter!aboratory Comparison Study: Methods for Volatile and
Semi-Volatile Compounds, Environmental Monitoring Systems Laboratory,
Office of Research and Development, Las Vegas, NV, EPA 600/4-84-027, 1984.
3. Christopher S. Hein, Paul J. Marsden, Arthur S. Shurtleff, "Evaluation of
Methods 3540 (Soxhlet) and 3550 (Sonication) for Evaluation of Appendix IX
Analytes form Solid Samples", S-CUBED, Report for EPA Contract 68-03-33-
75, Work Assignment No. 03, Document No. SSS-R-88-9436, October 1988.
3550A - 9 Revision 1
September 1994
-------
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METHOD 3550A
ULTRASONIC EXTRACTION
| Start j
~^
7.1 Prepare samples
ucing appropriate method
for the waete matrix
7.2 Determine the
percent dry weight
of the eample
7.5.2 Add anhydroue
sodium sulfate to
sample
7.5.2
le organic
concentration
expected to be
< 20 mg/kg?
7.3.1 Add eurrogate
etandarde to all
•ample*, epika*,
and blanke
7.5.3 Add eurrogate
standards to all
sample*, spikee,
and blanke
7.3.2 - 7.3.5
Sonicate sample at
least 3 times
I
7.5.4
Adjust
volume; disrupt
sample with tapered
microtip
ultrasonic
probe
7.3.7 Dry and
collect extract in
K-D concentrator
7.5.5 Is
further
concentration
required?
7.5.5 Filter
through glass wool
7.3.8 Concentrate
extract and collect
in K-D concentrator
o
3550A - 12
Revision 1
September 1994
-------
METHOD 3550A
continued
7.3.9 Add exchango
solvent;
concentrate extract
Yes
7.3.10 Use Method
3660 for cleanup
Yes
7.3.9 Is
a solvent
exchange
required?
7.3.10 Do
sulfur crystals
form?
7.3.11 Further
concentrate and/or
adjust volume
/ Cleanup or \
I analyze I
3550A - 13
Revision 1
September 1994
-------
METHOD 3580A
WASTE DILUTION
1.0 SCOPE AND APPLICATION
1.1 This method describes a solvent dilution of a non-aqueous waste
sample prior to cleanup and/or analysis. It is designed for wastes that may
contain organic chemicals at a concentration greater than 20,000 mg/kg and that
are soluble in the dilution solvent.
1.2 It is recommended that an aliquot of the diluted sample be cleaned
up. See this chapter, Organic Analytes, Section 4.2.2 (Cleanup).
2.0 SUMMARY OF METHOD
2.1 One gram of sample is weighed into a capped tube, and the sample is
diluted to 10.0 mL with an appropriate solvent.
3.0 INTERFERENCES
3.1 Refer to Method 3500.
4.0 APPARATUS AND MATERIALS
4.1 Glass scintillation vials: At least 20 mL, with Teflon or aluminum
foil lined screw-cap, or equivalent.
4.2 Spatula: Stainless steel or Teflon.
4.3 Balance: Capable of weighing 100 g to the nearest 0.01 g.
4.4 Vials and caps: 2 mL for GC autosampler.
4.5 Disposable pipets: Pasteur.
4.6 Test tube rack.
4.7 Pyrex glass wool.
4.8 Volumetric flasks, Class A: 10 mL (optional).
5.0 REAGENTS
5.1 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
3580A - 1 Revision 1
July 1992
-------
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.2 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.3 Hexane, C6HU - Pesticide quality or equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Samples consisting of multiphases must be prepared by the phase
separation method (Chapter Two) before extraction.
7.2 The sample dilution may be performed in a 10 mL volumetric flask.
If disposable glassware is preferred, the 20 mL scintillation vial may be
calibrated for use. Pipet 10.0 mL of extraction solvent into the scintillation
vial and mark the bottom of the meniscus. Discard this solvent.
7.3 Transfer approximately 1 g of each phase of the sample to separate
20 mL vials or 10 mL volumetric flasks (record weight to the nearest 0.1 g).
Wipe the mouth of the vial with a tissue to remove any sample material. Cap the
vial before proceeding with the next sample to avoid any cross-contamination.
7.4 Add 2.0 mL surrogate spiking solution to all samples and blanks. For
the sample in each analytical batch selected for spiking, add 2.0 mL of the
matrix spiking standard. For base/neutral-acid analysis, the amount added of the
surrogates and matrix spiking compounds should result in a final concentration
of 200 ng//iL of each base/neutral analyte and 400 ng/^L of each acid analyte in
the extract to be analyzed (assuming a 1 /uL injection). If Method 3640, Gel-
permeation cleanup, is to be used, add twice the volume of surrogates and matrix
spiking compounds since half the extract is lost due to loading of the GPC
column. See Method 3500 and the determinative method to be used for details on
the surrogate standard and matrix spiking solutions.
7.5 Immediately dilute to 10 mL with the appropriate solvent. For
compounds to be analyzed by GC/ECD, e.g., organochlorine pesticides and PCBs, the
dilution solvent should be hexane. For base/neutral and acid semivolatile
priority pollutants, use methylene chloride. If the dilution is to be cleaned
up by gel permeation chromatography (Method 3640), use methylene chloride as the
dilution solvent for all compounds.
7.6 Add 2.0 g of anhydrous sodium sulfate to the sample.
7.7 Cap and shake the sample for 2 min.
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7.8 Loosely pack disposable Pasteur pipets with 2-3 cm glass wool plugs.
Filter the extract through the glass wool and collect 5 mL of the extract in a
tube or vial.
7.9 The extract is ready for cleanup or analysis, depending on the extent
of interfering co-extractives.
8.0 QUALITY CONTROL
8.1 Any reagent blanks and matrix spike samples should be subjected to
exactly the same analytical procedures as those used on actual samples.
8.2 Refer to Chapter One for specific quality control procedures and
Method 3500 for extraction and sample preparation procedures.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
10.0 REFERENCES
10.1 None applicable.
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METHOD 3580A
WASTE DILUTION
7 3 Transfer 1 g o
each phase to
separate vials or
flasks
7 4 Add sur ragate
spiking solution to
all samples and
blanks
7 4 Add ma t rix
spiking standard to
sample selected for
spiking
7 5 Dilute with
appropriate solvent
7 1 Use phase
separation method
(Chapter 2)
7 6 Add anhydrous
ammonium sulfa Le
7 7 Cap and shake
7 8 Filler through
glass wool
Cleanup or analyze
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METHOD 5030A
PURGE-AND-TRAP
1.0 SCOPE AND APPLICATION
1.1 This method describes sample preparation and extraction for the
analysis of volatile organics by a purge-and-trap procedure. The gas
chromatographic determinative steps are found in Methods 8010, 8015, 8020, 8021
and 8030. Although applicable to Methods 8240 and 8260, the purge-and-trap
procedure is already incorporated into Methods 8240 and 8260.
1.2 Method 5030 can be used for most volatile organic compounds that have
boiling points below 200°C and are insoluble or slightly soluble in water.
Volatile water-soluble compounds can be included in this analytical technique;
however, quantitation limits (by GC or GC/MS) are approximately ten times higher
because of poor purging efficiency. The method is also limited to compounds that
elute as sharp peaks from a GC column packed with graphitized carbon lightly
coated with a carbowax or a coated capillary column. Such compounds include low
molecular weight halogenated hydrocarbons, aromatics, ketones, nitriles,
acetates, acrylates, ethers, and sulfides.
1.3 Water samples can be analyzed directly for volatile organic compounds
by purge-and-trap extraction and gas chromatography. Higher concentrations of
these analytes in water can be determined by direct injection of the sample into
the chromatographic system.
1.4 This method also describes the preparation of water-miscible liquids,
non-water-miscible liquids, solids, wastes, and soils/sediments for analysis by
the purge-and-trap procedure.
2.0 SUMMARY OF METHOD
2.1 The purge-and-trap process: An inert gas is bubbled through the
solution at ambient temperature, and the volatile components are efficiently
transferred from the aqueous phase to the vapor phase. The vapor is swept
through a sorbent column where the volatile components are adsorbed. After
purging is completed, the sorbent column is heated and backflushed with inert gas
to desorb the components onto a gas chromatographic column.
2.2 If the sample introduction technique in Section 2.1 is not
applicable, a portion of the sample is dispersed in methanol to dissolve the
volatile organic constituents. A portion of the methanolic solution is combined
with water in a specially designed purging chamber. It is then analyzed by
purge-and-trap GC following the normal water method.
3.0 INTERFERENCES
3.1 Impurities in the purge gas, and from organic compounds out-gassing
from the plumbing ahead of the trap, account for the majority of contamination
problems. The analytical system must be demonstrated to be free from
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contamination under the conditions of the analysis by running laboratory reagent
blanks. The use of non-TFE plastic coating, non-TFE thread sealants, or flow
controllers with rubber components in the purging device should be avoided.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly methylene chloride and fluorocarbons) through the septum seal of
the sample vial during shipment and storage. A trip blank prepared from
organic-free reagent water and carried through sampling and handling protocols
serves as a check on such contamination.
3.3 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are analyzed sequentially. Whenever an unusually
concentrated sample is analyzed, it should be followed by an analysis of
organic-free reagent water to check for cross-contamination. The trap and other
parts of the system are subject to contamination. Therefore, frequent bake-out
and purging of the entire system may be required.
3.4 The laboratory where volatile analysis is performed should be
completely free of solvents.
4.0 APPARATUS AND MATERIALS
4.1 Microsyringes - 10 jiL, 25 p.1, 100 /xL, 250 /LtL, 500 /iL, and 1,000 /uL.
These syringes should be equipped with a 20 gauge (0.006 in ID) needle having a
length sufficient to extend from the sample inlet to within 1 cm of the glass
frit in the purging device. The needle length will depend upon the dimensions
of the purging device employed.
4.2 Syringe valve - Two-way, with Luer ends (three each), if applicable
to the purging device.
4.3 Syringe - 5 ml, gas-tight with shutoff valve.
4.4 Analytical balance - 0.0001 g.
4.5 Top-loading balance - 0.1 g.
4.6 Glass scintillation vials - 20 ml, with screw-caps and Teflon liners
or glass culture tubes with screw-caps and Teflon liners.
4.7 Volumetric flasks, Class A - 10 mL and 100 ml_, with ground-glass
stoppers.
4.8 Vials - 2 ml, for GC autosampler.
4.9 Spatula - Stainless steel.
4.10 Disposable pipets - Pasteur.
4.11 Purge-and-trap device: The purge-and-trap device consists of three
separate pieces of equipment: the sample purger, the trap, and the desorber.
Several complete devices are commercially available.
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4.11.1 The recommended purging chamber is designed to accept 5
ml samples with a water column at least 3 cm deep. The gaseous headspace
between the water column and the trap must have a total volume of less
than 15 ml. The purge gas must pass through the water column as finely
divided bubbles with a diameter of less than 3 mm at the origin. The
purge gas must be introduced no more than 5 mm from the base of the water
column. The sample purger, illustrated in Figure 1, meets these design
criteria. Alternate sample purge devices may be used, provided equivalent
performance is demonstrated.
4.11.2 The trap must be at least 25 cm long and have an inside
diameter of at least 0.105 in. Starting from the inlet, the trap must
contain the following amounts of adsorbents: 1/3 of 2,6-diphenylene oxide
polymer, 1/3 of silica gel, and 1/3 of coconut charcoal. It is
recommended that 1.0 cm of methyl silicone-coated packing be inserted at
the inlet to extend the life of the trap (see Figures 2 and 3). If it is
not necessary to analyze for dichlorodifluoromethane or other fluoro-
carbons of similar volatility, the charcoal can be eliminated and the
polymer increased to fill 2/3 of the trap. If only compounds boiling
above 35°C are to be analyzed, both the silica gel and charcoal can be
eliminated and the polymer increased to fill the entire trap. Before
initial use, the trap should be conditioned overnight at 180°C by
backflushing with an inert gas flow of at least 20 mL/min. Vent the trap
effluent to the hood, not to the analytical column. Prior to daily use,
the trap should be conditioned for 10 min at 180°C with backflushing. The
trap may be vented to the analytical column during daily conditioning;
however, the column must be run through the temperature program prior to
analysis of samples.
4.11.3 The desorber should be capable of rapidly heating the
trap to 180°C for desorption. The polymer section of the trap should not
be heated higher than 180°C, and the remaining sections should not exceed
220°C during bake-out mode. The desorber design illustrated in Figures 2
and 3 meet these criteria.
4.11.4 The purge-and-trap device may be assembled as a separate
unit or may be coupled to a gas chromatograph, as shown in Figures 4
and 5.
4.11.5 Trap Packing Materials
4.11.5.1 2,6-Diphenylene oxide polymer - 60/80 mesh,
chromatographic grade (Tenax GC or equivalent).
4.11.5.2 Methyl silicone packing - OV-1 (3%) on
Chromosorb-W, 60/80 mesh or equivalent.
4.11.5.3 Silica gel - 35/60 mesh, Davison, grade 15 or
equivalent.
4.11.5.4 Coconut charcoal - Prepare from Barnebey Cheney,
CA-580-26 lot #M-2649, or equivalent, by crushing through 26 mesh
screen.
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4.12 Heater or heated oil bath - capable of maintaining the purging
chamber to within 1°C, over a temperature range from ambient to 100°C.
5.0 REAGENTS
5.1 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2 Methanol, CH3OH - Pesticide quality or equivalent. Store away from
other solvents.
5.3 Reagent Tetraglyme - Reagent tetraglyme is defined as tetraglyme in
which interference is not observed at the method detection limit of the compounds
of interest.
5.3.1 Tetraglyme (tetraethylene glycol dimethyl ether, Aldrich #17,
240-5 or equivalent), CgH18Or. Purify by treatment at reduced pressure in
a rotary evaporator. The tetraglyme should have a peroxide content of
less than 5 ppm as indicated by EM Quant Test Strips (available from
Scientific Products Co., Catalog No. P1126-8 or equivalent).
CAUTION: Glycol ethers are suspected carcinogens. All solvent
handling should be done in a hood while using proper
protective equipment to minimize exposure to liquid and
vapor.
Peroxides may be removed by passing the tetraglyme through a column
of activated alumina. The tetraglyme is placed in a round bottom flask
equipped with a standard taper joint, and the flask is affixed to a rotary
evaporator. The flask is immersed in a water bath at 90-100°C and a vacuum
is maintained at < 10 mm Hg for at least two hours using a two stage
mechanical pump. The vacuum system is equipped with an all glass trap,
which is maintained in a dry ice/methanol bath. Cool the tetraglyme to
ambient temperature and add 100 mg/L of 2,6-di-tert-butyl-4-methyl-phenol
to prevent peroxide formation. Store the tetraglyme in a tightly sealed
screw cap bottle in an area that is not contaminated by solvent vapors.
5.3.2 In order to demonstrate that all interfering volatiles have
been removed from the tetraglyme, an organic-free reagent water/tetraglyme
blank must be analyzed.
5.4 Polyethylene glycol, H(OCH2CH2)nOH. Free of interferences at the
detection limit of the analytes.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Refer to the introductory material to this chapter, Organic Analytes,
Section 4.1. Samples should be stored in capped bottles, with minimum headspace,
at 4°C or less.
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7.0 PROCEDURE
7.1 Initial calibration: Prior to using this introduction technique for
any GC method, the system must be calibrated. General calibration procedures are
discussed in Method 8000, while the specific determinative methods and Method
3500 give details on preparation of standards.
7.1.1 Assemble a purge-and-trap device that meets the specification
in Section 4.10. Condition the trap overnight at 180°C in the purge mode
with an inert gas flow of at least 20 mL/min. Prior to use, condition the
trap daily for 10 min while backflushing at 180°C with the column at 220°C.
7.1.2 Connect the purge-and-trap device to a gas chromatograph.
7.1.3 Prepare the final solutions containing the required
concentrations of calibration standards, including surrogate standards,
directly in the purging device. Add 5.0 ml of organic-free reagent water
to the purging device. The organic-free reagent water is added to the
purging device using a 5 ml glass syringe fitted with a 15 cm 20-gauge
needle. The needle is inserted through the sample inlet shown in
Figure 1. The internal diameter of the 14-gauge needle that forms the
sample inlet will permit insertion of the 20-gauge needle. Next, using a
10 jtL or 25 /it micro-syringe equipped with a long needle (Section 4.1),
take a volume of the secondary dilution solution containing appropriate
concentrations of the calibration standards. Add the aliquot of
calibration solution directly to the organic-free reagent water in the
purging device by inserting the needle through the sample inlet. When
discharging the contents of the micro-syringe, be sure that the end of the
syringe needle is well beneath the surface of the organic-free reagent
water. Similarly, add 10 /xL of the internal standard solution. Close the
2-way syringe valve at the sample inlet.
7.1.4 Carry out the purge-and-trap analysis procedure using the
specific conditions given in Table 1.
7.1.5 Calculate response factors or calibration factors for each
analyte of interest using the procedure described in Method 8000.
7.1.6 The average RF must be calculated for each compound. A system
performance check should be made before this calibration curve is used.
If the purge-and-trap procedure is used with Method 8010, the following
five compounds are checked for a minimum average response factor:
chloromethane; 1,1-dichloroethane; bromoform; 1,1,2,2-tetrachloroethane;
and chlorobenzene. The minimum acceptable average RF for these compounds
should be 0.300 (0.250 for bromoform). These compounds typically have RFs
of 0.4-0.6, and are used to check compound stability and to check for
degradation caused by contaminated lines or active sites in the system.
Examples of these occurrences are:
7.1.6.1 Chloromethane: This compound is the most likely
compound to be lost if the purge flow is too fast.
7.1.6.2 Bromoform: This compound is one of the compounds
most likely to be purged very poorly if the purge flow is too slow.
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Cold spots and/or active sites in the transfer lines may adversely
affect response.
7.1.6.3 Tetrachloroethane and 1,1-dichloroethane: These
compounds are degraded by contaminated transfer lines in purge-and-
trap systems and/or active sites in trapping materials.
7.2 On-going calibration: Refer to Method 8000 for details on continuing
calibration.
7.3 Sample preparation
7.3.1 Water samples
7.3.1.1 Screening of the sample prior to purge-and-trap
analysis will provide guidance on whether sample dilution is
necessary and will prevent contamination of the purge-and-trap
system. Two screening techniques that can be utilized are: the use
of an automated headspace sampler (modified Method 3810), interfaced
to a gas chromatograph (GC), equipped with a photo ionization
detector (PID), in series with an electrolytic conductivity detector
(HECD); and extraction of the sample with hexadecane (Method 3820)
and analysis of the extract on a GC with a FID and/or an ECD.
7.3.1.2 All samples and standard solutions must be allowed
to warm to ambient temperature before analysis.
7.3.1.3 Assemble the purge-and-trap device. The operating
conditions for the GC are given in Section 7.0 of the specific
determinative method to be employed.
7.3.1.4 Daily GC calibration criteria must be met (Method
8000) before analyzing samples.
7.3.1.5 Adjust the purge gas flow rate (nitrogen or
helium) to that shown in Table 1, on the purge-and-trap device.
Optimize the flow rate to provide the best response for
chloromethane and bromoform, if these compounds are analytes.
Excessive flow rate reduces chloromethane response, whereas
insufficient flow reduces bromoform response.
7.3.1.6 Remove the plunger from a 5 ml syringe and attach
a closed syringe valve. Open the sample or standard bottle, which
has been allowed to come to ambient temperature, and carefully pour
the sample into the syringe barrel to just short of overflowing.
Replace the syringe plunger and compress the sample. Open the
syringe valve and vent any residual air while adjusting the sample
volume to 5.0 ml. This process of taking an aliquot destroys the
validity of the liquid sample for future analysis; therefore, if
there is only one VOA vial, the analyst should fill a second syringe
at this time to protect against possible loss of sample integrity.
This second sample is maintained only until such time when the
analyst has determined that the first sample has been analyzed
properly. Filling one 20 ml syringe would allow the use of only one
5030A - 6 Revision 1
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syringe. If a second analysis is needed from a syringe, it must be
analyzed within 24 hr. Care must be taken to prevent air from
leaking into the syringe.
7.3.1.7 The following procedure is appropriate for
diluting purgeable samples. All steps must be performed without
delays until the diluted sample is in a gas-tight syringe.
7.3.1.7.1 Dilutions may be made in volumetric flasks
(10 ml to 100 ml). Select the volumetric flask that will
allow for the necessary dilution. Intermediate dilutions may
be necessary for extremely large dilutions.
7.3.1.7.2 Calculate the approximate volume of
organic-free reagent water to be added to the volumetric flask
selected and add slightly less than this quantity of organic-
free reagent water to the flask.
7.3.1.7.3 Inject the proper aliquot of samples from
the syringe prepared in Section 7.3.1.5 into the flask.
Aliquots of less than 1 ml are not recommended. Dilute the
sample to the mark with organic-free reagent water. Cap the
flask, invert, and shake three times. Repeat the above
procedure for additional dilutions.
7.3.1.7.4 Fill a 5 ml syringe with the diluted sample
as in Section 7.3.1.5.
7.3.1.8 Add 10.0 /uL of surrogate spiking solution (found
in each determinative method, Section 5.0) and, if applicable, 10 /xL
of internal standard spiking solution through the valve bore of the
syringe; then close the valve. The surrogate and internal standards
may be mixed and added as a single spiking solution. Matrix spiking
solutions, if indicated, should be added (10 /zL) to the sample at
this time.
7.3.1.9 Attach the syringe-syringe valve assembly to the
syringe valve on the purging device. Open the syringe valves and
inject the sample into the purging chamber.
7.3.1.10 Close both valves and purge the sample for the
time and at the temperature specified in Table 1.
7.3.1.11 At the conclusion of the purge time, attach the
trap to the chromatograph, adjust the device to the desorb mode, and
begin the gas chromatographic temperature program and GC data
acquisition. Concurrently, introduce the trapped materials to the
gas chromatographic column by rapidly heating the trap to 180°C
while backflushing the trap with inert gas between 20 and 60 mL/min
for the time specified in Table 1.
7.3.1.12 While the trap is being desorbed into the gas
chromatograph, empty the purging chamber. Wash the chamber with a
minimum of two 5 ml flushes of organic-free reagent water (or
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methanol followed by organic-free reagent water) to avoid carryover
of pollutant compounds into subsequent analyses.
7.3.1.13 After desorbing the sample, recondition the trap
by returning the purge-and-trap device to the purge mode. Wait 15
sec; then close the syringe valve on the purging device to begin gas
flow through the trap. The trap temperature should be maintained at
180°C for Methods 8010, 8020, 8021, 8240 and 8260 and 210°C for
Methods 8015 and 8030. Trap temperatures up to 220°C may be
employed. However, the higher temperatures will shorten the useful
life of the trap. After approximately 7 min, turn off the trap
heater and open the syringe valve to stop the gas flow through the
trap. When cool, the trap is ready for the next sample.
7.3.1.14 If the initial analysis of a sample or a dilution
of the sample has a concentration of analytes that exceeds the
initial calibration range, the sample must be reanalyzed at a higher
dilution. When a sample is analyzed that has saturated response
from a compound, this analysis must be followed by a blank organic-
free reagent water analysis. If the blank analysis is not free of
interferences, the system must be decontaminated. Sample analysis
may not resume until a blank can be analyzed that is free of
interferences.
7.3.1.15 All dilutions should keep the response of the
major constituents (previously saturated peaks) in the upper half of
the linear range of the curve. Proceed to Method 8000 and the
specific determinative method for details on calculating analyte
response.
7.3.2 Water-miscible liquids:
7.3.2.1 Water-miscible liquids are analyzed as water
samples after first diluting them at least 50-fold with organic-free
reagent water.
7.3.2.2 Initial and serial dilutions can be prepared by
pipetting 2 ml of the sample into a 100 mL volumetric flask and
diluting to volume with organic-free reagent water. Transfer
immediately to a 5 ml gas-tight syringe.
7.3.2.3 Alternatively, prepare dilutions directly in a 5
ml syringe filled with organic-free reagent water by adding at least
20 /iL, but not more than 100 /iL of liquid sample. The sample is
ready for addition of surrogate and, if applicable, internal and
matrix spiking standards.
7.3.3 Sediment/soil and waste samples: It is highly recommended
that all samples of this type be screened prior to the purge-and-trap GC
analysis. These samples may contain percent quantities of purgeable
organics that will contaminate the purge-and-trap system, and require
extensive cleanup and instrument downtime. See Section 7.3.1.1 for
recommended screening techniques. Use the screening data to determine
whether to use the low-concentration method (0.005-1 mg/kg) or the high-
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concentration method (>1 mg/kg).
7.3.3.1 Low-concentration method: This is designed for
samples containing individual purgeable compounds of <1 mg/kg. It
is limited to sediment/soil samples and waste that is of a similar
consistency (granular and porous). The low-concentration method is
based on purging a heated sediment/soil sample mixed with
organic-free reagent water containing the surrogate and, if
applicable, internal and matrix spiking standards. Analyze all
reagent blanks and standards under the same conditions as the
samples.
7.3.3.1.1 Use a 5 g sample if the expected
concentration is <0.1 mg/kg or a 1 g sample for expected
concentrations between 0.1 and 1 mg/kg.
7.3.3.1.2 The GC system should be set up as in
Section 7.0 of the specific determinative method. This should
be done prior to the preparation of the sample to avoid loss
of volatiles from standards and samples. A heated purge
calibration curve must be prepared and used for the
quantitation of all samples analyzed with the low-
concentration method. Follow the initial and daily
calibration instructions, except for the addition of a 40°C
purge temperature for Methods 8010, 8020, and 8021.
7.3.3.1.3 Remove the plunger from a 5 mL Luerlock type
syringe equipped with a syringe valve and fill until
overflowing with organic-free reagent water. Replace the
plunger and compress the reagent water to vent trapped air.
Adjust the volume to 5.0 ml. Add 10 p.1 each of surrogate
spiking solution and internal standard solution to the syringe
through the valve. (Surrogate spiking solution and internal
standard solution may be mixed together.) Matrix spiking
solutions, if indicated, should be added (10 juL) to the sample
at this time.
7.3.3.1.4 The sample (for volatile organics) consists
of the entire contents of the sample container. Do not
discard any supernatant liquids. Mix the contents of the
sample container with a narrow metal spatula. Weigh the
amount determined in Section 7.3.3.1.1 into a tared purge
device. Note and record the actual weight to the nearest 0.1
9-
7.3.3.1.5 Determination of sample % dry weight - In
certain cases, sample results are desired based on dry weight
basis. When such data is desired, a portion of sample for
this determination should be weighed out at the same time as
the portion used for analytical determination.
WARNING: The drying oven should be contained in a
hood or vented. Significant laboratory
contamination may result from a heavily
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contaminated hazardous waste sample.
7.3.3.1.5.1 Immediately after weighing the sample
for extraction, weigh 5-10 g of the sample into a tared
crucible. Determine the % dry weight of the sample by
drying overnight at 105°C. Allow to cool in a
desiccator before weighing:
% dry weight = q of dry sample x 100
g of sample
7.3.3.1.6 Add the spiked organic-free reagent water to
the purge device, which contains the weighed amount of sample,
and connect the device to the purge-and-trap system.
NOTE: Prior to the attachment of the purge device,
Sections 7.3.3.1.4 and 7.3.3.1.6 must be
performed rapidly and without interruption to
avoid loss of volatile organics. These steps
must be performed in a laboratory free of solvent
fumes.
7.3.3.1.7 Heat the sample to 40°C ± 1°C (Methods 8010,
8020 and 8021) or to 85°C ± 26C (Methods 8015 and 8030) and
purge the sample for the time shown in Table 1.
7.3.3.1.8 Proceed with the analysis as outlined in
Sections 7.3.1.11-7.3.1.15. Use 5 ml of the same organic-free
reagent water as in the reagent blank. If saturated peaks
occurred or would occur if a 1 g sample were analyzed, the
high-concentration method must be followed.
7.3.3.1.9 For matrix spike analysis of
low-concentration sediment/soils, add 10 \i\. of the matrix
spike solution to 5 ml of organic-free reagent water (Section
7.3.3.1.3 ). The concentration for a 5 g sample would be
equivalent to 50 jig/kg of each matrix spike standard.
7.3.3.2 High-concentration method: The method is based on
extracting the sediment/soil with methanol. A waste sample is
either extracted or diluted, depending on its solubility in
methanol. Wastes (i.e. petroleum and coke wastes) that are
insoluble in methanol are diluted with reagent tetraglyme or
polyethylene glycol (PEG). An aliquot of the extract is added to
organic-free reagent water containing surrogate and, if applicable,
internal and matrix spiking standards. This is purged at the
temperatures indicated in Table 1. All samples with an expected
concentration of >1.0 mg/kg should be analyzed by this method.
7.3.3.2.1 The sample (for volatile organics) consists
of the entire contents of the sample container. Do not
discard any supernatant liquids. Mix the contents of the
sample container with a narrow metal spatula. For
sediment/soil and waste that are insoluble in methanol, weigh
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4 g (wet weight) of sample Into a tared 20 ml vial. Use a
top-loading balance. Note and record the actual weight to 0.1
gram and determine the percent dry weight of the sample using
the procedure in Section 7.3.3.1.5. For waste that is soluble
in methanol, tetraglyme, or PEG, weigh 1 g (wet weight) into
a tared scintillation vial or culture tube or a 10 ml
volumetric flask. (If a vial or tube is used, it must be
calibrated prior to use. Pipet 10.0 ml of methanol into the
vial and mark the bottom of the meniscus. Discard this
solvent.)
7.3.3.2.2 For sediment/soil or solid waste, quickly
add 9.0 ml of appropriate solvent; then add 1.0 ml of the
surrogate spiking solution to the vial. For a solvent
miscible sample, dilute the sample to 10 ml with the
appropriate solvent after adding 1.0 ml of the surrogate
spiking solution. Cap and shake for 2 min.
NOTE: Sections 7.3.3.2.1 and 7.3.3.2.2 must be
performed rapidly and without interruption to
avoid loss of volatile organics. These steps
must be performed in a laboratory free from
solvent fumes.
7.3.3.2.3 Pipet approximately 1 ml of the extract into
a GC vial for storage, using a disposable pipet. The
remainder may be discarded. Transfer approximately 1 ml of
reagent methanol to a separate GC vial for use as the method
blank for each set of samples. These extracts may be stored
at 4°C in the dark, prior to analysis.
7.3.3.2.4 The GC system should be set up as in
Section 7.0 of the specific determinative method. This should
be done prior to the addition of the methanol extract to
organic-free reagent water.
7.3.3.2.5 Table 2 can be used to determine the volume
of methanol extract to add to the 5 ml of organic-free reagent
water for analysis. If a screening procedure was followed,
use the estimated concentration to determine the appropriate
volume. Otherwise, estimate the concentration range of the
sample from the low-concentration analysis to determine the
appropriate volume. If the sample was submitted as a high-
concentration sample, start with 100 /xL. All dilutions must
keep the response of the major constituents (previously
saturated peaks) in the upper half of the linear range of the
curve.
7.3.3.2.6 Remove the plunger from a 5.0 ml Luerlock
type syringe equipped with a syringe valve and fill until
overflowing with organic-free reagent water. Replace the
plunger and compress the water to vent trapped air. Adjust
the volume to 4.9 ml. Pull the plunger back to 5.0 ml to
allow volume for the addition of the sample extract and of
5030A - 11 Revision 1
July 1992
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standards. Add 10 jil_ of internal standard solution. Also add
the volume of methanol extract determined in Section 7.3.3.2.5
and a volume of methanol solvent to total 100 juL (excluding
methanol in standards).
7.3.3.2.7 Attach the syringe-syringe valve assembly to
the syringe valve on the purging device. Open the syringe
valve and inject the water/methanol sample into the purging
chamber.
7.3.3.2.8 Proceed with the analysis as outlined in the
specific determinative method. Analyze all reagent blanks on
the same instrument as that used for the samples. The
standards and blanks should also contain 100 jiL of methanol
to simulate the sample conditions.
7.3.3.2.9 For a matrix spike in the high-concentration
sediment/soil samples, add 8.0 ml of methanol, 1.0 ml of
surrogate spike solution and 1.0 ml of matrix spike solution.
Add a 100 /zL aliquot of this extract to 5 ml of water for
purging (as per Section 7.3.3.2.6).
7.4 Sample analysis:
7.4.1 The samples prepared by this method may be analyzed by Methods
8010, 8015, 8020, 8021, 8030, 8240, and 8260. Refer to these methods for
appropriate analysis conditions.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3500 for sample preparation procedures.
8.2 Before processing any samples, the analyst should demonstrate through
the analysis of a calibration blank that all glassware and reagents are
interference free. Each time a set of samples is extracted, or there is a change
in reagents, a method blank should be processed as a safeguard against chronic
laboratory contamination. The blanks should be carried through all stages of
the sample preparation and measurement.
8.3 Standard quality assurance practices should be used with this method.
Field duplicates should be collected to validate the precision of the sampling
technique. Laboratory replicates should be analyzed to validate the precision
of the analysis. Spiked samples should be carried through all stages of sample
preparation and measurement; they should be analyzed to validate the sensitivity
and accuracy of the analysis. If the spiked samples do not indicate sufficient
sensitivity to detect < 1 jig/g of the analytes in the sample, then the
sensitivity of the instrument should be increased, or the sample should be
subjected to additional cleanup.
5030A - 12 Revision 1
July 1992
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9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
5030A - 13 Revision 1
July 1992
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TABLE 1
PURGE-AND-TRAP OPERATING PARAMETERS
Analysis Method
8010
8015
8020/8021
8030
Purge gas
Purge gas flow rate
(mL/min)
Purge time (min)
Purge temperature (°C)
Desorb temperature (°C)
Backflush inert gas flow
(mL/min)
Desorb time (min)
Nitrogen or Nitrogen or
Heli urn
40
11.0 ± 0.1
Ambient
180
20-60
4
Helium
20
15.0 ± 0.1
85 + 2
180
20-60
1.5
Nitrogen or Nitrogen or
Helium Helium
40
11.0 ± 0.1
Ambient
180
20-60
4
20
15.0 + 0.1
85 ± 2
180
20-60
1.5
5030A - 14
Revision 1
July 1992
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TABLE 2
QUANTITY OF METHANOL EXTRACT REQUIRED FOR ANALYSIS OF
HIGH-CONCENTRATION SOILS/SEDIMENTS
Approximate Volume of
Concentration Range Methanol Extract8
500-10,000 M9/kg 100 /xL
1,000-20,000 /ig/kg so ML
5,000-100,000 jug/kg 10 ^i
25,000-500,000 jug/kg 100 /zL of 1/50 dilution b
Calculate appropriate dilution factor for concentrations exceeding this table.
"The volume of methanol added to 5 ml of water being purged should be kept
constant. Therefore, add to the 5 mL syringe whatever volume of methanol is
necessary to maintain a volume of 100 /iL added to the syringe.
Dilute an aliquot of the methanol extract and then take 100 jiL for analysis.
5030A - 15 Revision 1
July 1992
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Figure 1
Purging Chamber
OPTIONAL
FOAM TRAP
* loch 0. D.
10 mm Ot«t* Frit
Moftwm Awmitv
Inlti Jt Inch 0. 0.
Swnplf Intet
2-W«v Svnnoj
17 cm. 20 Gfluoj Syrinot
6 mm 0. 0. Rubber Stptum
-10 mm 0.0.
Inch 0. D.
1/16 Inch 0 0.
Stit:
Ftow Control
5030A - 16
Revision 1
July 1992
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Figure 2
Trap Packing and Construction for Method 8010
Packing Proctduri
Construction
GlauWOOl
Activatad
Charcoal
Grada 15
Silica Gal
3%OV-1
GlauWoei
7.7cm
Tanax 7.7 cm
8mm
711/Foot
Ratiitanca
Wira Wrapped
Solid
(Doubla Layar)
711/Foot
Rtiistanc*
Wira Wrapptd
Solid
(Singla Lavar)
8cm
Comprauion
Fining Nut
and Farrulai
Tharmoeouplt/
Controllar
Santer
Elactronic
Tamparatura
Control and
Pyromatar
Tubing 25 cm
0.105 In. 1.0.
0.125 In. 0.0.
Stainittt Staal
Trap Inlat
5030A - 17
Revision 1
July 1992
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Figure 3
Trap Packing and Construction for Methods 8020 and 8030
Packing Procedure
Construction
Glass Wool 5mm %
Tenax 23 cm
3%OV-1 1cm';
Glass Wool 5 mm
Compression Fining Nut
and Ferrules
14 Ft. ?n/Foot Resistance
Wire Wrapped Solid
Thermocouple/Controller Sensor
Electronic
Temperature
Control and
Pyrometer
Tubing 25 cm
0.105 In. 1.0.
0.125 In. O.D.
Stainless Steel
Trap Inlet
5030A - 18
Revision 1
July 1992
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Figure 4
Purge-and-Trap System
Purge-Sorb Mode
For Method 8010, 8020, and 8030
CARRIER GAS
FLOW CONTROL
PRESSURE
REGULATOR
UOWO INJECTON PORTS
COLUMN OVEN
CONFIRMATORY COLUMN
TO DETECTOR
PURGE OAS
FLOW CONTROL
13X MOLECULAR
SIEVE FILTER
ANALYTICAL COLUMN
OPTIONAL WORT COLUMN
SELECTION VALVE
PURGING
DEVICE
NOTE
ALL LINES BETWEEN TRAP
AND GC SHOULD BE HEATED
TO80-C
5030A - 19
Revision 1
July 1992
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Figure 5
Purge-and-Trap System
Desorb Mode
For Method 8010, 8020, and 8030
CARPJCftGAS
FLOW CONTROL
PRESSURE
REGULATOR
UOUtO INJECTION PORTS
COLUMN OVEN
OPTIONAL 4PORT COLUMN
SELECTION VALVE
PURGE GAS
ROW CONTROL
13X MOLECULAR
SIEVE FILTER
CONFIRMATORY COLUMN
TO DETECTOR
ANALYTICAL COLUMN
PURGING
DEVICE
NOTE:
ALL LINES BETWEEN TRAP
AND GC SHOULD BE HEATED
TO SOX.
5030A - 20
Revision 1
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METHOD 5030A
PURGE-AND-TRAP
Start
7 1 Calibrate
CC system
712 Assemble
purge-and-trap
device and
condi tion t rap
712 Connect
to gas
chroma tograph
I
7 1 3 P
fin
solut i
repare
ons
714 Corry out
purge-and- trap
analysis
715 Calculate
response or
calibration factors
for each analyte
(Method 8000)
7 1 6
Calculate
averafe RF
for each
compound
/ '
/
5030A - 21
Revision 1
July 1992
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METHOD 5030A
continued
7331
Prepare
samples and
set-up CC
sys tem
73314
Heigh sample
into tared
device
/
Low concentration
Soil/sediment
High concentration
Soil/sediment
water samples and
water-miscible wastes
I
73315
Neigh another
sample and
determine %
dry weight
7 3 3 1 6 Add
spiked reagent
water, connect
device to
sys tem
73317
Heat and
purge sample
/ • /
731 Screen
samples prior to
purge-and- trap
analysis, dilute
water miscible
liquids
731 Prepare
sample and
purg-and-trap
device
7317
Dilute
purgeable
samples
7 3 1 8 Add
surrogate and
internal spiking
solutions (if
indicated)
7332 Add
methano 1
ex t ract to
reagent water
for ana lysis
7 3 3 2 Set
up CC sys tem
73326 Fill
syringe wi th
reagent wa ter ,
vent air and
ad j us t vo 1 ume
7 3 3 2 6 Add
internal
s tanda rd and
methano 1
extract
Ana 1 y ze
acco rd ing t o
determinative
me thod
7319
Inject sample
into chamber ,
purge
7 3 1 11
Desorb trap
into CC
7 3 1 13
Recondi tion
trap and
start gas
flow
7 3 1 13 Stop
gas flow and
cool trap for
next sample
Analyze
acco r ding to
determinative
method
5030A - 22
Revision 1
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METHOD 5040A
ANALYSIS OF SORBENT CARTRIDGES FROM VOLATILE ORGANIC SAMPLING TRAIN (VOST):
GAS CHROMATOGRAPHY/MASS SPECTROMETRY TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 5040 was formerly Method 3720 in the Second Edition of this
manual.
1.2 This method covers the determination of volatile principal organic
hazardous constituents (POHCs), collected on Tenax and Tenax/charcoal sorbent
cartridges using a volatile organic sampling train, VOST (1). Much of the
description for purge-and-trap GC/MS analysis is described in Method 8240 of this
chapter. Because the majority of gas streams sampled using VOST will contain a
high concentration of water, the analytical method is based on the quantitative
thermal desorption of volatile POHCs from the Tenax and Tenax/charcoal traps and
analysis by purge-and-trap GC/MS. For the purposes of definition, volatile POHCs
are those POHCs with boiling points less than 100°C.
1.3 This method is applicable to the analysis of Tenax and Tenax/
charcoal cartridges used to collect volatile POHCs from wet stack gas effluents
from hazardous waste incinerators.
1.4 The sensitivity of the analytical method for a particular volatile
POHC depends on the level of interferences and the presence of detectable levels
of volatile POHCs in blanks. The desired target detection limit of the
analytical method is 0.1 ng/L (20 ng on a single pair of traps) for a particular
volatile POHC desorbed from either a single pair of Tenax and Tenax/charcoal
cartridges or by thermal desorption of up to six pairs of traps onto a single
pair of Tenax and Tenax/charcoal traps. The resulting single pair of traps is
then thermally desorbed and analyzed by purge-and-trap GC/MS.
1.5 This method is recommended for use only by experienced mass
spectroscopists or under the close supervision of such qualified persons.
2.0 SUMMARY OF METHOD
2.1 A schematic diagram of the analytical system is shown in Figure 1.
The contents of the sorbent cartridges are spiked with an internal standard and
thermally desorbed for 10 min at 180°C with organic-free nitrogen or helium gas
(at a flow rate of 40 mL/min), bubbled through 5 ml of organic-free reagent
water, and trapped on an analytical adsorbent trap. After the 10 min.
desorption, the analytical adsorbent trap is rapidly heated to 180°C, with the
carrier gas flow reversed so that the effluent flow from the analytical trap is
directed into the GC/MS. The volatile POHCs are separated by temperature
programmed gas chromatography and detected by low-resolution mass spectrometry.
The concentrations of volatile POHCs are calculated using the internal standard
technique.
5040A - 1 Revision 1
September 1994
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3.0 INTERFERENCES
3.1 Refer to Methods 3500 and 8240.
4.0 APPARATUS AND MATERIALS
4.1 Thermal desorption unit:
4.1.1 The thermal desorption unit (for Inside/Inside VOST
cartridges, use Supelco "clamshell" heater; for Inside/Outside VOST
cartridges, user-fabricated unit is required) should be capable of
thermally desorbing the sorbent resin tubes. It should also be capable of
heating the tubes to 180 + 10°C with flow of organic-free nitrogen or
helium through the tubes.
4.2 Purge-and-trap unit:
4.2.1 The purge-and-trap unit consists of three separate pieces of
equipment: the sample purger, trap, and the desorber. It should be
capable of meeting all requirements of Method 5030 for analysis of
purgeable organic compounds from water.
4.3 GC/MS system: As described in Method 8240.
5.0 REAGENTS
5.1 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2 Methanol, CH3OH - Pesticide grade, or equivalent.
5.3 Analytical trap reagents:
5.3.1 2,6-Diphenylene oxide polymer: Tenax (60/80 mesh), chromato-
graphic grade or equivalent.
5.3.2 Methyl silicone packing: 3% OV-1 on Chromosorb W (60/80 mesh)
or equivalent.
5.3.3 Silica gel: Davison Chemical (35/00 mesh), Grade 15, or
equivalent.
5.3.4 Charcoal: Petroleum-based (SKC Lot 104 or equivalent).
5.4 Stock standard solution:
5.4.1 Stock standard solutions will be prepared from pure standard
materials or purchased as certified solutions. The stock standards should
be prepared in methanol using assayed liquids or gases, as appropriate.
Because of the toxicity of some of the organohalides, primary dilutions of
these materials should be prepared in a hood. A NIOSH/MESA approved toxic
5040A - 2 Revision 1
September 1994
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gas respirator should be used when the analyst handles high concentrations
of such materials.
5.4.2 Fresh stock standards should be prepared weekly for volatile
POHCs with boiling points of <35°C. All other standards must be replaced
monthly, or sooner if comparison with check standards indicates a problem.
5.5 Secondary dilution standards:
5.5.1 Using stock standard solutions, prepare, in methanol,
secondary dilution standards that contain the compounds of interest,
either singly or mixed together. The secondary dilution standards should
be prepared at concentrations such that the desorbed calibration standards
will bracket the working range of the analytical system.
5.6 4-Bromofluorobenzene (BFB) standard:
5.6.1 Prepare a 25 ng//xL solution of BFB in methanol.
5.7 Deuterated benzene:
5.7.1 Prepare a 25 ng/^uL solution of benzene-d6 in methanol.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Refer to Method 0030, Chapter Ten.
6.2 Sample trains obtained from the VOST should be analyzed within 2-6
weeks of sample collection.
7.0 PROCEDURE
7.1 Assembly of PTD device:
7.1.1 Assemble a purge-and-trap desorption device (PTD) that meets
all the requirements of Method 5030 (refer to Figure 1).
7.1.2 Connect the thermal desorption device to the PTD device.
Calibrate the PTD-GC/MS system using the internal standard technique.
7.2 Internal standard calibration procedure:
7.2.1 This approach requires the use of deuterated benzene as the
internal standard for these analyses. Other internal standards may be
proposed for use in certain situations. The important criteria for
choosing a particular compound as an internal standard are that it be
similar in analytical behavior to the compounds of interest and that it
can be demonstrated that the measurement of the internal standard be
unaffected by method or matrix interferences. Other internal standards
that have been used are ethylbenzene-d10 and, l-2-dichloroethane-d4. One
adds 50 ng of BFB to all sorbent cartridges (in addition to one or more
5040A - 3 Revision 1
September 1994
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internal standards) to provide continuous monitoring of the GC/MS
performance relative to BFB.
7.2.2 Prepare calibration standards at a minimum of three
concentration levels for each analyte of interest.
7.2.3 The calibration standards are prepared by spiking a blank
Tenax or Tenax/charcoal trap with a methanolic solution of the calibration
standards (including 50 ng of the internal standard, such as deuterated
benzene), using the flash evaporation technique. The flash evaporation
technique requires filling the needle of a 5.0 /jL syringe with clean
methanol and drawing air into the syringe to the 1.0 fj,L mark. This is
followed by drawing a methanolic solution of the calibration standards
(containing 25 Aig/juL of the internal standard) to the 2.0 /iL mark. The
glass traps should be attached to the injection port of a gas
chromatograph while maintaining the injector temperature at 160°C. The
carrier gas flow through the traps should be maintained at about 50
mL/min.
7.2.4 After directing the gas flow through the trap, the contents of
the syringe should be slowly expelled through the gas chromatograph
injection port over about 15 sec. After 25 sec have elapsed, the gas flow
through the trap should be shut off, the syringe removed, and the trap
analyzed by the PTD-GC/MS procedure outlined in Method 8240. The total
flow of gas through the traps during addition of calibration standards to
blank cartridges, or internal standards to sample cartridges, should be 25
ml or less.
7.2.5 Analyze each calibration standard for both Tenax and Tenax/
charcoal cartridges according to Section 7.3. Tabulate the area response
of the characteristic ions of each analyte against the concentration of
the internal standard and calculate the response factor (RF) for each
compound, using Equation 1.
RF = AsC,s/AisCs (1)
where:
As = Area of the characteristic ion for the analyte to be
measured.
P(. = Area of the characteristic ion for the internal
•is
standard.
CIS = Amount (ng) of the internal standard.
Cs = Amount (ng) of the volatile POHC in calibration
standard.
If the RF value over the working range is a constant (<10% RSD), the
RF can be assumed to be invariant, and the average RF can be used for
calculations. Alternatively, the results can be used to plot a
calibration curve of response ratios, AS/AIS versus RF.
5040A - 4 Revision 1
September 1994
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7.2.6 The working calibration curve or RF must be verified on each
working day by the measurement of one or more of the calibration
standards. If the response varies by more than ±25% for any analyte, a
new calibration standard must be prepared and analyzed for that analyte.
7.3 The schematic of the PTD-GC/MS system is shown in Figure 1. The
sample cartridge is placed in the thermal desorption apparatus (for Inside/
Inside VOST cartridges, use Supelco "clamshell" heater; for Inside/Outside VOST
cartridges, user fabricated unit is required) and desorbed in the purge-and-trap
system by heating to 180°C for 10 min at a flow rate of 40 mL/min. The desorbed
components pass into the bottom of the water column, are purged from the water,
and collected on the analytical adsorbent trap. After the 10 min desorption
period, the compounds are desorbed from the analytical adsorbent trap into the
GC/MS system according to the procedures described in Method 8240.
7.4 Qualitative analysis
7.4.1 The qualitative identification of compounds determined by this
method is based on retention time, and on comparison of the sample mass
spectrum, after background correction, with characteristic ions in a
reference mass spectrum. The reference mass spectrum must be generated by
the laboratory using the conditions of this method. The characteristic
ions from the reference mass spectrum are defined to be the three ions of
greatest relative intensity, or any ions over 30% relative intensity if
less than three such ions occur in the reference spectrum. Compounds
should be identified as present when the criteria below are met.
7.4.1.1 The intensities of the characteristic ions of a
compound maximize in the same scan or within one scan of each other.
Selection of a peak by a data system target compound search routine,
where the search is based on the presence of a target
chromatographic peak containing ions specific for the target
compound at a compound-specific retention time, will be accepted as
meeting this criterion.
7.4.1.2 The RRT of the sample component is within
+ 0.06 RRT units of the RRT of the standard component.
7.4.1.3 The relative intensities of the characteristic
ions agree within 30% of the relative intensities of these ions in
the reference spectrum. (Example: For an ion with an abundance of
50% in the reference spectrum, the corresponding abundance in a
sample spectrum can range between 20% and 80%.)
7.4.1.4 Structural isomers that produce very similar mass
spectra should be identified as individual isomers if they have
sufficiently different GC retention times. Sufficient GC resolution
is achieved if the height of the valley between two isomer peaks is
less than 25% of the sum of the two peak heights. Otherwise,
structural isomers are identified as isomeric pairs.
7.4.1.5 Identification is hampered when sample components
are not resolved chromatographically and produce mass spectra
5040A - 5 Revision 1
September 1994
-------
containing ions contributed by more than one analyte. When gas
chromatographic peaks obviously represent more than one sample
component (i.e., a broadened peak with shoulder(s) or a valley
between two or more maxima), appropriate selection of analyte
spectra and background spectra is important. Examination of
extracted ion current profiles of appropriate ions can aid in the
selection of spectra, and in qualitative identification of
compounds. When analytes coelute (i.e., only one chromatographic
peak is apparent), the identification criteria can be met, but each
analyte spectrum will contain extraneous ions contributed by the
coeluting compound.
7.4.2 For samples containing components not associated with the
calibration standards, a library search may be made for the purpose of
tentative identification. The necessity to perform this type of
identification will be determined by the type of analyses being conducted.
Guidelines for making tentative identification are:
(1) Relative intensities of major ions in the reference spectrum
(ions > 10% of the most abundant ion) should be present in the sample
spectrum.
(2) The relative intensities of the major ions should agree within
+ 20%. (Example: For an ion with an abundance of 50% in the standard
spectrum, the corresponding sample ion abundance must be between 30 and
70%).
(3) Molecular ions present in the reference spectrum should be
present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the reference
spectrum should be reviewed for possible background contamination or
presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the sample
spectrum should be reviewed for possible subtraction from the sample
spectrum because of background contamination or coeluting peaks. Data
system library reduction programs can sometimes create these
discrepancies.
Computer generated library search routines should not use
normalization routines that would misrepresent the library or unknown
spectra when compared to each other. Only after visual comparison of the
sample with the nearest library searches will the mass spectral
interpretation specialist assign a tentative identification.
7.5 Quantitative analysis
7.5.1 When an analyte has been qualitatively identified,
quantitation should be based on the integrated abundance from the EICP of
the primary characteristic ion chosen for that analyte. If the sample
produces an interference for the primary characteristic ion, a secondary
characteristic ion should be used.
5040A - 6 Revision 1
September 1994
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7.5.1.1 Using the internal standard calibration procedure,
the amount of analyte in the sample cartridge is calculated using
the response factor (RF) determined in Section 7.2.5 and Equation 2.
Amount of POHC = AsCis/Ai8RF (2)
where:
As = Area of the characteristic ion for the analyte to be
measured.
Ais = Area for the characteristic ion of the internal
standard.
Cis = Amount (ng) of internal standard.
7.5.1.2 The choice of methods for evaluating data
collected using VOST for incinerator trial burns is a regulatory
decision. The procedures used extensively by one user are outlined
below.
7.5.1.3 The total amount of the POHCs of interest
collected on a pair of traps should be summed.
7.5.1.4 The observation of high concentrations of POHCs of
interest in blank cartridges indicates possible residual
contamination of the sorbent cartridges prior to shipment to and use
at the site. Data that fall in this category (especially data
indicating high concentrations of POHCs in blank sorbent cartridges)
should be qualified with regard to validity, and blank data should
be reported separately. The applicability of data of this type to
the determination of ORE is a regulatory decision. Continued
observation of high concentrations of POHCs in blank sorbent
cartridges indicates that procedures for cleanup, monitoring,
shipment, and storage of sorbent cartridges by a particular user be
investigated to eliminate this problem.
7.5.1.5 If any internal standard recoveries fall outside
the control limits established in Section 8.4, data for all analytes
determined for that cartridge(s) must be qualified with the
observation.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 0030 for sample preparation procedures.
8.2 Each laboratory that uses this method is required to operate a formal
quality control program. The minimum requirements of this program consist of an
initial demonstration of laboratory capability and the analysis of blank Tenax
and Tenax/charcoal cartridges spiked with the analytes of interest. The
laboratory is required to maintain performance records to define the quality of
5040A - 7 Revision 1
September 1994
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data that are generated. Ongoing performance checks must be compared with
established performance criteria to determine if results are within the expected
precision and accuracy limits of the method.
8.2.1 Before performing any analyses, the analyst must demonstrate
the ability to generate acceptable precision and accuracy with this
method. This ability is established as described in Section 7.2.
8.2.2 The laboratory must spike all Tenax and Tenax/charcoal
cartridges with the internal standard(s) to monitor continuing laboratory
performance. This procedure is described in Section 7.2.
8.3 To establish the ability to generate acceptable accuracy and
precision, the analyst must spike blank Tenax and Tenax/charcoal cartridges with
the analytes of interest at two concentrations in the working range.
8.3.1 The average response factor (Rf) and the standard deviation
(s) for each must be calculated.
8.3.2 The average recovery and standard deviation must fall within
the expected range for determination of volatile POHCs using this method.
The expected range for recovery of volatile POHCs using this method is 50-
150%.
8.4 The analyst must calculate method performance criteria for the
internal standard(s).
8.4.1 Calculate upper and lower control limits for method
performances using the average area response (A) and standard
deviation(s) for internal standard:
Upper Control Limit (UCL) = A + 3s
Lower Control Limit (LCL) = A - 3s
The UCL and LCL can be used to construct control charts that are
useful in observing trends in performance. The control limits must be
replaced by method performance criteria as they become available from the
U.S. EPA.
8.5 The laboratory is required to spike all sample cartridges (Tenax and
Tenax/charcoal) with internal standard.
8.6 Each day, the analyst must demonstrate through analysis of blank
Tenax and Tenax/charcoal cartridges and organic-free reagent water that
interferences from the analytical system are under control.
8.7 The daily GC/MS performance tests required for this method are
described in Method 8240.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
5040A - 8 Revision 1
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10.0 REFERENCES
1. Protocol for Collection and Analysis of Volatile POHC's Using VOST.
EPA/600/8-84-007, March 1984.
2. Validation of the Volatile Organic Sampling Train (VOST) Protocol.
Volumes I and II. EPA/600/4-86-014a, January 1986.
5040A - 9 Revision 1
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METHOD 5040A
ANALYSIS OF SORBENT CARTRIDGES FROM VOLATILE ORGANIC SAMPLING TRAIN
GAS CHROMATOGRAPHY/MASS SPECTROMETRY TECHNIQUE
(VOST)
( Start j
7.1.1 Assemble
purge and trap
desorption
device.
7.1.2 Connect
thermal
desorption
device;
calib. system.
7.2.1 Select
internal
standard.
7.2.3 Prepare
calibration
standards using
flash evaporat.
technique.
7.2.4 Direct
gas flow
through traps.
7.2.4 Expel
contents of
syringe through
GC injection
port.
7.2.4 Analyze
trap by P-T-D
GC/MS
procedure.
7.2.5 Analyze
•ach calib.
standard for
both cartridges
(aee 7.3).
7.2.5 Tabulate
area response
and calculate
response factor.
7.2.6 Verify
response
factor each
day.
7.3 Place
sample
cartridge in
desorp. apparatus;
desorb in P-T.
7.3 Desorb
into GC/MS
system.
7.4.1
Qualitatively
identify
volatile POHCs.
7.5.1 Use
primary
characteristic
ion for
quantitation.
7.6.1.1
Calculate
amount of analyte
in sample.
7.5.1.3 Sum
amount of POHCs
of interest for
each pair of
traps.
7.5.1.4 Examine
blank* data for
•igns of residual
contamination.
7.5.1.5 Compare
int. std.
recoveries to
Section 8.4
control limits.
Stop
5040A - 11
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METHOD 5041
PROTOCOL FOR ANALYSIS OF SORBENT CARTRIDGES FROM VOLATILE ORGANIC
SAMPLING TRAIN (VOST): WIDE-BORE CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 This method describes the analysis of volatile principal organic
hazardous constituents (POHCs) collected from the stack gas effluents of
hazardous waste incinerators using the VOST methodology (1). For a comprehensive
description of the VOST sampling methodology see Method 0030. The following
compounds may be determined by this method:
Compound Name
CAS No.'
Acetone
Acrylonitrile
Benzene
Bromodi chl oromethane
Bromoformb
Bromomethane0
Carbon disulfide
Carbon tetrachloride
Chlorobenzene
Chl orodi bromomethane
Chloroethanec
Chloroform
Chl oromethane0
Di bromomethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans- 1,2-Di chl oroethene
1 , 2-Di chl oropropane
ci s- 1 ,3-Di chl oropropene
trans-l,3-Dichloropropene
Ethyl benzene6
lodomethane
Methylene chloride
Styreneb
l,l,2,2-Tetrachloroethaneb
Tetrachloroethene
Toluene
67-64-1
107-13-1
71-43-2
75-27-4
75-25-2
74-83-9
75-15-0
56-23-5
108-90-7
124-48-1
75-00-3
67-66-3
74-87-3
74-95-3
75-35-3
107-06-2
75-35-4
156-60-5
78-87-5
10061-01-5
10061-02-6
100-41-4
74-88-4
75-09-2
100-42-5
79-34-5
127-18-4
108-88-3
(continued)
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Compound Name CAS No.a
1,1,1-Trichloroethane 71-55-6
1,1,2-Trichloroethane 79-00-5
Trichloroethene 79-01-6
Trichlorofluoromethane 75-69-4
l,2,3-Trichloropropaneb 96-18-4
Vinyl chloride0 75-01-4
Xylenesb
8 Chemical Abstract Services Registry Number.
b Boiling point of this compound is above 132°C. Method 0030 is not
appropriate for quantitative sampling of this analyte.
0 Boiling point of this compound is below 30°C. Special precautions must
be taken when sampling for this analyte by Method 0030. Refer to Sec. 1.3 for
discussion.
1.2 This method is most successfully applied to the analysis of non-polar
organic compounds with boiling points between 30°C and 100°C. Data are applied
to the calculation of destruction and removal efficiency (ORE), with limitations
discussed below.
1.3 This method may be applied to analysis of many compounds which boil
above 100°C, but Method 0030 is always inappropriate for collection of compounds
with boiling points above 132°C. All target analytes with boiling points greater
than 132°C are so noted in the target analyte list presented in Sec. 1.1. Use
of Method 0030 for collection of compounds boiling between 100°C and 132°C is
often possible, and must be decided based on case by case inspection of
information such as sampling method collection efficiency, tube desorption
efficiency, and analytical method precision and bias. An organic compound with
a boiling point below 30°C may break through the sorbent under the conditions
used for sample collection. Quantitative values obtained for compounds with
boiling points below 30°C must be qualified, since the value obtained represents
a minimum value for the compound if breakthrough has occurred. In certain cases,
additional QC measures may have been taken during sampling very low boilers with
Method 0030. This information should be considered during the data
interpretation stage.
When Method 5041 is used for survey analyses, values for compounds boiling
above 132°C may be reported and qualified since the quantity obtained represents
a minimum value for the compound. These minimum values should not be used for
trial burn ORE calculations or to prove insignificant risk.
1.4 The VOST analytical methodology can be used to quantitate volatile
organic compounds that are insoluble or slightly soluble in water. When
volatile, water soluble compounds are included in the VOST organic compound
analyte list, quantitation limits can be expected to be approximately ten times
5041 - 2 Revision 0
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higher than quantitation limits for water insoluble compounds (if the compounds
can be recovered at all) because the purging efficiency from water (and possibly
from Tenax-GC®) is poor.
1.5 Overall sensitivity of the method is dependent upon the level of
interferences encountered in the sample and the presence of detectable
concentrations of volatile POHCs in blanks. The target detection limit of this
method is 0.1 pg/m3 (ng/L) of flue gas, to permit calculation of a ORE equal to
or greater than 99.99% for volatile POHCs which may be present in the waste
stream at 100 ppm. The upper end of the range of applicability of this method
is limited by the dynamic range of the analytical instrumentation, the overall
loading of organic compounds on the exposed tubes, and breakthrough of the
volatile POHCs on the sorbent traps used to collect the sample. Table 1 presents
retention times and characteristic ions for volatile compounds which can be
determined by this method. Table 2 presents method detection limits for a range
of volatile compounds analyzed by the wide-bore VOST methodology.
1.6 The wide-bore VOST analytical methodology is restricted to use by,
or under the supervision of, analysts experienced in the use of sorbent media,
purge-and-trap systems, and gas chromatograph/mass spectrometers, and skilled in
the interpretation of mass spectra and their use as a quantitative tool.
2.0 SUMMARY OF METHOD
2.1 The sorbent tubes are thermally desorbed by heating and purging with
organic-free helium. The gaseous effluent from the tubes is bubbled through
pre-purged organic-free reagent water and trapped on an analytical sorbent trap
in a purge-and-trap unit (Figure 2). After desorption, the analytical sorbent
trap is heated rapidly and the gas flow from the analytical trap is directed to
the head of a wide-bore column under subambient conditions. The volatile organic
compounds desorbed from the analytical trap are separated by temperature
programmed high resolution gas chromatography and detected by continuously
scanning low resolution mass spectrometry (Figure 3). Concentrations of volatile
organic compounds are calculated from a multi-point calibration curve, using the
method of response factors.
3.0 INTERFERENCES
3.1 Sorbent tubes which are to be analyzed for volatile organic compounds
can be contaminated by diffusion of volatile organic compounds (particularly
Freon® refrigerants and common organic solvents) through the external container
(even through a Teflon® lined screw cap on a glass container) and the Swagelok®
sorbent tube caps during shipment and storage. The sorbent tubes can also be
contaminated if organic solvents are present in the analytical laboratory. The
use of blanks is essential to assess the extent of any contamination. Field
blanks must be prepared and taken to the field. The end caps of the tubes are
removed for the period of time required to exchange two pairs of traps on the
VOST sampling apparatus. The tubes are recapped and shipped and handled exactly
as the actual field samples are shipped and handled. At least one pair of field
blanks is included with each six pairs of sample cartridges collected.
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3.2 At least one pair of blank cartridges (one Tenax-GC®, one
Tenax-GC®/charcoal) shall be included with shipment of cartridges to a hazardous
waste incinerator site as trip blanks. These trip blanks will be treated like
field blanks except that the end caps will not be removed during storage at the
site. This pair of traps will be analyzed to monitor potential contamination
which may occur during storage and shipment.
3.3 Analytical system blanks are required to demonstrate that
contamination of the purge-and-trap unit and the gas chromatograph/mass
spectrometer has not occurred or that, in the event of analysis of sorbent tubes
with very high concentrations of organic compounds, no compound carryover is
occurring. Tenax® from the same preparation batch as the Tenax® used for field
sampling should be used in the preparation of the method (laboratory) blanks.
A sufficient number of cleaned Tenax® tubes from the same batch as the field
samples should be reserved in the laboratory for use as blanks.
3.4 Cross contamination can occur whenever low-concentration samples are
analyzed after high-concentration samples, or when several high-concentration
samples are analyzed sequentially. When an unusually concentrated sample is
analyzed, this analysis should be followed by a method blank to establish that
the analytical system is free of contamination. If analysis of a blank
demonstrates that the system is contaminated, an additional bake cycle should be
used. If the analytical system is still contaminated after additional baking,
routine system maintenance should be performed: the analytical trap should be
changed and conditioned, routine column maintenance should be performed (or
replacement of the column and conditioning of the new column, if necessary), and
bakeout of the ion source (or cleaning of the ion source and rods, if required).
After system maintenance has been performed, analysis of a blank is required to
demonstrate that the cleanliness of the system is acceptable.
3.5 Impurities in the purge gas and from organic compounds out-gassing
in tubing account for the majority of contamination problems. The analytical
system must be demonstrated to be free from contamination under the conditions
of the analysis by analyzing two sets of clean, blank sorbent tubes with organic-
free reagent purge water as system blanks. The analytical system is acceptably
clean when these two sets of blank tubes show values for the analytes which are
within one standard deviation of the normal system blank. Use of plastic
coatings, non-Teflon® thread sealants, or flow controllers with rubber
components should be avoided.
3.6 VOST tubes are handled in the laboratory to spike standards and to
position the tubes within the desorption apparatus. When sorbent media are
handled in the laboratory atmosphere, contamination is possible if there are
organic solvents in use anywhere in the laboratory. It is therefore necessary
to make daily use of system blanks to monitor the cleanliness of the sorbents and
the absence of contamination from the analytical system. A single set of system
blank tubes shall be exposed to normal laboratory handling procedures and
analyzed as a sample. This sample should be within one standard deviation of
normal VOST tube blanks to demonstrate lack of contamination of the sorbent
media.
3.7 If the emission source has a high concentration of non-target organic
compounds (for example, hydrocarbons at concentrations of hundreds of ppm), the
5041 - 4 Revision 0
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presence of these non-target compounds will interfere with the performance of the
VOST analytical methodology. If one or more of the compounds of interest
saturates the chromatographic and mass spectrometric instrumentation, no
quantitative calculations can be made and the tubes which have been sampled under
the same conditions will yield no valid data for any of the saturated compounds.
In the presence of a very high organic loading, even if the compounds of interest
are not saturated, the instrumentation is so saturated that the linear range has
been surpassed. When instrument saturation occurs, it is possible that compounds
of interest cannot even be identified correctly because a saturated mass
spectrometer may mis-assign masses. Even if compounds of interest can be
identified, accurate quantitative calculations are impossible at detector
saturation. No determination can be made at detector saturation, even if the
target compound itself is not saturated. At detector saturation, a negative bias
will be encountered in analytical measurements and no accurate calculation can
be made for the Destruction and Removal Efficiency if analytical values may be
biased negatively.
3.8 The recoveries of the surrogate compounds, which are spiked on the
VOST tubes immediately before analysis, should be monitored carefully as an
overall indicator of the performance of the methodology. Since the matrix of
stack emissions is so variable, only a general guideline for recovery of 50-150%
can be used for surrogates. The analyst cannot use the surrogate recoveries as
a guide for correction of compound recoveries. The surrogates are valuable only
as a general indicator of correct operation of the methodology. If surrogates
are not observed or if recovery of one or more of the surrogates is outside the
50-150% range, the VOST methodology is not operating correctly. The cause of the
failure in the methodology is not obvious. The matrix of stack emissions
contains large amounts of water, may be highly acidic, and may contain large
amounts of target and non-target organic compounds. Chemical and surface
interactions may be occurring on the tubes. If recoveries of surrogate compounds
are extremely low or surrogate compounds cannot even be identified in the
analytical process, then failure to observe an analyte may or may not imply that
the compound of interest has been removed from the emissions with a high degree
of efficiency (that is, the Destruction and Removal Efficiency for that analyte
is high).
4.0 APPARATUS AND MATERIALS
4.1 Tube desorption apparatus: Acceptable performance of the methodology
requires: 1) temperature regulation to ensure that tube temperature during
desorption is regulated to 180°C + 10°; 2) good contact between tubes and the
heating apparatus to ensure that the sorbent bed is thoroughly and uniformly
heated to facilitate desorption of organic compounds; and 3) gas-tight
connections to the ends of the tubes to ensure flow of desorption gas through the
tubes without leakage during the heating/desorption process. A simple clamshell
heater which will hold tubes which are 3/4" in outer diameter will perform
acceptably as a desorption apparatus.
4.2 Purge-and-trap device: The purge-and-trap device consists of three
separate pieces of equipment: a sample purge vessel, an analytical trap, and a
desorber. Complete devices are commercially available from a variety of sources,
or the separate components may be assembled. The cartridge thermal desorption
5041 - 5 Revision 0
September 1994
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apparatus is connected to the sample purge vessel by 1/8" Teflon® tubing
(unheated transfer line). The tubing which connects the desorption chamber to
the sample purge vessel should be as short as is practical.
4.2.1 The sample purge vessel is required to hold 5 ml of organic-
free reagent water, through which the gaseous effluent from the VOST tubes
is routed. The water column should be at least 3 cm deep. The gaseous
headspace between the water column and the analytical trap must have a
total volume of less than 15 ml. The purge gas must pass through the
water column as finely divided bubbles with a diameter of less than 3 mm
at the origin. The sample purger shown in Figure 4 meets these
requirements. Alternate sample purging vessels may be used if equivalent
performance is demonstrated.
4.2.2 The analytical trap must be at least 25 cm and have an
internal diameter of at least 0.105 in. The analytical trap must contain
the following components:
2,6-diphenylene oxide polymer: 60/80 mesh, chromatograph grade
(Tenax-GC®, or equivalent)
methyl silicone packing: OV-1 (3%) on Chromosorb-W 60/80
mesh, or equivalent
silica gel: 35/60 mesh, Davison grade 15 or
equivalent
coconut charcoal: prepare from Barneby Cheney,
CA-580-26, or equivalent, by
crushing through 26 mesh
screen.
The proportions are: 1/3 Tenax-GC®, 1/3 silica gel, and 1/3
charcoal, with approximately 1.0 cm of methyl silicone packing. The
analytical trap should be conditioned for four hours at 180°C with gas flow
(10 mL/min) prior to use in sample analysis. During conditioning, the
effluent of the trap should not be vented to the analytical column. The
thermal desorption apparatus is connected to the injection system of the
mass spectrometer by a transfer line which is heated to 100°C.
4.2.3 The desorber must be capable of rapidly heating the analytical
trap to 180°C for desorption. The polymer section of the trap should not
exceed 180°C, and the remaining sections should not exceed 220°C, during
bake-out mode.
4.3 Gas chromatograph/mass spectrometer/data system:
4.3.1 Gas chromatograph: An analytical system complete with a
temperature programmable oven with sub-ambient temperature capabilities
and all required accessories, including syringes, analytical columns, and
gases.
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4.3.2 Chromatographic column: 30 m x 0.53 mm ID wide-bore fused
silica capillary column, 3 /urn film thickness, DB-624 or equivalent.
4.3.3 Mass spectrometer: capable of scanning from 35-260 amu every
second or less, using 70 eV (nominal) electron energy in the electron
ionization mode and producing a mass spectrum that meets all of the
criteria in Table 3 when 50 ng of 4-bromofluorobenzene (BFB) is injected
into the water in the purge vessel.
4.3.4 Gas chromatograph/mass spectrometer interface: Any gas
chromatograph to mass spectrometer interface that gives acceptable
calibration points at 50 ng or less per injection of each of the analytes,
and achieves the performance criteria for 4-bromofluorobenzene shown in
Table 3, may be used. If a glass jet separator is used with the wide-bore
column, a helium make-up flow of approximately 15 ml, introduced after the
end of the column and prior to the entrance of the effluent to the
separator, will be required for optimum performance.
4.3.5 Data system: A computer system that allows the continuous
acquisition and storage on machine readable media of all mass spectra
obtained throughout the duration of the Chromatographic program must be
interfaced to the mass spectrometer. The computer must have software that
allows searching any gas chromatographic/mass spectrometric data file for
ions of a specified mass and plotting such ion abundances versus time or
scan number. This type of plot is defined as an Extracted Ion Current
Profile (EICP). Software must also be available that allows the
integration of the ion abundances in any EICP between specified time or
scan number limits. The most recent version of the EPA/NIST Mass Spectral
Library should also be available.
4.4 Wrenches: 9/16", 1/2", 7/16", and 5/16".
4.5 Teflon® tubing: 1/8" diameter.
4.6 Syringes: 25 ^uL syringes (2), 10 /j,l syringes (2).
4.7 Fittings: 1/4" nuts, 1/8" nuts, 1/16" nuts, 1/4" to 1/8" union, 1/4"
to 1/4" union, 1/4" to 1/16" union.
4.8 Adjustable stand to raise the level of the desorption unit, if
required.
4.9 Volumetric flasks: 5 ml, class A with ground glass stopper.
4.10 Injector port or equivalent, heated to 180°C for loading standards
onto VOST tubes prior to analysis.
4.11 Vials: 2 ml, with Teflon® lined screw caps or crimp tops.
4.12 Syringe: 5 ml, gas-tight with shutoff valve.
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5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2.1 It is advisable to maintain the stock of organic-free reagent
water generated for use in the purge-and-trap apparatus with a continuous
stream of inert gas bubbled through the water. Continuous bubbling of the
inert gas maintains a positive pressure of inert gas above the water as a
safeguard against contamination.
5.3 Methanol, CH3OH. Pesticide quality or equivalent. To avoid
contamination with other laboratory solvents, it is advisable to maintain a
separate stock of methanol for the preparation of standards for VOST analysis and
to regulate the use of this methanol very carefully.
5.4 Stock standard solutions - Can be prepared from pure standard
materials or can be purchased as certified solutions. Stock standard solutions
must be prepared in high purity methanol. All preparation of standards should
take place in a hood, both to avoid contamination and to ensure safety of the
analyst preparing the standards.
5.4.1 Place about 4 ml of high purity methanol in a 5 ml volumetric
flask. Allow the flask to stand, unstoppered, for about 10 min, or until
all alcohol wetted surfaces have dried.
5.4.1.1 Add appropriate volumes of neat liquid chemicals
or certified solutions, using a syringe of the appropriate volume.
Liquid which is added to the volumetric flask must fall directly
into the alcohol without contacting the neck of the flask. Gaseous
standards can be purchased as methanol solutions from several
commercial vendors.
5.4.1.2 Dilute to volume with high purity methanol,
stopper, and then mix by inverting the flask several times. Calcu-
late concentration by the dilution of certified solutions or neat
chemicals.
5.4.2 Transfer the stock standard solution into a Teflon® sealed
screw cap bottle. An amber bottle may be used. Store, with minimal
headspace, at -10°C to -20°C, and protect from light.
5.4.3 Prepare fresh standards every two months for gases. Reactive
compounds such as styrene may need to be prepared more frequently. All
other standards must be replaced after six months, or sooner if comparison
with check standards indicates a problem.
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5.5 Secondary dilution standards: Using stock standard solutions,
prepare, in high purity methanol, secondary dilution standards containing the
compounds of interest, either singly or mixed together. Secondary dilution
standards must be stored with minimal headspace and should be checked frequently
for signs of degradation or evaporation, especially just prior to preparing
calibration standards from them.
5.6 Surrogate standards: The recommended surrogates are toluene-d8,
4-bromofluorobenzene, and l,2-dichloroethane-d4. Other compounds may be used as
surrogate compounds, depending upon the requirements of the analysis. Surrogate
compounds are selected to span the elution range of the compounds of interest.
Isotopically labeled compounds are selected to preclude the observation of the
same compounds in the stack emissions. More than one surrogate is used so that
surrogate measurements can still be made even if analytical interferences with
one or more of the surrogate compounds are encountered. However, at least three
surrogate compounds should be used to monitor the performance of the methodology.
A stock surrogate compound solution in high purity methanol should be prepared
as described in Sec. 5.4, and a surrogate standard spiking solution should be
prepared from the stock at a concentration of 250 jug/10 ml in high purity
methanol. Each pair of VOST tubes (or each individual VOST tube, if the tubes
are analyzed separately) must be spiked with 10 juL of the surrogate spiking
solution prior to GC/MS analysis.
5.7 Internal standards: The recommended internal standards are
bromochloromethane, 1,4-difluorobenzene, and chlorobenzene-d5. Other compounds
may be used as internal standards as long as they have retention times similar
to the compounds being analyzed by GC/MS. The internal standards should be
distributed through the chromatographic elution range. Prepare internal standard
stock and secondary dilution standards in high purity methanol using the
procedures described in Sees. 5.2 and 5.3. The secondary dilution standard
should be prepared at a concentration of 25 mg/L of each of the internal standard
compounds. Addition of 10 fj,l of this internal standard solution to each pair
of VOST tubes (or to each VOST tube, if the tubes are analyzed individually) is
the equivalent of 250 ng total.
5.8 4-Bromofluorobenzene (BFB) standard: A standard solution containing
25 ng/juL of BFB in high purity methanol should be prepared for use as a tuning
standard.
5.9 Calibration standards: Calibration standards at a minimum of five
concentrations will be required from the secondary dilution of stock standards
(see Sees. 5.2 and 5.3). A range of concentrations for calibration can be
obtained by use of different volumes of a 50 mg/L methanol solution of the
calibration standards. One of the concentrations used should be at a
concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations found
in field samples but should not exceed the linear range of the GC/MS analytical
system (a typical range for a calibration would be 10, 50, 100, 350, and 500 ng,
for example). Each calibration standard should contain each analyte for
detection by this method. Store calibration standards for one week only in a
vial with no headspace.
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5.10 Great care must be taken to maintain the integrity of all standard
solutions. All standards of volatile compounds in methanol must be stored at
-10° to -20°C in amber bottles with Teflon® lined screw caps or crimp tops. In
addition, careful attention must be paid to the use of syringes designated for
a specific purpose or for use with only a single standard solution since cross
contamination of volatile organic standards can occurs very readily.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See Method 0030 for the VOST Sampling Methodology.
6.2 VOST samples are collected on paired cartridges. The first of the
pair of sorbent cartridges is packed with approximately 1.6 g of Tenax-GC®
resin. The second cartridge of the pair is packed with Tenax-GC® and petroleum
based charcoal (3:1 by volume; approximately 1 g of each). In sampling, the
emissions gas stream passes through the Tenax-GC® layer first and then through
the charcoal layer. The Tenax-GC® is cleaned and reused; charcoal is not reused
when tubes are prepared. Sorbent is cleaned and the tubes are packed. The tubes
are desorbed and subjected to a blank check prior to being sent to the field.
When the tubes are used for sampling (see Figure 5 for a schematic diagram of the
Volatile Organic Sampling Train (VOST)), cooling water is circulated to the
condensers and the temperature of the cooling water is maintained near 0°C. The
end caps of the sorbent cartridges are placed in a clean, screw capped glass
container during sample collection.
6.3 After the apparatus is leak checked, sample collection is
accomplished by opening the valve to the first condenser, turning on the pump,
and sampling at a rate of 1 liter/min for 20 minutes. The volume of sample for
any pair of traps should not exceed 20 liters. An alternative set of conditions
for sample collection requires sampling at a reduced flow rate, where the overall
volume of sample collected is 5 liters at a rate of 0.25 L/min for 20 minutes.
The 20 minute period is required for collecting an integrated sample.
6.4 Following collection of 20 liters of sample, the train is leak
checked a second time at the highest pressure drop encountered during the run to
minimize the chance of vacuum desorption of organics from the Tenax®.
6.5 The train is returned to atmospheric pressure and the two sorbent
cartridges are removed. The end caps are replaced and the cartridges are placed
in a suitable environment for storage and transport until analysis. The sample
is considered invalid if the leak test does not meet specifications.
6.6 A new pair of cartridges is placed in the VOST, the VOST is leak
checked, and the sample collection process is repeated until six pairs of traps
have been exposed.
6.7 All sample cartridges are kept in coolers on cold packs after
exposure and during shipment. Upon receipt at the laboratory, the cartridges are
stored in a refrigerator at 4°C until analysis.
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7.0 PROCEDURE
7.1 Recommended operating conditions for cartridge desorber,
purge-and-trap unit, and gas chromatograph/mass spectrometer using the wide-bore
column are:
Cartridge Desorption Oven
Desorb Temperature
Desorb Time
Desorption Gas Flow
Desorption/Carrier Gas
Purge-and-Trap Concentrator
Analytical Trap Desorption Flow
Purge Temperature
Purge Time
Analytical Trap Desorb Temperature
Analytical Trap Desorb Time
Gas Chromatoqraph
Column
Carrier Gas Flow
Makeup Gas Flow
Injector Temperature
Transfer Oven Temperature
Initial Temperature
Initial Hold Time
Program Rate
Final Temperature
Final Hold Time
Mass Spectrometer
Manifold Temperature
Scan Rate
Mass Range
Electron Energy
Source Temperature
180°C
11 minutes
40 mL/min
Helium, Grade 5.0
2.5 mL/min helium
Ambient
11 minutes
180°C
5 minutes
DB-624, 0.53 mm ID x 30 m thick
film (3 jum) fused silica capillary,
or equivalent
15 mL/min
15 mL/min
200°C
240°C
5°C
2 minutes
6°C/min
240°C
1 minute, or until elution ceases
105°C
1 sec/cycle
35-260 amu
70 eV (nominal)
According to
specifications
manufacturer's
7.2 Each GC/MS system must be hardware tuned to meet the criteria in
Table 3 for a 50 ng injection of 4-bromofluorobenzene (2 juL injection of the BFB
standard solution into the water of the purge vessel). No analyses may be
initiated until the criteria presented in Table 3 are met.
7.3 Assemble a purge-and-trap device that meets the specifications in
Method 5030. Condition the analytical trap overnight at 180°C in the purge mode,
with an inert gas flow of at least 20 mL/min. Prior to use each day, condition
the trap for 10 minutes by backflushing at 180°C, with the column at 220°C.
7.4 Connect the purge-and-trap device to a gas chromatograph.
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7.5 Assemble a VOST tube desorption apparatus which meets the
requirements of Sec. 4.1.
7.6 Connect the VOST tube desorption apparatus to the purge-and-trap
unit.
7.7 Calibrate the instrument using the internal standard procedure, with
standards and calibration compounds spiked onto cleaned VOST tubes for
calibration.
7.7.1 Compounds in methanolic solution are spiked onto VOST tubes
using the flash evaporation technique. To perform flash evaporation, the
injector of a gas chromatograph or an equivalent piece of equipment is
required.
7.7.1.1 Prepare a syringe with the appropriate volume of
methanolic standard solution (either surrogates, internal standards,
or calibration compounds).
7.7.1.2 With the injector port heated to 180°C, and with
an inert gas flow of 10 mL/min through the injector port, connect
the paired VOST tubes (connected as in Figure 1, with gas flow in
the same direction as the sampling gas flow) to the injector port;
tighten with a wrench so that there is no leakage of gas. If
separate tubes are being analyzed, an individual Tenax® or
Tenax®/charcoal tube is connected to the injector.
7.7.1.3 After directing the gas flow through the VOST
tubes, slowly inject the first standard solution over a period of 25
seconds. Wait for 5 sec before withdrawing the syringe from the
injector port.
7.7.1.4 Inject a second standard (if required) over a
period of 25 seconds and wait for 5 sec before withdrawing the
syringe from the injector port.
7.7.1.5 Repeat the sequence above as required until all of
the necessary compounds are spiked onto the VOST tubes.
7.7.1.6 Wait for 30 seconds, with gas flow, after the last
spike before disconnecting the tubes. The total time the tubes are
connected to the injector port with gas flow should not exceed 2.5
minutes. Total gas flow through the tubes during the spiking
process should not exceed 25 ml to prevent break through of adsorbed
compounds during the spiking process. To allow more time for
connecting and disconnecting tubes, an on/off valve may be installed
in the gas line to the injector port so that gas is not flowing
through the tubes during the connection/disconnection process.
7.8 Prepare the purge-and-trap unit with 5 ml of organic-free reagent
water in the purge vessel.
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7.9 Connect the paired VOST tubes to the gas lines in the tube desorption
unit. The tubes must be connected so that the gas flow during desorption will
be opposite to the flow of gas during sampling: i.e., the tube desorption gas
passes through the charcoal portion of the tube first. An on/off valve may be
installed in the gas line leading to the tube desorption unit in order to prevent
flow of gas through the tubes during the connection process.
7.10 Initiate tube desorption/purge and heating of the VOST tubes in the
desorption apparatus.
7.11 Set the oven of the gas chromatograph to subambient temperatures by
cooling with liquid nitrogen.
7.12 Prepare the GC/MS system for data acquisition.
7.13 At the conclusion of the tube/water purge time, attach the analytical
trap to the gas chromatograph, adjust the purge-and-trap device to the desorb
mode, and initiate the gas chromatographic program and the GC/MS data
acquisition. Concurrently, introduce the trapped materials to the gas
chromatographic column by rapidly heating the analytical trap to 180°C while
backflushing the trap with inert gas at 2.5 mL/min for 5 min. Initiate the
program for the gas chromatograph and simultaneously initiate data acquisition
on the GC/MS system.
7.14 While the analytical trap is being desorbed into the gas
chromatograph, empty the purging vessel. Wash the purging vessel with a minimum
of two 5 ml flushes of organic-free reagent water (or methanol followed by
organic-free reagent water) to avoid carryover of analytes into subsequent
analyses.
7.15 After the sample has been desorbed, recondition the analytical trap
by employing a bake cycle on the purge-and-trap unit. The analytical trap may
be baked at temperatures up to 220°C. However, extensive use of high
temperatures to recondition the trap will shorten the useful life of the
analytical trap. After approximately 11 minutes, terminate the trap bake and
cool the trap to ambient temperatures in preparation for the next sample. This
procedure is a convention for reasonable samples and should be adequate if the
concentration of contamination does not saturate the analytical system. If the
organic compound concentration is so high that the analytical system is saturated
beyond the point where even extended system bakeout is not sufficient to clean
the system, a more extensive system maintenance must be performed. To perform
extensive system maintenance, the analytical trap is replaced and the new trap
is conditioned. Maintenance is performed on the GC column by removing at least
one foot from the front end of the column. If the chromatography does not
recover after column maintenance, the chromatographic column must be replaced.
The ion source should be baked out and, if the bakeout is not sufficient to
restore mass spectrometric peak shape and sensitivity, the ion source and the
quadrupole rods must be cleaned.
7.16 Initial calibration for the analysis of VOST tubes: It is essential
that calibration be performed in the mode in which analysis will be performed.
If tubes are being analyzed as pairs, calibration standards should be prepared
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on paired tubes. If tubes are being analyzed individually, a calibration should
be performed on individual Tenax® only tubes and Tenax®/charcoal tubes.
7.16.1 Prepare the calibration standards by spiking VOST tubes
using the procedure described in Sec. 7.7.1. Spike each pair of VOST
tubes (or each of the individual tubes) immediately before analysis.
Perform the calibration analyses in order from low concentration to high
to minimize the compound carryover. Add 5.0 ml of organic-free reagent
water to the purging vessel. Initiate tube desorb/purge according to the
procedure described above.
7.16.2 Tabulate the area response of the characteristic primary
ions (Table 1) against concentration for each target compound, each
surrogate compound, and each internal standard. The first criterion for
quantitative analysis is correct identification of compounds. The
compounds must elute within + 0.06 retention time units of the elution
time of the standard analyzed on the same analytical system on the day of
the analysis. The analytes should be quantitated relative to the closest
eluting internal standard, according to the scheme shown in Table 4.
Calculate response factors (RF) for each compound relative to the internal
standard shown in Table 4. The internal standard selected for the
calculation of RF is the internal standard that has a retention time
closest to the compound being measured. The RF is calculated as follows:
RF = (Ax/Cis)/(Als/Cx)
where:
Ax = area of the characteristic ion for the compound being
measured.
Ais = area of the characteristic ion for the specific internal
standard.
C,s = concentration of the specific internal standard.
Cx = concentration of the compound being measured.
7.16.3 The average RF must be calculated for each compound. A
system performance check should be made before the calibration curve is
used. Five compounds (the System Performance Check Compounds, or SPCCs)
are checked for a minimum average response factor. These compounds are
chloromethane, 1,1-dichloroethane, bromoform, 1,1,2,2-tetrachloroethane,
and chlorobenzene. The minimum acceptable average RF for these compounds
should be 0.300 (0.250 for bromoform). These compounds typically have RFs
of 0.4 - 0.6, and are used to check compound instability and check for
degradation caused by contaminated lines or active sites in the system.
Examples of these occurrences are:
7.16.3.1 Chloromethane: This compound is the most likely
compound to be lost if the purge flow is too fast.
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7.16.3.2 Bromoform: This compound is one of the compounds
most likely to be purged very poorly if the purge flow is too slow.
Cold spots and/or active sites in transfer lines may adversely
affect response. Response of the primary quantitation ion (m/z 173)
is directly affected by the tuning for 4-bromofluorobenzene at the
ions of masses 174 and 176. Increasing the ratio of ions 174 and 176
to mass 95 (the base peak of the mass spectrum of
bromofluorobenzene) may improve bromoform response.
7.16.3.3 1,1,2,2-Tetrachloroethane and 1,1-dichloroethane:
These compounds are degraded by contaminated transfer lines in
purge-and-trap systems and/or active sites in trapping materials.
7.16.4 Using the response factors from the initial calibration,
calculate the percent relative standard deviation (%RSD) for the
Calibration Check Compounds (CCCs).
%RSD = (SD/X) x 100
where:
%RSD
RF
SD
percent relative standard deviation
individual RF measurement
mean of 5 initial RFs for a compound (the 5 points
over the calibration range)
standard deviation of average RFs for a compound,
where SD is calculated:
SD =
N
(RFi-RF)
jn
The %RSD for each individual CCC should be less than 30 percent.
This criterion must be met in order for the individual calibration to be
valid. The CCCs are: 1,1-dichloroethene, chloroform, 1,2-dichloropropane,
toluene, ethyl benzene, and vinyl chloride.
7.17 Daily GC/MS Calibration
7.17.1 Prior to the analysis of samples, purge 50 ng of the
4-bromofluorobenzene standard. The resultant mass spectrum for the BFB
must meet all of the criteria given in Table 3 before sample analysis
begins. These criteria must be demonstrated every twelve hours of
operation.
7.17.2 The initial calibration curve (Sec. 7.16) for each compound
of interest must be checked and verified once every twelve hours of
analysis time. This verification is accomplished by analyzing a
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calibration standard that is at a concentration near the midpoint
concentration for the working range of the GC/MS and checking the SPCC
(Sec. 7.16.3) and CCC (Sec. 7.16.4).
7.17.3 System Performance Check Compounds (SPCCs): A system
performance check must be made each twelve hours of analysis. If the SPCC
criteria are met, a comparison of response factors is made for all
compounds. This is the same check that is applied during the initial
calibration. If the minimum response factors are not achieved, the system
must be evaluated, and corrective action must be taken before analysis is
allowed to begin. The minimum response factor for volatile SPCCs is 0.300
(0.250 for bromoform). If these minimum response factors are not
achieved, some possible problems may be degradation of the standard
mixture, contamination of the injector port, contamination at the front
end of the analytical column, and active sites in the column or
chromatographic system. If the problem is active sites at the front end
of the analytical column, column maintenance (removal of approximately 1
foot from the front end of the column) may remedy the problem.
7.17.4 Calibration Check Compounds: After the system performance
check has been met, CCCs listed in Sec. 7.16.4 are used to check the
validity of the initial calibration. Calculate the percent difference
using the following equation:
(RF, - RFC) x 100
% Difference =
where:
RFi = average response factor from initial calibration
RFC = response factor from current calibration check standard.
If the percent difference for any compound is greater than 20, the
laboratory should consider this a warning limit. Benzene, toluene, and
styrene will have problems with response factors if Tenax® decomposition
occurs (either as a result of sampling exposure or temperature
degradation), since these compounds are decomposition products of Tenax®.
If the percent difference for each CCC is less than 25%, the initial
calibration is assumed to be valid. If the criterion of percent
difference less than 25% is not met for any one CCC, corrective action
MUST be taken. Problems similar to those listed under SPCCs could affect
this criterion. If a source of the problem cannot be determined after
corrective action is taken, a new five-point calibration curve MUST be
generated. The criteria for the CCCs MUST be met before quantitative
analysis can begin.
7.17.5 Internal standard responses and retention times in the
check calibration standard must be evaluated immediately after or during
data acquisition. If the retention time for any internal standard changes
by more than 30 seconds from the last check calibration (12 hr), the
chromatographic system must be inspected for malfunctions and corrections
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must be made, as required. A factor which may influence the retention
times of the internal standards on sample tubes is the level of overall
organic compound loading on the VOST tubes. If the VOST tubes are very
highly loaded with either a single compound or with multiple organic
compounds, retention times for standards and compounds of interest will be
affected. If the area for the primary ion of any of the internal
standards changes by a factor of two (-50% to +100%) from the last daily
calibration check, the gas chromatograph and mass spectrometer should be
inspected for malfunctions and corrections must be made, as appropriate.
If the level of organic loading of samples is high, areas for the primary
ions of both compounds of interest and standards will be adversely
affected. Calibration check standards should not be subject to variation,
since the concentrations of organic compounds on these samples are set to
be within the linear range of the instrumentation. If instrument
malfunction has occurred, analyses of samples performed under conditions
of malfunction may be invalidated.
7.18 GC/MS Analysis of Samples
7.18.1 Set up the cartridge desorption unit, purge-and-trap
unit, and GC/MS as described above.
7.18.2 BFB tuning criteria and daily GC/MS calibration check
criteria must be met before analyzing samples.
7.18.3 Adjust the helium purge gas flow rate (through the
cartridges and purge vessel) to approximately 40 mL/min. Optimize the
flow rate to provide the best response for chloromethane and bromoform, if
these compounds are analytes. A flow rate which is too high reduces the
recovery of chloromethane, and an insufficient gas flow rate reduces the
recovery of bromoform.
7.18.4 The first analysis performed after the tuning check and
the calibration or daily calibration check is a method blank. The method
blank consists of clean VOST tubes (both Tenax® and Tenax®/charcoal) which
are spiked with surrogate compounds and internal standards according to
the procedure described in Sec. 7.7.1. The tubes which are used for the
method blanks should be from the same batch of sorbent as the sorbent used
for the field samples. After the tubes are cleaned and prepared for
shipment to the field, sufficient pairs of tubes should be retained from
the same batch in the laboratory to provide method blanks during the
analysis.
7.18.5 The organic-free reagent water for the purge vessel for
the analysis of each of the VOST samples should be supplied from the
laboratory inventory which is maintained with constant bubbling of inert
gas to avoid contamination.
7.18.6 If the analysis of a pair of VOST tubes has a
concentration of analytes that exceeds the initial calibration range, no
reanalysis of desorbed VOST tubes is possible. An additional calibration
point can be added to bracket the higher concentration encountered in the
samples so that the calibration database encompasses six or more points.
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Alternatively, the data may be flagged in the report as "extrapolated
beyond the upper range of the calibration." The use of the secondary ions
shown in Table 1 is permissible only in the case of interference with the
primary quantitation ion. Use of secondary ions to calculate compound
concentration in the case of saturation of the primary ion is not an
acceptable procedure, since a negative bias of an unpredictable magnitude
is introduced into the quantitative data when saturation of the mass
spectrum of a compound is encountered. If high organic loadings, either
of a single compound or of multiple compounds, are encountered, it is
vital that a method blank be analyzed prior to the analysis of another
sample to demonstrate that no compound carryover is occurring. If
concentrations of organic compounds are sufficiently high that carryover
problems are profound, extensive bakeout of the purge-and-trap unit will
be required. Complete replacement of the contaminated analytical trap,
with the associated requirement for conditioning the new trap, may also be
required for VOST samples which show excessive concentrations of organic
compounds. Other measures which might be required for decontamination of
the analytical system include bakeout of the mass spectrometer,
replacement of the filament of the mass spectrometer, cleaning of the ion
source of the mass spectrometer, and/or (depending on the nature of the
contamination) maintenance of the chromatographic column or replacement of
the chromatographic column, with the associated requirement for
conditioning the new chromatographic column.
7.19 Data Interpretation
7.19.1 Qualitative analysis:
7.19.1.1 The qualitative identification of compounds
determined by this method is based on retention time, and on
comparison of the sample mass spectrum, after background correction,
with characteristic ions in a reference mass spectrum. The
reference mass spectrum must be generated by the laboratory using
the conditions of this method. The characteristic ions from the
reference mass spectrum are defined to be the three ions of greatest
relative intensity, or any ions over 30% relative intensity if less
than three such ions occur in the reference spectrum. Compounds
should be identified as present when the criteria below are met.
7.19.1.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographic peak containing ions
specific for the target compound at a compound specific
retention time will be accepted as meeting this criterion.
7.19.1.1.2 The RRT of the sample component is + 0.06
RRT units of the RRT of the standard component.
7.19.1.1.3 The relative intensities of the
characteristic ions agree within 30% of the relative
intensities of these ions in the reference spectrum.
(Example: For an ion with an abundance of 50% in the
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reference spectrum, the corresponding abundance in a sample
spectrum can range between 20% and 80%.)
7.19.1.1.4 Structural isomers that produce very similar
mass spectra should be identified as individual isomers if
they have sufficiently different GC retention times.
Sufficient GC resolution is achieved if the height of the
valley between two isomer peaks is less than 25% of the sum of
the two peak heights. Otherwise, structural isomers are
identified as isomeric pairs.
7.19.1.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce
mass spectra containing ions contributed by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
appropriate selection of analyte spectra and background
spectra is important. Examination of extracted ion current
profiles of appropriate ions can aid in the selection of
spectra, and in qualitative identification of compounds. When
analytes coelute (i.e., only one chromatographic peak is
apparent), the identification criteria can be met, but each
analyte spectrum will contain extraneous ions contributed by
the coeluting compound.
7.19.1.2 For samples containing components not associated
with the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the type of analyses
being conducted. Guidelines for making tentative identification
are:
(1) Relative intensities of major ions in the reference
spectrum (ions > 10% of the most abundant ion) should be present in
the sample spectrum.
(2) The relative intensities of the major ions should agree
within + 20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
between 30 and 70%).
(3) Molecular ions present in the reference spectrum should
be present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the
reference spectrum should be reviewed for possible background
contamination or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the
sample spectrum should be reviewed for possible subtraction from the
sample spectrum because of background contamination or coeluting
peaks. Data system library reduction programs can sometimes create
these discrepancies.
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Computer generated library search routines should not use
normalization routines that would misrepresent the library or
unknown spectra when compared to each other. Only after visual
comparison of sample with the nearest library searches will the mass
spectral interpretation specialist assign a tentative
identification.
7.19.2 Quantitative analysis:
7.19.2.1 When a compound has been identified, the
quantitative analysis of that compound will be based on the
integrated abundance from the extracted ion current profile of the
primary characteristic ion for that compound (Table 1). In the
event that there is interference with the primary ion so that
quantitative measurements cannot be made, a secondary ion may be
used.
NOTE: Use of a secondary ion to perform quantitative
calculations in the event of the saturation of the
primary ion is not an acceptable procedure because of
the unpredictable extent of the negative bias which is
introduced. Quantitative calculations are performed
using the internal standard technique. The internal
standard used to perform quantitative calculations shall
be the internal standard nearest the retention time of
a given analyte (see Table 4).
7.19.2.2 Calculate the amount of each identified analyte
from the VOST tubes as follows:
Amount (ng) = (AsCis)/(AisRF)
where:
As = area of the characteristic ion for the analyte to be
measured.
Ais = area of the characteristic ion of the internal standard.
Cis = amount (ng) of the internal standard.
7.19.2.3 The choice of methods for evaluating data
collected using the VOST methodology for incinerator trial burns is
a regulatory decision. Various procedures are used to decide
whether blank correction should be performed and how blank
correction should be performed. Regulatory agencies to which VOST
data are submitted also vary in their preferences for data which are
or which are not blank corrected.
7.19.2.4 The total amount of the POHCs of interest
collected on a pair of traps should be summed.
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7.19.2.5 The occurrence of high concentrations of analytes
on method blank cartridges indicates possible residual contamination
of sorbent cartridges prior to shipment and use at the sampling
site. Data with high associated blank values must be qualified with
respect to validity, and all blank data should be reported
separately. No blank corrections should be made in this case.
Whether or not data of this type can be applied to the determination
of destruction and removal efficiency is a regulatory decision.
Continued observation of high concentrations of analytes on blank
sorbent cartridges indicates that procedures for cleanup and quality
control for the sampling tubes are inadequate. Corrective action
MUST be applied to tube preparation and monitoring procedures to
maintain method blank concentrations below detection limits for
analytes.
7.19.2.6 Where applicable, an estimate of concentration for
noncalibrated components in the sample may be made. The formulae
for quantitative calculations presented above should be used with
the following modifications: The areas Ax and Ais should be from the
total ion chromatograms, and the Response Factor for the
noncalibrated compound should be assumed to be 1. The nearest
eluting internal standard free from interferences in the total ion
chromatogram should be used to determine the concentration. The
concentration which is obtained should be reported indicating: (1)
that the value is an estimate; and (2) which internal standard was
used.
7.19.2.7 If any internal standard recoveries fall outside
the control limits established in Sec. 8.4, data for all analytes
determined for that cartridge(s) must be qualified with the
observation. Report results without correction for surrogate
compound recovery data. When duplicates are analyzed, report the
data obtained with the sample results.
8.0 QUALITY CONTROL
8.1 Each laboratory that uses these methods is required to operate a
formal quality control program. The minimum quality control requirements are
specified in Chapter One. In addition, this program should consist of an initial
demonstration of laboratory capability and an ongoing analysis of check samples
to evaluate and document data quality. The laboratory must maintain records to
document the quality of the data generated. Ongoing data quality checks are
compared with established performance criteria to determine if the results of
analyses meet the performance characteristics of the method. When sample
analyses indicate atypical method performance, a quality control check standard
(spiked method blank) must be analyzed to confirm that the measurements were
performed in an in-control mode of instrument operation.
8.2 Before processing any samples, the analyst should demonstrate,
through the analysis of a method blank (laboratory blank sorbent tubes, reagent
water purge) that interferences from the analytical system, glassware, sorbent
tube preparation, and reagents are under control. Each time a new batch of
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sorbent tubes is analyzed, a method blank should be processed as a safeguard
against chronic laboratory contamination. Blank tubes which have been carried
through all the stages of sorbent preparation and handling should be used in the
analysis.
8.3 The experience of the analyst performing the GC/MS analyses is
invaluable to the success of the analytical methods. Each day that the analysis
is performed, the daily calibration check standard should be evaluated to
determine if the chromatographic and tube desorption systems are operating
properly. Questions that should be asked are: Do the peaks look normal? Is the
system response obtained comparable to the response from previous calibrations?
Careful examination of the chromatogram of the calibration standard can indicate
whether column maintenance is required or whether the column is still usable,
whether there are leaks in the system, whether the injector septum requires
replacing, etc. If changes are made to the system (such as change of a column),
a calibration check must be carried out and a new multipoint calibration must be
generated.
8.4 Required instrument quality control is found in the following
sections:
8.4.1 The mass spectrometer must be tuned to meet the specifications
for 4-bromofluorobenzene in Sec. 7.2 (Table 3).
8.4.2 An initial calibration of the tube desorption/purge-and-trap/
GC/MS must be performed as specified in Sec. 7.7.
8,4.3 The GC/MS system must meet the SPCC criteria specified in Sec.
7.16.3 and the CCC criteria in Sec. 7.16.4 each twelve hours of instrument
operation.
8.5 To establish the ability to generate acceptable accuracy and
precision, the analyst must perform the following operations.
8.5.1 A quality control (QC) check sample concentrate is required
containing each analyte at a concentration of 10 mg/L in high purity
methanol. The QC check sample concentrate may be prepared from pure
standard materials or purchased as certified solutions. If the QC check
sample concentrate is prepared by the laboratory, the QC check sample
concentrate must be prepared using stock standards prepared independently
from the stock standards used for calibration.
8.5.2 Spike four pairs of cleaned, prepared VOST tubes with 10 /zL
of the QC check sample concentrate and analyze these spiked VOST tubes
according to the method beginning in Sec. 7.0.
8.5.3 Calculate the average recovery (X) in ng and the standard
deviation of the recovery (s) in ng for each analyte using the results of
the four analyses.
8.5.4 The average recovery and standard deviation must fall within
the expected range for determination of volatile organic compounds using
the VOST analytical methodology. The expected range for recovery of
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volatile organic compounds using this method is 50-150%. Standard
deviation will be compound dependent, but should, in general, range from
15 to 30 ng. More detailed method performance criteria must be generated
from historical records in the laboratory or from interlaboratory studies
coordinated by the Environmental Protection Agency. Since the additional
steps of sorbent tube spiking and desorption are superimposed upon the
methodology of Method 8260, direct transposition of Method 8260 criteria
is questionable. If the recovery and standard deviation for all analytes
meet the acceptance criteria, the system performance is acceptable and the
analysis of field samples may begin. If any individual standard deviation
exceeds the precision limit or any individual recovery falls outside the
range for accuracy, then the system performance is unacceptable for that
analyte.
NOTE: The large number of analytes listed in Table 1 presents a
substantial probability that one or more will fail at least
one of the acceptance criteria when all analytes for this
method are determined.
8.5.5 When one or more of the analytes tested fails at least one of
the acceptance criteria, the analyst must proceed according to one of the
alternatives below.
8.5.5.1 Locate and correct the source of any problem with
the methodology and repeat the test for all the analytes beginning
with Sec. 8.5.2.
8.5.5.2 Beginning with Sec. 8.5.2, repeat the test only
for those analytes that have failed to meet acceptance criteria.
Repeated failure, however, will confirm a general problem either
with the measurement system or with the applicability of the
methodology to the particular analyte (especially if the analyte in
question is not listed in Table 1). If the problem is identified as
originating in the measurement system, locate and correct the source
of the problem and repeat the test for all compounds of interest
beginning with Sec. 8.5.2.
8.6 To determine acceptable accuracy and precision limits for surrogate
standards, the following procedure should be performed.
8.6.1 For each sample analyzed, calculate the percent recovery of
each surrogate compound in the sample.
8.6.2 Once a minimum of thirty samples has been analyzed, calculate
the average percent recovery (p) and the standard deviation of the percent
recovery (s) for each of the surrogate compounds.
8.6.3 Calculate the upper and lower control limits for method
performance for each surrogate standard. This calculation is performed as
follows:
Upper Control Limit (UCL) = p + 3s
Lower Control Limit (LCL) = p - 3s
5041 - 23 Revision 0
September 1994
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For reference, the comparable control limits for recovery of the
surrogate compounds from water and soil in Method 8240 are:
4-Bromofluorobenzene Water: 86-115% Soil: 74-121%
l,2-Dichloroethane-d4 Water: 76-114% Soil: 70-121%
Toluene-d8 Water: 88-110% Soil: 81-117%
The control limits for the VOST methodology would be expected to be
similar, but exact data are not presently available. Individual laboratory
control limits can be established by the analysis of replicate samples.
8.6.4 If surrogate recovery is not within the limits established by
the laboratory, the following procedures are required: (1) Verify that
there are no errors in calculations, preparation of surrogate spiking
solutions, and preparation of internal standard spiking solutions. Also,
verify that instrument performance criteria have been met. (2) Recalculate
the data and/or analyze a replicate sample, if replicates are available.
(3) If all instrument performance criteria are met and recovery of
surrogates from spiked blank VOST tubes (analysis of a method blank) is
acceptable, the problem is due to the matrix. Emissions samples may be
highly acidic and may be highly loaded with target and non target organic
compounds. Both of these conditions will affect the ability to recover
surrogate compounds which are spiked on the field samples. The surrogate
compound recovery is thus a valuable indicator of the interactions of
sampled compounds with the matrix. If surrogates spiked immediately
before analysis cannot be observed with acceptable recovery, the
implications for target organic analytes which have been sampled in the
field must be assessed very carefully. If chemical or other interactions
are occurring on the exposed tubes, the failure to observe an analyte may
not necessarily imply that the Destruction and Removal Efficiency for that
analyte is high.
8.7 It is recommended that the laboratory adopt additional quality
assurance practices for use with this method. The specific practices that are
most productive depend upon the needs of the laboratory and the nature of the
samples analyzed. Field duplicates may be analyzed to assess the precision of
the environmental measurements. When doubt exists over the identification of a
peak on the chromatogram, confirmatory techniques such as gas chromatography with
a dissimilar column or a different ionization mode using a mass spectrometer may
be used, if replicate samples showing the same compound are available. Whenever
possible, the laboratory should analyze standard reference materials and
participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 The method detection limit (MDL) is defined in Chapter One. The MDL
concentrations listed in Table 2 were obtained using cleaned blanked VOST tubes
and reagent water. Similar results have been achieved with field samples. The
MDL actually achieved in a given analysis will vary depending upon instrument
sensitivity and the effects of the matrix. Preliminary spiking studies indicate
that under these conditions, the method detection limit for spiked compounds in
extremely complex matrices may be larger by a factor of 500-1000.
5041 - 24 Revision 0
September 1994
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10.0 REFERENCES
1. Protocol for Collection and Analysis of Volatile POHCs Using VOST.
EPA/600/8-84-007, March, 1984.
2. Validation of the Volatile Organic Sampling Train (VOST) Protocol.
Volumes I and II. EPA/600/4-86-014A, January, 1986.
3. U. S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for
Analysis of Pollutants Under the Clean Water Act, Method 624," October 26,
1984.
4. Bellar, T. A., and J. J. Lichtenberg, J. Amer. Water Works Assoc., 66(12),
739-744, 1974.
5. Bellar, T. A., and J. J. Lichtenberg, "Semi-Automated Headspace Analysis
of Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds," in Van Hall, ed., Measurement of Organic Pollutants in Water
and Wastewater, ASTM STP 686, pp 108-129, 1979.
5041 - 25 Revision 0
September 1994
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TABLE 1.
RETENTION TIMES AND CHARACTERISTIC IONS FOR VOLATILE COMPOUNDS
WHICH CAN BE ANALYZED BY METHOD 5041
Retention
Compound Time (min)
Acetone
Acrylonitrile
Benzene
Bromochl oromethane
Bromodichloromethane
4-Bromof 1 uorobenzene
Bromoform
Bromome thane
Carbon disulfide
Carbon tetrachloride
Chlorobenzene
Chi orodi bromomethane
Chloroethane
Chloroform
Chi oromethane
Di bromomethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans-l,2-Dichloroethene
1,2-Dichloropropane
cis-l,3-Dichloropropene
trans-l,3-Dichloropropene
1,4-Difl uorobenzene
Ethyl benzene
lodomethane
Methylene chloride
Styrene
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
1,1,1 -Tr i chl oroethane
1 , 1 , 2 -Tri chl oroethane
Trichloroethene
Trichlorofluoromethane
1 , 2 , 3-Tr i chl oropropane
Vinyl chloride
Xylenes*
7.1
8.6
13.3
12.0
16.0
23.4
22.5
4.1
7.1
12.6
20.5
19.3
4.2
12.2
3.0
15.4
10.0
13.3
6.4
8.6
15.2
17.0
18.2
14.2
21.1
7.0
8.1
22.3
24.0
18.6
17.4
12.4
18.4
14.5
5.1
24.0
3.2
22.2
Primary Ion
Mass
43
53
78
128
83
95
173
94
76
117
112
129
64
83
50
93
63
62
96
96
63
75
75
114
106
142
84
104
83
164
92
97
97
130
101
75
62
106
Secondary Ion(s)
Mass(es)
58
52, 51
52, 77
49, 130, 51
85, 129
174, 176
171, 175, 252
96, 79
78
119, 121
114, 77
208, 206
66, 49
85, 47
52, 49
174, 95
65, 83
64, 98
61, 98
61, 98
62, 41
77, 39
77, 39
63, 88
91
127, 141
49, 51, 86
78, 103
85, 131, 133
129, 131, 166
91, 65
99, 117
83, 85, 99
95, 97, 132
103, 66
110, 77, 61
64, 61
91
The retention time given is for m- and p-xylene, which coelute on the wide-bore
column. o-Xylene elutes approximately 50 seconds later.
5041 - 26
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September 1994
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TABLE 2.
PRELIMINARY METHOD DETECTION LIMITS AND BOILING POINTS
FOR VOLATILE ORGANICS ANALYZED BY METHOD 5041*
Compound
Chloromethane
Bromomethane
Vinyl chloride
Chl oroethane
Methyl ene chloride
Acetone
Carbon disulfide
1,1-Dichloroethene
1,1-Dichloroethane
trans-l,2-Dichloroethene
Chloroform
1,2-Dichloroethane
1,1,1 -Tri chl oroethane
Carbon tetrachloride
Bromodi chloromethane
1,1, 2, 2 -Tetrachl oroethane**
1 ,2-Dichloropropane
trans - 1 , 3 -Di chl oropropene
Trichloroethene
Di bromochl oromethane
1,1,2-Trichloroethane
Benzene
cis-1, 3 -Di chl oropropene
Bromoform**
Tetrachl oroethene
Toluene
Chlorobenzene^
Ethylbenzene"*
Styrene**
Tri chl orof 1 uoromethane
lodomethane
Acrylonitrile
Dibromomethane
1 ,2,3-Trichloropropane**
total Xylenes**
CAS Number
74-87-3
74-83-9
75-01-4
75-00-3
75-09-2
67-64-1
75-15-0
75-35-4
75-35-3
156-60-5
67-66-3
107-06-2
71-55-6
56-23-5
75-27-4
79-34-5
78-87-5
10061-02-6
79-01-6
124-48-1
79-00-5
71-43-2
10061-01-5
75-25-2
127-18-4
108-88-3
108-90-7
100-41-4
100-42-5
75-69-4
74-88-4
107-13-1
74-95-3
96-18-4
Detection
Limit, ng
58
26
14
21
9
35
11
14
12
11
11
13
8
8
11
23
12
17
11
21
26
26
27
26
11
15
15
21
46
17
9
13
14
37
22
Boiling
Point, °C
-24
4
-13
13
40
56
46
32
57
48
62
83
74
77
88
146
95
112
87
122
114
80
112
150
121
111
132
136
145
24
43
78
97
157
138-144
* The method detection limit (MDL) is defined as the minimum concentration of a
substance that can be measured and reported with 99% confidence that the analyte
concentration is greater than zero and is determined from analysis of a sample in
a given matrix containing the analyte. The detection limits cited above were
determined according to Title 40 CFR, Part 136, Appendix B, using standards spiked
onto clean VOST tubes. Since clean VOST tubes were used, the values cited above
represent the best that the methodology can achieve. The presence of an emissions
matrix will affect the ability of the methodology to perform at its optimum level.
** Not appropriate for quantitative sampling by Method 0030.
5041 - 27
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TABLE 3.
KEY ION ABUNDANCE CRITERIA FOR 4-BROMOFLUOROBENZENE
Mass Ion Abundance Criteria
50 15 to 40% of mass 95
75 30 to 60% of mass 95
95 base peak, 100% relative abundance
96 5 to 9% of mass 95
173 less than 2% of mass 174
174 greater than 50% of mass 95
175 5 to 9% of mass 174
176 greater than 95%, but less than 101% of mass 174
177 5 to 9% of mass 176
5041 - 28 Revision 0
September 1994
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TABLE 4.
VOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
Bromochloromethane
Acetone
Acrylonitrile
Bromomethane
Carbon disulfide
Chloroethane
Chloroform
Chloromethane
1,1-Dichloroethane
1,2-Dichloroethane
l,2-Dichloroethane-d4 (surrogate)
1,1-Dichloroethene
Trichloroethene
trans-l,2-Dichloroethene
lodomethane
Methylene chloride
Trichlorofluoromethane
Vinyl chloride
1,4-Di f1uorobenzene
Benzene
Bromodi chloromethane
Bromoform
Carbon tetrachloride
Chlorodi bromomethane
Dibromomethane
1,2-Dichloropropane
cis-l,3-Dichloropropene
trans-1,3-Di chloropropene
1,1,1-Trichloroethane
1,1,2-Trichloroethane
Chlorobenzene-d5
4-Bromofluorobenzene (surrogate)
Chlorobenzene
Ethyl benzene
Styrene
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
Toluene-dg (surrogate)
1,2,3-Trichloropropane
Xylenes
5041 - 29
Revision 0
September 1994
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Figure 4. Sample Purge Vessel
5041 - 33
Revision 0
September 1994
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METHOD 5041
PROTOCOL FOR ANALYSIS OF SORBENT CARTRIDGES FROM VOLATILE ORGANIC
SAMPLING TRAIN: WIDE-BORE CAPILLARY COLUMN TECHNIQUE
1 Start j
1
r
7.1 Conditions for
cartridge
desorption oven,
purge-and-trap
concentrator, GC,
and MS.
1
r
7.2 Daily, tune
the GC/MS with
BFB and check
calibration curve
(see Section 7.17).
>
r
7.3 - 7.6
Assemble the
system.
^
r
7.7.1 Calibrate the
instrument system
using the internal std.
procedure. Stds. and
calibration compounds
are apiked into cleaned
VOST tubes using the
flash evaporation
technique.
1
r
7.8 Prep the
purge-and-trap
unit with 5 ml
organic-free
reagent water.
1
7.9 C
paired
tubas
gas lir
desor
r
onnect
VOST
to the
tee for
ption.
7.10 Initiate
tube desorption/
purge and
heating.
J
r
7.11 Set the GC
oven to subambient
temperature
with liquid
nitrogen.
1
r
7.12 Prep the
GC/MS system
for data
aquisition.
J
r
7.13 After the tube/
water purge time,
attach the
analytical trap to
the GC/MS for
desorption.
^
r
7.14 Wash purging
vassal with two
5 mL flushes of
organic-free
reagent water.
^
r
7.15 Recondition the
analytical trap by
making it cut at
temps up to 220 C for
11 min. Trap replacemer
may be necessary
if the analytical trap
is saturated beyond
cleanup.
i
r
7.16.1 Prep
calibration stds.
as in 7.7.1. Add
water to vessel
and desorb.
7.16.2
Tabulate the
area response
of all compounds
of interest.
7.16.3
Calculate the
average RF for
each compound
of interest.
7.16.4 Calculate
the %RSD
for the CCCs.
The %RSD must
be <30%.
7.18 GC/MS
analysis of
samples.
7.19.1 Qualitative
analysis of data
and ident. guidelines
of compounds.
7.19.2 Quantitative
analysis of data for
the compounds of
interest.
( Stop J
5041 - 35
Revision 0
September 1994
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4.2 SAMPLE PREPARATION METHODS
4.2.2 CLEANUP
The following methods are included in this section:
Method 3600B: Cleanup
Method 3610A: Alumina Column Cleanup
Method 3611A: Alumina Column Cleanup and Separation of
Petroleum Wastes
Method 3620A: Florisil Column Cleanup
Method 3630B: Silica Gel Cleanup
Method 3640A: Gel-Permeation Cleanup
Method 3650A: Acid-Base Partition Cleanup
Method 3660A: Sulfur Cleanup
Method 3665: Sulfuric Acid/Permanganate Cleanup
FOUR - 9 Revision 2
September 1994
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METHOD 3600B
CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Method 3600 provides general guidance on selection of cleanup methods
that are appropriate for the target analytes of interest. Cleanup methods are
applied to the extracts prepared by one of the extraction methods, to eliminate
sample interferences. The following table lists the cleanup methods and provides
a brief description of the type of cleanup.
SW-846 CLEANUP METHODS
Method # Method Name Cleanup Type
3610 Alumina Cleanup Adsorption
3611 Alumina Cleanup & Separation Adsorption
for Petroleum Waste
3620 Florisil Cleanup Adsorption
3630 Silica Gel Cleanup Adsorption
3640 Gel-Permeation Cleanup Size-Separation
3650 Acid-Base Partition Cleanup Acid-Base Partitioning
3660 Sulfur Cleanup Oxidation/Reduction
3665 Sulfuric Acid/Permanganate Oxidation/Reduction
Cleanup
1.2 The purpose of applying a cleanup method to an extract is to remove
interferences and high boiling material that may result in: (1) errors in
quantitation (data may be biased low because of analyte adsorption in the
injection port or front of the GC column or biased high because of overlap with
an interference peak); (2) false positives because of interference peaks falling
within the analyte retention time window; (3) false negatives caused by shifting
the analyte outside the retention time window; (4) rapid deterioration of
expensive capillary columns; and, (5) instrument downtime caused by cleaning and
rebuilding of detectors and ion sources. Most extracts of soil and waste require
some degree of cleanup, whereas, cleanup for water extracts may be unnecessary.
Highly contaminated extracts (e.g. sample extracts of oily waste or soil
containing oily residue) often require a combination of cleanup methods. For
example, when analyzing for organochlorine pesticides and PCBs, it may be
necessary to use gel permeation chromatography (GPC), to eliminate the high
boiling material and a micro alumina or Florisil column to eliminate
interferences with the analyte peaks on the GC/ECD.
3600B - 1 Revision 2
September 1994
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1.3 The following techniques have been applied to extract purification:
adsorption chromatography; partitioning between immiscible solvents; gel
permeation chromatography; oxidation of interfering substances with acid, alkali,
or oxidizing agents. These techniques may be used individually or in various
combinations, depending on the extent and nature of the co-extractives.
1.3.1 Adsorption column chromatography - Alumina (Methods 3610
and 3611), Florisil (Method 3620), and silica gel (Method 3630) are useful
for separating analytes of a relatively narrow polarity range away from
extraneous, interfering peaks of a different polarity. These are
primarily used for cleanup of a specific chemical group of relatively
non-polar analytes, i.e., organochlorine pesticides, polynuclear aromatic
hydrocarbons (PAHs), nitrosamines, etc.. Solid phase extraction
cartridges have been added as an option.
1.3.2 Acid-base partitioning (Method 3650) - Useful for
separating acidic or basic organics from neutral organics. It has been
applied to analytes such as the chlorophenoxy herbicides and phenols. It
is very useful for separating the neutral PAHs from the acidic phenols
when analyzing a site contaminated with creosote and pentachlorophenol.
1.3.3 Gel permeation chromatography (GPC) (Method 3640) - The
most universal cleanup technique for a broad range of semivolatile
organics and pesticides. It is capable of separating high
molecular-weight, high boiling material from the sample analytes. It has
been used successfully for all the semivolatile base, neutral, and acid
compounds associated with the EPA Priority Pollutant and the Superfund
Target Compound list prior to GC/MS analysis for semivolatiles and
pesticides. GPC may not be applicable to elimination of extraneous peaks
on a chromatogram which interfere with the analytes of interest. It is,
however, useful for the removal of high boiling materials which would
contaminate injection ports and column heads, prolonging column life,
stabilizing the instrument, and reducing column reactivity.
1.3.4 Sulfur cleanup (Method 3660) - Useful in eliminating
sulfur from sample extracts, which may cause chromatographic interference
with analytes of interest.
1.4 Several of the methods are also useful for fractionation of complex
mixtures of analytes. Use the solid phase extraction cartridges in Method 3630
(Silica Gel) for separating the PCBs away from most organochlorine pesticides.
Method 3611 (Alumina) is for the fractionation of aliphatic, aromatic and polar
analytes. Method 3620 (Florisil) provides fractionation of the organochlorine
pesticides.
1.5 Cleanup capacity is another factor that must be considered in
choosing a cleanup technique. The adsorption methods (3610, 3620, and 3630)
provide the option of using standard column chromatography techniques or solid
phase extraction cartridges. The decision process in selecting between the
different options available generally depends on the amount of interferences/high
boiling material in the sample extract and the degree of cleanup required by the
determinative method. The solid phase extraction cartridges require less elution
solvent and less time, however, their cleanup capacity is drastically reduced
when comparing a 0.5 g or 1.0 g Florisil cartridge to a 20 g standard Florisil
3600B - 2 Revision 2
September 1994
-------
column. The same factor enters into the choice of the 70 g gel permeation column
specified in Method 3640 versus a high efficiency column.
1.6 Table 1 indicates the recommended cleanup techniques for the
indicated groups of compounds. This information can also be used as guidance for
compounds that are not listed. Compounds that are chemically similar to these
groups of compounds should behave similarly when taken through the cleanup
procedure,however, this must be demonstrated by determining recovery of standards
taken through the method.
2.0 SUMMARY OF METHOD
2.1 Refer to the specific cleanup method for a summary of the procedure.
3.0 INTERFERENCES
3.1 Analytical interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware. All of these
materials must be routinely demonstrated to be free of interferences, under the
conditions of the analysis, by running laboratory reagent blanks.
3.2 More extensive procedures than those outlined in the methods may be
necessary for reagent purification.
4.0 APPARATUS AND MATERIALS
4.1 Refer to the specific cleanup method for apparatus and materials
needed.
5.0 REAGENTS
5.1 Refer to the specific cleanup method for the reagents needed.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Prior to using the cleanup procedures, samples normally undergo
solvent extraction. Chapter Two, Section 2.0, may be used as a guide for
choosing the appropriate extraction procedure based on the physical composition
of the waste and on the analytes of interest in the matrix (see also Method 3500
for a general description of the extraction technique). For some organic
liquids, extraction prior to cleanup may not be necessary.
3600B - 3 Revision 2
September 1994
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7.2 Most soil/sediment and waste sample extracts will require some degree
of cleanup. The extract is then analyzed by one of the determinative methods.
If interferences still preclude analysis for the analytes of interest, additional
cleanup may be required.
7.3 Many of the determinative methods specify cleanup methods that should
be used when determining particular analytes (e.g. Method 8061, gas
chromatography of phthalate esters, recommends using either Method 3610 (Alumina
column cleanup) or Method 3620 (Florisil column cleanup) if interferences prevent
analysis. However, the experience of the analyst may prove invaluable in
determining which cleanup methods are needed. As indicated in Section 1.0 of
this method, many matrices may require a combination of cleanup procedures in
order to ensure proper analytical determinations.
7.4 Guidance for cleanup is specified in each of the methods that follow.
The amount of extract cleanup required prior to the final determination depends
on the concentration of interferences in the sample, the selectivity of both the
extraction procedure and the determinative method and the required detection
limit.
7.5 Following cleanup, the sample is concentrated to whatever volume is
required in the determinative method. Analysis follows as specified in the
determinative procedure.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 The analyst must demonstrate that the compounds of interest are
being quantitatively recovered by the cleanup technique before the cleanup is
applied to actual samples. For sample extracts that are cleaned up, the
associated quality control samples (e.g. spikes, blanks, replicates, and
duplicates) must also be processed through the same cleanup procedure.
8.3 The analysis using each determinative method (GC, GC/MS, HPLC)
specifies instrument calibration procedures using stock standards. It is
recommended that cleanup also be performed on a series of the same type of
standards to validate chromatographic elution patterns for the compounds of
interest and to verify the absence of interferences from reagents.
9.0 METHOD PERFORMANCE
9.1 Refer to the specific cleanup method for performance data.
10.0 REFERENCES
10.1 Refer to the specific cleanup method.
3600B - 4 Revision 2
September 1994
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TABLE 1.
RECOMMENDED CLEANUP TECHNIQUES FOR INDICATED GROUPS OF COMPOUNDS
Analyte Group
Determinative8
Method
Cleanup
Method Options
Phenols
Phthalate esters
Nitrosamines
Organochlorine pesticides & PCBs
PCBs
Nitroaromatics and cyclic ketones
Polynuclear aromatic hydrocarbons
Chlorinated hydrocarbons
Organophosphorus pesticides
Chlorinated herbicides
Semivolatile organics
Petroleum waste
PCDDs and PCDFs by LR/MS
PCDDs and PCDFs by HR/MS
N-methyl carbamate pesticides
8040
8060/8061
8070
8080/8081
8080/8081
8090
8100/8310
8120/8121
8140/8141
8150/8151
8250/8270
8250/8270
8280
8290
8318
3630b, 3640, 3650, 8040°
3610, 3620, 3640
3610, 3620, 3640
3620, 3640, 3660
3665
3620, 3640
3611, 3630, 3640
3620, 3640
3620
8150d, 8151d, 3620
3640, 3650, 3660
3611, 3650
8280
8290
8318
The GC/MS Methods, 8250 and 8270, are also appropriate determinative methods
for all analyte groups, unless lower detection limits are required.
b Cleanup applicable to derivatized phenols.
0 Method 8040 includes a derivatization technique followed by GC/ECD analysis,
if interferences are encountered using GC/FID.
d Methods 8150 and 8151 incorporate an acid-base cleanup step as an integral
part of the methods.
3600B - 5
Revision 2
September 1994
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METHOD 3600B
CLEANUP
Start
\
7.1
Do solvent
extraction
1
7.2
Analyze analyte
by a determinative
method from Sec. 4.3
7.2 Are
analytes
undeterminable
due to
nterferenoe?
7.3
Use cleanup method
specified for the
determinative method
7.5
Concentrate sample
to required volume
3600B - 6
Revision 2
September 1994
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METHOD 3610A
ALUMINA COLUMN CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Scope: Alumina is a highly porous and granular form of aluminum
oxide. It is available in three pH ranges (basic, neutral, and acidic) for use
in column chromatography. It is used to separate analytes from interfering
compounds of a different chemical polarity.
1.2 General Applications (Gordon and Ford):
1.2.1 Basic (B) pH (9-10): USES: Basic and neutral compounds stable
to alkali, alcohols, hydrocarbons, steroids, alkaloids, natural pigments.
DISADVANTAGES: Can cause polymerization, condensation, and dehydration
reactions; cannot use acetone or ethyl acetate as eluants.
1.2.2 Neutral (N): USES: Aldehydes, ketones, quinones, esters,
lactones, glycoside. DISADVANTAGES: Considerably less active than the
basic form.
1.2.3 Acidic (A) pH (4-5): USES: Acidic pigments (natural and
synthetic), strong acids (that otherwise chemisorb to neutral and basic
alumina).
1.2.4 Activity grades: Acidic, basic, or neutral alumina can be
prepared in various activity grades (I to V), according to the Brockmann
scale, by addition of water to Grade 1 (prepared by heating at 400-450°C
until no more water is lost). The Brockmann scale (Gordon and Ford, p.
374) is reproduced below:
Water added (wt. %): 03 6 10 15
Activity grade: I II III IV V
RF (p-aminoazobenzene): 0.0 0.13 0.25 0.45 0.55
1.3 Specific applications: This method includes guidance for cleanup of
sample extracts containing phthalate esters and nitrosamines. For alumina column
cleanup of petroleum wastes, see Method 3611, Alumina Column Cleanup of Petroleum
Wastes.
2.0 SUMMARY OF METHOD
2.1 The column is packed with the required amount of adsorbent, topped
with a water adsorbent, and then loaded with the sample to be analyzed. Elution
of the analytes is effected with a suitable solvent(s), leaving the interfering
compounds on the column. The eluate is then concentrated (if necessary).
3610A - 1 Revision 1
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3.0 INTERFERENCES
3.1 A reagent blank should be performed for the compounds of interest
prior to the use of this method. The level of interferences must be below the
method detection limit before this method is performed on actual samples.
3.2 More extensive procedures than those outlined in this method may be
necessary for reagent purification.
4.0 APPARATUS AND MATERIALS
4.1 Chromatography column: 300 mm x 10 mm ID, with Pyrex glass wool at
bottom and a Teflon stopcock.
NOTE: Fritted glass discs are difficult to decontaminate after
highly contaminated extracts have been passed through.
Columns without frits may be purchased. Use a small pad of
Pyrex glass wool to retain the adsorbent. Prewash the glass
wool pad with 50 mL of acetone followed by 50 mL of elution
solvent prior to packing the column with adsorbent.
4.2 Beakers: 500 mL.
4.3 Reagent bottle: 500 mL.
4.4 Muffle furnace.
4.5 Kuderna-Danish (K-D) apparatus:
4.5.1 Concentrator tube: 10 mL, graduated (Kontes K-570050-1025 or
equivalent). A ground-glass stopper is used to prevent evaporation of
extracts.
4.5.2 Evaporation flask: 500 mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.5.3 Snyder column: Three ball macro (Kontes K-503000-0121 or
equivalent).
4.5.4 Snyder column: Two ball micro (Kontes K-569001-0219 or
equivalent).
4.5.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.6 Boiling chips: Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.7 Water bath: Heated, with concentric ring cover, capable of
temperature control (±5°C). The bath should be used in a hood.
4.8 Vials: Glass, 2 mL capacity, with Teflon lined screw caps or crimp
tops.
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4.9 Erlenmeyer flasks: 50 and 250 mL
5.0 REAGENTS
5.1 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2 Sodium sulfate: Sodium sulfate (granular, anhydrous), Na2S04.
Purify by heating at 400°C for 4 hours in a shallow tray, or by precleaning the
sodium sulfate with methylene chloride. If the sodium sulfate is precleaned with
methylene chloride, a method blank must be analyzed, demonstrating that there is
no interference from the sodium sulfate.
5.3 Eluting solvents:
5.3.1 Diethyl Ether, C2H5OC2H5. Pesticide quality or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl alcohol preservative must be
added to each liter of ether.
5.3.2 Methanol, CH3OH - Pesticide quality or equivalent.
5.3.3 Pentane, CH3(CH2)3CH3 - Pesticide quality or equivalent.
5.3.4 Hexane, C6H14 - Pesticide quality or equivalent.
5.3.5 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.4 Alumina;
5.4.1 For cleanup of phthalate extracts: Alumina-Neutral, activity
Super I, W200 series (ICN Life Sciences Group, No. 404583, or equivalent).
To prepare for use, place 100 g of alumina into a 500 ml beaker and heat
for approximately 16 hr at 400°C. After heating, transfer to a 500 ml
reagent bottle. Tightly seal and cool to room temperature. When cool,
add 3 ml of organic-free reagent water. Mix thoroughly by shaking or
rolling for 10 min and let it stand for at least 2 hr. Keep the bottle
sealed tightly.
5.4.2 For cleanup of nitrosamine extracts: Alumina-Basic, activity
Super I, W200 series (ICN Life Sciences Group, No. 404571, or equivalent).
To prepare for use, place 100 g of alumina into a 500 mL reagent bottle
and add 2 mL of organic-free reagent water. Mix the alumina preparation
thoroughly by shaking or rolling for 10 min and let it stand for at least
2 hr. The preparation should be homogeneous before use. Keep the bottle
sealed tightly to ensure proper activity.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
3610A - 3 Revision 1
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7.0 PROCEDURE
7.1 Phthalate esters:
7.1.1 Reduce the sample extract volume to 2 ml prior to cleanup.
The extract solvent must be hexane.
7.1.2 Place approximately 10 g of alumina into a 10 mm ID
chromatographic column. Tap the column to settle the alumina and add 1-2
cm of anhydrous sodium sulfate to the top.
7.1.3 Pre-elute the column with 40 ml of hexane. The rate for all
elutions should be about 2 mL/min. Discard the eluate and, just prior to
exposure of the sodium sulfate layer to the air, quantitatively transfer
the 2 ml sample extract onto the column using an additional 2 ml of hexane
to complete the transfer. Just prior to exposure of the sodium sulfate
layer to the air, add 35 ml of hexane and continue the elution of the
column. Discard this hexane eluate.
7.1.4 Next, elute the column with 140 ml of 20% ethyl ether in
hexane (v/v) into a 500 mL K-D flask equipped with a 10 mL concentrator
tube. Concentrate the collected fraction using the Kuderna-Danish
technique. No solvent exchange is necessary. Adjust the volume of the
cleaned up extract to whatever volume is required (10.0 ml for Method
8060) and analyze. Compounds that elute in this fraction are as follows:
Bis(2-ethylhexyl) phthalate
Butyl benzyl phthalate
Di-n-butyl phthalate
Diethyl phthalate
Dimethyl phthalate
Di-n-octyl phthalate.
7.2 Nitrosamines:
7.2.1 Reduce the sample extract to 2 ml prior to cleanup.
7.2.2 Diphenylamine, if present in the original sample extract, must
be separated from the nitrosamines if N-nitrosodiphenylamine is to be
determined by this method.
7.2.3 Place approximately 12 g of the alumina preparation into a 10
mm ID chromatographic column. Tap the column to settle the alumina and
add 1-2 cm of anhydrous sodium sulfate to the top.
7.2.4 Pre-elute the column with 10 ml of ethyl ether/pentane
(3:7)(v/v). Discard the eluate (about 2 ml) and, just prior to exposure
of the sodium sulfate layer to the air, quantitatively transfer the 2 ml
sample extract onto the column using an additional 2 ml of pentane to
complete the transfer.
7.2.5 Just prior to exposure of the sodium sulfate layer to the air,
add 70 ml of ethyl ether/pentane (3:7)(v/v). Discard the first 10 ml of
eluate. Collect the remainder of the eluate in a 500 mL K-D flask
3610A - 4 Revision 1
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equipped with a 10 ml concentrator tube. This fraction contains N-
nitroso-di-n-propylamine.
7.2.6 Next, elute the column with 60 ml of ethyl ether/pentane
(l:l)(v/v), collecting the eluate in a second 500 mL K-D flask equipped
with a 10 ml concentrator tube. Add 15 mL of methanol to the K-D flask.
This fraction will contain N-nitrosodimethylamine, most of the N-nitroso-
di-n-propylamine, and any diphenylamine that is present.
7.2.7 Concentrate both fractions using the Kuderna-Danish Technique
(if necessary), using pentane to prewet the Snyder column. When the
apparatus is cool, remove the Snyder column and rinse the flask and its
lower joint into the concentrator tube with 1-2 ml of pentane. Adjust the
final volume to whatever is required in the appropriate determinative
method (Section 4.3 of this chapter). Analyze the fractions.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3600 for cleanup procedures.
8.2 The analyst should demonstrate that the compounds of interest are
being quantitatively recovered before applying this method to actual samples.
8.3 For sample extracts that are cleaned up using this method, the
associated quality control samples must also be processed through this cleanup
method.
9.0 METHOD PERFORMANCE
9.1 Performance data are not available.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
3610A - 5 Revision 1
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METHOD 3610A
ALUMINA COLUMN CLEANUP
711 Reduce
volume of
sample
extract
7.1 2 Put
alumina in
column, add
anhydrous
sodium sulfate
7 1 3
Preclute
column with
hexane
713 Transfer
sample ex tract
to column,
elute column
with hexane.
714 Elute column
with ethyl
ether/hexane
Collect eluate in
flask
7 2 1 Reduce
volume of
sample
extract
7 2 3 Put
alumina in
column, add
anhydrous
sodium sulfate
724 Preelute
column with ethyl
ether/pentane
Transfer sample
extract to column,
add pentane
725 Elute column
with ethyl
ether/pentane
Collect eluate in
flask
7 2 6 Elute column
with ethyl
ether/pentane
Collect eluate in
second flask, add
methanol
714
Concentrate
collected
fraction,
adjust volume
Analyze by
appropriate
determinative
method
7 2 ?
Concentrate
both fractions,
ad jus t volume
3610A - 6
Revision 1
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METHOD 3611A
ALUMINA COLUMN CLEANUP AND SEPARATION OF PETROLEUM WASTES
1.0 SCOPE AND APPLICATION
1.1 Method 3611 was formerly Method 3570 in the Second Edition of this
manual.
1.2 Specific application: This method includes guidance for separation
of petroleum wastes into aliphatic, aromatic, and polar fractions.
2.0 SUMMARY OF METHOD
2.1 The column is packed with the required amount of adsorbent, topped
with a water adsorbent, and then loaded with the sample to be analyzed. Elution
of the analytes is effected with a suitable solvent(s), leaving the interfering
compounds on the column. The eluate is then concentrated (if necessary).
3.0 INTERFERENCES
3.1 A reagent blank should be performed for the compounds of interest
prior to the use of this method. The level of interferences must be below the
method detection limit before this method is performed on actual samples.
3.2 More extensive procedures than those outlined in this method may be
necessary for reagent purification.
3.3 Caution must be taken to prevent overloading of the chromatographic
column. As the column loading for any of these types of wastes approaches
0.300 g of extractable organics, separation recoveries will suffer. If
overloading is suspected, an aliquot of the base-neutral extract prior to cleanup
may be weighed and then evaporated to dryness. A gravimetric determination on
the aliquot will indicate the weight of extractable organics in the sample.
3.4 Mixtures of petroleum wastes containing predominantly polar solvents,
i.e., chlorinated solvents or oxygenated solvents, are not appropriate for this
method.
4.0 APPARATUS AND MATERIALS
4.1 Chromatography column: 300 mm x 10 mm ID, with Pyrex glass wool at
bottom and a Teflon stopcock.
NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without
frits may be purchased. Use a small pad of Pyrex glass wool to
retain the adsorbent. Prewash the glass wool pad with 50 mL of
acetone followed by 50 mL of elution solvent prior to packing the
column with adsorbent.
3611A - 1 Revision 1
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4.2 Beakers: 500 ml.
4.3 Reagent bottle: 500 ml.
4.4 Muffle furnace.
4.5 Kuderna-Danish (K-D) apparatus:
4.5.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025 or
equivalent). A ground-glass stopper is used to prevent evaporation of
extracts.
4.5.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.5.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.5.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.5.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.6 Boiling chips: Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.7 Water bath: Heated with concentric ring cover, capable of
temperature control (±5°C). The bath should be used in a hood.
4.8 Erlenmeyer flasks: 50 and 250 mL.
5.0 REAGENTS
5.1 Sodium sulfate: (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.2 Eluting solvents:
5.2.1 Methanol, CH3OH - Pesticide quality or equivalent.
5.2.2 Hexane, C6HU - Pesticide quality or equivalent.
5.2.3 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.3 Alumina: Neutral 80-325 MCB chromatographic grade or equivalent.
Dry alumina overnight at 130°C prior to use.
3611A - 2 Revision 1
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 It is suggested that Method 3650, Acid-Base Partition Cleanup, be
performed on the sample extract prior to alumina cleanup.
7.2 Place approximately 10 g of alumina into a chromatographic column,
tap to settle the alumina, and add 1 cm of anhydrous sodium sulfate to the top.
7.3 Pre-elute the column with 50 mL of hexane. Discard the eluate and,
just prior to exposure of the sodium sulfate layer to the air, quantitatively
transfer the 1 mL sample extract onto the column using an additional 1 mL of
hexane to complete the transfer. To avoid overloading the column, it is
suggested that no more than 0.300 g of extractable organics be placed on the
column (see Section 3.3).
7.4 Just prior to exposure of the sodium sulfate to the air, elute the
column with a total of 15 mL of hexane. If the extract is in 1 mL of hexane, and
if 1 mL of hexane was used as a rinse, then 13 mL of additional hexane should be
used. Collect the effluent in a 50 mL flask and label this fraction
"base/neutral aliphatics." Adjust the flow rate to 2 mL/min.
7.5 Elute the column with 100 mL of methylene chloride and collect the
effluent in a 250 mL flask. Label this fraction "base/neutral aromatics."
7.6 Elute the column with 100 mL of methanol and collect the effluent in
a 250 mL flask. Label this fraction "base/neutral polars."
7.7 Concentrate the extracts (if necessary) by the standard K-D technique
to the volume (1-10 mL) required in the appropriate determinative method (Chapter
Four). Analyze the fractions containing the analytes of interest.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3600 for cleanup procedures.
8.2 The analyst should demonstrate that the compounds of interest are
being quantitatively recovered before applying this method to actual samples.
8.3 For sample extracts that are cleaned up using this method, the
associated quality control samples must also be processed through this cleanup
method.
3611A - 3 Revision 1
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9.0 METHOD PERFORMANCE
9.1 The precision and accuracy of the method will depend upon the overall
performance of the sample preparation and analysis.
9.2 Rag oil is an emulsion consisting of crude oil, water, and soil
particles. It has a density greater than crude oil and less than water. This
material forms a layer between the crude oil and water when the crude oil is
allowed to gravity separate at the refinery. A rag oil sample was analyzed by
a number of laboratories according to the procedure outlined in this method. The
results of these analyses by GC/MS for selected components in the rag oil are
presented in Table 1. Reconstructed ion chromatograms from the GC/MS analyses
are included as Figures 1 and 2.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
3611A - 4 Revision 1
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Table 1.
RESULTS OF ANALYSIS FOR SELECTED COMPONENTS IN RAG OIL
Mean Standard
Analyte Cone. (mg/kg)a Deviation %RSD
Naphthalene 216 42 19
Fluorene 140 66 47
Phenanthrene 614 296 18
2-Methylnaphthalene 673 120 18
Dibenzothiophene 1084 286 26
Methylphenanthrene 2908 2014 69
Methyldibenzothiophene 2200 1017 46
Average Surrogate Recovery
Nitrobenzene-d5 58.6 11
Terphenyl-du 83.0 2.6
Phenol-d6 80.5 27.6
Naphthalene-d8 64.5 5.0
a Based on five determinations from three laboratories.
b Percent Relative Standard Deviation.
3611A - 5 Revision 1
July 1992
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Figure 1
3611A - 6
Revision 1
July 1992
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Figure 2
ts
XI
II
u
fO
V
3611A - 7
Revision 1
July 1992
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METHOD 3611A
ALUMINA COLUMN CLEANUP AND SEPARATION OF PETROLEUM WASTES
Start
7 1 Cleanup
using Method
3650
7 2 Add alumina
to
chroma tographic
column
7 2 Add
anhydrous
sodium sulfate
to top of
column
7 3 Preelute
column with
hexane
7 3
Quantitatively
add extract to
column
7 4 Elute
"base-neutral
al iphatics "
fraction wi th
hexi
me
7 5 Elute
"base-neutral
ar omatics "
fraction with
CH2C12
7 6 Elute
"base-neutral
polars"
fraction wi th
met Hanoi
7
7
Concent ra te
ex tracts
s^ "^
/ Analyze using
/ appropriate
method
3611A - 8
Revision 1
July 1992
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METHOD 3620A
FLORISIL COLUMN CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Florisil, a registered trade name of the Floridin Co., is a magnesium
silicate with acidic properties. It is used for general column chromatography
as a cleanup procedure prior to sample analysis by gas chromatography.
1.2 General applications: Cleanup of pesticide residues and other
chlorinated hydrocarbons; the separation of nitrogen compounds from hydrocarbons;
the separation of aromatic compounds from aliphatic-aromatic mixtures; and
similar applications for use with fats, oils, and waxes (Floridin).
Additionally, Florisil is considered good for separations with steroids, esters,
ketones, glycerides, alkaloids, and some carbohydrates (Gordon and Ford).
1.3 Specific applications: This method includes guidance for cleanup of
sample extracts containing the following analyte groups: phthalate esters;
nitrosamines; organochlorine pesticides; nitroaromatics; haloethers; chlorinated
hydrocarbons; and organophosphorus pesticides.
2.0 SUMMARY OF METHOD
2.1 The column is packed with the required adsorbent, topped with a water
adsorbent, and then loaded with the sample to be analyzed. Elution is effected
with a suitable solvent(s) leaving the interfering compounds on the column. The
eluate is then concentrated (if necessary).
3.0 INTERFERENCES
3.1 A reagent blank should be performed for the compounds of interest
prior to the use of this method. The level of interferences must be below the
method detection limit before this method is performed on actual samples.
3.2 More extensive procedures than those outlined in this method may be
necessary for reagent purification.
4.0 APPARATUS AND MATERIALS
4.1 Beaker - 500 mL.
4.2 Chromatographic column - 300 mm long x 10 mm ID or 400 mm long x
20 mm ID, as specified in Section 7.0; with Pyrex glass wool at bottom and a
Teflon stopcock.
NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without
frits may be purchased. Use a small pad of Pyrex glass wool to
retain the adsorbent. Prewash the glass wool pad with 50 mL of
3620A - 1 Revision 1
July 1992
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acetone followed by 50 ml of elution solvent prior to packing the
column with adsorbent.
4.3 Kuderna-Danish (K-D) apparatus.
4.3.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025 or
equivalent). Ground-glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 ml (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Muffle furnace.
4.5 Reagent bottle - 500 ml.
4.6 Water bath - Heated, with concentric ring cover, capable of
temperature control (±5°C). The bath should be used in a hood.
4.7 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.8 Erlenmeyer flasks - 50 and 250 ml.
4.9 Top-loading balance - 0.01 g.
5.0 REAGENTS
5.1 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2 Florisil - Pesticide residue (PR) grade (60/100 mesh); purchase
activated at 1250°F (677°C), stored in glass containers with ground-glass
stoppers or foil-lined screw caps.
5.2.1 Deactivation of Florisil - for cleanup of phthalate esters.
To prepare for use, place 100 g of Florisil into a 500 ml beaker and heat
for approximately 16 hr at 40°C. After heating, transfer to a 500 ml
reagent bottle. Tightly seal and cool to room temperature. When cool add
3 ml of organic-free reagent water. Mix thoroughly by shaking or rolling
for 10 min and let stand for at least 2 hr. Keep the bottle sealed
tightly.
5.2.2 Activation of Florisil - for cleanup of nitrosamines,
3620A - 2 Revision 1
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organochlorine pesticides and PCBs, nitroaromatics, haloethers,
chlorinated hydrocarbons, and organophosphorus pesticides. Just before
use, activate each batch at least 16 nr at 130°C in a glass container
loosely covered with aluminum foil. Alternatively, store the Florisil in
an oven at 130°C. Cool the Florisil before use in a desiccator. (Florisil
from different batches or sources may vary in adsorptive capacity. To
standardize the amount of Florisil which is used, the use of lauric acid
value is suggested. The referenced procedure determines the adsorption
from hexane solution of lauric acid (mg) per g of Florisil. The amount of
Florisil to be used for each column is calculated by dividing 110 by this
ratio and multiplying by 20 g (Mills).
5.3 Sodium sulfate (granular, anhydrous), Na2S04 - Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.4 Eluting solvents
5.4.1 Diethyl ether, C2H5OC^H5 - Pesticide quality or equivalent.
Must be free of peroxides, as indicated by test strips (EM Quant or
equivalent). Procedures recommended for removal of peroxides are provided
with the test strips. After cleanup, 20 ml ethyl alcohol preservative
must be added to each liter of ether.
5.4.2 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.4.3 Hexane, C6H14 - Pesticide quality or equivalent.
5.4.4 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.4.5 Pentane, CH3(CH2)3CH3 - Pesticide quality or equivalent.
5.4.6 Petroleum ether (boiling range 30-60°C) - Pesticide quality or
equivalent.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Phthalate esters
7.1.1 Reduce the sample extract volume to 2 mL prior to cleanup.
The extract solvent must be hexane.
7.1.2 Place approximately 10 g of deactivated Florisil (Section
5.1.1) into a 10 mm ID chromatographic column. Tap the column to settle
the Florisil and add approximately 1 cm of anhydrous sodium sulfate to the
3620A - 3 Revision 1
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top.
7.1.3 Preelute the column with 40 ml of hexane. The rate for all
elutions should be about 2 mL/min. Discard the eluate and, just prior to
exposure of the sodium sulfate layer to the air, quantitatively transfer
the 2 ml sample extract onto the column using an additional 2 ml of hexane
to complete the transfer. Just prior to exposure of the sodium sulfate
layer to the air, add 40 ml of hexane and continue the elution of the
column. Discard this hexane eluate.
7.1.4 Next, elute the column with 100 ml of 20% ethyl ether in
hexane (v/v) into a 500 ml K-D flask equipped with a 10 ml concentrator
tube. Concentrate the collected fraction as needed. No solvent exchange
is necessary. Adjust the volume of the cleaned-up extract to whatever
volume is required (10 ml for Method 8060) and analyze by gas
chromatography. Compounds that elute in this fraction are:
Bis(2-ethylhexyl) phthalate
Butyl benzyl phthalate
Di-n-butyl phthalate
Diethyl phthalate
Dimethyl phthalate
Di-n-octyl phthalate
7.2 Nitrosamines
7.2.1 Reduce the sample extract volume to 2 ml prior to cleanup.
7.2.2 Add a weight of activated Florisil (nominally 22 g)
predetermined by calibration (Section 5.1.2) into a 20 mm ID
chromatographic column. Tap the column to settle the Florisil and add
about 5 mm of anhydrous sodium sulfate to the top.
7.2.3 Pre-elute the column with 40 ml of ethyl ether/pentane (15:85)
(v/v). Discard the eluate and, just prior to exposure of the sodium
sulfate layer to the air, quantitatively transfer the 2 ml sample extract
onto the column using an additional 2 ml of pentane to complete the
transfer.
7.2.4 Elute the column with 90 ml of ethyl ether/pentane (15:85)
(v/v) and discard the eluate. This fraction will contain the
diphenylamine, if it is present in the extract.
7.2.5 Next, elute the column with 100 ml of acetone/ethyl ether
(5:95) (v/v) into a 500 ml K-D flask equipped with a 10 ml concentrator
tube. This fraction will contain all of the nitrosamines listed in the
scope of the method.
7.2.6 Add 15 ml of methanol to the collected fraction, concentrate
as needed using pentane to prewet the K-D column and set the water bath at
70 to 75°C. When the apparatus is cool, remove the Snyder column and rinse
the flask and its lower joint into the concentrator tube with 1 to 2 ml of
pentane.
3620A - 4 Revision 1
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7.3 Organochlorine pesticides, haloethers, and organophosphorus
pesticides (see Tables 1 and 2 for fractionation patterns of compounds tested)
7.3.1 Reduce the sample extract volume to 10 ml prior to cleanup.
The extract solvent must be hexane.
7.3.2 Add a weight of activated Florisil (nominally 20 g),
predetermined by calibration (Section 5.1.2), to a 20 mm ID
chromatographic column. Settle the Florisil by tapping the column. Add
anhydrous sodium sulfate to the top of the Florisil to form a layer 1 to
2 cm deep. Add 60 ml of hexane to wet and rinse the sodium sulfate and
Florisil. Just prior to exposure of the sodium sulfate to air, stop the
elution of the hexane by closing the stopcock on the chromatographic
column. Discard the eluate.
7.3.3 Adjust the sample extract volume to 10 ml with hexane and
transfer it from the K-D concentrator tube to the Florisil column. Rinse
the tube twice with 1-2 ml hexane, adding each rinse to the column.
7.3.4 Place a 500 ml K-D flask and clean concentrator tube under the
chromatographic column. Drain the column into the flask until the sodium
sulfate layer is nearly exposed. Elute the column with 200 ml of 6% ethyl
ether in hexane (v/v) (Fraction 1) using a drip rate of about 5 mL/min.
All of the haloethers are in this fraction. Remove the K-D flask and set
aside for later concentration. Elute the column again, using 200 ml of
15% ethyl ether in hexane (v/v) (Fraction 2), into a second K-D flask.
Perform a third elution using 200 ml of 50% ethyl ether in hexane (v/v)
(Fraction 3), and a final elution with 200 ml of 100% ethyl ether
(Fraction 4), into separate K-D flasks.
7.3.5 If necessary, concentrate the eluates by standard K-D
techniques using the water bath at about 85°C (75°C for Fraction 4).
Adjust the final volume to whatever volume is required (1-10 ml).
7.4 Nitroaromatics and isophorone
7.4.1 Reduce the sample extract volume to 2 ml prior to cleanup.
7.4.2 Add a weight of activated Florisil (nominally 10 g)
predetermined by calibration (Section 5.1.2) into a 10 mm ID
chromatographic column. Tap the column to settle the Florisil and add
about 1 cm of anhydrous sodium sulfate to the top.
7.4.3 Pre-elute the column with methylene chloride/hexane (1:9)
(v/v) at about 2 mL/min. Discard the eluate and, just prior to exposure
of the sodium sulfate layer to the air, quantitatively transfer the sample
extract onto the column using an additional 2 ml of hexane to complete the
transfer. Just prior to exposure of the sodium sulfate layer to the air,
add 30 ml of methylene chloride/hexane (1:9) (v/v) and continue the
elution of the column. Discard the eluate.
7.4.4 Elute the column with 90 ml of ethyl ether/pentane (15:85)
(v/v) and discard the eluate. This fraction will contain the
diphenylamine, if it is present in the extract.
3620A - 5 Revision 1
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7.4.5 Next, elute the column with 100 mL of acetone/ethyl ether
(5:95) (v/v) into a 500 ml K-D flask equipped with a 10 ml concentrator
tube, this fraction will contain all of the nitrosamines listed in the
scope of the method.
7.4.6 Add 15 mL of methanol to the collected fraction, concentrate
using pentane to prewet the K-D column, and set the water bath at 70 to
75°C. When the apparatus is cool, remove the Snyder column and rinse the
flask and its lower joint into the concentrator tube with 1 to 2 ml of
pentane.
7.4.7 Next, elute the column with 30 ml of acetone/methylene
chloride (1:9) (v/v) into a 500 ml K-D flask equipped with a 10 ml
concentrator tube. Concentrate the collected fraction, while exchanging
the solvent to hexane. To exchange the solvent, reduce the elution
solvent to about 10 mL. Add 50 mL of hexane, a fresh boiling chip, and
return the reassembled K-D apparatus to the hot water bath. Adjust the
final volume of the cleaned-up extract to whatever volume is required (1-
10 mL). Compounds that elute in this fraction are:
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Isophorone
Nitrobenzene.
7.5 Chlorinated hydrocarbons
7.5.1 Reduce the sample extract volume to 2 mL prior to cleanup.
The extract solvent must be hexane.
7.5.2 Add a weight of activated Florisil (nominally 12 g)
predetermined by calibration (Section 5.1.2) into a 10 mm ID
chromatographic column. Tap the column to settle the Florisil and add
about 1 to 2 cm of anhydrous sodium sulfate to the top.
7.5.3 Preelute the column with 100 mL of petroleum ether. Discard
the eluate and, just prior to exposure of the sodium sulfate layer to the
air, quantitatively transfer the sample extract to the column by
decantation and subsequent petroleum ether washings. Discard the eluate.
Just prior to exposure of the sodium sulfate layer to the air, begin
eluting the column with 200 mL of petroleum ether and collect the eluate
in a 500 mL K-D flask equipped with a 10 mL concentrator tube. This
fraction should contain all of the chlorinated hydrocarbons:
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachlorobutadiene
Hexachlorocyclopentadi ene
Hexachloroethane
1,2,4-Tri chlorobenzene.
3620A - 6 Revision 1
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7.5.4 Concentrate the fraction, using hexane to prewet the column.
When the apparatus is cool, remove the Snyder column and rinse the flask
and its lower joint into the concentrator tube with hexane. Adjust the
final volume of the cleaned-up extract to whatever volume is required
(1-10 ml).
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3600 for cleanup procedures.
8.2 The analyst should demonstrate that the compounds of interest are
being quantitatively recovered before applying this method to actual samples.
8.3 For sample extracts that are cleaned up using this method, the
associated quality control samples should also be processed through this cleanup
method.
9.0 METHOD PERFORMANCE
9.1 Table 1 indicates the distribution of chlorinated pesticides, PCB's,
and haloethers in various Florisil column fractions.
9.2 Table 2 indicates the distribution of organophosphorus pesticides in
various Florisil column fractions.
10.0 REFERENCES
1. Gordon, A.J. and R.A. Ford, The Chemist's Companion: A Handbook of
Practical Data, Techniques, and References (New York: John Wiley & Sons,
Inc.), pp. 372, 374, and 375, 1972.
2. Floridin of ITT System, Florisil: Properties, Application, Bibliography,
Pittsburgh, Pennsylvania, 5M381DW.
3. Mills, P.A., "Variation of Florisil Activity; Simple Method for Measuring
Absorbent Capacity and its use in Standardizing Florisil Columns," Journal
of the Association of Official Analytical Chemists, 51, 29, 1968.
4. U.S. Food and Drug Association, Pesticides Analytical Manual (Volume 1),
July 1985.
5. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
3620A - 7 Revision 1
July 1992
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TABLE 1
DISTRIBUTION OF CHLORINATED PESTICIDES. PCBs.
AND HALOETHERS INTO FLORISIL COLUMN FRACTIONS
Parameter
Percent Recovery by Fraction8
1
Aldrin
a-BHC
B-BHC
y-BHC
5-BHC
Chlordane
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Haloethers
Heptachlor
Heptachlor epoxide
Toxaphene
PCB-1016
PCB-1221
PCB-1232
PCB-1242
PCB-1248
PCB-1254
PCB-1260
100
100
97
98
100
100
99
98
100
0 100
37 64
0 7
0 0
4 96
0 68
R
100
100
96
97
97
95 4
97
103
90
95
91
106
26
Eluant composition:
Fraction 1 - 6% ethyl ether in hexane
Fraction 2 - 15% ethyl ether in hexane
Fraction 3 - 50% ethyl ether in hexane
R = Recovered (no percent recovery data presented).
SOURCE: U.S. EPA and FDA data.
3620A - 8
Revision 1
July 1992
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R
NR
V
ND
TABLE 2
DISTRIBUTION OF ORGANOPHOSPHORUS PESTICIDES
INTO FLORISIL COLUMN FRACTIONS
Percent Recovery by Fraction8
Parameter
Azinphos methyl
Bolstar (Sulprofos)
Chlorpyrifos
Coumaphos
Demeton
Diazinon
Dichlorvos
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfothion
Fenthion
Malathion
Merphos
Mevinphos
Monochrotophos
Naled
Parathion
Parathion methyl
Phorate
Ronnel
1
ND
>80
NR
100
NR
ND
25-40
V
ND
R
V
ND
ND
NR
0-62
>80
Stirophos (Tetrachlorvinphos) ND
Sulfotepp
TEPP
Tokuthion (Prothiofos)
Trichloronate
Eluant composition:
V
ND
>80
>80
Fraction 1 -
Fraction 2 -
Fraction 3 -
Fraction 4 -
2
ND
NR
100
NR
ND
>80
V
ND
R
5
V
ND
ND
NR
100
100
ND
V
ND
200 mL
200 ml
200 ml
200 mL
3 4
20 80
ND ND
NR
NR
ND ND
V
ND ND
95
V
ND ND
ND ND
NR
ND ND
ND ND
of 6% ethyl ether in
of 15% ethyl ether in
of 50% ethyl ether in
of 100% ethyl ether
hexane
hexane
hexane
Recovered (no percent recovery information presented) (U.S. FDA)
Not recovered (U.S. FDA).
Variable recovery (U.S. FDA).
Not determined.
SOURCE: U.S. EPA and FDA data.
3620A - 9
Revision 1
July 1992
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METHOD 3620A
FLORISIL COLUMN CLEANUP
711 Reduce volume
of sample extract
to 2 mL
Phthalate Esters
7 1 2 Place
Florisil into
chromatographic
column; add
anhydrous sodium
sulfate
7 1 3 Preelute
column with hexane,
transfer sample
extract, add hexane
7 1 4 Elute column
with ethyl ether in
hexane
721 Reduce volume
of sample extract
to 2 mL
7 2 2 Put Florisil
into
chromatographic
column; add
anhydrous sodium
sulfate
7 2 3 Preelute
column with ethyl
ether/pentane,
transfer extract,
add pentane
7 2 4 Elute column
with ethyl
ether/pentane
7 1 4 Concentrate
fraction, adjust
volume, analyze
7 2 5 Elute column
with acetone/ethyl
ether into flask
3620A - 10
Revision 1
July 1992
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METHOD 3620A
continued
Organochl or me
pesticides, haloethers
and organophosphorous
731 Reduce volume
of sample ex tract
to 2 mL
troa romatics
and is ophorone
Chi orinated
hydrocarbons
Ana 1yze by CC
741 Reduce volume
of sample ex tract
to 2 mL
Fl onail
tographic
n, add
* sodium
e then
discard
ate
ust sample
volume ,
to column
Lh hexane
in col umn ,
olumn 4
o separate
iks
icent ra te
ad jus t
jme
7 S 1 Reduce volume
of sample ex tract
to 2 mL
7 5 2
Place
Florisil in
chroma tographic
col urn. add
anhydrous s oaium
sulfate
7 5 3 P
*eel u te
col umn with
pet r o leum ether ,
t ransf er sampl e
ex t rac t to col umn ,
discard
ei -a te
7 5 4 Concentrate
i rac tion
aQ jus t
final vo 1 ume
•~\
7 4 2 Put
3 lur t
chroma tc
col umr
anhydrous
sul;
7 4 3 Tt
sample ext
col umn
methy
chl o r ide/
discard
7 4 4 Elut
wit
ace t one/m
chloride
solvent t
7 4 4 Con
f racti on ,
fina 1 v
2 6 Add melhanol
to f raction;
concentrate
add
anafer
.ract on
i, add
excnange
3620A - 11
Revision 1
July 1992
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METHOD 3630B
SILICA GEL CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Silica gel is a regenerative adsorbent of amorphous silica with
weakly acidic properties. It is produced from sodium silicate and sulfuric acid.
Silica gel can be used in column chromatography for the separation of analytes
from interfering compounds of a different chemical polarity. It may be used
activated, after heating to 150 - 160°C, or deactivated with up to 10% water.
1.2 This method includes guidance for standard column cleanup of sample
extracts containing polynuclear aromatic hydrocarbons, derivatized phenolic
compounds, organochlorine pesticides, and PCBs as Aroclors.
1.3 This method also provides cleanup procedures using solid-phase
extraction cartridges for pentafluorobenzyl bromide-derivatized phenols,
organochlorine pesticides, and PCBs as Aroclors. This technique also provides
the best separation of PCBs from most single component organochlorine pesticides.
When only PCBs are to be measured, this method can be used in conjunction with
sulfuric acid/permanganate cleanup (Method 3665).
1.4 Other analytes may be cleaned up using this method if the analyte
recovery meets the criteria specified in Sec. 8.0.
2.0 SUMMARY OF METHOD
2.1 This method provides the option of using either standard column
chromatography techniques or solid-phase extraction cartridges. Generally, the
standard column chromatography techniques use larger amounts of adsorbent and,
therefore, have a greater cleanup capacity.
2.2 In the standard column cleanup protocol, the column is packed with
the required amount of adsorbent, topped with a water adsorbent, and then loaded
with the sample to be analyzed. Elution of the analytes is accomplished with a
suitable sol vent(s) that leaves the interfering compounds on the column. The
eluate is then concentrated (if necessary).
2.3 The cartridge cleanup protocol uses silica solid-phase extraction
cartridges packed with 1 g or 2 g of adsorbent. Each cartridge is solvent washed
immediately prior to use. Aliquots of sample extracts are loaded onto the
cartridges, which are then eluted with suitable solvent(s). A vacuum manifold
is required to obtain reproducible results. The collected fractions may be
further concentrated prior to gas chromatographic analysis.
2.4 The appropriate gas chromatographic method is listed at the end of
each technique. Analysis may also be performed by gas chromatography/mass
spectrometry (Method 8270).
3630B - 1 Revision 2
September 1994
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3.0 INTERFERENCES
3.1 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All these materials must be demonstrated to be free from
interferences under the conditions of the analysis, by analyzing reagent blanks.
See Sec. 8 for guidance on a reagent blank check.
3.2 Phthalate ester contamination may be a problem with certain
cartridges The more inert the column and/or cartridge material (i.e., glass or
Teflon), the less problem with phthalates. Phthalates create interference
problems for all method analytes, not just the phthalate esters themselves.
3.3 More extensive procedures than those outlined in this method may be
necessary for reagent purification.
4.0 APPARATUS AND MATERIALS
4.1 Chromatographic column - 250 mm long x 10 mm ID; with Pyrex glass
wool at bottom and a Teflon stopcock.
NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without
frits may be purchased. Use a small pad of Pyrex glass wool to
retain the adsorbent. Prewash the glass wool pad with 50 ml of
acetone followed by 50 ml of elution solvent prior to packing the
column with adsorbent.
4.2 Beakers - 500 ml.
4.3 Vials - 2, 10, 25 ml, glass with Teflon lined screw-caps or crimp
tops.
4.4 Muffle furnace.
4.5 Reagent bottle - 500 ml.
4.6 Erlenmeyer flasks - 50 and 250 ml.
4.7 Vacuum manifold: VacElute Manifold SPS-24 (Analytichem
International), Visiprep (Supelco, Inc.) or equivalent, consisting of glass
vacuum basin, collection rack and funnel, collection vials, replaceable stainless
steel delivery tips, built-in vacuum bleed valve and gauge. The system is
connected to a vacuum pump or water aspirator through a vacuum trap made from a
500 ml sidearm flask fitted with a one-hole stopper and glass tubing.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
3630B - 2 Revision 2
September 1994
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provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Silica gel for chromatography columns.
5.3.1 Silica Gel for Phenols and Polynuclear Aromatic Hydrocarbons:
100/200 mesh desiccant (Davison Chemical grade 923 or equivalent). Before
use, activate for at least 16 hr. at 130°C in a shallow glass tray, loosely
covered with foil.
5.3.2 Silica Gel for Organochlorine pesticides/PCBs: 100/200 mesh
desiccant (Davison Chemical grade 923 or equivalent). Before use,
activate for at least 16 hr. at 130°C in a shallow glass tray, loosely
covered with foil. Deactivate it to 3.3% with reagent water in a 500 ml
glass jar. Mix the contents thoroughly and allow to equilibrate for 6
hours. Store the deactivated silica gel in a sealed glass jar inside a
desiccator.
5.4 Silica cartridges: 40 fj,m particles, 60 A pores. The cartridges with
which this method was developed consist of 6 mL serological-grade polypropylene
tubes, with the 1 g of silica held between two polyethylene or stainless steel
frits with 20 /xm pores. 2 g silica cartridges are also used in this method, and
0.5 g cartridges are available. The compound elution patterns must be verified
when cartridges other than the specified size are used.
5.5 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. A method blank must be analyzed in order to demonstrate that
there is no interference from the sodium sulfate.
5.6 Eluting solvents
5.6.1 Cyclohexane, C6H12 - Pesticide quality or equivalent.
5.6.2 Hexane, C6H14 - Pesticide quality or equivalent.
5.6.3 2-Propanol, (CH3)2CHOH - Pesticide quality or equivalent.
5.6.4 Toluene, C6H5CH3 - Pesticide quality or equivalent.
5.6.5 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.6.6 Pentane, C5H12 - Pesticide quality or equivalent.
5.6.7 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.6.8 Diethyl Ether, C2H5OC2H5. Pesticide quality or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
3630B - 3 Revision 2
September 1994
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test strips. After cleanup, 20 ml of ethanol preservative must be added
to each liter of ether.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes, Sec.
4.1.
7.0 PROCEDURE
7.1 General Guidance
7.1.1 The procedure contains two cleanup options for the derivatized
phenols and organochlorine pesticides/Aroclors, but only one technique for
the polynuclear aromatic hydrocarbons (PAHs) (standard column
chromatography). Cleanup techniques by standard column chromatography for
all analytes are found in Sec. 7.2. Cleanup techniques by solid-phase
cartridges for derivatized phenols and PAHs are found in Sec. 7.3. The
standard column chromatography techniques are packed with a greater amount
of silica gel adsorbent and, therefore, have a greater cleanup capacity.
A rule of thumb relating to cleanup capacity is that 1 g of sorbent
material will remove 10 to 30 mg of total interferences. (However,
capacity is also dependent on the sorbent retentiveness of the
interferences.) Therefore, samples that exhibit a greater degree of
sample interference should be cleaned up by the standard column technique.
However, both techniques have limits on the amount of interference that
can be removed. If the interference is caused by high boiling material,
then Method 3640 should be used prior to this method. If the interference
is caused by relatively polar compounds of the same boiling range as the
analytes, then multiple column or cartridge cleanups may be required. If
crystals of sulfur are noted in the extract, then Method 3660 should be
utilized prior to this method. The cartridge cleanup techniques are often
faster and use less solvent, however they have less cleanup capacity.
7.1.2 Allow the extract to reach room temperature if it was in cold
storage. Inspect the extracts visually to ensure that there are no
particulates or phase separations and that the volume is as stated in the
accompanying documents. Verify that the solvent is compatible with the
cleanup procedures. If crystals of sulfur are visible or if the presence
of sulfur is suspected, proceed with Method 3660.
7.1.3 If the extract solvent is methylene chloride, for most cleanup
techniques, it must be exchanged to hexane. (For the PAHs, exchange to
cyclohexane as per Sec. 7.2.1). Follow the standard Kuderna-Danish
concentration technique provided in each extraction method. The volume of
methylene chloride should have been reduced to 1 - 2 mL. Add 40 ml of
hexane, a fresh boiling chip and repeat the concentration as written. The
final volume required for the cleanup techniques is normally 2 ml.
3630B - 4 Revision 2
September 1994
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7.2 Standard Column Cleanup Techniques
7.2.1 Polynuclear aromatic hydrocarbons
7.2.1.1 Before the silica gel cleanup technique can be
utilized, the extract solvent must be exchanged to cyclohexane. The
exchange is performed by adding 4 ml of cyclohexane following
reduction of the sample extract to 1-2 mL using the macro Snyder
column. Attach the two ball micro Snyder column and reduce the
volume to 2 mL.
CAUTION: When the volume of solvent is reduced below 1 ml,
semivolatile analytes may be lost. If the
extract goes to dryness, the extraction must be
repeated.
7.2.1.2 Prepare a slurry of 10 g of activated silica gel
(Sec. 5.3.1) in methylene chloride and place this into a 10 mm ID
chromatographic column. Tap the column to settle the silica gel and
elute the methylene chloride. Add 1 to 2 cm of anhydrous sodium
sulfate to the top of the silica gel.
7.2.1.3 Pre-elute the column with 40 ml of pentane. The
rate for all elutions should be about 2 mL/min. Discard the eluate
and, just prior to exposure of the sodium sulfate layer to the air,
transfer the 2 ml cyclohexane sample extract onto the column using
an additional 2 mL cyclohexane to complete the transfer. Just prior
to exposure of the sodium sulfate layer to the air, add 25 ml of
pentane and continue the elution of the column. Discard this
pentane eluate.
7.2.1.4 Next, elute the column with 25 mL of methylene
chloride/pentane (2:3)(v/v) into a 500 mL K-D flask equipped with a
10 mL concentrator tube. Concentrate the collected fraction to
whatever volume is required (1-10 mL). Proceed with HPLC (Method
8310) or GC analysis (Method 8100). Validated components that elute
in this fraction are:
Acenaphthene
Acenaphthylene
Anthracene
Benzo(a)anthracene
Benzo(a)pyrene
Benzo(b)fluoranthene
Benzo(g,h,i)pery1ene
Benzo(k)fluoranthene
Chrysene
Dibenzo(a,h)anthracene
Fluoranthene
Fluorene
Indeno(l,2,3-cd)pyrene
Naphthalene
Phenanthrene
Pyrene
3630B - 5 Revision 2
September 1994
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7.2.2 Derivatized Phenols
7.2.2.1 This silica gel cleanup procedure is performed on
sample extracts that have undergone pentafluorobenzyl bromide
derivatization, as described in Method 8040. The sample extract
must be in 2 ml of hexane at this point.
7.2.2.2 Place 4.0 g of activated silica gel (Sec. 5.3.1)
into a 10 mm ID chromatographic column. Tap the column to settle
the silica gel and add about 2 g of anhydrous sodium sulfate to the
top of the silica gel.
7.2.2.3 Pre-elute the column with 6 mL of hexane. The
rate for all elutions should be about 2 mL/min. Discard the eluate
and, just prior to exposure of the sodium sulfate layer to the air,
pipet onto the column 2 mL of the hexane solution that contains the
derivatized sample or standard. Elute the column with 10.0 mL of
hexane and discard the eluate.
7.2.2.4 Elute the column, in order, with 10.0 ml of 15%
toluene in hexane (Fraction 1); 10.0 mL of 40% toluene in hexane
(Fraction 2); 10.0 mL of 75% toluene in hexane (Fraction 3); and
10.0 mL of 15% 2-propanol in toluene (Fraction 4). All elution
mixtures are prepared on a volume:volume basis. Elution patterns
for the phenolic derivatives are shown in Table 1. Fractions may be
combined, as desired, depending upon the specific phenols of
interest or level of interferences. Proceed with GC analysis
(Method 8040).
7.2.3 Organochlorine Pesticides and Aroclors
7.2.3.1 Transfer a 3 g portion of deactivated silica gel
(Sec. 5.3.2) into a 10 mm ID glass chromatographic column and top it
with 2 to 3 cm of anhydrous sodium sulfate.
7.2.3.2 Add 10 mL of hexane to the top of the column to
wet and rinse the sodium sulfate and silica gel. Just prior to
exposure of the sodium sulfate layer to air, stop the hexane eluate
flow by closing the stopcock on the chromatographic column. Discard
the eluate.
7.2.3.3 Transfer the sample extract (2 mL in hexane) onto
the column. Rinse the extract vial twice with 1 to 2 mL of hexane
and add each rinse to the column. Elute the column with 80 mL of
hexane (Fraction I) at a rate of about 5 mL/min. Remove the
collection flask and set it aside for later concentration. Elute
the column with 50 mL of hexane (Fraction II) and collect the
eluate. Perform a third elution with 15 mL of methylene chloride
(Fraction III). The elution patterns for the organochlorine
pesticides, Aroclor-1016, and Aroclor-1260 are shown in Table 2.
7.2.3.4 Prior to gas chromatographic analysis, the
extraction solvent must be exchanged to hexane. Fractions may be
combined, as desired, depending upon the specific
3630B - 6 Revision 2
September 1994
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pesticides/Aroclors of interest or level of interferences. If
mixtures of Aroclors and pesticides are expected, it is best to
analyze Fraction I separately, since it contains the Aroclors
separated from most pesticides. Proceed with GC analysis as per
Method 8081.
7.3 Cartridge Cleanup Techniques
7.3.1 Cartridge Set-up and Conditioning
7.3.1.1 Arrange the 1 g silica cartridges (2 g for phenol
cleanup) on the manifold in the closed-valve position. Other size
cartridges may be used, however the data presented in the Tables are
all based on 1 g cartridges for pesticides/Aroclors and 2 g
cartridges for phenols. Therefore, supporting recovery data must be
developed for other sizes. Larger cartridges will probably require
larger volumes of elution solvents.
7.3.1.2 Turn on the vacuum pump and set pump vacuum to 10
inches (254 mm) of Hg. Do not exceed the manufacturer's
recommendation for manifold vacuum. Flow rates can be controlled by
opening and closing cartridge valves.
7.3.1.3 Condition the cartridges by adding 4 ml of hexane
to each cartridge. Slowly open the cartridge valves to allow hexane
to pass through the sorbent beds to the lower frits. Allow a few
drops per cartridge to pass through the manifold to remove all air
bubbles. Close the valves and allow the solvent to soak the entire
sorbent bed for 5 minutes. Do not turn off the vacuum.
7.3.1.4 Slowly open cartridge valves to allow the hexane
to pass through the cartridges. Close the cartridge valves when
there is still at least 1 mm of solvent above the sorbent bed. Do
not allow cartridges to become dry. If cartridges go dry, repeat
the conditioning step.
7.3.2 Derivatized Phenols
7.3.2.1 Reduce the sample extract volume to 2 ml prior to
cleanup. The extract solvent must be hexane and the phenols must
have undergone derivatization by pentafluorobenzyl bromide, as per
Method 8040.
7.3.2.2 Transfer the extract to the 2 g cartridge that has
been conditioned as described in Sec. 7.3.1. Open the cartridge
valve to allow the extract to pass through the cartridge bed at
approximately 2 mL/minute.
7.3.2.3 When the entire extract has passed through the
cartridges, but before the cartridge becomes dry, rinse the sample
vials with an additional 0.5 ml of hexane, and add the rinse to the
cartridges to complete the quantitative transfer.
3630B - 7 Revision 2
September 1994
-------
7.3.2.4 Close the cartridge valve and turn off the vacuum
after the solvent has passed through, ensuring that the cartridge
never gets dry.
7.3.2.5 Place a 5 ml vial or volumetric flask into the
sample rack corresponding to the cartridge position. Attach a
solvent-rinsed stainless steel solvent guide to the manifold cover
and align with the collection vial.
7.3.2.6 Add 5 mL of hexane to the cartridge. Turn on the
vacuum pump and adjust the pump pressure to 10 inches (254 mm) of
Hg. Allow the solvent to soak the sorbent bed for 1 minute or less.
Slowly open the cartridge valve, and collect the eluate (this is
Fraction 1, and should be discarded).
NOTE: If cartridges smaller than 2 g are used, then Fraction
1 cannot be discarded, since it contains some of the
phenols.
7.3.2.7 Close the cartridge valve, replace the collection
vial, and add 5 ml of toluene/hexane (25/75, v/v) to the cartridge.
Slowly open the cartridge valve and collect the eluate into the
collection vial. This is Fraction 2, and should be retained for
analysis.
7.3.2.8 Adjust the final volume of the eluant to a known
volume which will result in analyte concentrations appropriate for
the project requirements (normally 1-10 ml). Table 3 shows
compound recoveries for 2 g silica cartridges. The cleaned up
extracts are ready for analysis by Method 8040.
7.3.3 Organochlorine Pesticides/Aroclors
NOTE: The silica cartridge procedure is appropriate when
polychlorinated biphenyls are known to be present.
7.3.3.1 Reduce the sample extract volume to 2 ml prior to
cleanup. The extract solvent must be hexane.
7.3.3.2 Use the 1 g cartridges conditioned as described in
Sec. 7.3.1.
7.3.3.3 Transfer the extract to the cartridge. Open the
cartridge valve to allow the extract to pass through the cartridge
bed at approximately 2 mL/minute.
7.3.3.4 When the entire extract has passed through the
cartridges, but before the cartridge becomes dry, rinse the sample
vials with an additional 0.5 ml of solvent, and add the rinse to the
cartridges to complete the quantitative transfer.
7.3.3.5 Close the cartridge valve and turn off the vacuum
after the solvent has passed through, ensuring that the cartridge
never goes dry.
3630B - 8 Revision 2
September 1994
-------
7.3.3.6 Place a 5 ml vial or volumetric flask into the
sample rack corresponding to the cartridge position. Attach a
solvent-rinsed stainless steel solvent guide to the manifold cover
and align with the collection vial.
7.3.3.7 Add 5 mL of hexane to the cartridge. Turn on the
vacuum pump and adjust the pump pressure to 10 inches (254 mm) of
Hg. Allow the solvent to soak the sorbent bed for 1 minute or less.
Slowly open the cartridge valve and collect the eluate into the
collection vial (Fraction 1).
7.3.3.8 Close the cartridge valve, replace the collection
vial, and add 5 ml of diethyl ether/hexane (50/50, v/v) to the
cartridge. Slowly open the cartridge valve and collect the eluate
into the collection vial (Fraction 2).
7.3.3.9 Adjust the final volume of each of the two
fractions to a known volume which will result in analyte
concentrations appropriate for the project requirements (normally 1
- 10 mL). The fractions may be combined prior to final adjustment
of volume, if analyte fractionation is not required. Table 4 shows
compound recoveries for 1 g silica cartridges. The cleaned up
extracts are ready for analysis by Method 8081.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3600 for cleanup procedures.
8.2 A reagent blank (consisting of the elution solvents) must be passed
through the column or cartridge and checked for the compounds of interest, prior
to the use of this method. This same performance check is required with each new
lot of adsorbent or cartridges. The level of interferences must be below the
method detection limit before this method is performed on actual samples.
8.3 The analyst must demonstrate that the compounds of interest are being
quantitatively recovered before applying this method to actual samples. See the
attached Tables for acceptable recovery data. For compounds that have not been
tested, recovery must be > 85%.
8.3.1 Before any samples are processed using the solid-phase
extraction cartridges, the efficiency of the cartridge must be verified.
A recovery check must be performed using standards of the target analytes
at known concentration. Only lots of cartridges that meet the recovery
criteria for the spiked compounds can be used to process the samples.
8.3.2 A check should also be performed on each individual lot of
cartridges and for every 300 cartridges of a particular lot.
8.4 For sample extracts that are cleaned up using this method, the
associated quality control samples should also be processed through this cleanup
method.
3630B - 9 Revision 2
September 1994
-------
9.0 METHOD PERFORMANCE
9.1 Table 1 provides performance information on the fractionation of
phenolic derivatives using standard column chromatography.
9.2 Table 2 provides performance information on the fractionation of
organochlorine pesticides/Aroclors using standard column chromatography.
9.3 Table 3 shows recoveries of derivatized phenols obtained using 2 g
silica cartridges.
9.4 Table 4 shows recoveries and fractionation of organochlorine
pesticides obtained using 1 g silica cartridges.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures
for the Analysis of Pollutants Under the Clean Water Act; Final Rule
and Interim Final Rule and Proposed Rule," October 26, 1984.
2. U.S EPA "Evaluation of Sample Extract Cleanup Using Solid-Phase
Extraction Cartridges," Project Report, December 1989.
3630B - 10 Revision 2
September 1994
-------
TABLE 1
SILICA GEL FRACTIONATION OF PFBB DERIVATIVES
Percent Recovery by Fraction8
Parameter 123
2-Chlorophenol
2-Nitrophenol
Phenol
2,4-Dimethylphenol
2,4-Dichlorophenol
2,4,6-Trichlorophenol
4-Chl oro -3 -methyl phenol
Pentachlorophenol
4-Nitrophenol
90
90
95
95
50 50
84
75 20
1
9
10
7
1
14
1
90
90
a Eluant composition:
Fraction 1 - 15% toluene in hexane.
Fraction 2 - 40% toluene in hexane.
Fraction 3 - 75% toluene in hexane.
Fraction 4 - 15% 2-propanol in toluene.
Data from Reference 1 (Method 604)
3630B - 11 Revision 2
September 1994
-------
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-------
TABLE 3
PERCENT RECOVERIES AND ELUTION PATTERNS FOR 18
PHENOLS FROM 2 g SILICA CARTRIDGES3
Fraction 2
Average Percent
Compound Recovery RSD
Phenol
2-Methyl phenol
3 -Methyl phenol
4-Methyl phenol
2, 4-Dimethyl phenol
2-Chlorophenol
2,6-Dichlorophenol
4-Chl oro-3-methyl phenol
2,4-Dichlorophenol
2,4,6-Trichlorophenol
2 , 3 , 6-Tri chl orophenol
2,4,5-Trichlorophenol
2 , 3 , 5-Tri chl orophenol
2,3,5,6-Tetrachlorophenol
2,3,4,6-Tetrachlorophenol
2, 3, 4-Tri chl orophenol
2,3,4, 5-Tetrachl orophenol
Pentachl orophenol
74.1
84.8
86.4
82.7
91.8
88.5
90.4
94.4
94.5
97.8
95.6
92.3
92.3
97.5
97.0
72.3
95.1
96.2
5.2
5.2
4.4
5.0
5.6
5.0
4.4
7.1
7.0
6.6
7.1
8.2
8.2
5.3
6.1
8.7
6.8
8.8
a Silica cartridges (Supelco, Inc.) were used; each cartridge was conditioned
with 4 mL of hexane prior to use. Each experiment was performed in duplicate
at three spiking concentrations (0.05 ptg, 0.2 jug, and 0.4 y,q per compound per
cartridge). Fraction 1 was eluted with 5 mL hexane and was discarded.
Fraction 2 was eluted with 5 mL toluene/hexane (25/75, v/v).
Data from Reference 2
3630B - 14 Revision 2
September 1994
-------
TABLE 4
PERCENT RECOVERIES AND ELUTION PATTERNS FOR 17 ORGANOCHLORINE
PESTICIDES AND AROCLORS FROM 1 g SILICA CARTRIDGES3
Compound
Fraction 1
Average Percent
Recovery RSD
Fraction 2
Average Percent
Recovery RSD
alpha-BHC
gamma -BHC
beta-BHC
Heptachlor
delta-BHC
Aldrin
Heptachlor epoxide
Endosulfan I
4,4'-DDE
Dieldrin
Endrin
4,4'-DDD
Endosulfan II
4,4'-DDT
Endrin aldehyde
Endosulfan sulfate
4,4'-Methoxychlor
Aroclor 1016
Aroclor 1221
Aroclor 1232
Aroclor 1242
Aroclor 1248
Aroclor 1254
Aroclor 1264
0
0
0
97.3 1.3
0
95.9 1.0
0
0
99.9 1.7
0
0
10.7 41
0
94.1 2.0
0
0
0
124
93.5
118
116
114
108
112
98.7
94.8
94.3
0
90.8
0
97.9
102
0
92.3
117
92.4
96.0
0
59.7
97.8
98.0
2.3
1.9
3.0
2.5
2.1
2.3
2.0
2.6
3.3
2.2
2.6
2.1
2.4
a Silica cartridges (Supelco, Inc. lot SP0161) were used; each cartridge was
conditioned with 4 mL hexane prior to use. The organochlorine pesticides were
tested separately from PCBs. Each organochlorine pesticides experiment was
performed in duplicate, at three spiking concentrations (0.2 ;ug, 1.0 /xg, and
2.0 fj,g per compound per cartridge). Fraction 1 was eluted with 5 mL of
hexane, Fraction 2 with 5 mL of diethyl ether/hexane (50/50, v/v). PCBs were
spiked at 10 ng per cartridge and were eluted with 3 mL of hexane. The values
given for PCBs are the percent recoveries for a single determination.
Data from Reference 2
3630B - 15
Revision 2
September 1994
-------
METHOD 3630B
SILICA GEL CLEANUP
oc
PCBs
y/Com
> 10-30 mg (
I ^\,nt.r
7.2
Colurr
Standard
n Cleanup.
Pesticide
ft. Phenols
7.I.N.
emrationN.
"' J
ferences./
( StSrt )
/ 7.1.1 N.
, .. ,( An«|vte \
N. Type. /
j/T\
VJ
I
\_/ 7.3 Cartridge
Cleanup.
y/Analyte N^
I C Type. .? 1
Denvatized >^ / OC Pesticides
Phenols ^*S ft. PCBs
1 +
7.2.2.1 Do PFBB
derivatization on
sample extract
(8040).
4
7.2.2.2 Place
activated silica gel
in chromatographic
column; add
anhydroua NajSO*.
4
7.2.2.3 Preelute
column with hexane;
pipet hexane
solution onto column;
alute.
1
7.2.2.4 Elute column
with specified
solvents.
4
Analyze
by GC
(Method I
V. 8040). }
7.2.3.1 Deactivate
silica gel, prepare
column.
1
7.2.3.2 Elute the
GC column
with hexane.
V
7.2.3.3 Transfer
extract onto column
snd elute with
specified solvents.
^
7.3.4 Exchange the
elution solvent
to hexane (Section
7.1.3).
|
Analyze
by GC
I Method
N. 8081. >
i
7.3.1 Cartridge
Set-up ft.
Conditioning.
X'Analyte \^
|—C TVP.. >— |
Derivatized >. / OC Peaticides
Phenols N./ ft PCBs
1 1
7.3.2.1 Do PFBB
derivatization on
sample extract
(8040).
4
7.3.2.3 ft 7.3.2.4
Transfer extract
to cartridge.
\r
7.3.2.6 ft 7.3.2.7
Rinse cartridge
with hexane ft
discard.
1
7.3.2.8 Elute
cartridge with
toluene/he xane.
^f
Analyze by
GC Method
8040 or
GC/MS
\ Method /
\8270./
7.3.3.1 Exchange
solvent to
hexane.
^ r
7.3.3.3 & 7.3.3.4
Transfer extract
to cartridge.
4
7.3.3.6 & 7.3.3.7
Elute cartridge
with hexane as
Fraction 1.
1
7.3.3.8 Elute
cartridge with
ether/hexane as
Fraction II.
V
Analyze
each fraction
by GC
Method
\*™./
3630B - 16
Revision I
September 1994
-------
METHOD 3630B
(continued)
7.2 Standard
Column Cleanup.
7.2.1.1 Exchange
extract solvent to
cyclohexane during
K-D procedure.
7.2.1.2 Prepare
slurry activated
silica gel, prepare
column.
7.2.1.3 Preelute
column with
pentane, transfer
extract onto column
and elute with
pentane.
7.2.1.4 Elute
column with
CH2CI2 /pentane;
concentrate
collected fraction;
adjust volume.
Analyze
by GC Method
8100 or
GC/MS
Method
8270.
3630B - 17
Revision 2
September 1994
-------
METHOD 3640A
GEL-PERMEATION CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Gel-permeation chromatography (GPC) is a size exclusion cleanup
procedure using organic solvents and hydrophobic gels in the separation of
synthetic macromolecules (1). The packing gel is porous and is characterized by
the range or uniformity (exclusion range) of that pore size. In the choice of
gels, the exclusion range must be larger than the molecular size of the molecules
to be separated (2). A cross-linked divinylbenzene-styrene copolymer (SX-3 Bio
Beads or equivalent) is specified for this method.
1.2 General cleanup application - GPC is recommended for the elimination
from the sample of lipids, polymers, copolymers, proteins, natural resins and
polymers, cellular components, viruses, steroids, and dispersed high-molecular-
weight compounds (2). GPC is appropriate for both polar and non-polar analytes,
therefore, it can be effectively used to cleanup extracts containing a broad
range of analytes.
1.3 Specific application - This method includes guidance for cleanup of
sample extracts containing the following analytes from the RCRA Appendix VIII and
Appendix IX lists:
Compound Name CAS No.'
Acenaphthene 83-32-9
Acenaphthylene 208-96-8
Acetophenone 98-86-2
2-Acetylaminofluorene 53-96-3
Aldrin 309-00-2
4-Aminobiphenyl 92-67-1
Aniline 62-53-3
Anthracene 120-12-7
Benomyl 17804-35-2
Benzenethiol 108-98-5
Benzidine 92-87-5
Benz(a)anthracene 56-55-3
Benzo(b)fluoranthene 205-99-2
Benzo(a)pyrene 50-32-8
Benzo(ghi)perylene 191-24-2
Benzo(k)fluoranthene 207-08-9
Benzoic acid 65-85-0
Benzotrichloride 98-07-7
Benzyl alcohol 100-51-6
Benzyl chloride 100-44-7
alpha-BHC 319-84-6
beta-BHC 319-85-7
3640A - 1 Revision 1
September 1994
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Compound Name
gamma-BHC
delta-BHC
4-Bromophenyl phenyl ether
Butyl benzyl phthalate
2-sec-butyl -4,6-dinitrophenol (Dinoseb)
Carbazole
Carbendazim
alpha-Chlordane
gamma-Chlordane
4-Chloro-3-methyl phenol
4-Chloroaniline
Chi orobenzi late
Bi s (2-chl oroethoxy )methane
Bis(2-chloroethyl) ether
Bis (2-chl oroisopropyl) ether
2-Chloronaphthalene
2-Chlorophenol
4-Chlorophenol
3-Chlorophenol
4-Chlorophenyl phenyl ether
3-Chloropropionitrile
Chrysene
2-Cresol
3-Cresol
4-Cresol
Cyclophosphamide
ODD
DDE
DDT
Di-n-butyl phthalate
Dial late
Dibenzo(a,e)pyrene
Dibenzo(a,i)pyrene
Dibenz(a,j)acridine
Dibenz (a, h) anthracene
Dibenzofuran
Dibenzothiophene
l,2-Dibromo-3-chloropropane
1,2-Dibromoethane
trans- l,4-Dichloro-2-butene
cis-l,4-Dichloro-2-butene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
3, 3 '-Dichl orobenzi dine
2,6-Dichlorophenol
2,4-Dichlorophenoxyacetic acid (2,4-D)
2,4-Dichlorophenol
CAS No.a
58-89-9
319-86-8
101-55-3
85-68-7
88-85-7
86-74-8
10605-21-7
5103-71-9
5566-34-7
59-50-7
106-47-8
510-15-6
111-91-1
111-44-4
108-60-1
91-58-7
95-57-8
106-48-9
108-43-0
7005-72-3
542-76-7
218-01-9
95-48-7
108-39-4
106-44-5
50-18-0
72-54-8
72-55-9
50-29-3
84-74-2
2303-16-4
192-65-4
189-55-9
224-42-0
53-70-3
132-64-9
132-65-0
96-12-8
106-93-4
110-57-6
1476-11-5
95-50-1
106-46-7
541-73-1
91-94-1
87-65-0
94-75-7
120-83-2
3640A - 2
Revision 1
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Compound Name
2,4-Dichlorotoluene
l,3-Dichloro-2-propanol
Dieldrin
Diethyl phthalate
Dimethoate
Dimethyl phthalate
p-Di methyl ami noazobenzene
7, 12-Dimethyl -benz(a)anthracene
2,4-Dimethylphenol
3,3-Dimethylbenzidine
4,6-Dinitro-o-cresol
1,3-Dinitrobenzene
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Diphenylamine
Diphenyl ether
1 , 2 -Di phenyl hydrazi ne
Disulfoton
Endosulfan sulfate
Endosulfan I
Endosulfan II
Endrin
Endrin aldehyde
Endrin ketone
Ethyl methane sulfonate
Ethyl methacrylate
Bis(2-ethylhexyl) phthalate
Famphur
Fluorene
Fluoranthene
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachl orobutadi ene
Hexachl orocycl opentadi ene
Hexachloroethane
Hexachl oropropene
Indeno(l,2,3-cd)pyrene
Isodrin
Isophorone
cis-Isosafrole
trans-Isosafrole
Kepone
Malononitrile
Merphos
Methoxychlor
3-Methylcholanthrene
CAS No.a
95-73-8
96-23-1
60-57-1
84-66-2
60-51-5
131-11-3
60-11-7
57-97-6
105-67-9
119-93-7
534-52-1
99-65-0
51-28-5
121-14-2
606-20-2
122-39-4
101-84-8
122-66-7
298-04-4
1031-07-8
959-98-8
33213-65-9
72-20-8
7421-93-4
53494-70-5
62-50-0
97-63-2
117-81-7
52-85-7
86-73-7
206-44-0
76-44-8
1024-57-3
118-74-1
87-68-3
77-47-4
67-72-1
1888-71-7
193-39-5
465-73-6
78-59-1
17627-76-8
4043-71-4
143-50-0
109-77-3
150-50-5
72-43-5
56-49-5
3640A - 3
Revision 1
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Compound Name
2-Methyl naphthal ene
Methyl parathion
4,4'-Methylene-bis(2-chloroaniline)
Naphthalene
1,4-Naphthoquinone
2-Naphthylamine
1-Naphthylamine
5-Nitro-o-toluidine
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Nitrobenzene
2-Nitrophenol
4-Nitrophenol
N-Nitrosodi-n-butylamine
N-Nitrosodiethanolamine
N-Nitrosodi ethyl ami ne
N-Nitrosodi methyl ami ne
N-Ni trosodi phenyl ami ne
N-Nitrosodi-n-propylamine
N-Ni trosomethyl ethyl ami ne
N-Ni trosomorphol ine
N-Nitrosopiperidine
N-Nitrosopyrolidine
Di-n-octyl phthalate
Parathion
Pentachl orobenzene
Pentachl oroethane
Pentachloronitrobenzene (PCNB)
Pentachl orophenol
Phenacetin
Phenanthrene
Phenol
1 , 2- Phenyl enedi ami ne
Phorate
2-Picoline
Pronamide
Pyrene
Resorcinol
Safrole
1,2,4, 5 -Tetrachl orobenzene
2,3,5,6-Tetrachloronitrobenzene
2,3,5, 6 -Tetrachl orophenol
2,3,4,6-Tetrachlorophenol
Tetraethyl dithiopyrophosphate (Sulfotep)
Thiosemicarbazide
2-Toluidine
4-Toluidine
CAS No.a
91-57-6
298-00-0
101-14-4
91-20-3
130-15-4
91-59-8
134-32-7
99-55-8
88-74-4
99-09-2
100-01-6
98-95-3
79-46-9
100-02-7
924-16-3
1116-54-7
55-18-5
62-75-9
86-30-6
621-64-7
10595-95-6
59-89-2
100-75-4
930-55-2
117-84-0
56-38-2
608-93-5
76-01-7
82-68-8
87-86-5
62-44-2
85-01-8
108-95-2
95-54-5
298-02-2
109-06-8
23950-58-5
129-00-0
108-46-3
94-59-7
95-94-3
117-18-0
935-95-5
58-90-2
3689-24-5
79-19-6
106-49-0
95-53-4
3640A - 4
Revision 1
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Compound Name CAS No.'
Thiourea, l-(o-chlorophenyl) 5344-82-1
Toluene-2,4-diamine 95-80-7
1,2,3-Trichlorobenzene 87-61-6
1,2,4-Trichlorobenzene 120-82-1
2,4,6-Trichlorophenol 88-06-2
2,4,5-Trichlorophenol 95-95-4
2,4,5-Trichlorophenoxyacetic acid (2,4,5-T) 93-76-5
2,4,5-Trichlorophenoxypropionic acid (2,4,5-TP) 93-72-1
Warfarin 81-81-2
a Chemical Abstract Services Registry Number.
Table 1 presents average percent recovery and percent RSD data for these
analytes, as well as the retention volumes of each analyte on a single GPC
system. Retention volumes vary from column to column. Figure 1 provides
additional information on retention volumes for certain classes of compounds.
The data for the semivolatiles were determined by GC/MS, whereas, the pesticide
data were determined by GC/ECD or GC/FPD. Compounds not amenable to GC were
determined by HPLC. Other analytes may also be appropriate for this cleanup
technique, however, recovery through the GPC should be >70%.
1.4 Normally, this method is most efficient for removing high boiling
materials that condense in the injection port area of a gas chromatograph (GC)
or the front of the GC column. This residue will ultimately reduce the
chromatographic separation efficiency or column capacity because of adsorption
of the target analytes on the active sites. Pentachlorophenol is especially
susceptible to this problem. GPC, operating on the principal of size exclusion,
will not usually remove interference peaks that appear in the chromatogram since
the molecular size of these compounds is relative similar to the target analytes.
Separation cleanup techniques, based on other molecular characteristics (i.e.,
polarity), must be used to eliminate this type of interference.
2.0 SUMMARY OF METHOD
2.1 The column is packed with the required amount of preswelled
absorbent, and is flushed with solvent for an extended period. The column is
calibrated and then loaded with the sample extract to be cleaned up. Elution is
effected with a suitable solvent(s) and the product is then concentrated.
3.0 INTERFERENCES
3.1 A reagent blank should be analyzed for the compound of interest prior
to the use of this method. The level of interferences must be below the
estimated quantitation limits (EQLs) of the analytes of interest before this
method is performed on actual samples.
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3.2 More extensive procedures than those outlined in this method may be
necessary for reagent purification.
4.0 APPARATUS
4.1 Gel-permeation chromatography system - GPC Autoprep Model 1002 A
or B, or equivalent, Analytical Biochemical Laboratories, Inc. Systems that
perform very satisfactorily have also been assembled from the following
components - an HPLC pump, an auto sampler or a valving system with sample loops,
and a fraction collector. All systems, whether automated or manual, must meet
the calibration requirements of Sec. 7.2.2.
4.1.1 Chromatographic column - 700 mm x 25 mm ID glass column. Flow
is upward. (Optional) To simplify switching from the UV detector during
calibration to the GPC collection device during extract cleanup, attach a
double 3-way valve (Rheodyne Type 50 Teflon Rotary Valve #10-262 or
equivalent) so that the column exit flow can be shunted either to the UV
flow-through cell or to the GPC collection device.
4.1.2 Guard column - (Optional) 5 cm, with appropriate fittings to
connect to the inlet side of the analytical column (Supelco 5-8319 or
equivalent).
4.1.3 Bio Beads (S-X3) - 200-400 mesh, 70 g (Bio-Rad Laboratories,
Richmond, CA, Catalog 152-2750 or equivalent). An additional 5 g of Bio
Beads are required if the optional guard column is employed. The quality
of Bio Beads may vary from lot to lot because of excessive fines in some
lots. The UV chromatogram of the Calibration solution should be very
similar to that in Figure 2, and the backpressure should be within 6-
10 psi. Also, the gel swell ratio in methylene chloride should be in the
range of 4.4 - 4.8 mL/g. In addition to fines having a detrimental effect
on chromatography, they can also pass through the column screens and
damage the valve.
4.1.4 Ultraviolet detector - Fixed wavelength (254 nm) with a semi-
prep flow-through cell.
4.1.5 Strip chart recorder, recording integrator or laboratory data
system.
4.1.6 Syringe - 10 mL with Luerlok fitting.
4.1.7 Syringe filter assembly, disposable - Bio-Rad "Prep Disc"
sample filter assembly #343-0005, 25 mm, and 5 micron filter discs or
equivalent. Check each batch for contaminants. Rinse each filter
assembly (prior to use) with methylene chloride if necessary.
4.2 Analytical balance - 0.0001 g.
4.3 Volumetric flasks, Class A - 10 mL to 1000 mL
4.4 Graduated cylinders
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5.0 REAGENTS
5.1 Methylene chloride, CH2C12. Pesticide quality or equivalent.
5.1.1 Some brands of methylene chloride may contain unacceptably
high levels of acid (HC1). Check the pH by shaking equal portions of
methylene chloride and water, then check the pH of the water layer.
5.1.1.1 If the pH of the water layer is < 5, filter the
entire supply of solvent through a 2 in. x 15 in. glass column
containing activated basic alumina. This column should be
sufficient for processing approximately 20-30 liters of solvent.
Alternatively, find a different supply of methylene chloride.
5.2 Cyclohexane, C6H12. Pesticide quality or equivalent.
5.3 n-Butyl chloride, CHaC^C^CHjCl. Pesticide quality or equivalent.
5.4 GPC Calibration Solution. Prepare a calibration solution in
methylene chloride containing the following analytes (in elution order):
Compound mg/L
corn oil 25,000
bis(2-ethylhexyl) phthalate 1,000
methoxychlor 200
perylene 20
sulfur 80
NOTE: Sulfur is not very soluble in methylene chloride, however, it is
soluble in warm corn oil. Therefore, one approach is to weigh out
the corn oil, warm it and transfer the weighed amount of sulfur into
the warm corn oil. Mix it and then transfer into a volumetric flask
with methylene chloride, along with the other calibration compounds.
Store the calibration solution in an amber glass bottle with a Teflon lined
screw-cap at 4°C, and protect from light. (Refrigeration may cause the corn oil
to precipitate. Before use, allow the calibration solution to stand at room
temperature until the corn oil dissolves.) Replace the calibration standard
solution every 6 months, or more frequently if necessary.
5.5 Corn Oil Spike for Gravimetric Screen. Prepare a solution of corn
oil in methylene chloride (5 g/100 ml).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes, Sec.
4.1.
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7.0 PROCEDURE
7.1 It is very important to have consistent laboratory temperatures
during an entire GPC run, which could be 24 hours or more. If temperatures are
not consistent, retention times will shift, and the dump and collect times
determined by the calibration standard will no longer be appropriate. The ideal
laboratory temperature to prevent outgassing of the methylene chloride is 72°F.
7.2 GPC Setup and Calibration
7.2.1 Column Preparation
7.2.1.1 Weigh out 70 g of Bio Beads (SX-3). Transfer them
to a quart bottle with a Teflon lined cap or a 500 ml separatory
funnel with a large bore stopcock, and add approximately 300 ml of
methylene chloride. Swirl the container to ensure the wetting of
all beads. Allow the beads to swell for a minimum of 2 hours.
Maintain enough solvent to sufficiently cover the beads at all
times. If a guard column is to be used, repeat the above with 5 g
of Bio Beads in a 125 ml bottle or a beaker, using 25 ml of
methylene chloride.
7.2.1.2 Turn the column upside down from its normal
position, and remove the inlet bed support plunger (the inlet
plunger is longer than the outlet plunger). Position and tighten
the outlet bed support plunger as near the end as possible, but no
closer than 5 cm (measured from the gel packing to the collar).
7.2.1.3 Raise the end of the outlet tube to keep the
solvent in the GPC column, or close the column outlet stopcock if
one is attached. Place a small amount of solvent in the column to
minimize the formation of air bubbles at the base of poured column
packing.
7.2.1.4 Swirl the bead/solvent slurry to get a homogeneous
mixture and, if the wetting was done in a quart bottle, quickly
transfer it to a 500 ml separatory funnel with a large bore
stopcock. Drain the excess methylene chloride directly into the
waste beaker, and then start draining the slurry into the column by
placing the separatory funnel tip against the column wall. This
will help to minimize bubble formation. Swirl occasionally to keep
the slurry homogeneous. Drain enough to fill the column. Place the
tubing from the column outlet into a waste beaker below the column,
open the stopcock (if attached) and allow the excess solvent to
drain. Raise the tube to stop the flow and close the stopcock when
the top of the gel begins to look dry. Add additional methylene
chloride to just rewet the gel.
7.2.1.5 Wipe any remaining beads and solvent from the
inner walls of the top of the column with a laboratory tissue.
Loosen the seal slightly on the other plunger assembly (long
plunger) and insert it into the column. Make the seal just tight
3640A - 8 Revision 1
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enough so that any beads on the glass surface will be pushed
forward, but loose enough so that the plunger can be pushed forward.
CAUTION: Do not tighten the seal if beads are between the
seal and the glass surface because this can
damage the seal and cause leakage.
7.2.1.6 Compress the column as much as possible without
applying excessive force. Loosen the seal and gradually pull out
the plunger. Rinse and wipe off the plunger. Slurry any remaining
beads and transfer them into the column. Repeat Sec. 7.2.1.5 and
reinsert the plunger. If the plunger cannot be inserted and pushed
in without allowing beads to escape around the seal, continue
compression of the beads without tightening the seal, and loosen and
remove the plunger as described. Repeat this procedure until the
plunger is successfully inserted.
7.2.1.7 Push the plunger until it meets the gel, then
compress the column bed about four centimeters.
7.2.1.8 Pack the optional 5 cm column with approximately
5 g of preswelled beads (different guard columns may require
different amounts). Connect the guard column to the inlet of the
analytical column.
7.2.1.9 Connect the column inlet to the solvent reservoir
(reservoir should be placed higher than the top of the column) and
place the column outlet tube in a waste container. Placing a
restrictor in the outlet tube will force air out of the column more
quickly. A restrictor can be made from a piece of capillary
stainless steel tubing of 1/16" OD x 10/1000" ID x 2". Pump
methylene chloride through the column at a rate of 5 mL/min for one
hour.
7.2.1.10 After washing the column for at least one hour,
connect the column outlet tube, without the restrictor, to the inlet
side of the UV detector. Connect the system outlet to the outlet
side of the UV detector. A restrictor (same size as in Sec.
7.2.1.9) in the outlet tube from the UV detector will prevent bubble
formation which causes a noisy UV baseline. The restrictor will not
effect flow rate. After pumping methylene chloride through the
column for an additional 1-2 hours, adjust the inlet bed support
plunger until approximately 6-10 psi backpressure is achieved. Push
the plunger in to increase pressure or slowly pull outward to reduce
pressure.
7.2.1.11 When the GPC column is not to be used for several
days, connect the column outlet line to the column inlet to prevent
column drying and/or channeling. If channeling occurs, the gel must
be removed from the column, reswelled, and repoured as described
above. If drying occurs, methylene chloride should be pumped
through the column until the observed column pressure is constant
and the column appears wet. Always recalibrate after column drying
has occurred to verify retention volumes have not changed.
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7.2.2 Calibration of the GPC Column
7.2.2.1 Using a 10 ml syringe, load sample loop #1 with
calibration solution (Sec. 5.6). With the ABC automated system, the
5 ml sample loop requires a minimum of 8 ml of the calibration
solution. Use a firm, continuous pressure to push the sample onto
the loop. Switch the valve so that GPC flow is through the UV flow-
through cell.
7.2.2.2 Inject the calibration solution and obtain a UV
trace showing a discrete peak for each component. Adjust the
detector and/or recorder sensitivity to produce a UV trace similar
to Figure 2 that meets the following requirements. Differences
between manufacturers' cell volumes and detector sensitivities may
require a dilution of the calibration solution to achieve similar
results. An analytical flow-through detector cell will require a
much less concentrated solution than the semi-prep cell, and
therefore the analytical cell is not acceptable for use.
7.2.2.3 Following are criteria for evaluating the UV
chromatogram for column condition.
7.2.2.3.1 Peaks must be observed, and should be
symmetrical, for all compounds in the calibration solution.
7.2.2.3.2 Corn oil and phthalate peaks must exhibit
>85% resolution.
7.2.2.3.3 Phthalate and methoxychlor peaks must
exhibit >85% resolution.
7.2.2.3.4 Methoxychlor and perylene peaks must exhibit
>85% resolution.
7.2.2.3.5 Perylene and sulfur peaks must not be
saturated and must exhibit >90% baseline resolution.
7.2.2.3.6 Nitroaromatic compounds are particularly
prone to adsorption. For example, 4-nitrophenol recoveries
may be low due to a portion of the analyte being discarded
after the end of the collection time. Columns should be
tested with the semivolatiles matrix spiking solution. GPC
elution should continue until after perylene has eluted, or
long enough to recover at least 85% of the analytes, whichever
time is longer.
7.2.2.4 Calibration for Semivolatiles - Using the
information from the UV trace, establish appropriate collect and
dump time periods to ensure collection of all target analytes.
Initiate column eluate collection just before elution of
bis(2-ethylhexyl) phthalate and after the elution of the corn oil.
Stop eluate collection shortly after the elution of perylene.
Collection should be stopped before sulfur elutes. Use a "wash"
time of 10 minutes after the elution of sulfur. Each laboratory is
3640A - 10 Revision 1
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required to establish its specific time sequences. See Figure 2 for
general guidance on retention time. Figure 1 illustrates retention
volumes for different classes of compounds.
7.2.2.5 Calibration for Organochlorine Pesticides/PCBs -
Determine the elution times for the phthalate, methoxychlor,
perylene, and sulfur. Choose a dump time which removes >85% of the
phthalate, but collects >95% of the methoxychlor. Stop collection
after the elution of perylene, but before sulfur elutes.
7.2.2.6 Verify the flow rate by collecting column eluate
for 10 minutes in a graduated cylinder and measure the volume, which
should be 45-55 ml (4.5-5.5 mL/min). If the flow rate is outside of
this range, corrective action must be taken, as described above.
Once the flow rate is within the range of 4.5-5.5 mL/min, record the
column pressure (should be 6-10 psi) and room temperature. Changes
in pressure, solvent flow rate, and temperature conditions can
affect analyte retention times, and must be monitored. If the flow
rate and/or column pressure do not fall within the above ranges, a
new column should be prepared. A UV trace that does not meet the
criteria in Sec. 7.2.2.3 would also indicate that a new column
should be prepared. It may be necessary to obtain a new lot of Bio
Beads if the column fails all the criteria.
7.2.2.7 Reinject the calibration solution after
appropriate collect and dump cycles have been set, and the solvent
flow and column pressure have been established.
7.2.2.7.1 Measure and record the volume of collected
GPC eluate in a graduated cylinder. The volume of GPC eluate
collected for each sample extract processed may be used to
indicate problems with the system during sample processing.
7.2.2.7.2 The retention times for bis(2-ethylhexyl)
phthalate and perylene must not vary more than +5% between
calibrations. If the retention time shift is >5%, take
corrective action. Excessive retention time shifts are caused
by:
7.2.2.7.2.1 Poor 1 aboratory temperature control or
system leaks.
7.2.2.7.2.2 An unstabilized column that requires
pumping methylene chloride through it for several more
hours or overnight.
7.2.2.7.2.3 Excessive laboratory temperatures,
causing outgassing of the methylene chloride.
7.2.2.8 Analyze a GPC blank by loading 5 ml of methylene
chloride into the GPC. Concentrate the methylene chloride that
passes through the system during the collect cycle using a Kuderna-
Danish (KD) evaporator. Analyze the concentrate by whatever
detectors will be used for the analysis of future samples. Exchange
3640A - 11 Revision 1
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the solvent, if necessary. If the blank exceeds the estimated
quantitation limit of the analytes, pump additional methylene
chloride through the system for 1-2 hours. Analyze another GPC
blank to ensure the system is sufficiently clean. Repeat the
methylene chloride pumping, if necessary.
7.3 Extract Preparation
7.3.1 Adjust the extract volume to 10.0 ml. The solvent extract
must be primarily methylene chloride. All other solvents, e.g.
1:1 methylene chloride/acetone, must be concentrated to 1 ml (or as low as
possible if a precipitate forms) and diluted to 10.0 ml with methylene
chloride. Thoroughly mix the extract before proceeding.
7.3.2 Filter the extract through a 5 micron filter disc by attaching
a syringe filter assembly containing the filter disc to a 10 ml syringe.
Draw the sample extract through the filter assembly and into the 10 ml
syringe. Disconnect the filter assembly before transferring the sample
extract into a small glass container, e.g. a 15 ml culture tube with a
Teflon lined screw cap. Alternatively, draw the extract into the syringe
without the filter assembly. Attach the filter assembly and force the
extract through the filter and into the glass container. The latter is
the preferred technique for viscous extracts or extracts with a lot of
solids. Particulate larger than 5 microns may scratch the valve, which
may result in a system leak and cross-contamination of sample extracts in
the sample loops. Repair of the damaged valve is quite expensive.
NOTE: Viscosity of a sample extract should not exceed the viscosity
of 1:1 water/glycerol. Dilute samples that exceed this
viscosity.
7 A Screening the Extract
7.4.1 Screen the extract to determine the weight of dissolved
residue by evaporating a 100 juL aliquot to dryness and weighing the
residue. The weight of dissolved residue loaded on the GPC column cannot
exceed 0.500 g. Residues exceeding 0.500 g will very likely result in
incomplete extract cleanup and contamination of the GPC switching valve
(which results in cross-contamination of sample extracts).
7.4.1.1 Transfer 100 /zL of the filtered extract from
Sec. 7.3.2 to a tared aluminum weighing dish.
7.4.1.2 A suggested evaporation technique is to use a heat
lamp. Set up a 250 watt heat lamp in a hood so that it is
8 + 0.5 cm from a surface covered with a clean sheet of aluminum
foil. Surface temperature should be 80-100°C (check temperature by
placing a thermometer on the foil and under the lamp). Place the
weighing dish under the lamp using tongs. Allow it to stay under
the lamp for 1 min. Transfer the weighing dish to an analytical
balance or a micro balance and weigh to the nearest 0.1 mg. If the
residue weight is less than 10 mg/100 jLtL, then further weighings are
not necessary. If the residue weight is greater than 10 mg/100 /zL,
3640A - 12 Revision 1
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then determine if constant weight has been achieved by placing the
weighing dish and residue back under the heat lamp for 2 or more
additional 0.5 min. intervals. Reweigh after each interval.
Constant weight is achieved when three weights agree within +10%.
7.4.1.3 Repeat the above residue analysis on a blank and
a spike. Add 100 fj,l of the same methylene chloride used for the
sample extraction to a weighing dish and determine residue as above.
Add 100 jitL of a corn oil spike (5 g/100 mL) to another weighing dish
and repeat the residue determination.
7.4.2 A residue weight of 10 mg/100 /zL of extract represents 500 mg
in 5 ml of extract. Any sample extracts that exceed the 10 mg/100 /zL
residue weight must be diluted so that the 5 ml loaded on the GPC column
does not exceed 0.500 g. When making the dilution, keep in mind that a
minimum volume of 8 ml is required when loading the ABC GPC unit.
Following is a calculation that may be used to determine what dilution is
necessary if the residue exceeds 10 mg.
Y ml taken =
for dilution
Example:
Y ml taken =
for dilution
10 ml final
volume
10 ml final
volume
10 mq maximum
X mg of residue
10 mq maximum
15 mg of residue
Y ml taken for dilution = 6.7 ml
Therefore, taking 6.7 ml of sample extract from Sec. 7.3.2, and
diluting to 10 ml with methylene chloride, will result in 5 ml of diluted
extract loaded on the GPC column that contains 0.500 g of residue.
NOTE: This dilution factor must be included in the final calculation
of analyte concentrations. In the above example, the dilution
factor is 1.5.
7.5 GPC Cleanup
7.5.1 Calibrate the GPC at least once per week following the
procedure outlined in Sees. 7.2.2 through 7.2.2.6. Ensure that UV trace
requirements, flow rate and column pressure criteria are acceptable.
Also, the retention time shift must be <5% when compared to retention
times in the last calibration UV trace.
7.5.1.1 If these criteria are not met, try cleaning the
column by loading one or more 5 ml portions of butyl chloride and
running it through the column. Butyl chloride or 9:1 (v/v)
methylene chloride/methanol removes the discoloration and
particulate that may have precipitated out of the methylene chloride
extracts. Backflushing (reverse flow) with methylene chloride to
dislodge particulates may restore lost resolution. If a guard
column is being used, replace it with a new one. This may correct
3640A - 13
Revision 1
September 1994
-------
the problem. If column maintenance does not restore acceptable
performance, the column must be repacked with new Bio Beads and
calibrated.
7.5.2 Draw a minimum of 8 ml of extract (diluted, if necessary, and
filtered) into a 10 ml syringe.
7.5.3 Attach the syringe to the turn lock on the injection port.
Use firm, continuous pressure to push the sample onto the 5-mL sample
loop. If the sample is difficult to load, some part of the system may be
blocked. Take appropriate corrective action. If the back pressure is
normal (6-10 psi), the blockage is probably in the valve. Blockage may be
flushed out of the valve by reversing the inlet and outlet tubes and
pumping solvent through the tubes. (This should be done before sample
loading.)
NOTE: Approximately 2 ml of the extract remains in the lines between
the injection port and the sample loop; excess sample also
passes through the sample loop to waste.
7.5.4 After loading a loop, and before removing the syringe from the
injection port, index the GPC to the next loop. This will prevent loss of
sample caused by unequal pressure in the loops.
7.5.5 After loading each sample loop, wash the loading port with
methylene chloride in a PTFE wash bottle to minimize cross-contamination.
Inject approximately 10 ml of methylene chloride to rinse the common
tubes.
7.5.6 After loading all the sample loops, index the GPC to the 00
position, switch to the "RUN" mode and start the automated sequence.
Process each sample using the collect and dump cycle times established in
Sec. 7.2.2.
7.5.7 Collect each sample in a 250 ml Erlenmeyer flask, covered with
aluminum foil to reduce solvent evaporation, or directly into a Kuderna-
Danish evaporator. Monitor sample volumes collected. Changes in sample
volumes collected may indicate one or more of the following problems:
7.5.7.1 Change in solvent flow rate, caused by channeling
in the column or changes in column pressure.
7.5.7.2 Increase in column operating pressure due to the
absorption of particles or gel fines onto either the guard column or
the analytical column gel, if a guard column is not used.
7.5.7.3 Leaks in the system or significant variances in
room temperature.
7.6 Concentrate the extract by the standard K-D technique (see any of the
extraction methods, Sec. 4.2.1 of this chapter). See the determinative methods
(Chapter Four, Sec. 4.3) for the final volume.
3640A - 14 Revision 1
September 1994
-------
7.7 It should be remembered that only half of the sample extract is
processed by the GPC (5 ml of the 10 ml extract is loaded onto the GPC column),
and thus, a dilution factor of 2 (or 2 multiplied by any dilution factor in Sec.
7.4.2) must be used for quantitation of the sample in the determinative method.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 3600 for specific quality control
procedures.
8.2 The analyst should demonstrate that the compound(s) of interest are
being quantitatively recovered before applying this method to actual samples.
8.3 For sample extracts that are cleaned up using this method, the
associated quality control samples must also be processed through this cleanup
method.
9.0 METHOD PERFORMANCE
9.1 Refer to Table 1 for single laboratory performance data.
10.0 REFERENCES
1. Wise, R.H.; Bishop, D.F.; Williams, R.T.; Austern, B.M. "Gel Permeation
Chromatography in the GC/MS Analysis of Organics in Sludges"; U.S. EPA
Municipal Environmental Research Laboratory: Cincinnati, Ohio 45268.
2. Czuczwa, J.; Alford-Stevens, A. "Optimized Gel Permeation Chromatographic
Cleanup for Soil, Sediment, Waste and Waste Oil Sample Extracts for GC/MS
Determination of Semivolatile Organic Pollutants, JAOAC, submitted April
1989.
3. Marsden, P.J.; Taylor, V.; Kennedy, M.R. "Evaluation of Method 3640 Gel
Permeation Cleanup"; Contract No. 68-03-3375, U.S. Environmental
Protection Agency, Cincinnati, Ohio, pp. 100, 1987.
3640A - 15 Revision 1
September 1994
-------
TABLE 1
GPC RECOVERY AND RETENTION VOLUMES FOR RCRA
APPENDIX VIII ANALYTES
Compound
Acenaphthene
Acenaphthylene
Acetophenone
2-Acetylaminofluorene
Aldn'n
4-Aminobiphenyl
Aniline
Anthracene
Benomyl
Benzenethiol
Benzidine
Benz(a)anthracene
Benzo(b)fluoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Benzo(k)fluoranthene
Benzoic acid
Benzotrichloride
Benzyl alcohol
Benzyl chloride
alpha-BHC
beta-BHC
gamma- BHC
delta-BHC
4-Bromophenyl phenyl ether
Butyl benzyl phthalate
2-sec-butyl-4,6-dinitrophenol (Dinoseb)
Carbazole
Carbendazim
alpha-Chlordane
gamma-Chlordane
4-Chloro-3-methyl phenol
4-Chloroaniline
Chi orobenzi late
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
2-Chloronaphthalene
2-Chlorophenol
3-Chlorophenol
4-Chlorophenol
4-Chlorophenyl phenyl ether
3-Chloropropionitrile
Chrysene
2-Cresol
% Rec1
97
72
94
97
99
96
93
89
131
92
95
100
93
93
90
91
66
93
95
99
84
94
93
102
93
104
103
99
131
97
93
87
88
92
89
76
83
89
90
86
87
98
80
102
91
% RSD2
2
10
7
2
9
7
4
2
8
11
5
3
5
3
6
4
7
7
17
4
13
9
4
7
1
3
18
5
8
2
2
1
3
5
1
2
2
1
1
3
2
2
5
1
1
Ret. Vol.3 (ml
196-235
196-235
176-215
156-195
196-215
176-215
196-235
196-235
146-195
196-235
176-215
196-235
196-235
196-235
196-235
196-235
176-195
176-215
176-215
176-215
196-215
196-215
196-215
216-255
176-215
136-175
176-195
196-255
146-195
196-235
196-215
196-255
196-235
176-235
156-195
156-215
156-195
196-235
196-215
196-215
196-215
176-215
176-215
196-235
196-215
3640A - 16
Revision 1
September 1994
-------
TABLE 1 (continued)
Compound
3-Cresol
4-Cresol
Cyclophosphamide
ODD
DDE
DDT
Di-n-butyl phthalate
Dial! ate
Dibenzo(a,e)pyrene
Dibenzo(a,i)pyrene
Dibenz(a, j)acridine
Dibenz( a, h) anthracene
Dibenzofuran
Dibenzothiophene
l,2-Dibromo-3-chloropropane
1,2-Dibromoethane
trans-l,4-Dichloro-2-butene
cis-l,4-Dichloro-2-butene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1 ,4-Dichlorobenzene
3,3'-Dichlorobenzidine
2,6-Dichlorophenol
2,4-Dichlorophenoxyacetic acid (2,4-D)
2,4-Dichlorophenol
2,4-Dichlorotoluene
l,3-Dichloro-2-propanol
Dieldrin
Diethyl phthalate
Dimethoate
3,3'-Dimethoxybenzidinea
Dimethyl phthalate
p-Dimethylaminoazobenzene
7, 12-Dimethyl -benz(a)anthracene
2,4-Dimethylphenol
3, 3' -Dimethyl benzi dine
4,6-Dinitro-o-cresol
1,3-Dinitrobenzene
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Diphenylamine
Diphenyl ether
1 , 2-Di phenyl hydrazine
Disulfoton
Endosulfan sulfate
Endosulfan I
% Rec1
70
88
114
94
94
96
104
97
94
99
117
92
94
94
83
121
107
106
81
81
81
98
86
80
87
70
73
100
103
79
15
100
96
77
93
93
100
99
118
93
101
95
67
92
81
94
99
%RSD2
3
2
10
4
2
6
3
6
10
8
9
5
1
3
2
8
6
6
1
1
1
3
3
NA
2
9
13
5
3
15
11
1
1
1
2
2
1
2
7
4
2
6
12
1
15
2
8
Ret. Vol.3 (ml
196-215
196-215
146-185
196-235
196-235
176-215
136-175
156-175
216-235
216-235
176-195
196-235
176-235
196-235
176-215
196-215
176-195
176-215
196-235
196-235
196-235
176-215
196-215
76-215
96-215
196-235
176-215
196-215
136-195
146-185
156-195
156-195
176-215
176-215
176-215
156-215
156-195
156-195
176-195
156-195
156-175
176-235
196-215
176-215
146-165
176-195
176-215
3640A - 17
Revision 1
September 1994
-------
TABLE 1 (continued)
Compound
Endosulfan II
Endrin
Endrin aldehyde
Endrin ketone
Ethyl methane sulfonate
Ethyl methacrylate
Bis(2-ethylhexyl) phthalate
Famphur
Fluorene
Fluoranthene
Heptachlor
Heptachlor epoxide
Hexachl orobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachl oroethane
Hexachl oropropene
Indeno(l,2,3-cd)pyrene
Isodrin
Isophorone
cis-Isosafrole
trans-Isosafrole
Kepone
Malononitrile
Merphos
Methoxychlor
3-Methylchol anthrene
2-Methyl naphthal ene
Methyl parathion
4,4'-Methylene-bis(2-chloroaniline)
Naphthalene
1,4-Naphthoquinone
2-Naphthylamine
1-Naphthylamine
5-Nitro-o-toluidine
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Nitrobenzene
2-Nitrophenol
4-Nitrophenol
N-Nitroso-di-n-butylamine
N-Nitrosodiethanolamine
N-Nitrosodi ethyl ami ne
N-Nitrosodimethylamine
N-Nitrosodiphenylamine
N-Nitrosodi-n-propylamine
% Rec1
92
95
97
94
62
126
101
99
95
94
85
91
108
86
89
85
91
79
98
68
90
88
102
111
93
94
74
67
84
96
95
73
94
96
77
96
96
103
86
95
77
89
104
94
86
99
85
%RSD2
6
6
1
4
7
7
1
NA
1
1
2
11
2
2
3
1
2
13
5
7
4
16
NA
9
12
6
12
6
13
1
7
7
8
6
2
8
2
8
2
3
3
4
3
2
13
2
4
Ret. Vol.3 (ml
196-215
196-215
176-215
176-215
176-235
176-195
120-145
126-165
176-235
196-235
195-215
156-195
196-235
176-215
176-215
196-235
196-235
216-255
196-235
156-195
176-215
156-195
196-235
156-195
126-165
156-195
176-195
196-215
146-185
176-215
196-215
176-215
196-235
196-235
176-195
176-215
176-215
176-215
176-195
176-195
196-215
156-175
146-185
156-175
156-195
156-195
156-175
3640A - 18
Revision 1
September 1994
-------
TABLE 1 (continued)
Compound
N-Ni trosomethyl ethyl ami ne
N-Nitrosomorphol ine
N-Nitrosopiperidine
N-Nitrosopyrol idine
Di-n-octyl phthalate
Parathion
Pentachl orobenzene
Pentachl oroethane
Pentachl oronitrobenzene (PCNB)
Pentachl orophenol
Phenacetin
Phenanthrene
Phenol
1,2-Phenylenediamine
Phorate
2-Picoline
Pronamide
Pyrene
Resorcinol
Safrole
Streptozotocin8
1,2, 4, 5-Tetrachl orobenzene
2, 3, 5, 6-Tetrachloro -nitrobenzene
2,3,4 , 6-Tetrachl orophenol
2,3,5,6-Tetrachlorophenol
Tetraethyl dithiopyrophosphate (Sulfotep)
Thiosemicarbazide
2-Toluidine
4-Toluidine
Thiourea, l-(o-chlorophenyl)
Toluene-2,4-diamine
1,2, 3 -Trichl orobenzene
1,2, 4 -Trichl orobenzene
2, 4, 5-Trichl orophenol
2, 4, 6-Trichl orophenol
2,4,5-Trichlorophenoxyacetic acid (2,4,5-T)
2,4,5-Trichlorophenoxypropionic acid
Warfarin
% Rec1
83
86
84
92
83
109
95
74
91
102
100
94
83
91
74
99
105
98
70
93
6
96
85
95
96
89
74
92
87
75
69
87
89
77
95
71
67
94
%RSD2
7
4
4
1
4
14
2
1
8
1
3
2
2
1
NA
14
15
2
6
1
48
2
9
1
7
14
3
3
8
11
7
1
1
1
1
23
NA
2
Ret. Vol.3 (ml
156-175
156-195
156-195
156-175
120-156
146-170
196-235
196-235
156-195
196-215
156-195
196-235
156-195
196-215
116-135
156-215
156-195
215-235
196-215
176-215
225-245
196-235
176-215
196-215
196-215
116-135
146-185
176-235
176-235
166-185
176-215
196-235
196-235
216-235
216-235
156-235
216-215
166-185
NA = Not applicable, recovery presented as the average of two determinations.
a Not an appropriate analyte for this method.
1 The percent recovery is based on an average of three recovery values.
2 The % relative standard deviation is determined from three recovery values.
3 These Retention Volumes are for guidance only as they will differ from column to
column and from system to system.
3640A - 19
Revision 1
September 1994
-------
Figure 1
GPC RETENTION VOLUME OF CLASSES OF ANALYTES
W//////////////,
W/////////////A
PHTHALATB -»•
onOANOPHOSPHATE
PESTICIDES
CORN
PAH'*
CHLOROBENZENES
NITR03AMINE3, NITROAROMATICS
AROMATIC AMINES
NITROPHCNOL3
CHLOROPHENOL3
OROANOCHLORINE
PESTICIDES/PCS'*
HERBICIDES
-------
Figure 2
UV CHROMATOGRAM OF THE CALIBRATION SOLUTION
Injection
5 fflls
on column
— 0 minaces
Corn oil
25 mg/nL
Bis(2-ethylhesyl) -phthaiate
1.0 mg/niL
Methoxychlor
0.2 ng/mL
Perylene
0.02 mg/mL
Sulfur
0.08 mg/oL — ","~
15 minuces
"' 30 minutes
45 minutes
700 am X25 am col-
70 g Bio-Beads SX
Bed length » 490
CH.C12 at 5.0 uL
254 nu
1-1.: ~ "1_ "_:.._"......"_! 60 minutes
* f _, .
mn« __
3640A - 21
Revision 1
September 1994
-------
METHOD 3640A
GEL-PERMEATION CLEANUP
7 1 Ensure ambient temp, consistent
throughout GPC run.
7.2 GPC Setup and Calibration
7 2.1 Column Preparation
I
7.2.1.1 Place Bio Beads and MeCI
in a container. Swirl and
allow beads to swell.
I
7 2.1.2 Remove column inlet bed
support plunger. Position and tighten
outlet bed support plunger to column end.
7 2.1 3 Ensure GPC column outlet
contains solvent. Place small amount
solvent in column to minimize
bubble formation.
7.2.1.4 Transfer bead mixture into
sep. funnel. Drain excess solvent;
drain beads into column. Keep
beads wet throughout.
7.2 1 5 Loosen seal on opposite
plunger assembly, insert into column.
1
7 2.1.6 Compress column Slurry
remaining beads and repeat Section
7 2.1.5 and column compression.
7.2.1.7 Compress column bed
approximately four cm.
7 2.1.8 Pack option 5 cm. guard
column w/ roughly 5 gm.
preswelled beads
7 2.1.9 Connect column inlet to
solvent reservoir. Pump MeCI at
5 ml/mm for 1 hr
I
7.2.1.10 Connect column outlet to
UV-Vis detector Place restrictor
at detector outlet. Run MeCI for
additional 1 -2 hrs. Compress
column bed to provide 6-10 psi
backpressure.
7.2.1.11 Connect outlet line to column
inlet when column not in use. Repack
column when channeling is observed.
Assure consistent backpressure when
beads are rewetted after drying.
3640A - 22
Revision 1
September 1994
-------
METHOD 3640A
continued
7 2.2 Calibration of the GPC column
7 2.2.1 Load sample loop with
calibration solution.
I
7 2.2.2 Inject calibration soln.; adjust
recorder or detector sensitivity
to produce similar UV trace as Fig 2
7 2.2.3 Evaluation criteria for
UV chromatogram.
7.2 2.4 Calibration for Semivolatiles
Use information from UV trace to
obtain collect and dump times.
Initiate collection before bis(2-ethylhexyl)
phthalate, stop after perylene. Stop run
before sulfur e/utes.
7 2.2.5 Calibration for Organochlonne
Pesticides/PCBs
Choose dump time which removes
> 85% phthalate, but collects at
times > 95% methoxychlor Stop
collection between perylene and
sulfur elution
I
7 2.2.6 Verify column flow rate and
backpressure Correct
inconsistencies when criteria
are not met.
7.2.2.7 Reinject calibration soln when
collect and dump cycles are set,
and column criteria are met
7 2.2.7 1 Measure and record
volume of GPC eluate
7.2.2.7 2 Correct for retention time
shifts of > W- 5% for
bis(2-ethylhexyl) phthalate
and perylene.
7.2.2.8 Inject and analyze GPC blank
for column cleanliness Pump
through MeCI as column wash.
3640A - 23
Revision 1
September 1994
-------
METHOD 3640A
continued
V
7 3 Extract Preparation
*
7.3.1 Adjust extract volume to 10 ml.
Primary solvent should be MeCI.
*
7.3.2 Filter extract through 5 micron filter
disc/syringe assembly into small
glass container.
*
7.4 Screening the Extract
*
7.4.1 Screen extract by determining
residue wt. of 100 uL aliquot.
*
7.4.1 .1 Transfer 100 uL of filtered
extract from Section 7.3.2 to tared
aluminum weighing dish.
*
7.4.1 .2 Evaporate extract solvent under
heating lamp. Weigh residue to nearest
0 1 mg.
*
7.4.1 .3 Repeat residue analysis of Section
7412 w/blank and spike sample.
*
7 .4 2 Use dilution example to determine
necessary dilution when residue
wts. > 10mg.
1 '
7.5 GPC Cleanup
*
7.5.1 Calibrate GPC weekly. Assure
column criteria. UV trace, retention
time shift criteria are met.
*
7.5.1 .1 Clean column w/butyl chloride
loadings, or replacement of
guard column.
*
7.5.2 Draw 8 ml extract into syringe.
*
7.5.3 Load sample into injection loop.
I
7.5.4 Index GPC to next loop to
prevent sample loss.
1
7.5.5 Wash sample port w/MeCI
between sample loadings.
*
7 5.6 At end of loadings, index GPC to
00, switch to "RUN" mode, start
automated sequence.
*
7.5 7 Collect sample into aluminum foil
covered Erlenmeyer flask or into
Kuderna- Danish evaporator
i
7.6 Concentrate extract by std.
Kuderna-Danish technique.
*
7.7 Note dilution factor of GPC method
into final determinations.
3640A - 24
Revision 1
September 1994
-------
METHOD 3650A
ACID-BASE PARTITION CLEANUP
1.0 SCOPE AND APPLICATION
manual.
1.1 Method 3650 was formerly Method 3530 in the second edition of this
1.2 Method 3650 is a liquid-liquid partitioning cleanup method to
separate acid analytes, e.g. organic acids and phenols, from base/neutral
analytes, e.g. amines, aromatic hydrocarbons, and halogenated organic compounds,
using pH adjustment. It may be used for cleanup of petroleum waste prior to
analysis or further cleanup (e.g., alumina cleanup). The following compounds can
be separated by this method:
Compound Name
CAS No.a
Fraction
Benz ( a ) anthracene
Benzo(a)pyrene
Benzo(b)fluoranthene
Chlordane
Chlorinated dibenzodioxins
2-Chlorophenol
Chrysene
Creosote
Cresol(s)
Dichlorobenzene(s)
Dichlorophenoxyacetic acid
2, 4-Dimethyl phenol
Dinitrobenzene
4,6-Dinitro-o-cresol
2,4-Dinitrotoluene
Heptachlor
Hexachlorobenzene
Hexachl orobutadi ene
Hexachloroethane
Hexachl orocyclopentadi ene
Naphthalene
Nitrobenzene
4-Nitrophenol
Pentachlorophenol
Phenol
Phorate
2-Picoline
Pyridine
Tetrachlorobenzene(s)
Tetrachlorophenol (s)
Toxaphene
Trichlorophenol(s)
2,4,5-TP (Silvex)
56-55-3
50-32-8
205-99-2
57-74-9
95-57-8
218-01-9
8001-58-9
94-75-7
105-67-9
25154-54-5
534-52-1
121-14-2
76-44-8
118-74-1
87-68-3
67 -.72-1
77-47-4
91-20-3
98-95-3
100-02-7
87-86-5
108-95-2
298-02-2
109-06-8
110-86-1
8001-35-2
93-72-1
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Acid
Base-neutral
Base-neutral and Acid
Acid
Base-neutral
Acid
Acid
Base-neutral
Acid
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Acid
Acid
Acid
Base-neutral
Base-neutral
Base-neutral
Base-neutral
Acid
Base-neutral
Acid
Acid
Chemical Abstract Services Registry Number.
3650A - 1
Revision 1
July 1992
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2.0 SUMMARY OF METHOD
2.1 The solvent extract from a prior solvent extraction method is shaken
with water that is strongly basic. The acid analytes partition into the aqueous
layer, whereas, the basic and neutral compounds stay in the organic solvent. The
base/neutral fraction is concentrated and is then ready for further cleanup, if
necessary, or analysis. The aqueous layer is acidified and extracted with an
organic solvent. This extract is concentrated (if necessary) and is then ready
for analysis of the acid analytes.
3.0 INTERFERENCES
3.1 More extensive procedures than those outlined in this method may be
necessary for reagent purification.
3.2 A method blank must be run for the compounds of interest prior to
use of the method. The interferences must be below the method detection limit
before this method is applied to actual samples.
4.0 APPARATUS AND MATERIALS
4.1 Drying column - 20 mm ID Pyrex chromatographic column with Pyrex
glass wool at bottom, or equivalent.
NOTE: Fritted glass discs are difficult to clean after highly contaminated
extracts have been passed through them. Columns without frits are
recommended. Use a small pad of Pyrex glass wool to retain the
adsorbent. Prewash the glass wool pad with 50 ml of acetone
followed by 50 ml of elution solvent prior to packing the column
with adsorbent.
4.2 Kuderna-Danish (K-D) apparatus
4.2.1 Concentrator tube - 10 ml graduated (Kontes K570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of the
extracts.
4.2.2 Evaporation flask - 500 ml (K-570001-0500 or equivalent).
Attach to concentrator tube with springs, clamps, or equivalent.
4.2.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Snyder column - Two ball micro (Kontes K569001-0219 or
equivalent).
tops.
4.2.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Vials - Glass, 2 ml capacity with Teflon lined screw-caps or crimp
4.4 Water bath - Heated, concentric ring cover, temperature control of
± 2°C. Use this bath in a hood.
3650A - 2 Revision 1
July 1992
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4.5 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.6 pH indicator paper - pH range including the desired extraction pH.
4.7 Separatory funnel - 125 mi.
4.8 Erlenmeyer flask - 125 ml.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests.
Unless otherwise indicated, it is intended that all inorganic reagents shall
conform to the specifications of the Committee on Analytical Reagents of the
American Chemical Society, where such specifications are available. Other grades
may be used, provided it is first ascertained that the reagent is of sufficiently
high purity to permit its use without lessening the accuracy of the
determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium hydroxide, NaOH, (ION) - Dissolve 40 g of sodium hydroxide
in 100 ml of organic-free reagent water.
5.4 Sulfuric acid, H?S04, (1:1 v/v in water) - Slowly add 50 ml H2S04 to
50 ml of organic-free reagent water.
5.5 Sodium sulfate (granular, anhydrous), Na2SO, - Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.6 Solvents:
5.6.1 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.6.2 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.6.3 Methanol, CH3OH - Pesticide quality or equivalent.
5.6.4 Diethyl Ether, C2H5OC2H5 - Pesticide quality or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl alcohol preservative must be
added to each liter of ether.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
3650A - 3 Revision 1
July 1992
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7.0 PROCEDURE
7.1 Place 10 ml of the solvent extract from a prior extraction procedure
into a 125 ml separatory funnel.
7.2 Add 20 mL of methylene chloride to the separatory funnel.
7.3 Slowly add 20 ml of prechilled organic-free reagent water which has
been previously adjusted to a pH of 12-13 with ION sodium hydroxide.
7.4 Seal and shake the separatory funnel for at least 2 minutes with
periodic venting to release excess pressure.
NOTE: Methylene chloride creates excessive pressure very rapidly;
therefore, initial venting should be done immediately after the
separatory funnel has been sealed and shaken once. The separatory
funnel should be vented into a hood to prevent unnecessary exposure
of the analyst to the organic vapor.
7.5 Allow the organic layer to separate from the aqueous phase for a
minimum of 10 minutes. If the emulsion interface between layers is more than
one-third the size of the solvent layer, the analyst must employ mechanical
techniques to complete the phase separation. The optimum technique depends upon
the sample, and may include stirring, filtration of the emulsion through glass
wool, centrifugation, or other physical methods.
7.6 Separate the aqueous phase and transfer it to a 125 ml Erlenmeyer
flask. Repeat the extraction two more times using 20 ml aliquots of dilute
sodium hydroxide (pH 12-13). Combine the aqueous extracts.
7.7 Water soluble organic acids and phenols will be primarily in the
aqueous phase. Base/neutral analytes will be in the methylene chloride. If the
analytes of interest are only in the aqueous phase, discard the methylene
chloride and proceed to Section 7.8. If the analytes of interest are only in the
methylene chloride, discard the aqueous phase and proceed to Section 7.10.
7.8 Externally cool the 125 mL Erlenmeyer flask with ice while adjusting
the aqueous phase to a pH of 1-2 with sulfuric acid (1:1). Quantitatively
transfer the cool aqueous phase to a clean 125 mL separatory funnel. Add 20 mL
of methylene chloride to the separatory funnel and shake for at least 2 minutes.
Allow the methylene chloride to separate from the aqueous phase and collect the
methylene chloride in an Erlenmeyer flask.
7.9 Add 20 mL of methylene chloride to the separatory funnel and extract
at pH 1-2 a second time. Perform a third extraction in the same manner combining
the extracts in the Erlenmeyer flask.
7.10 Assemble a Kuderna-Danish (K-D) concentrator (if necessary) by
attaching a 10 mL concentrator tube to a 500 mL evaporation flask.
7.11 Dry both acid and base/neutral fractions by passing them through a
drying column containing about 10 cm of anhydrous sodium sulfate. Collect the
dried fractions in K-D concentrators. Rinse the Erlenmeyer flasks which
3650A - 4 Revision 1
July 1992
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contained the solvents and the columns with 20 ml of methylene chloride to
complete the quantitative transfer.
7.12 Concentrate both acid and base/neutral fractions as follows: Add
one or two boiling chips to the flask and attach a three ball macro-Snyder
column. Prewet the Snyder column by adding about 1 ml of methylene chloride to
the top of the column. Place the K-D apparatus on a hot water bath (80-90°C) so
that the concentrator tube is partially immersed in the warm water. Adjust the
vertical position of the apparatus and the water temperature as required to
complete the concentration in 15-20 minutes. At the proper rate of distillation,
the balls of the column will actively chatter but the chambers will not flood.
When the apparent volume of liquid reaches 1 ml, remove the K-D apparatus from
the water bath and allow it to cool. Remove the Snyder column and rinse the
flask and its lower joints into the concentrator tube with 1-2 ml of methylene
chloride. Concentrate the extract to the final volume using either the micro-
Snyder column technique (7.12.1) or nitrogen blowdown technique (7.12.2).
7.12.1 Micro-Snyder Column Technique
7.12.1.1 Add another one or two boiling chips to the
concentrator tube and attach a two ball micro-Snyder column. Prewet
the column by adding 0.5 ml of methylene chloride to the top of the
column. Place the K-D apparatus in a hot water bath (80-90°C) so
that the concentrator tube is partially immersed in the hot water.
Adjust the vertical position of the apparatus and the water
temperature as required to complete the concentration in 5-10
minutes. At the proper rate of distillation the balls of the column
will actively chatter but the chambers will not flood. When the
apparent volume of the liquid reaches 0.5 ml, remove the K-D
apparatus and allow it to cool. Remove the Snyder column and rinse
the flask and its lower joints into the concentrator tube with 0.2
ml of methylene chloride. Adjust the final volume to 1 ml with
methylene chloride.
7.12.2 Nitrogen Blowdown Technique
7.12.2.1 Place the concentrator tube in a warm water bath
(35°C) and evaporate the solvent volume to 1.0-2.0 ml using a gentle
stream of clean, dry nitrogen (filtered through a column of
activated carbon).
CAUTION: Do not use plasticized tubing between the carbon
trap and the sample.
7.12.2.2 The internal wall of the concentrator tube must be
rinsed down several times with the appropriate solvent during the
operation. During evaporation, the tube solvent level must be
positioned to avoid condensation water. Under normal procedures,
the extract must not be allowed to become dry.
CAUTION: When the volume of solvent is reduced below 1 ml,
semivolatile analytes may be lost.
7.13 The acid fraction is now ready for analysis. If the base/neutral
3650A - 5 Revision 1
July 1992
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fraction requires further cleanup by the alumina column cleanup for petroleum
waste (Method 3611), the solvent may have to be changed to hexane. If a solvent
exchange is required, momentarily remove the Snyder column, add approximately 5
ml of the exchange solvent and a new boiling chip, and reattach the Snyder
column. Concentrate the extract as described in Section 7.12.1.1, raising the
temperature of the water bath, if necessary, to maintain proper distillation.
When the apparent volume again reaches 1 ml, remove the K-D apparatus from the
water bath and allow it to drain and cool for at least 10 minutes. Repeat the
exchange 2 more times. If no further cleanup of the base/neutral extract is
required, it is also ready for analysis.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for general quality control procedures and
Method 3600 for cleanup procedures.
8.2 The analyst must demonstrate that the compounds of interest are
being quantitatively recovered before applying this method to actual samples.
8.3 For samples that are cleaned using this method, the associated
quality control samples must be processed through this cleanup method.
9.0 METHOD PERFORMANCE
9.1 Refer to the determinative methods for performance data.
10.0 REFERENCES
1. Test Methods; Methods for Organic Chemical Analysis of Municipal and
Industrial Wastewater; U.S. Environmental Protection Agency. Office of
Research and Development. Environmental Monitoring and Support Laboratory.
ORD Publication Offices of Center for Environmental Research Information:
Cincinnati, OH, 1982; EPA-600/4-82-057.
3650A - 6 Revision 1
July 1992
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METHOD 3650A
ACID-BASE PARTITION CLEANUP
START
7 1 Place extract
or organic liquid
waste into
sapara to ry funnel
7 2 Add mathylena
chlorxde
7 3 Add prechilled
dilute 3odium
hydroxide
7 4 Seal and shake
separatory funnel
7 5 Allow
separation of
or gam c layer from
aqueous phase
— >
7 5 Complete phase
mechanical
techniques
7 6 Transfer
aqueous phase to
fJasV, repeat
extraction twice,
combine aqueous
ex t racts
7 7 Discard aqueous
phase
Aqueous
7 10 Assemolc
apparaLus
K-D
7 7 Discard organic
phase
7 8 Adjust pH with
JulJfuric acid, trans
fer aqueous phase to
clean scparatory fun
nel . add ntethylene
chloride. shake,
allow phase separa-
tion, collee t solven
phase in flask
7 9 Perform 2 mote
ex tractions,
combine all
extracts
3650A - 7
Revision 1
July 1992
-------
METHOD 3650A
(Continued)
711 Dry extracts,
collect extracts in
K-D concen t ra t o r,
rinse flask with
methylene chloride
7 12 Concent rate
both f tac Lions
' 14 LKchange
solvent
Ana 1y ze
fractions by
appropriate
determinative
me t h o d
3650A - 8
Revision 1
July 1992
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METHOD 3660A
SULFUR CLEANUP
1.0 SCOPE AND APPLICATION
1.1 Elemental sulfur is encountered in many sediment samples (generally
specific to different areas in the country), marine algae, and some industrial
wastes. The solubility of sulfur in various solvents is very similar to the
organochlorine and organophosphorus pesticides. Therefore, the sulfur
interference follows along with the pesticides through the normal extraction and
cleanup techniques. In general, sulfur will usually elute entirely in Fraction
1 of the Florisil cleanup (Method 3620).
1.2 Sulfur will be quite evident in gas chromatograms obtained from
electron capture detectors, flame photometric detectors operated in the sulfur
or phosphorous mode, and Coulson electrolytic conductivity detectors in the
sulfur mode. If the gas chromatograph is operated at the normal conditions for
pesticide analysis, the sulfur interference can completely mask the region from
the solvent peak through Aldrin.
1.3 Three techniques for the elimination of sulfur are detailed within
this method: (1) the use of copper powder; (2) the use of mercury; and (3) the
use of tetrabutylammonium sulfite. Tetrabutylammonium sulfite causes the least
amount of degradation of a broad range of pesticides and organic compounds, while
copper and mercury may degrade organophosphorus and some organochlorine
pesticides.
2.0 SUMMARY OF METHOD
2.1 The sample to undergo cleanup is mixed with either copper, mercury,
or tetrabutylammonium (TBA) sulfite. The mixture is shaken and the extract is
removed from the sulfur cleanup reagent.
3.0 INTERFERENCES
3.1 Removal of sulfur using copper:
3.1.1 The copper must be very reactive. Therefore, all oxides of
copper must be removed so that the copper has a shiny, bright appearance.
3.1.2 The sample extract must be vigorously agitated with the
reactive copper for at least one minute.
4.0 APPARATUS AND MATERIALS
4.1 Mechanical shaker or mixer - Vortex Genie or equivalent.
4.2 Pipets, disposable - Pasteur type.
3660A - 1 Revision 1
July 1992
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4.3 Centrifuge tubes, calibrated - 12 ml.
4.4 Glass bottles or vials - 10 mL and 50 mL, with Teflon-lined screw
caps or crimp tops.
4.5 Kuderna-Danish (K-D) apparatus.
4.5.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.5.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.5.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.5.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.5.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Nitric acid, HN03, dilute.
5.4 Solvents
5.4.1 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.4.2 Hexane, C6HU - Pesticide quality or equivalent.
5.4.3 2-Propanol, CH3CH(OH)CH3 - Pesticide quality or equivalent.
5.5 Copper powder - Remove oxides by treating with dilute nitric acid,
rinse with organic-free reagent water to remove all traces of acid, rinse with
acetone and dry under a stream of nitrogen. (Copper, fine granular Mallinckrodt
4649 or equivalent).
5.6 Mercury, triple distilled.
5.7 Tetrabutylammonium (TBA) sulfite reagent
3660A - 2 Revision 1
July 1992
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5.7.1 Tetrabutylammonium hydrogen sulfate, [CH3(CH2)3]4NHS04.
5.7.2 Sodium sulfite, Na2S03.
5.7.3 Prepare reagent by dissolving 3.39 g tetrabutylammonium
hydrogen sulfate in 100 ml organic-free reagent water. To remove
impurities, extract this solution three times with 20 ml portions of
hexane. Discard the hexane extracts, and add 25 g sodium sulfite to the
water solution. Store the resulting solution, which is saturated with
sodium sulfite, in an amber bottle with a Teflon-lined screw cap. This
solution can be stored at room temperature for at least one month.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Removal of sulfur using copper
7.1.1 Concentrate the sample to exactly 1.0 mL or other known
volume. Perform concentration using the Kuderna-Danish (K-D) Technique
(Method 3510, Sections 7.10.1 through 7.10.4).
CAUTION; When the volume of solvent is reduced below 1 mL,
semi volatile analytes may be lost.
7.1.2 If the sulfur concentration is such that crystallization
occurs, centrifuge to settle the crystals, and carefully draw off the
sample extract with a disposable pi pet leaving the excess sulfur in the K-
D tube. Transfer 1.0 mL of the extract to a calibrated centrifuge tube.
7.1.3 Add approximately 2 g of cleaned copper powder (to the 0.5 mL
mark) to the centrifuge tube. Mix for at least 1 min on the mechanical
shaker.
7.1.4 Separate the extract from the copper by drawing off the
extract with a disposable pipet and transfer to a clean vial. The volume
remaining still represents 1.0 mL of extract.
NOTE: This separation is necessary to prevent further degradation of
the pesticides.
7.2 Removal of sulfur using mercury
NOTE: Mercury is a highly toxic metal. All operations involving mercury
should be performed in a hood. Prior to using mercury, it is
recommended that the analyst become acquainted with proper handling
and cleanup techniques associated with this metal.
7.2.1 Concentrate the sample extract to exactly 1.0 mL or other
3660A - 3 Revision 1
July 1992
-------
known volume. Perform concentration using the Kuderna-Danish (K-D)
Technique (Method 3510, Sections 7.10.1 through 7.10.4).
CAUTION: When the volume of solvent is reduced below 1 ml,
semi volatile analytes may be lost.
7.2.2 Pipet 1.0 ml of the extract into a clean concentrator tube or
Teflon-sealed vial.
7.2.3 Add one to three drops of mercury to the vial and seal.
Agitate the contents of the vial for 15-30 sec. Prolonged shaking (2 hr)
may be required. If so, use a mechanical shaker.
7.2.4 Separate the sample from the mercury by drawing off the
extract with a disposable pipet and transfer to a clean vial.
7.3 Removal of sulfur using TBA sulfite
7.3.1 Concentrate the sample extract to exactly 1.0 ml or other
known volume. Perform concentration using the Kuderna-Danish (K-D)
Technique (Method 3510, Sections 7.10.1 through 7.10.4).
CAUTION: When the volume of solvent is reduced below 1 ml,
semivolatile analytes may be lost.
7.3.2 Transfer 1.0 ml of the extract to a 50 ml clear glass bottle
or vial with a Teflon-lined screw-cap. Rinse the concentrator tube with
1 ml of hexane, adding the rinsings to the 50 ml bottle.
7.3.3 Add 1.0 mL TBA sulfite reagent and 2 ml 2-propanol, cap the
bottle, and shake for at least 1 min. If the sample is colorless or if
the initial color is unchanged, and if clear crystals (precipitated sodium
sulfite) are observed, sufficient sodium sulfite is present. If the
precipitated sodium sulfite disappears, add more crystalline sodium
sulfite in approximately 0.100 g portions until a solid residue remains
after repeated shaking.
7.3.4 Add 5 ml organic free reagent water and shake for at least 1
min. Allow the sample to stand for 5-10 min. Transfer the hexane layer
(top) to a concentrator tube and concentrate the extract to approximately
1.0 ml with the micro K-D Technique (Section 7.3.5) or the Nitrogen
Slowdown Technique (Section 7.3.6). Record the actual volume of the final
extract.
7.3.5 Micro-Snyder Column Technique
7.3.5.1 Add another one or two clean boiling chips to the
concentrator tube and attach a two ball micro-Snyder column. Prewet
the column by adding about 0.5 ml of hexane to the top of the
column. Place the K-D apparatus in a hot water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete the concentration in 5-10 minutes. At the
proper rate of distillation the balls of the column will actively
3660A - 4 Revision 1
July 1992
-------
chatter, but the chambers will not flood. When the apparent volume
of liquid reaches 0.5 ml, remove the K-D apparatus from the water
bath and allow it to drain and cool for at least 10 minutes. Remove
the Snyder column and rinse the flask and its lower joints with
about 0.2 mL of solvent and add to the concentrator tube. Adjust
the final volume to approximately 1.0 mL with hexane.
7.3.6 Nitrogen Slowdown Technique
7.3.6.1 Place the concentrator tube in a warm water bath
(approximately 35°C) and evaporate the solvent volume to 1.0-2.0 ml,
using a gentle stream of clean, dry nitrogen (filtered through a
column of activated carbon).
CAUTION: Do not use plasticized tubing between the carbon
trap and the sample.
7.3.6.2 The internal wall of the tube must be rinsed down
several times with the appropriate solvent during the operation.
During evaporation, the solvent level in the tube must be positioned
to prevent water from condensing into the sample (i.e., the solvent
level should be below the level of the water bath). Under normal
operating conditions, the extract should not be allowed to become
dry.
CAUTION: When the volume of solvent is reduced below 1 mL,
semi volatile analytes may be lost.
7.4 Analyze the cleaned up extracts by gas chromatography (see the
determinative methods, Section 4.3 of this chapter).
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 3600 for cleanup procedures.
8.2 All reagents should be. checked prior to use to verify that
interferences do not exist.
9.0 METHOD PERFORMANCE
9.1 Table 1 indicates the effect of using copper and mercury to remove
sulfur on the recovery of certain pesticides.
10.0 REFERENCES
1. Loy, E.W., private communication.
2. Goerlitz, D.F. and L.M. Law, Bulletin for Environmental Contamination and
Toxicology, 6, 9 (1971).
3660A - 5 Revision 1
July 1992
-------
3. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, Revision, July 1985.
3660A - 6 Revision 1
July 1992
-------
Table 1.
EFFECT OF MERCURY AND COPPER ON PESTICIDES
Percent Recovery3 using:
Pesticide Mercury Copper
Aroclor 1254
Lindane
Heptachlor
Aldrin
Heptachlor epoxide
DDE
DDT
BHC
Dieldrin
Endrin
Chi orobenzi late
Malathion
Diazinon
Parathion
Ethion
Trithion
97.10
75.73
39.84
95.52
69.13
92.07
78.78
81.22
79.11
70.83
7.14
0.00
0.00
0.00
0.00
0.00
104.26
94.83
5.39
93.29
96.55
102.91
85.10
98.08
94.90
89.26
0.00
0.00
0.00
0.00
0.00
0.00
a Percent recoveries cited are averages based on duplicate analyses for all
compounds other than for Aldrin and BHC. For Aldrin, four and three
determinations were averaged to obtain the result for mercury and copper,
respectively. Recovery of BHC using copper is based on one analysis.
3660A - 7 , Revision 1
July 1992
-------
METHOD 3660A
SULFUR CLEANUP
7 1 1
Concentrate
sample
ex tract
7 1 2
Centrifuge
and draw off
sample
ex tract
712
Transfer
extract to
centrifuge
tube
i
721
Concentrate
sample
extract
722 Pipet
extract into
concentrator
tube or vial
7 2 3 Add
mercury,
agitate
1 L
741
Concentrate
sample
extract
732
Transfer
extract to
centrifuge
tube.
733 Add
TBA-sulfite
and
2-propanol,
agitate
3660A - 8
Revision 1
July 1992
-------
METHOD 3660A
continued
1^1
7 1 3 Add
copper
powder, mix
7 2 4
Separate
sample from
mercury
7 1 4
Separate
ex tract from
copper
7 3 3 Add
more sodium
sulfite,
shake
7 3 4 Add
reagent
wa ter, shake,
concent rate
extract
Analyze extract
using appropriate
determinative
procedure
3660A - 9
Revision 1
July 1992
-------
METHOD 3665
SULFURIC ACID/PERMANGANATE CLEANUP
1.0 SCOPE AND APPLICATION
1.1 This method is suitable for the rigorous cleanup of sample extracts
prior to analysis for polychlorinated biphenyls. This method should be used
whenever elevated baselines or overly complex chromatograms prevent accurate
quantitation of PCBs. This method cannot be used to cleanup extracts for other
target analytes, as it will destroy most organic chemicals including the
pesticides Aldrin, Dieldrin, Endrin, Endosulfan (I and II), and Endosulfan
sulfate.
2.0 SUMMARY OF METHOD
2.1 An extract is solvent exchanged to hexane, then the hexane is
sequentially treated with (1) concentrated sulfuric acid and, if necessary, (2)
5% aqueous potassium permanganate. Appropriate caution must be taken with these
corrosive reagents.
2.2 Blanks and replicate analysis samples must be subjected to the same
cleanup as the samples associated with them.
2.3 It is important that all the extracts be exchanged to hexane before
initiating the following treatments.
3.0 INTERFERENCES
3.1 This technique will not destroy chlorinated benzenes, chlorinated
naphthalenes (Halowaxes), and a number of chlorinated pesticides.
4.0 APPARATUS
4.1 Syringe or Class A volumetric pipet, glass; 1.0, 2.0 and 5.0 mL.
4.2 Vials -1,2 and 10 mL, glass with Teflon lined screw caps or crimp
tops.
4.3 Kuderna-Danish (K-D) apparatus.
4.3.1 Concentrator tube - 10 mL graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
3665 - 1 Revision 0
September 1994
-------
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Vortex mixer.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sulfuric acid/Water, H2S04/H20, (1:1, v/v).
5.4 Hexane, C6H14 - Pesticide grade or equivalent.
5.5 Potassium permanganate, KMn04, 5 percent aqueous solution (w/v).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes, Sec.
4.1.
7.0 PROCEDURE
7.1 Sulfuric acid cleanup
7.1.1 Using a syringe or a volumetric pipet, transfer 1.0 or 2.0 mL
of the hexane extract to a 10 mL vial and, in a fume hood, carefully add
5 mL of the 1:1 sulfuric acid/water solution.
7.1.2 The volume of hexane extract used depends on the requirements
of the GC autosampler used by the laboratory. If the autosampler
functions reliably with 1 mL of sample volume, 1.0 mL of extract should be
used. If the autosampler requires more than 1 mL of sample volume, 2.0 mL
of extract should be used.
CAUTION: Make sure that there is no exothermic reaction nor
evolution of gas prior to proceeding.
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7.1.3 Cap the vial tightly and vortex for one minute. A vortex must
be visible in the vial.
CAUTION: Stop the vortexing immediately if the vial leaks, AVOID
SKIN CONTACT, SULFURIC ACID BURNS.
7.1.4 Allow the phases to separate for at least 1 minute. Examine
the top (hexane) layer; it should not be highly colored nor should it have
a visible emulsion or cloudiness.
7.1.5 If a clean phase separation is achieved, proceed to
Sec. 7.1.8.
7.1.6 If the hexane layer is colored or the emulsion persists for
several minutes, remove the sulfuric acid layer from the vial and dispose
of it properly. Add another 5 ml of the clean 1:1 sulfuric acid/water.
NOTE: Do not remove any hexane at this stage of the procedure.
7.1.7 Vortex the sample for one minute and allow the phases to
separate.
7.1.8 Transfer the hexane layer to a clean 10 ml vial.
7.1.9 Add an additional 1 ml of hexane to the sulfuric acid layer,
cap and shake. This second extraction is done to ensure quantitative
transfer of the PCBs and Toxaphene.
7.1.10 Remove the second hexane layer and combine with the
hexane from Sec. 7.1.8.
7.2 Permanganate cleanup
7.2.1 Add 5 ml of the 5 percent aqueous potassium permanganate
solution to the combined hexane fractions from 7.1.10.
CAUTION: Make sure that there is no exothermic reaction nor
evolution of gas prior to proceeding.
7.2.2 Cap the vial tightly and vortex for 1 minute. A vortex must
be visible in the vial.
CAUTION: Stop the vortexing immediately if the vial leaks. AVOID
SKIN CONTACT, POTASSIUM PERMANGANATE BURNS.
7.2.3 Allow the phases to separate for at least 1 minute. Examine
the top (hexane) layer, it should not be highly colored nor should it have
a visible emulsion or cloudiness.
7.2.4 If a clean phase separation is achieved, proceed to
Sec. 7.2.7.
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7.2.5 If the hexane layer is colored or the emulsion persists for
several minutes, remove the permanganate solution from the vial via a
glass pipette and dispose of it properly. Add another 5 ml of the clean
aqueous permanganate solution.
NOTE: Do not remove any hexane at this stage of the procedure.
7.2.6 Vortex the sample and allow the phases to separate.
7.2.7 Transfer the hexane layer to a clean 10 ml vial.
7.2.8 Add an additional 1 ml of hexane to the permanganate layer,
cap the vial securely and shake. This second extraction is done to ensure
quantitative transfer of the PCBs and Toxaphene.
7.2.9 Remove the second hexane layer and combine with the hexane
from Sec. 7.2.7.
7.3 Final preparation
7.3.1 Reduce the volume of the combined hexane layers to the
original volume (1 or 2 ml) using the Kuderna-Danish Technique
(Sec. 7.3.1.1).
7.3.1.1 Add one or two clean boiling chips to the flask
and attach a three ball Snyder column. Prewet the Snyder column by
adding about 1 ml of hexane to the top of the column. Place the K-D
apparatus on a hot water bath (15-20°C above the boiling point of
the solvent) so that the concentrator tube is partially immersed in
the hot water and the entire lower rounded surface of the flask is
bathed with hot vapor. Adjust the vertical position of the
apparatus and the water temperature, as required, to complete the
concentration in 10-20 minutes. At the proper rate of distillation
the balls of the column will actively chatter, but the chambers will
not flood. When the apparent volume of liquid reaches 1-2 mL,
remove the K-D apparatus from the water bath and allow it to drain
and cool for at least 10 minutes.
7.3.1.2 Remove the Snyder column and rinse the flask and
its lower joints into the concentrator tube with 1-2 mL of hexane.
The extract may be further concentrated by using either the micro
Snyder column technique (Sec. 7.3.2) or nitrogen blowdown technique
(Sec. 7.3.3).
7.3.2 Micro Snyder Column Technique
7.3.2.1 Add another one or two clean boiling chips to the
concentrator tube and attach a two ball micro Snyder column. Prewet
the column by adding about 0.5 mL of hexane to the top of the
column. Place the K-D apparatus in a hot water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete the concentration in 5-10 minutes. At the
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proper rate of distillation the balls of the column will actively
chatter, but the chambers will not flood. When the apparent volume
of liquid reaches 0.5 ml, remove the K-D apparatus from the water
bath and allow it to drain and cool for at least 10 minutes. Remove
the Snyder column and rinse the flask and its lower joints with
about 0.2 ml of hexane and add to the concentrator tube. Adjust the
final volume to 1.0-2.0 ml, as required, with hexane.
7.3.3 Nitrogen Slowdown Technique
7.3.3.1 Place the concentrator tube in a warm water bath
(approximately 35°C) and evaporate the solvent volume to the
required level using a gentle stream of clean, dry nitrogen
(filtered through a column of activated carbon).
CAUTION: Do not use plasticized tubing between the carbon
trap and the sample.
7.3.3.2 The internal wall of the tube must be rinsed down
several times with the appropriate solvent during the operation.
During evaporation, the solvent level in the tube must be positioned
to prevent water from condensing into the sample (i.e., the solvent
level should be below the level of the water bath). Under normal
operating conditions, the extract should not be allowed to become
dry.
7.3.4 Remove any remaining organochlorine pesticides from the
extracts using Florisil Column Cleanup (Method 3620) or Silica Gel Cleanup
(Method 3630).
7.3.5 The extracts obtained may now be analyzed for the target
analytes using the appropriate organic technique(s) (see Sec. 4.3 of this
Chapter). If analysis of the extract will not be performed immediately,
stopper the concentrator tube and store in a refrigerator. If the extract
will be stored longer than 2 days, it should be transferred to a vial with
a Teflon lined screw cap or crimp top, and labeled appropriately.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
9.0 METHOD PERFORMANCE
9.1 No performance data are currently available.
10.0 REFERENCES
None required.
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METHOD 3665
SULFURIC ACID/PERMANGANATE CLEANUP
Start
7.1.1 Carefully
combine hexane
with 1:1
H2S04/H20
solution.
7.1.2
Transfer the
appropriate
volume to
vial.
7.1.3 - 7.1.4
Cap. vortex
and allow
phase
separation.
7.1.8
Transfer
hexane layer
to clean vial.
7.1.6 Remove
and dispose
H2SO4 solution,
add clean HZS04
solution.
7.1.7 Cap,
vortex, and
allow phase
separation.
7.1.9 Add
hexane to
H2SO4 layer,
cap and shake.
7.1.10
Combine two
hexane layers.
7.2.1 Add
KMn04
solution.
7.2.2 - 7.2.3
Cap, vortex,
and allow
phase
separation.
/ 7.2.4 Is N.
/ phase \ No w
V. separation )~~
N. clean? /
[Yes
7..
Tra
hexan
to cle
1
2.7
isfer
an vial.
f
7.2.8 Add
hexane to
KMn04 layer.
cap and shake.
>
r
7.2.9 Combine
two hexane
layers.
w
7.2.5 Remove
and dispose
KMnQ4 aotution,
add clean KMnO4
solution.
V
7.2.6 Cap
vortex and
separation.
7.3.1 - 7.3.3
Reduce volumn
using K-D
and/or nitrogen
btowdown tech.
V
7.3.4 Use
Method 3620 or
Method 3630 to
further remove
contaminants.
7.3.5 Stopper
and
refrigerate
for further
analysis.
Stop
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4.3 DETERMINATION OF ORGANIC ANALYTES
4.3.1 GAS CHROMATOGRAPHIC METHODS
The following methods are included in this section:
Method 8000A:
Method 8010B:
Method 8011:
Method 8015A:
Method 8020A:
Method 8021A:
Method 8030A:
Method 8031:
Method 8032:
Method 8040A:
Method 8060:
Method 8061:
Method 8070:
Method 8080A:
Method 8081:
Method 8090:
Method 8100:
Method 8110:
Method 8120A:
Method 8121:
Method 8140:
Method 8141A:
Method 8150B:
Method 8151:
Gas Chromatography
Halogenated Volatile Organics by Gas
Chromatography
1,2-Dibromoethane and l,2-Dibromo-3-chloropropane
by Microextraction and Gas Chromatography
Nonhalogenated Volatile Organics by Gas
Chromatography
Aromatic Volatile Organics by Gas Chromatography
Halogenated Volatiles by Gas Chromatography Using
Photoionization and Electrolytic Conductivity
Detectors in Series: Capillary Column Technique
Acrolein and Acrylonitrile by Gas Chromatography
Acrylonitrile by Gas Chromatography
Acrylamide by Gas Chromatography
Phenols by Gas Chromatography
Phthalate Esters
Phthalate Esters by Capillary Gas Chromatography
with Electron Capture Detection (GC/ECD)
Nitrosamines by Gas Chromatography
Organochlorine Pesticides and Polychlorinated
Biphenyls by Gas Chromatography
Organochlorine Pesticides and PCBs as Aroclors by
Gas Chromatography: Capillary Column Technique
Nitroaromatics and Cyclic Ketones
Polynuclear Aromatic Hydrocarbons
Haloethers by Gas Chromatography
Chlorinated Hydrocarbons by Gas Chromatography
Chlorinated Hydrocarbons by Gas Chromatography:
Capillary Column Technique
Organophosphorus Pesticides
Organophosphorus Compounds by Gas Chromatography:
Capillary Column Technique
Chlorinated Herbicides by Gas Chromatography
Chlorinated Herbicides by GC Using Methylation or
Pentafluorobenzylation Derivatization: Capillary
Column Technique
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METHOD 8000A
GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Gas chromatography is a quantitative technique useful for the
analysis of organic compounds capable of being volatilized without being
decomposed or chemically rearranged. Gas chromatography (GC), also known as
vapor phase chromatography (VPC), has two subcategories distinguished by: gas-
solid chromatography (GSC), and gas-liquid chromatography (GLC) or gas-liquid
partition chromatography (GLPC). This last group is the most commonly used,
distinguished by type of column adsorbent or packing.
1.2 The chromatographic methods are recommended for use only by, or under
the close supervision of, experienced residue analysts.
2.0 SUMMARY OF METHOD
2.1 Each organic analytical method that follows provides a recommended
technique for extraction, cleanup, and occasionally, derivatization of the
samples to be analyzed. Before the prepared sample is introduced into the GC,
a procedure for standardization must be followed to determine the recovery and
the limits of detection for the analytes of interest. Following sample
introduction into the GC, analysis proceeds with a comparison of sample values
with standard values. Quantitative analysis is achieved through integration of
peak area or measurement of peak height.
3.0 INTERFERENCES
3.1 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
sample syringe or purging device must be rinsed out between samples with water
or solvent. Whenever an unusually concentrated sample is encountered, it should
be followed by an analysis of a solvent blank or of water to check for cross
contamination. For volatile samples containing large amounts of water-soluble
materials, suspended solids, high boiling compounds or high organohalide
concentrations, it may be necessary to wash out the syringe or purging device
with a detergent solution, rinse it with distilled water, and then dry it in a
105°C oven between analyses.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph - Analytical system complete with gas chromatograph
suitable for on-column injections and all required accessories, including
detectors, column supplies, recorder, gases, and syringes. A data system for
measuring peak height and/or peak areas is recommended.
4.2 Gas chromatographic columns - See the specific determinative method.
Other packed or capillary (open-tubular) columns may be used if the requirements
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of Section 8.6 are met.
5.0 REAGENTS
5.1 See the specific determinative method for the reagents needed.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Extraction - Adhere to those procedures specified in the referring
determinative method.
7.2 Cleanup and separation - Adhere to those procedures specified in the
referring determinative method.
7.3 The recommended gas chromatographic columns and operating conditions
for the instrument are specified in the referring determinative method.
7.4 Calibration
7.4.1 Establish gas chromatographic operating parameters equivalent
to those indicated in Section 7.0 of the determinative method of interest.
Prepare calibration standards using the procedures indicated in
Section 5.0 of the determinative method of interest. Calibrate the
chromatographic system using either the external standard technique
(Section 7.4.2) or the internal standard technique (Section 7.4.3).
7.4.2 External standard calibration procedure
7.4.2.1 For each analyte of interest, prepare calibration
standards at a minimum of five concentrations by adding volumes of
one or more stock standards to a volumetric flask and diluting to
volume with an appropriate solvent. One of the external standards
should be at a concentration near, but above, the method detection
limit. The other concentrations should correspond to the expected
range of concentrations found in real samples or should define the
working range of the detector.
7.4.2.2 Inject each calibration standard using the
technique that will be used to introduce the actual samples into the
gas chromatograph (e.g. 2-5 pi injections, purge-and-trap, etc.).
Tabulate peak height or area responses against the mass injected.
The results can be used to prepare a calibration curve for each
analyte. Alternatively, for samples that are introduced into the
gas chromatograph using a syringe, the ratio of the response to the
amount injected, defined as the calibration factor (CF), can be
calculated for each analyte at each standard concentration. If the
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percent relative standard deviation (%RSD) of the calibration factor
is less than 20% over the working range, linearity through the
origin can be assumed, and the average calibration factor can be
used in place of a calibration curve.
Calibration factor -
* For multiresponse pesticides/PCBs, use the total area of
all peaks used for quantitation.
7.4.2.3 The working calibration curve or calibration
factor must be verified on each working day by the injection of one
or more calibration standards. The frequency of verification is
dependent on the detector. Detectors, such as the electron capture
detector, that operate in the sub-nanogram range are more
susceptible to changes in detector response caused by GC column and
sample effects. Therefore, more frequent verification of
calibration is necessary. The flame ionization detector is much
less sensitive and requires less frequent verification. If the
response for any analyte varies from the predicted response by more
than + 15%, a new calibration curve must be prepared for that
analyte. For methods 8010, 8020, and 8030, see Table 3 in each
method for calibration and quality control acceptance criteria.
R! - R2
Percent Difference = x 100
RI
where:
R, = Calibration Factor from first analysis.
R2 = Calibration Factor from succeeding analyses.
7.4.3 Internal standard calibration procedure
7.4.3.1 To use this approach, the analyst must select one
or more internal standards that are similar in analytical behavior
to the compounds of interest. The analyst must further demonstrate
that the measurement of the internal standard is not affected by
method or matrix interferences. Due to these limitations, no
internal standard applicable to all samples can be suggested.
7.4.3.2 Prepare calibration standards at a minimum of five
concentrations for each analyte of interest by adding volumes of one
or more stock standards to a volumetric flask. To each calibration
standard, add a known constant amount of one or more internal
standards and dilute to volume with an appropriate solvent. One of
the standards should be at a concentration near, but above, the
method detection limit. The other concentrations should correspond
to the expected range of concentrations found in real samples or
should define the working range of the detector.
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7.4.3.3 Inject each calibration standard using the same
introduction technique that will be applied to the actual samples
(e.g. 2 to 5 /xL injection, purge-and-trap, etc.). Tabulate the peak
height or area responses against the concentration of each compound
and internal standard. Calculate response factors (RF) for each
compound as follows:
RF - (AsCis)/(AisCs)
where:
As = Response for the analyte to be measured.
Ais = Response for the internal standard.
Cjs = Concentration of the internal standard, /xg/L.
Cs = Concentration of the analyte to be measured,
M9/L.
If the RF value over the working range is constant (< 20%
RSD), the RF can be assumed to be invariant, and the average RF can
be used for calculations. Alternatively, the results can be used to
plot a calibration curve of response ratios, As/Ajs versus RF.
7.4.3.4 The working calibration curve or RF must be
verified on each working day by the measurement of one or more
calibration standards. The frequency of verification is dependent
on the detector. Detectors, such as the electron capture detector,
that operate in the sub-nanogram range are more susceptible to
changes in detector response caused by GC column and sample effects.
Therefore, more frequent verification of calibration is necessary.
The flame ionization detector is much less sensitive and requires
less frequent verification. If the response for any analyte varies
from the predicted response by more than + 15%, a new calibration
curve must be prepared for that compound. For methods 8010, 8020,
and 8030, see Table 3 in each method for calibration and quality
control acceptance criteria.
7.5 Retention time windows
7.5.1 Before establishing windows, make sure the GC system is within
optimum operating conditions. Make three injections of all single
component standard mixtures and multiresponse products (i.e. PCBs)
throughout the course of a 72 hour period. Serial injections over less
than a 72 hour period result in retention time windows that are too tight.
7.5.2 Calculate the standard deviation of the three retention times
(use any function of retention time; including absolute retention time, or
relative retention time) for each single component standard. For
multiresponse products, choose one major peak from the envelope and
calculate the standard deviation of the three retention times for that
peak. The peak chosen should be fairly immune to losses due to
degradation and weathering in samples.
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7.5.2.1 Plus or minus three times the standard deviation
of the retention times for each standard will be used to define the
retention time window; however, the experience of the analyst should
weigh heavily in the interpretation of chromatograms. For
multiresponse analytes (i.e. PCBs), the analyst should use the
retention time window, but should primarily rely on pattern
recognition.
7.5.2.2 In those cases where the standard deviation for a
particular standard is zero, the laboratory must substitute the
standard deviation of a close eluting, similar compound to develop
a valid retention time window.
7.5.3 The laboratory must calculate retention time windows for each
standard on each GC column and whenever a new GC column is installed. The
data must be retained by the laboratory.
7.6 Gas chromatographic analysis
7.6.1 Introduction of organic compounds into the gas chromatograph
varies depending on the volatility of the compound. Volatile organics are
primarily introduced by purge-and-trap (Method 5030). However, there are
limited applications (in Method 5030) where direct injection is
acceptable. Use of Method 3810 or 3820 as a screening technique for
volatile organic analysis may be valuable with some sample matrices to
prevent overloading and contamination of the GC systems. Semivolatile
organics are introduced by direct injection.
7.6.2 The appropriate detector(s) is given in the specific method.
7.6.3 Samples are analyzed in a set referred to as an analysis
sequence. The sequence begins with instrument calibration followed by
sample extracts interspersed with multi-concentration calibration
standards. The sequence ends when the set of samples has been injected or
when qualitative and/or quantitative QC criteria are exceeded.
7.6.4 Direct Injection - Inject 2-5 juL of the sample extract using
the solvent flush technique, if the extract is manually injected. Smaller
volumes (1.0 /xL) can be injected, and the solvent flush technique is not
required, if automatic devices are employed. Record the volume injected
to the nearest 0.05 p.1 and the resulting peak size in area units or peak
height.
7.6.5 If the responses exceed the linear range of the system, dilute
the extract and reanalyze. It is recommended that extracts be diluted so
that all peaks are on scale. Overlapping peaks are not always evident
when peaks are off scale. Computer reproduction of chromatograms,
manipulated to ensure all peaks are on scale over a 100-fold range, are
acceptable if linearity is demonstrated. Peak height measurements are
recommended over peak area integration when overlapping peaks cause errors
in area integration.
7.6.6 If peak detection is prevented by the presence of
interferences, further cleanup is required.
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7.6.7 Examples of chromatograms for the compounds of interest are
frequently available in the referring analytical method.
7.6.8 Calibrate the system immediately prior to conducting any
analyses (see Section 7.4). A mid-concentration standard must also be
injected at intervals specified in the method and at the end of the
analysis sequence. The calibration factor for each analyte to be
quantitated, must not exceed a 15% difference when compared to the initial
standard of the analysis sequence. When this criterion is exceeded,
inspect the GC system to determine the cause and perform whatever
maintenance is necessary (see Section 7.7) before recalibrating and
proceeding with sample analysis. All samples that were injected after the
standard exceeding the criterion must be reinjected to avoid errors in
quantitation, if the initial analysis indicated the presence of the
specific target analytes that exceeded the criterion.
7.6.9 Establish daily retention time windows for each analyte. Use
the retention time for each analyte from Section 7.6.8 as the midpoint of
the window for that day. The daily retention time window equals the
midpoint ± three times the standard deviation determined in Section 7.5.
7.6.9.1 Tentative identification of an analyte occurs when
a peak from a sample extract falls within the daily retention time
window. Normally, confirmation is required: on a second GC column,
by GC/MS if concentration permits, or by other recognized
confirmation techniques. Confirmation may not be necessary if the
composition of the sample matrix is well established by prior
analyses.
7.6.9.2 Validation of GC system qualitative performance:
Use the mid-concentration standards interspersed throughout the
analysis sequence (Section 7.6.8) to evaluate this criterion. If
any of the standards fall outside their daily retention time window,
the system is out of control. Determine the cause of the problem
and correct it (see Section 7.7). All samples that were injected
after the standard exceeding the criteria must be reinjected to
avoid false negatives and possibly false positives.
7.7 Suggested chromatography system maintenance - Corrective measures may
require any one or more of the following remedial actions.
7.7.1 Packed columns - For instruments with injection port traps,
replace the demister trap, clean, and deactivate the glass injection port
insert or replace with a cleaned and deactivated insert. Inspect the
injection end of the column and remove any foreign material (broken glass
from the rim of the column or pieces of septa). Replace the glass wool
with fresh deactivated glass wool. Also, it may be necessary to remove
the first few millimeters of the packing material if any discoloration is
noted, also swab out the inside walls of the column if any residue is
noted. If these procedures fail to eliminate the degradation problem, it
may be necessary to deactivate the metal injector body (described in
Section 7.7.3) and/or repack/replace the column.
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7.7.2 Capillary columns - Clean and deactivate the glass injection
port insert or replace with a cleaned and deactivated insert. Break off
the first few inches, up to one foot, of the injection port side of the
column. Remove the column and solvent backflush according to the
manufacturer's instructions. If these procedures fail to eliminate the
degradation problem, it may be necessary to deactivate the metal injector
body and/or replace the column.
7.7.3 Metal injector body - Turn off the oven and remove the
analytical column when the oven has cooled. Remove the glass injection
port insert (instruments with off-column injection or Grob). Lower the
injection port temperature to room temperature. Inspect the injection
port and remove any noticeable foreign material.
7.7.3.1 Place a beaker beneath the injector port inside
the GC oven. Using a wash bottle, serially rinse the entire inside
of the injector port with acetone and then toluene; catching the
rinsate in the beaker.
7.7.3.2 Prepare a solution of deactivating agent (Sylon-CT
or equivalent) following manufacturer's directions. After all metal
surfaces inside the injector body have been thoroughly coated with
the deactivation solution, serially rinse the injector body with
toluene, methanol, acetone, and nexane. Reassemble the injector and
replace the GC column.
7.8 Calculations
7.8.1 External standard calibration - The concentration of each
analyte in the sample may be determined by calculating the amount of
standard purged or injected, from the peak response, using the calibration
curve or the calibration factor determined in Section 7.4.2. The
concentration of a specific analyte is calculated as follows:
Aqueous samples
Concentration (/ig/L) » [(Ax)(A)(Vt)(D)]/[(As)(V1)(V8)]
where:
Ax = Response for the analyte in the sample, units may be in
area counts or peak height.
A = Amount of standard injected or purged, ng.
As = Response for the external standard, units same as for
V
V, = Volume of extract injected, piL. For purge-and-trap
analysis, V1 is not applicable and therefore = 1.
D = Dilution factor, if dilution was made on the sample
prior to analysis. If no dilution was made, D = 1,
dimensionless.
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Vt - Volume of total extract, /uL. For purge-and-trap
analysis, Vt is not applicable and therefore = 1.
V8 = Volume of sample extracted or purged, ml.
Nonaqueous samples
Concentration (Mg/kg) - I(AJl)(A)(Vt)(D)]/[(AB)(Vl)(W)]
where:
W = Weight of sample extracted or purged, g. The wet weight
or dry weight may be used, depending upon the specific
applications of the data.
Ax, A8, A, Vt, D, and V, have the same definition as for aqueous
samples when a solid sample is purged (e.g., low concentration soil) for
volatile organic analysis or for semivolatile organic and pesticide
extracts. When the nonaqueous sample is extracted for purge and trap
analysis, V, « volume of methanol extract added to reagent water for purge
and trap analysis.
7.8.2 Internal standard calibration - For each analyte of interest,
the concentration of that analyte in the sample is calculated as follows:
Aqueous samples
Concentration (Mg/L) = [(Ax)(Cis)(D)]/[(A1s)(RF)(Vs)]
where:
Ax • Response of the analyte being measured, units may be in
area counts or peak height.
Cjs = Amount of internal standard added to extract or volume
purged, ng.
D = Dilution factor, if a dilution was made on the sample
prior to analysis. If no dilution was made, D = 1,
dimensionless.
Ajs = Response of the internal standard, units same as Ax.
RF = Response factor for analyte, as determined in Section
7.4.3.3.
Vs = Volume of water extracted or purged, ml.
Nonaqueous samples
Concentration (/tg/kg) = [(As)(Cis)(D)]/[(Ais)(RF)(W8)]
8000A - 8 Revision 1
July 1992
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where:
Ws = Weight of sample extracted, g. Either a dry weight or
wet weight may be used, depending upon the specific
application of the data.
As, Cjs, D, Ajs, and RF have the same definition as for aqueous
samples.
8.0 QUALITY CONTROL
8.1 Each laboratory that uses these methods is required to operate a
formal quality control program. The minimum requirements of this program consist
of an initial demonstration of laboratory capability and an ongoing analysis of
spiked samples to evaluate and document quality data. The laboratory should
maintain records to document the quality of the data generated. Ongoing data
quality checks are compared with established performance criteria to determine
if the results of analyses meet the performance characteristics of the method.
When results of sample spikes indicate atypical method performance, a quality
control check standard should be analyzed to confirm that the measurements were
performed in an in-control mode of operation.
8.2 Before processing any samples, the analyst should demonstrate,
through the analysis of a reagent blank, that interferences from the analytical
system, glassware, and reagents are under control. Each time a set of samples
is extracted or there is a change in reagents, an organic-free reagent water
blank should be processed as a safeguard against chronic laboratory
contamination. The blank samples should be carried through all stages of the
sample preparation and measurement steps.
8.3 For each analytical batch (up to 20 samples), a reagent blank, matrix
spike, and duplicate or matrix spike duplicate should be analyzed (the frequency
of the spikes may be different for different monitoring programs). The blank and
spiked samples should be carried through all stages of the sample preparation and
measurement steps.
8.4 The experience of the analyst performing gas chromatography is
invaluable to the success of the methods. Each day that analysis is performed,
the daily calibration sample should be evaluated to determine if the
chromatographic system is operating properly. Questions that should be asked
are: Do the peaks look normal?; Is the response obtained comparable to the
response from previous calibrations? Careful examination of the standard
chromatogram can indicate whether the column is still good, the injector is
leaking, the injector septum needs replacing, etc. If any changes are made to
the system (e.g. column changed), recalibration of the system should take place.
8.5 Required instrument QC
8.5.1 Step 7.4 requires that the %RSD vary by < 20% when comparing
calibration factors to determine if a five point calibration curve is
linear.
8000A - 9 Revision 1
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8.5.2 Section 7.4 sets a limit of + 15% difference when comparing
daily response of a given analyte versus the initial response. For
Methods 8010, 8020, and 8030, follow the guidance on limits specified in
Section 7.4.3.4. If the limit is exceeded, a new standard curve should be
prepared unless instrument maintenance corrects the problem for that
particular analyte.
8.5.3 Step 7.5 requires the establishment of retention time windows.
8.5.4 Section 7.6.8 sets a limit of + 15% difference when comparing
the response from the continuing calibration standard of a given analyte
versus any succeeding standards analyzed during an analysis sequence.
8.5.5 Step 7.6.9.2 requires that all succeeding standards in an
analysis sequence should fall within the daily retention time window
established by the first standard of the sequence.
8.6 To establish the ability to generate acceptable accuracy and
precision, the analyst should perform the following operations.
8.6.1 A quality control (QC) check sample concentrate is required
containing each analyte of interest. The QC check sample concentrate may
be prepared from pure standard materials, or purchased as certified
solutions. If prepared by the laboratory, the QC check sample concentrate
should be made using stock standards prepared independently from those
used for calibration.
8.6.1.1 The concentration of the QC check sample
concentrate is highly dependent upon the analytes being
investigated. Therefore, refer to Method 3500, Section 8.0 for the
required concentration of the QC check sample concentrate.
8.6.2 Preparation of QC check samples
8.6.2.1 Volatile organic analytes (Methods 8010, 8020, and
8030) - The QC check sample is prepared by adding 200 p.1 of the QC
check sample concentrate (Step 8.6.1) to 100 ml of water.
8.6.2.2 Semivolatile organic analytes (Methods 8040, 8060,
8070, 8080, 8090, 8100, 8110, and 8120) - The QC check sample is
prepared by adding 1.0 ml of the QC check sample concentrate (Step
8.6.1) to each of four 1-L aliquots of water.
8.6.3 Four aliquots of the well-mixed QC check sample are analyzed
by the same procedures used to analyze actual samples (Section 7.0 of each
of the methods). For volatile organics, the preparation/analysis process
is purge-and-trap/gas chromatography. For semivolatile organics, the QC
check samples should undergo solvent extraction (see Method 3500) prior to
chromatographic analysis.
8.6.4 Calculate the average recovery (x) in M9/L> and the standard
deviation of the recovery (s) in M9/L, for each analyte of interest using
the four results.
8000A - 10 Revision 1
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8.6.5 For each analyte compare s and x with the corresponding
acceptance criteria for precision and accuracy, respectively, given the QC
Acceptance Criteria Table at the end of each of the determinative methods.
If s and x for all analytes of interest meet the acceptance criteria, the
system performance is acceptable and analysis of actual samples can_begin.
If any individual s exceeds the precision limit or any individual x falls
outside the range for accuracy, then the system performance is
unacceptable for that analyte.
NOTE: The large number of analytes in each of the QC Acceptance
Criteria Tables present a substantial probability that one or
more will fail at least one of the acceptance criteria when
all analytes of a given method are determined.
8.6.6 When one or more of the analytes tested fail at least one of
the acceptance criteria, the analyst should proceed according to Step
8.6.6.1 or 8.6.6.2.
8.6.6.1 Locate and correct the source of the problem and
repeat the test for all analytes of interest beginning with
Step 8.6.2.
8.6.6.2 Beginning with Step 8.6.2, repeat the test only
for those analytes that failed to meet criteria. Repeated failure,
however, will confirm a general problem with the measurement system.
If this occurs, locate and correct the source of the problem and
repeat the test for all compounds of interest beginning with
Step 8.6.2.
8.7 The laboratory should, on an ongoing basis, analyze a reagent blank
and a matrix spiked duplicate for each analytical batch (up to a maximum of 20
samples/batch) to assess accuracy. For soil and waste samples where detectable
amounts of organics are present, replicate samples may be appropriate in place
of spiked duplicates. For laboratories analyzing one to ten samples per month,
at least one spiked sample per month is required.
8.7.1 The concentration of the spike in the sample should be
determined as follows:
8.7.1.1 If, as in compliance monitoring, the concentration
of a specific analyte in the sample is being checked against a
regulatory concentration limit, the spike should be at that limit,
or 1 to 5 times higher than the background concentration determined
in Step 8.7.2, whichever concentration would be larger.
8.7.1.2 If the concentration of a specific analyte in a
water sample is not being checked against a limit specific to that
analyte, the spike should be at the same concentration as the QC
reference sample (Step 8.6.2) or 1 to 5 times higher than the
background concentration determined in Step 8.7.2, whichever
concentration would be larger. For other matrices, the recommended
spiking concentration is 20 times the EQL.
8000A - 11 Revision 1
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8.7.1.3 For semivolatile organics, it may not be possible
to determine the background concentration levels prior to spiking
(e.g. maximum holding times will be exceeded). If this is the case,
the spike concentration should be (1) the regulatory concentration
limit, if any; or, if none (2) the larger of either 5 times higher
than the expected background concentration or the QC reference
sample concentration (Step 8.6.2). For other matrices, the
recommended spiking concentration is 20 times the EQL.
8.7.2 Analyze one unspiked and one spiked sample aliquot to
determine percent recovery of each of the spiked compounds.
8.7.2.1 Volatile organics - Analyze one 5-mL sample
aliquot to determine the background concentration (B) of each
analyte. If necessary, prepare a new QC reference sample
concentrate (Step 8.6.1) appropriate for the background
concentration in the sample. Spike a second 5-mL sample aliquot
with 10 fj.1 of the QC reference sample concentrate and analyze it to
determine the concentration after spiking (A) of each analyte.
Calculate each percent recovery (p) as 100(A - B)%/T, where T is the
known true value of the spike.
8.7.2.2 Semivolatile organics - Analyze one sample aliquot
(extract of 1-L sample) to determine the background concentration
(B) of each analyte. If necessary, prepare a new QC reference
sample concentrate (Step 8.6.1) appropriate for the background
concentration in the sample. Spike a second 1-L sample aliquot with
1.0 mL of the QC reference sample concentrate and analyze it to
determine the concentration after spiking (A) of each analyte.
Calculate each percent recovery (p) as 100(A - B)%/T, where T is the
known true value of the spike.
8.7.3 Compare the percent recovery (p) for each analyte in a water
sample with the corresponding criteria presented in the QC Acceptance
Criteria Table found at the end of each of the determinative methods.
These acceptance criteria were calculated to include an allowance for
error in measurement of both the background and spike concentrations,
assuming a spike to background ratio of 5:1. This error will be accounted
for to the extent that the analyst's spike to background ratio approaches
5:1. If spiking was performed at a concentration lower than the QC
reference sample concentration (Step 8.6.2), the analyst should use either
the QC acceptance criteria presented in the Tables, or optional QC
acceptance criteria calculated for the specific spike concentration. To
calculate optional acceptance criteria for the recovery of an analyte:
(1) Calculate accuracy (x') using the equation found in the Method
Accuracy and Precision as a Function of Concentration Table (appears at
the end of each determinative method), substituting the spike
concentration (T) for C; (2) calculate overall_precision (S') using the
equation in the same Table, substituting x' for x; (3) calculate the range
for recovery at the spike concentration as (lOOx'/T) + 2.44(100S'/T)%.
8.7.4 If any individual p falls outside the designated range for
recovery, that analyte has failed the acceptance criteria. A check
standard containing each analyte that failed the criteria should be
8000A - 12 Revision 1
July 1992
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analyzed as described in Step 8.8.
8.8 If any analyte in a water sample fails the acceptance criteria for
recovery in Step 8.7, a QC reference standard containing each analyte that failed
should be prepared and analyzed.
NOTE: The frequency for the required analysis of a QC reference standard
will depend upon the number of analytes being simultaneously tested,
the complexity of the sample matrix, and the performance of the
laboratory. If the entire list of analytes given in a method should
be measured in the sample in Step 8.7, the probability that the
analysis of a QC check standard will be required is high. In this
case, the QC check standard should be routinely analyzed with the
spiked sample.
8.8.1 Preparation of the QC check sample - For volatile organics,
add 10 fj.1 of the QC check sample concentrate (Step 8.6.1 or 8.7.2) to 5
ml of water. For semivolatile organics, add 1.0 ml of the QC check sample
concentrate (Step 8.6.1 or 8.7.2) to 1 L of water. The QC check sample
needs only to contain the analytes that failed criteria in the test in
Step 8.7. Prepare the QC check sample for analysis following the
guidelines given in Method 3500 (e.g. purge-and-trap, extraction, etc.).
8.8.2 Analyze the QC check sample to determine the concentration
measured (A) of each analyte. Calculate each percent recovery (ps) as
100(A/T)%, where T is the true value of the standard concentration.
8.8.3 Compare the percent recovery (ps) for each analyte with the
corresponding QC acceptance criteria found in the appropriate Table in
each of the methods. Only analytes that failed the test in Step 8.7 need
to be compared with these criteria. If the recovery of any such analyte
falls outside the designated range, the laboratory performance for that
analyte is judged to be out of control, and the problem should be
immediately identified and corrected. The result for that analyte in the
unspiked sample is suspect and may not be reported for regulatory
compliance purposes.
8.9 As part of the QC program for the laboratory, method accuracy for
each matrix studied should be assessed and records should be maintained. After
the analysis of five spiked samples (of the same matrix type) as in Step 8.7,
calculate the average percent recovery (p) and the standard deviation of the
percent recovery (s ). Express the accuracy assessment as a percent recovery
interval from p - 2s to p + 2s . If p = 90% and s = 10%, for example, the
accuracy interval is expressed as 70-110%. Update the accuracy assessment for
each analyte on a regular basis (e.g. after each five to ten new accuracy
measurements).
8.10 Calculate surrogate control limits as follows:
8.10.1 For each sample analyzed, calculate the percent recovery
of each surrogate in the sample.
8.10.2 Calculate the average percent recovery (p) and standard
deviation of the percent recovery (s) for each of the surrogates when
8000A - 13 Revision 1
July 1992
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surrogate data from 25 to 30 samples for each matrix is available.
8.10.3 For a given matrix, calculate the upper and lower
control limit for method performance for each surrogate standard. This
should be done as follows:
Upper Control Limit (UCL) = p + 3s
Lower Control Limit (LCL) = p - 3s
8.10.4 For aqueous and soil matrices, these laboratory
established surrogate control limits should, if applicable, be compared
with the control limits in Tables A and B of Methods 8240 and 8270,
respectively. The limits given in these methods are multi-laboratory
performance based limits for soil and aqueous samples, and therefore, the
single-laboratory limits established in Step 8.10.3 should fall within
those given in Tables A and B for these matrices.
8.10.5 If recovery is not within limits, the following is
required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are a
problem or flag the data as "estimated concentration."
8.10.6 At a minimum, each laboratory should update surrogate
recovery limits on a matrix-by-matrix basis, annually.
8.11 It is recommended that the laboratory adopt additional quality
assurance practices for use with this method. The specific practices that are
most productive depend upon the needs of the laboratory and the nature of the
samples. Field duplicates may be analyzed to assess the precision of the
environmental measurements. When doubt exists over the identification of a peak
on the chromatogram, confirmatory techniques such as gas chromatography with a
dissimilar column, specific element detector, or mass spectrometer should be
used. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 The method detection limit (MDL) is defined as the minimum
concentration of a substance that can be measured and reported with 99%
confidence that the value is above zero. The MDL concentrations listed in the
referring analytical methods were obtained using water. Similar results were
achieved using representative wastewaters. The MDL actually achieved in a given
analysis will vary depending on instrument sensitivity and matrix effects.
8000A - 14 Revision 1
July 1992
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9.2 Refer to the determinative method for specific method performance
information.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
2. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, July 1985, Revision.
8000A - 15 Revision 1
July 1992
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METHOD 8000A
GAS CHROMATOGRAPHY
Start
X^ s
7 1 Refer to
determinative
procedure for
extraction
procedure
recommendation
Internal Standard External Standard
7431 Select
internal standards
having behavior
similar to
compounds of
interest
1
7 2 Refer to
determinative
procedure for
cleanup and
preparation
procedure
recommendation*
7432 Prepare
calibration
s tandards
1
741 Establish
chroma togr a phi c
condi tions
/ • /
7433 Inject
cal ibra tion
standards ,
calculate RF
1
7434 Verify
working calibration
curve or RF each
day
S 1 4 l >v
< f Calibration ) »
^v technique? /
~l 5 Calculate
windows
7 4,2.1 Prepare
cal ibration
standards for each
compound of
interest
i
7422 Inject
t d d
1 fa t ' ^ '*
or calculate
calibration factor
1
7423 Verify
working cal ibration
curve each day
8000A - 16
Revision 1
July 1992
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METHOD 8000A
continued
Semivolatiles
7 6 1 If
necessary.
screen sample*
by Method 3810
or 3820
761 Introduce
compounds into CC
by purge-and-trap
or direct infection
(Method 5030)
761 Introduce
compounds into
CC by direct
injection
764 Inject
samples using
sol vent flush
technique,
record volume
765 Dilute
extract and
reanalyze
7 6 6 Do
fur ther
cleanup
6 8 Calibrate
system
immedia tely
prior to
analyses
7 6 9 Establish
daily retention
time windows
for each
analyte
7 7 Perform
chroma tography
sys tern
maintenance, if
needed
7 8 Calculate
concentration of
each analyte, using
appropriate formula
for matrix and type
of standard
Stop
8000A - 17
Revision 1
July 1992
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METHOD 8010B
HALOGENATED VOLATILE ORGANICS BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8010 is used to determine the concentration of various
volatile halogenated organic compounds. The following compounds can be
determined by this method:
Appropriate Technique
Compound Name
Allyl chloride
Benzyl chloride
Bis(2-chloroethoxy)methane
Bis(2-chloroisopropyl ) ether
Bromoacetone
Bromobenzene
Bromodi chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
Chloroethane
2-Chloroethanol
2-Chloroethyl vinyl ether
Chloroform
1-Chlorohexane
Chl oromethane
Chloromethyl methyl ether
Chloroprene
4-Chlorotoluene
Di bromochl oromethane
1 , 2-Di bromo-3-chl oropropane
Dibromomethane
1, 2-Di chlorobenzene
1, 3 -Di chlorobenzene
1, 4 -Di chlorobenzene
l,4-Dichloro-2-butene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
tr ans-1, 2-Di chl oroethene
Dichloromethane
1,2-Dichloropropane
l,3-Dichloro-2-propanol
ci s- 1 ,3-Di chl oropropene
trans-l,3-Dichloropropene
Epichlorhydrin
CAS No.a
107-05-1
100-44-7
111-91-1
39638-32-9
598-31-2
108-86-1
75-27-4
75-25-2
74-83-9
56-23-5
108-90-7
75-00-3
107-07-03
110-75-8
67-66-3
544-10-5
74-87-3
107-30-2
126-99-8
106-43-4
124-48-1
96-12-8
74-95-3
95-50-1
541-73-1
106-46-7
764-41-0
75-71-8
75-34-3
107-06-2
75-35-4
156-60-5
75-09-2
78-87-5
96-23-1
10061-01-5
10061-02-6
106-89-8
Purge-and-Trap
b
PP
PP
b
PP
b
b
b
b
b
b
b
PP
b
b
pc
b
PP
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
PP
b
b
PP
Direct
Injection
b
b
pc
b
b
b
b
b
b
b
b
b
b
b
b
pc
b
pc
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
8010B - 1
Revision 2
September 1994
-------
Appropriate Technique
Direct
Compound Name CAS No.a Purge-and-Trap Injection
Ethylene dibromide
Methyl iodide
1,1,2,2-Tetrachloroethane
1,1,1 , 2-Tetrachl oroethane
Tetrachl oroethene
1,1,1 -Tr i chl oroethane
1,1, 2 -Trichl oroethane
Trichloroethene
Tr i chl orof 1 uoromethane
1,2,3-Trichloropropane
Vinyl Chloride
106-93-4
74-88-4
79-34-5
630-20-6
127-18-4
71-55-6
79-00-5
79-01-6
75-69-4
96-18-4
75-01-4
b
PP
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
a Chemical Abstract Services Registry Number
b Adequate response using this technique
pp Poor purging efficiency, resulting in high EQLs
pc Poor chromatographic performance.
1.2 Table 1 indicates compounds that may be analyzed by this method and
lists the method detection limit for each compound in organic-free reagent water.
Table 2 lists the estimated quantitation limit for other matrices.
2.0 SUMMARY OF METHOD
2.1 Method 8010 provides gas chromatographic conditions for the
detection of halogenated volatile organic compounds. Samples can be introduced
into the GC using direct injection or purge-and-trap (Method 5030). Ground water
samples must be analyzed using Method 5030. A temperature program is used in the
gas chromatograph to separate the organic compounds. Detection is achieved by
a electrolytic conductivity detector (HECD).
2.2 The method provides an optional gas chromatographic column that may
be helpful in resolving the analytes from co-eluting non-target compounds and for
analyte confirmation.
3.0 INTERFERENCES
3.1 Refer to Methods 5030 and 8000.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly chl orofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A trip blank prepared from
organic-free reagent water and carried through sampling and subsequent storage
and handling can serve as a check on such contamination.
8010B - 2 Revision 2
September 1994
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4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph - analytical system complete with gas
chromatograph suitable for on-column injections or purge-and-trap sample
introduction and all required accessories, including detector, analytical
columns, recorder, gases, and syringes. A data system for measuring peak
heights and/or peak areas is recommended.
4.1.2 Columns
4.1.2.1 Column 1 - 8 ft x 0.1 in. ID stainless steel or
glass column packed with 1% SP-1000 on Carbopack-B 60/80 mesh or
equivalent.
4.1.2.2 Column 2 - 6 ft x 0.1 in. ID stainless steel or
glass column packed with chemically bonded n-octane on Porasil-C
100/120 mesh (Durapak) or equivalent.
4.1.3 Detector - Electrolytic conductivity (HECD).
4.2 Sample introduction apparatus, refer to Method 5030 for the
appropriate equipment for sample introduction purposes.
4.3 Syringes, 5 ml Luerlok glass hypodermic and a 5 ml, gas-tight with
shutoff valve.
4.4 Volumetric flask, Class A, Appropriate sizes with ground glass
stoppers.
4.5 Microsyringe, 10 and 25 juL with a 0.006 in. ID needle (Hamilton 702N
or equivalent) and a 100 //L.
4.6 Analytical balance - 0.0001 g.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH. Pesticide quality or equivalent. Store away from
other solvents.
8010B - 3 Revision 2
September 1994
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5.4 Stock standards - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standards in
methanol using assayed liquids or gases, as appropriate. Because of the toxicity
of some of the organohalides, primary dilutions of these materials should be
prepared in a hood.
5.4.1 Place about 9.8 ml of methanol in a 10 ml tared ground glass
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 minutes until all alcohol-wetted surfaces have dried. Weigh the
flask to the nearest 0.0001 g.
5.4.2 Add the assayed reference material, as described below.
5.4.2.1 Liquids. Using a 100 /LtL syringe, immediately add
two or more drops of assayed reference material to the flask; then
reweigh. The liquid must fall directly into the alcohol without
contacting the neck of the flask.
5.4.2.2 Gases. To prepare standards for any compounds
that boil below 30°C (e.g. bromomethane, chloroethane,
chloromethane, dichlorodifluoromethane, trichlorofluoromethane,
vinyl chloride), fill a 5 ml valved gas-tight syringe with the
reference standard to the 5.0 ml mark. Lower the needle to 5 mm
above the methanol meniscus. Slowly introduce the reference
standard above the surface of the liquid. The heavy gas rapidly
dissolves in the methanol. This may also be accomplished by using
a lecture bottle equipped with a Hamilton Lecture Bottle Septum
(#86600). Attach Teflon tubing to the side-arm relief valve and
direct a gentle stream of gas into the methanol meniscus.
5.4.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.4.4 Transfer the stock standard solution into a bottle with a
Teflon lined screw-cap. Store, with minimal headspace, at -10°C to -20°C
and protect from light.
5.4.5 Prepare fresh stock standards for gases weekly or sooner if
comparison with check standards indicates a problem. Reactive compounds
such as 2-chloroethyl vinyl ether may need to be prepared more frequently.
All other standards must be replaced after six months. Both gas and
liquid standards must be monitored closely by comparison to the initial
calibration curve and by comparison to QC check standards. It may be
necessary to replace the standards more frequently if either check exceeds
a 20% drift.
5.4.6 Optionally calibration using a certified gaseous mixture can
be accomplished daily utilizing commercially available gaseous analyte
8010B - 4 Revision 2
September 1994
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mixture of bromomethane, chloromethane, chloroethane, vinyl chloride,
dichlorodifluoromethane and trichlorofluoromethane in nitrogen. These
mixtures of documented quality are stable for as long as six months
without refrigeration. (VOA-CYL III, RESTEK Corporation, Cat. #20194 or
equivalent).
5.5 Secondary dilution standards. Using stock standard solutions,
prepare secondary dilution standards in methanol, as needed, containing the
compounds of interest, either singly or mixed together. The secondary dilution
standards should be prepared at concentrations such that the aqueous calibration
standards prepared in Sec. 5.6 will bracket the working range of the analytical
system. Secondary dilution standards should be stored with minimal headspace for
volatiles and should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards from them.
5.6 Calibration standards. Prepare calibration standards in
organic-free reagent water from the secondary dilution of the stock standards,
at a minimum of five concentrations. One of the concentrations should be at a
concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of the concentrations
found in real samples or should define the working range of the GC. Each
standard should contain each analyte for detection by this method (e.g. some or
all of the compounds listed in Table 1 may be included). In order to prepare
accurate aqueous standard solutions, the following precautions must be observed.
5.6.1 Do not inject more than 20 /uL of alcoholic standards into
100 mL of water.
5.6.2 Use a 25 /^L Hamilton 702N microsyringe or equivalent
(variations in needle geometry will adversely affect the ability to
deliver reproducible volumes of methanolic standards into water).
5.6.3 Rapidly inject the alcoholic standard into the filled
volumetric flask. Remove the needle as fast as possible after injection.
5.6.4 Mix aqueous standards by inverting the flask three times only.
5.6.5 Fill the sample syringe from the standard solution contained
in the expanded area of the flask (do not use any solution contained in
the neck of the flask).
5.6.6 Never use pipets to dilute or transfer samples or aqueous
standards.
5.6.7 Aqueous standards are not stable and should be discarded after
one hour, unless properly sealed and stored. The aqueous standards can
be stored up to 24 hours, if held in sealed vials with zero headspace.
5.7 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
8010B - 5 Revision 2
September 1994
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internal standard can be suggested that is applicable to all samples. The
compounds recommended for use as surrogate spikes (Sec. 5.8) have been used
successfully as internal standards, because of their generally unique retention
times.
5.7.1 Prepare calibration standards at a minimum of five
concentrations for each analyte of interest as described in Sec. 5.6.
5.7.2 Prepare a spiking solution containing each of the internal
standards using the procedures described in Sees. 5.4 and 5.5. It is
recommended that the secondary dilution standard be prepared at a
concentration of 15 ng/pL of each internal standard compound. The
addition of 10 juL of this standard to 5.0 ml of sample or calibration
standard would be equivalent to 30 /ug/L.
5.7.3 Analyze each calibration standard according to Sec. 7.0,
adding 10 juL of internal standard spiking solution directly to the
syringe.
5.8 Surrogate standards - The analyst should monitor both the
performance of the analytical system and the effectiveness of the method in
dealing with each sample matrix by spiking each sample, standard, and
organic-free reagent water blank with surrogate halocarbons. A combination of
bromochloromethane, bromochlorobenzene and bromofluorobenzene is recommended to
encompass the range of temperature program used in this method. From stock
standard solutions prepared as in Sec. 5.4, add a volume to give 750 /j,g of each
surrogate to 45 ml of organic-free reagent water contained in a 50 ml volumetric
flask, mix, and dilute to volume for a concentration of 15 ng//zL. Add 10 /xL of
this surrogate spiking solution directly into the 5 ml syringe with every sample
and reference standard analyzed. If the internal standard calibration procedure
is used, the surrogate compounds may be added directly to the internal standard
spiking solution (Sec. 5.7.2).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Volatile compounds are introduced into the gas chromatograph using
either direct injection or purge-and-trap (Method 5030). Method 5030 may be used
directly on ground water samples or low-concentration contaminated soils and
sediments. For medium-concentration soils or sediments, methanolic extraction,
as described in Method 5030, may be necessary prior to purge-and-trap analysis.
7.2 Gas chromatographic conditions (Recommended)
7.2.1 Column 1:
Helium flow rate = 40 mL/min
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Temperature program:
Initial temperature = 45°C, hold for 3 minutes
Program = 45°C to 220°C at 8°C/nnn
Final temperature = 220°C, hold for 15 minutes.
7.2.2 Column 2:
Helium flow rate = 40 mL/min
Temperature program:
Initial temperature = 50°C, hold for 3 minutes
Program = 50°C to 170°C at 6°C/min
Final temperature = 170°C, hold for 4 minutes.
7.3 Calibration. The procedure for internal or external calibration may
be used. Refer to Method 8000 for a description of each of these procedures. Use
Table 1 and Table 2 for guidance on selecting the lowest point on the calibration
curve.
7.3.1 Calibration must take place using the same sample introduction
method that will be used to analyze actual samples (see Sec. 7.4.1).
7.4 Gas chromatographic analysis
7.4.1 Introduce volatile compounds into the gas chromatograph using
either Method 5030 (purge-and-trap) or the direct injection method (see
Sec. 7.4.1.1). If the internal standard calibration technique is used,
add 10 juL of internal standard to the sample prior to purging.
7.4.1.1 In very limited applications (e.g. aqueous process
wastes) direct injection of the sample onto the GC column with a
10 jiiL syringe may be appropriate. The detection limit is very high
(approximately 10,000 M9/L) therefore, it is only permitted where
concentrations in excess of 10,000 p.g/1 are expected or for water-
soluble compounds that do not purge. The system must be calibrated
by direct injection (bypassing the purge-and-trap device).
7.4.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
7.4.3 Table 1 summarizes the estimated retention times on the two
columns for a number of organic compounds analyzable using this method.
An example of the separation achieved by Column 1 is shown in Figure 1.
7.4.4 Record the sample volume purged or injected and the resulting
peak sizes (in area units or peak heights).
7.4.5 Refer to Method 8000 for guidance on calculation of
concentration.
7.4.6 If analytical interferences are suspected, or for the purpose
of confirmation, analysis using the second GC column is recommended.
8010B - 7 Revision 2
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7.4.7 If the response for a peak is off-scale, i.e., beyond the
calibration range of the standards, prepare a dilution of the sample with
organic-free reagent water. The dilution must be performed on a second
aliquot of the sample which has been properly sealed and stored prior to
use.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 8000 for gas chromatographic procedures. Quality control to ensure the
proper operation of the purge-and-trap device is covered in Method 5030.
8.2 Quality control required to validate the GC system operation is
found in Method 8000.
8.2.1 The quality control check sample concentrate (Method 8000)
should contain each analyte of interest at a concentration of 10 mg/L in
methanol.
8.2.2 Table 3 indicates the calibration and QC acceptance criteria,
for water samples, for this method. Table 4 gives method accuracy and
precision as functions of concentration, for water samples, for the
analytes of interest. The contents of both Tables should be used to
evaluate a laboratory's ability to perform and generate acceptable data
by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if recovery is within limits (limits established by performing
QC procedure outlined in Method 8000).
8.3.1 If recovery is not within limits, the following is required:
• Check to be sure that there are no errors in
calculations, surrogate solutions and internal standards. Also,
check instrument performance.
• Recalculate the data and/or re-analyze the sample if
any of the above checks reveal a problem.
• Re-extract and re-analyze the sample if none of the
above are a problem or flag the data as "estimated concentration".
9.0 METHOD PERFORMANCE
9.1 This method was tested by 20 laboratories using organic-free reagent
water, drinking water, surface water, and three industrial wastewaters spiked at
six concentrations over the range 8.0-500 jug/L. Single operator precision,
overall precision, and method accuracy were found to be directly related to the
concentration of the analyte, and essentially independent of the sample matrix.
Linear equations to describe these relationships are presented in Table 4.
8010B - 8 Revision 2
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9.2 The accuracy and precision obtained will be determined by the sample
matrix, sample introduction technique, and by the calibration procedure used.
9.3 The method detection limits reported in Table 1 were generated under
optimum analytical conditions by an Agency contractor (Ref. 6) as guidance, and
may not be readily achievable by all laboratories at all times.
10.0 REFERENCES
1. Bellar, T.A.; Lichtenberg, J.J. vL. Amer. Water Works Assoc. 1974, 66(12),
pp. 739-744.
2. Bellar, T.A.; Lichtenberg, J.J., Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds, Measurement of Organic Pollutants in Water and Wastewater; Van
Hall, Ed.; ASTM STP 686, pp 108-129, 1979.
3. "Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters: Category 11 - Purgeables and Category 12 -
Acrolein, Acrylonitrile, and Dichlorodifluoromethane"; report for EPA
Contract 68-03-2635.
4. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act: Final Rule and Interim
Final Rule and Proposed Rule", October 26, 1984.
5. "EPA Method Validation Study 23, Method 601 (Purgeable Halocarbons)";
report for EPA Contract 68-03-2856.
6. Gebhart, J.E., S.V. Lucas, S.J. Naber, A.M. Berry, T.H. Danison and H.M.
Burkholder, "Validation of SW-846 Methods 8010, 8015, and 8020"; Report
for EPA Contract 68-03-1760, Work Assignment 2-15; US EPA, EMSL-
Cincinnati, 1987.
8010B - 9 Revision 2
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
FOR HALOGENATED VOLATILE ORGANICS
Compound
ATlyl chloride*^
Benzyl chloride*'0
Bis(2-chloroethoxy)methane*
Bis(2-chloroisopropyl ) ether*
Bromobenzene
Bromodichl oromethane
Bromoform*
Bromomethane*
Carbon tetrachloride*
Chloroacetaldehyde*
Chlorobenzene*
Chloroethane
Chloroform*
1-Chlorohexane
2-Chloroethyl^vinyl ether*
Chloromethane*
Chloromethyl methyl ether*
4-Chlorotoluene
Di bromochl oromethane
l,2-Dibromo-3-chloropropane*
Dibromomethane*
1 , 2-Di chl orobenzene*
1,3-Dichlorobenzene*
1, 4 -Di chl orobenzene*
l,4-Dichloro-2-butene*
Di chl orodi f 1 uoromethane*'d
1,1-Dichloroethane*
1, 2-Di chloroethane*
1, 1-Dichloroethene*
trans -1, 2-Di chl proethene*
Di chl oromethane*
1 , 2-Di chl oropropane*
trans-l,3-Dichloropropene*
Ethylene dibromide
1,1,2 , 2 -Tetrachl oroethane*
1,1, 1,2 -Tetrachl oroethane*
Tetrachloroethene*
1 , 1 , 1-Tri chl oroethane^
1 , 1 , 2-Tri chl oroethane*
CAS
Registry
Number
107-05-1
100-44-7
111-91-1
39638-32-9
108-86-1
75-27-4
75-25-2
74-83-9
56-23-5
107-20-0
108-90-7
75-00-3
67-66-3
544-10-5
110-75-8
74-87-3
107-30-2
106-43-4
124-48-1
96-12-8
74-95-3
95-50-1
541-73-1
106-46-7
764-41-0
75-71-8
75-34-3
107-06-2
75-35-4
156-60-5
75-09-2
78-87-5
10061-02-5
106-93-4
79-34-5
630-20-6
127-18-4
71-55-6
79-00-5
Retention Time
(minutes)
Column 1 Column 2
10.17
30.29
38.60
34.79
29.05
15.44
21.12
2.90
14.58
(b)
25.49
5.18
12.62
26.26
19.23
1.40
8.88
34.46
18.22
28.09
13.83
37.96
36.88
38.64
23.45
3.68
11.21
13.14
10.04
11.97
7.56
16.69
16.976
19.59
23.12
21.10
23.05
14.48
18.27
(b)
(b)
(b)
(b)
(b)
14.62
19.17
7.05
11.07
(b)
18.83
8.68
12.08
(b)
(b)
5.28
(b)
(b)
16.62
(b)
14.92
23.52
22.43
22.33
(b)
(b)
12.57
15.35
7.72
9.38
10.12
16.62
16.60
(b)
(b)
21.70
14.97
13.10
18.07
Method
Detection
Limit3
(M9/L)
(b)
(b)
(b)
(b)
(b)
0.002
0.020
0.030
0.003
(b)
0.001
0.008
0.002
(b)
0.130
0.010
(b)
(b)
(b)
0.030
(b)
(b)
(b)
(b)
(b)
(b)
0.002
0.002
0.003
0.002
(b)
(b)
0.340
(b)
0.010
(b)
0.001
0.003
0.007
8010B - 10
Revision 2
September 1994
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TABLE 1.
Continued
Compound
CAS
Registry
Number
Retention Time
(minutes)
Column 1 Column 2
Method
Detection
Limit3
(M9/L)
Trichloroethene
Trichlorofluoromethane*
1,2,3-Trichloropropane*
Vinyl Chloride*
79-01-6
75-69-4
96-18-4
75-01-4
17.40
9.26
22.95
3.25
13.12
(b)
(b)
5.28
0.001
(b)
(b)
0.006
a =
b =
* =
c =
d =
e =
Using purge-and-trap method (Method 5030). See Sec. 9.3.
Not determined
Appendix VIII compounds
Demonstrated very erratic results
See Sec. 4.10.2 of Method 5030
material
Estimated retention time
when tested by purge-and-trap
for guidance on selection of trapping
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQL)
FOR VARIOUS MATRICES3
Matrix
Factor
Ground water 10
Low-concentration soil 10
Water miscible liquid waste 500
High-concentration soil and sludge 1250
Non-water miscible waste 1250
EQL = [Method detection limit (see Table 1)] X [Factor found in
this table]. For non-aqueous samples, the factor is on a wet-
weight basis. Sample EQLs are highly matrix-dependent. The EQLs
listed herein are provided for guidance and may not always be
achievable.
8010B - 11
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TABLE 3.
CALIBRATION AND QC ACCEPTANCE CRITERIA3
Range Limit
for Q for S
Analyte
Bromodi chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
Chloroethane
2-Chloroethylvinyl ether
Chloroform
Chl oromethane
Di bromochl oromethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1 , 4-Di chl orobenzene
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans-l,2-Dichloroethene
Dichloromethane
1,2-Dichloropropane
cis-l,3-Dichloropropene
trans- 1,3-Di chl oropropene
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
1,1,1-Tri chl oroethane
1, 1, 2 -Tri chl oroethane
Trichloroethene
Trichlorofluoromethane
Vinyl chloride
(M9/L) (M9/L)
15.2-24.8
14.7-25.3
11.7-28.3
13.7-26.3
14.4-25.6
15.4-24.6
12.0-28.0
15.0-25.0
11.9-28.1
13.1-26.9
14.0-26.0
9.9-30.1
13.9-26.1
16.8-23.2
14.3-25.7
12.6-27.4
12.8-27.2
15.5-24.5
14.8-25.2
12.8-27.2
12.8-27.2
9.8-30.2
14.0-26.0
14.2-25.8
15.7-24.3
15.4-24.6
13.3-26.7
13.7-26.3
4.3
4.7
7.6
5.6
5.0
4.4
8.3
4.5
7.4
6.3
5.5
9.1
5.5
3.2
5.2
6.6
6.4
4.0
5.2
7.3
7.3
9.2
5.4
4.9
3.9
4.2
6.0
5.7
Q = Concentration measured in QC check sampl
Range
for x
(M9/L)
10.7-32.0
5.0-29.3
3.4-24.5
11.8-25.3
10.2-27.4
11.3-25.2
4.5-35.5
12.4-24.0
D-34.9
7.9-35.1
1.7-38.9
6.2-32.6
11.5-25.5
11.2-24.6
13.0-26.5
10.2-27.3
11.4-27.1
7.0-27.6
10.1-29.9
6.2-33.8
6.2-33.8
6.6-31.8
8.1-29.6
10.8-24.8
9.6-25.4
9.2-26.6
7.4-28.1
8.2-29.9
e, in /ig/L.
S = Standard deviation of four recovery measurements, in
x = Average recovery
P, Ps = Percent recovery
D = Detected; result
3 r ** 4 + r\ v< •! •» -P^/-.m /in rco D->
Range
P, Ps
(%)
42-172
13-159
D-144
43-143
38-150
46-137
14-186
49-133
D-193
24-191
D-208
7-187
42-143
47-132
51-147
28-167
38-155
25-162
44-156
22-178
22-178
8-184
26-162
41-138
39-136
35-146
21-156
28-163
M9/L.
for four recovery measurements, in /^g/L.
measured.
must be greater
v+ T3C fnv Ma+kn
than zero.
A CHI on
A utnva /*a1/~ii1
a + a/H accnminn
a QC check sample concentration of 20 M9/L.
8010B - 12
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TABLE 4.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION3
Analyte
Bromodi chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
Chloroethane
2-Chloroethyl vinyl etherb
Chloroform
Chl oromethane
Di bromochl oromethane
1, 2 -Di chlorobenzene
1,3-Dichlorobenzene
1, 4 -Di chlorobenzene
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans-l,2-Dichloroethene
Di chl oromethane
l,2-Dichloropropaneb
cis-l,3-Dichloropropeneb
trans- 1,3-Di chl oropropeneb
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
1,1,1-Tri chl oroethane
1 , 1 , 2-Tri chl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
Vinyl chloride
Accuracy, as
recovery, x'
(M9/L)
1.12C-1.02
0.96C-2.05
0.76C-1.27
0.98C-1.04
l.OOC-1.23
0.99C-1.53
l.OOC
0.93C-0.39
0.77C+0.18
0.94C+2.72
0.93C+1.70
0.95C+0.43
0.93C-0.09
0.95C-1.08
1.04C-1.06
0.98C-0.87
0.97C-0.16
0.91C-0.93
l.OOC
l.OOC
l.OOC
0.95C+0.19
0.94C+0.06
0.90C-0.16
0.86C+0.30
0.87C+0.48
0.89C-0.07
0.97C-0.36
Single analyst
precision, s '
(M9/U
0.11X+0.04
0.12X+0.58
0.28X+0.27
0.15X+0.38
0.15X-0.02
0.14X-0.13
0.20X
0.13X+0.15
0.28X-0.31
0.11X+1.10
0.20X+0.97
0.14X+2.33
0.15X+0.29
0.08X+0.17
0.11X+0.70
0.21X-0.23
0.11X+1.46
0.11X+0.33
0.13X
0.18X
0.18X
0.14X+2.41
0.14X+0.38
0.15X+0.04
0.13X-0.14
0.13X-0.03
0.15X+0.67
0.13X+0.65
Overall
precision,
S' (jug/L)
0.20X+1.00
0.21X+2.41
0.36X+0.94
0.20X+0.39
0.18X+1.21
0.17X+0.63
0.35X
0.19X-0.02
0.52X+1.31
0.24X+1.68
0.13X+6.13
0.26X+2.34
0.20X+0.41
0.14X+0.94
0.15X+0.94
0.29X-0.04
0.17X+1.46
0.21X+1.43
0.23X
0.32X
0.32X
0.23X+2.79
0.18X+2.21
0.20X+0.37
0.19X+0.67
0.23X+0.30
0.26X+0.91
0.27X+0.40
x' = Expected recovery for one or more measurements of a sample containing
a concentration of C, in /ug/L.
sr'= Expected single analyst standard deviation of measurements at an
average concentration of x, in ng/l.
S' = Expected interlaboratory standard deviation of measurements at an
average concentration found of x, in jug/L.
C = True value for the concentration, in M9/L.
X = Average recovery found for measurements of samples containing a
concentration of C, in M9/L.
a From 40 CFR Part 136 for Method 601.
b
Estimates based upon the performance in a single laboratory.
8010B - 13
Revision 2
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FIGURE 1.
GAS CHROMATOGRAM OF HALOGENATED VOLATILE ORGANICS
Column:
Program:
Detector:
1X SP-1000 on Carbopack-B
45°C-3 Minutes, 8'C/Hinute to 220°C
Hall 700-A Electrolytic Conductivity
14
20
n 74
HfTf MTHW TMf MHIUTUI
8010B - 14
Revision 2
September 1994
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METHOD 8010B
HALOGENATED VOLATILE ORGANICS BY GAS CHROMATOGRAPHY
Start
7.1 Introduce compounds
into gas chromatograph
by direct injection or
purge-and-trap
(Method 5030)
7.2 Set gas
chromatograph
condition.
7.3 Calibrate
(refer to Method 8000)
7.4.1 Introduce
volatile compounds
into gas chromatograph
by purge-and-trap or
direct injection.
7.4.2 Follow Method
8000 for analysis
sequence, etc.
7.4.4 Record volume
purged or injected
and peak sizes.
7.4.5 Calculate
concentration
(refer to Method 8000)
7.4.6 Are
analytical
interferences
suspected?
7.4.7 Is
response for
a peak
off-scale?
7.4.6 Analyze using
second GC column.
7.4.7 Dilute second
aliquot of sample.
8010B - 15
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METHOD 8011
1,2-DIBROMOETHANE AND 1.2-DIBROMO-3-CHLOROPROPANE
BY MICROEXTRACTION AND GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 This method is applicable to the determination of the following
compounds in drinking water and ground water:
Compound Name CAS No.a
1,2-Dibromoethane (EDB) 106-93-4
l,2-Dibromo-3-chloropropane (DBCP) 96-12-8
8 Chemical Abstract Services Registry Number.
1.2 For compounds and matrices other than those listed in Section 1.1,
the laboratory must demonstrate the usefulness of the method by collecting
precision and accuracy data on actual samples and provide qualitative
confirmation of results by gas chromatography/mass spectrometry (GC/MS).
1.3 The experimentally determined method detection limits (MDL) for EDB
and DBCP were calculated to be 0.01 /Lig/L. The method has been shown to be
useful for these analytes over a concentration range of approximately 0.03 to 200
jiig/L. Actual detection limits are highly dependent upon the characteristics of
the gas chromatographic system, sample matrix, and calibration.
1.4 This method is restricted to use by or under the Supervision of
analysts experienced in the use of gas chromatography and in the interpretation
of gas chromatograms. Each analyst must demonstrate the ability to generate
acceptable results with this method using the procedure described in Section 8.2.
1.5 1,2-Dibromoethane and l,2-Dibromo-3-chloropropane have been
tentatively classified as known or suspected human or mammalian carcinogens.
Pure standard materials and stock standard solutions of these compounds should
be handled in a hood. A NIOSH/MESA approved toxic gas respirator should be worn
when the analyst handles high concentrations of these toxic compounds.
2.0 SUMMARY OF METHOD
2.1 Thirty five ml of sample are extracted with 2 mL of hexane. Two fiL
of the extract are then injected into a gas chromatograph equipped with a
linearized electron capture detector for separation and analysis. Aqueous matrix
spikes are extracted and analyzed in an identical manner as the samples in order
to compensate for possible extraction losses.
2.2 The extraction and analysis time is 30 to 50 minutes per sample
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depending upon the analytical conditions chosen. See Table 1 and Figure 1.
2.3 Confirmatory evidence is obtained using a different column (Table 1).
3.0 INTERFERENCES
3.1 Impurities contained in the extracting solvent (hexane) usually
account for the majority of the analytical problems. Reagent blanks should be
analyzed for each new bottle of hexane before use. Indirect daily checks on the
hexane are obtained by monitoring the reagent blanks. Whenever an interference
is noted in the method or instrument blank, the laboratory should reanalyze the
hexane. Low level interferences generally can be removed by distillation or
column chromatography, however, it is generally more economical to obtain a new
source of hexane solvent. Interference-free hexane is defined as containing less
than 0.01 /ug/L of the analytes. Protect interference-free hexane by storing it
in an area known to be free of organochlorine solvents.
3.2 Several instances of accidental sample contamination have been
attributed to diffusion of volatile organics through the septum seal into the
sample bottle during shipment and storage. Trip blanks must be used to monitor
for this problem.
3.3 This 1 iquid/1 iquid extraction technique extracts a wide boil ing range
of non-polar organic compounds and, in addition, extracts some polar organic
compounds.
3.4 EDB at low concentrations may be masked by very high concentrations
of dibromochloromethane (DBCM), a common chlorinated drinking water contaminant,
when using the confirmation column.
4.0 APPARATUS AND MATERIALS
4.1 Microsyringe - 10, 25, and 100 juL with a 2 in. x 0.006 in. needle
(Hamilton 702N or equivalent).
4.2 Gas Chromatograph
4.2.1 The GC must be capable of temperature programming and should
be equipped with a linearized electron capture detector and a capillary
column splitless injector.
4.2.2 Columns
4.2.2.1 Column A - 0.32 mm ID x 30 m fused silica
capillary with dimethyl silicone mixed phase (Durawax-DX 3, 0.25 /xm
film, or equivalent).
4.2.2.2 Column B (confirmation column) - 0.32 mm ID x 30 m
fused silica capillary with methyl polysiloxane phase (DB-1, 0.25 jum
film, or equivalent).
4.3 Volumetric flasks, Class A - 10 mL.
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4.4 Glass bottles - 15 mL, with Teflon lined screw caps or crimp tops.
4.5 Analytical balance - 0.0001 g.
4.6 Graduated cylinder - 50 mL.
4.7 Transfer pi pet.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Hexane, C6H14 - UV grade (Burdick and Jackson #216 or equivalent).
5.4 Methyl alcohol, CH3OH - Demonstrated to be free of analytes.
5.5 Sodium chloride, NaCl - Pulverize a batch of NaCl and place it in a
muffle furnace at room temperature. Increase the temperature to 400°C for
30 minutes. Store in a capped bottle.
5.6 1,2-Dibromoethane (99%), C2H4Br2, (Aldrich Chemical Company, or
equivalent).
5.7 l,2-Dibromo-3-chloropropane (99.4%), C3H5Br2Cl, (AMVAC Chemical
Corporation, Los Angeles, California, or equivalent).
5.8 Stock standards - These solutions may be purchased as certified
solutions or prepared from pure standards using the following procedures:
5.8.1 Place about 9.8 mL of methanol into a 10 mL ground glass
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 minutes and weigh to the nearest 0.0001 g.
5.8.2 Use a 25 ptL syringe and immediately add two or more drops
(» 10 juL) of standard to the flask. Be sure that the standard falls
directly into the alcohol without contacting the neck of the flask.
5.8.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard.
5.8.4 Store stock standards in 15 mL bottles equipped with Teflon
lined screw-caps or crimp tops. Stock standards are stable for at least
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four weeks when stored at 4°C and away from light.
5.9 Intermediate standard - Use stock standards to prepare an
intermediate standard that contains both analytes in methanol. The intermediate
standard should be prepared at a concentration that can be easily diluted to
prepare aqueous calibration standards that will bracket the working concentration
range. Store the intermediate standard with minimal headspace and check
frequently for signs of deterioration or evaporation, especially just before
preparing calibration standards. The storage time described for stock standards
also applies to the intermediate standard.
5.10 Quality control (QC) reference sample - Prepare a QC reference sample
concentrate at 0.25 mg/L of both analytes from standards from a different source
than the standards used for the stock standard.
5.11 Check standard - Add an appropriate volume of the intermediate
standard to an aliquot of organic-free reagent water in a volumetric flask. Do
not add more than 20 /zL of an alcoholic intermediate standard to the water or
poor precision will result. Use a 25 /iL microsyringe and rapidly inject the
alcoholic intermediate standard into the expanded area of the almost filled
volumetric flask. Remove the needle as quickly as possible after injection. Mix
by inverting the flask several times. Discard the contents contained in the neck
of the flask. Aqueous calibration standards should be prepared every 8 hours.
6.0 SAMPLE COLLECTION, PRESERVATION, AND STORAGE
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Recommended Chromatographic Conditions
Two gas chromatography columns are recommended. Column A is a highly
efficient column that provides separations for EDB and DBCP without interferences
from trihalomethanes. Column A should be used as the primary analytical column
unless routinely occurring analytes are not adequately resolved. Column B is
recommended for use as a confirmatory column when GC/MS confirmation is not
available. Retention times for EDB and DBCP on these columns are presented in
Table 1.
Column A:
Injector temperature: 200°C.
Detector temperature: 290°C.
Carrier gas (Helium) Linear velocity: 25 cm/sec.
Temperature program:
Initial temperature: 40°C, hold for 4 min.
Program: 40°C to 190°C at 8°C/min.
Final temperature: 190°C, hold for 25 min., or
until all expected analytes
have eluted.
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See Figure 1 for a sample chromatogram and Table 1 for retention data.
Column B:
Injector temperature: 200°C.
Detector temperature: 290°C.
Carrier gas (Helium) Linear velocity: 25 cm/sec.
Temperature program:
Initial temperature: 40°C, hold for 4 min.
Program: 40°C to 270°C at 10°C/nrin.
Final temperature: 270°C, hold for 10 min., or
until all expected analytes
have eluted.
See Table 1 for retention data.
7.2 Calibration
7.2.1 Prepare at least five calibration standards. One should
contain EDB and DBCP at a concentration near, but greater than, the method
detection limit (Table 1) for each compound. The others should be at
concentrations that bracket the range expected in the samples. For
example, if the MDL is 0.01 M9/U and a sample expected to contain
approximately 0.10 pg/L is to be analyzed, aqueous calibration standards
should be prepared at concentrations of 0.03 ;ug/L, 0.05 /xg/L, 0.10
0.15 M9/L, and 0.20
7.2.2 Analyze each calibration standard and tabulate peak height or
area response versus the concentration in the standard. Prepare a
calibration curve for each compound. Alternatively, if the ratio of
response to concentration (calibration factor) is a constant over the
working range (< 10% relative standard deviation), linearity can be
assumed and the average ratio or calibration factor can be used in place
of a calibration curve.
7.3 Sample preparation
7.3.1 Remove samples and standards from storage and allow them to
reach room temperature.
7.3.2 For samples and field blanks contained in 40 ml bottles,
remove the container cap. Discard a 5 ml volume using a 5 ml transfer
pipet. Replace the container cap and weigh the container with contents to
the nearest 0.1 g and record this weight for subsequent sample volume
determination.
7.3.3 For calibration standards, check standards, QC reference
samples, and blanks, measure a 35 mL volume using a 50 ml graduated
cylinder and transfer it to a 40 mL sample container.
7.4 Extraction
7.4.1 Remove the container cap and add 7 g of NaCl to all samples.
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7.4.2 Recap the sample container and dissolve the NaCl by shaking by
hand for about 20 seconds.
7.4.3 Remove the cap and using a transfer pipet, add 2.0 ml of
hexane. Recap and shake vigorously by hand for 1 minute. Allow the water
and hexane phases to separate. If stored at this stage, keep the
container upside down.
7.4.4 Remove the cap and carefully transfer a sufficient amount
(0.5-1.0 mL) of the hexane layer into a vial using a disposable glass
pipet.
7.4.5 Transfer the remaining hexane phase, being careful not to
include any of the water phase, into a second vial. Reserve this second
vial at 4°C for reanalysis if necessary.
7.5 Analysis
7.5.1 Transfer the first sample vial to an autosampler set up to
inject 2.0 p.1 portions into the gas chromatograph for analysis.
Alternately, 2 /uL portions of samples, blanks and standards may be
manually injected, using the solvent flush technique, although an auto
sampler is strongly recommended.
7.6 Determination of sample volume
7.6.1 For samples and field blanks, remove the cap from the sample
container. Discard the remaining sample/hexane mixture. Shake off the
remaining few drops using short, brisk wrist movements. Reweigh the empty
container with original cap and calculate the net weight of sample by
difference to the nearest 0.1 g. This net weight is equivalent to the
volume of water extracted.
7.7 Calculations
7.7.1 Identify EDB and DBCP in the sample chromatogram by comparing
the retention time of the suspect peak to retention times generated by the
calibration standards and the check standard.
7.7.2 Use the calibration curve or calibration factor to directly
calculate the uncorrected concentration (Cj) of each analyte in the sample
(e.g. calibration factor x response).
7.7.3 Calculate the sample volume (Vs) as equal to the net sample
weight:
Vs (mL) = gross weight (grams) - bottle tare (grams)
7.7.4 Calculate the corrected sample concentration as:
Concentration (jLtg/L) = C, x 35
Vs
7.7.5 Report the results for the unknown samples in fj.g/1. Round the
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results to the nearest 0.01 /ig/L or two significant figures.
8.0 QUALITY CONTROL
8.1 Each laboratory that uses this method is required to operate a formal
quality control program.
8.1.1 The laboratory must make an initial determination of the
method detection limits and demonstrate the ability to generate acceptable
accuracy and precision with this method. This is established as described
in Section 8.2.
8.1.2 In recognition of laboratory advances that are occurring in
chromatography, the laboratory is permitted certain options to improve the
separations or lower the cost of measurements. Each time such a
modification is made to the method, the analyst is required to repeat the
procedure in Section 7.1 and 8.2.
8.1.3 The laboratory must analyze a reagent and calibration blank to
demonstrate that interferences from the analytical system are under
control every twenty samples or per analytical batch, whichever is more
frequent.
8.1.4 The laboratory must, on an ongoing basis, demonstrate through
the analyses of QC reference samples and check standards that the
operation of the measurement system is in control. The frequency of the
check standard analyses is equivalent to 5% of all samples or every
analytical batch, whichever is more frequent. On a weekly basis, the QC
reference sample must be run.
8.2 To establish the ability to achieve low detection limits and generate
acceptable accuracy and precision, the analyst must perform the following
operations:
8.2.1 Prepare seven samples each at a concentration of 0.03 jig/L.
8.2.2 Analyze the samples according to the method beginning in
Section 7.0.
8.2.3 Calculate the average concentration (X) in ^g/L and the
standard deviation of the concentrations (s) in iig/L, for each analyte
using the seven results. Then calculate the MDL at 99% confidence level
for seven replicates as 3.143s.
8.2.4 For each analyte in an aqueous matrix sample, X must be
between 60% and 140% of the true value. Additionally, the MDL may not
exceed the 0.03 jig/L spiked concentration. If both analytes meet the
acceptance criteria, the system performance is acceptable and analysis of
actual samples can begin. If either analyte fails to meet a criterion,
repeat the test. It is recommended that the laboratory repeat the MDL
determination on a regular basis.
8.3 The laboratory must demonstrate on a frequency equivalent to 5% of
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the sample load or once per analytical batch, whichever is more frequent, that
the measurement system is in control by analyzing a check standard of both
analytes at 0.25 jig/L.
8.3.1 Prepare a check standard (0.25 [ig/L) by diluting the
intermediate standard with water to 0.25 jig/L.
8.3.2 Analyze the sample according to Section 7.0 and calculate the
recovery for each analyte. The recovery must be between 60% and 140% of
the expected value for aqueous matrices. For non-aqueous matrices, the
U.S. EPA will set criteria after more interlaboratory data are gathered.
8.3.3 If the recovery for either analyte falls outside the
designated range, the analyte fails the acceptance criteria. A second
calibration verification standard containing each analyte that failed must
be analyzed. Repeated failure, however, will confirm a general problem
with the measurement system. If this occurs, locate and correct the
source of the problem and repeat the test.
8.4 On a weekly basis, the laboratory must demonstrate the ability to
analyze a QC reference sample.
8.4.1 Prepare a QC reference sample at 0.10 ng/L by diluting the QC
reference sample concentrate (Section 5.9).
8.4.2 For each analyte in an aqueous matrix, the recovery must be
between 60% and 140% of the expected value. When either analyte fails the
test, the analyst must repeat the test only for that analyte which failed
to meet the criteria. Repeated failure, however, will confirm a general
problem with the measurement system or faulty samples and/or standards.
If this occurs, locate and correct the source of the problem and repeat
the test. For non-aqueous matrices, the U.S. EPA will set criteria after
more interlaboratory data are gathered.
8.5 Instrument performance - Check the performance of the entire
analytical system daily using data gathered from analyses of blanks, standards,
and replicate samples.
8.5.1 Peak tailing significantly in excess of that shown in the
chromatogram (Figure 1) must be corrected. Tailing problems are generally
traceable to active sites on the GC column or to the detector operation.
8.5.2 Check the precision between replicate analyses. A properly
operating system should perform with an average relative standard
deviation of less than 10%. Poor precision is generally traceable to
pneumatic leaks, especially at the injection port.
9.0 METHOD PERFORMANCE
9.1 Method detection limits are presented in Table 1. Single laboratory
accuracy and precision at several concentrations in tap water are presented in
Table 2.
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9.2 In a preservation study extending over a 4 week period, the average
percent recoveries and relative standard deviations presented in Table 3 were
observed for organic-free reagent water (acidified), tap water and ground water.
The results for acidified and non-acidified samples were not significantly
different.
10.0 REFERENCES
1. Optimization of Liquid-Liquid Extraction Methods for Analysis of Organics
in Water, EPA-600/S4-83-052, 1984.
2. Henderson, J.E.; Peyton, G.R.; Glaze, W.H. Identification and Analysis of
Organic Pollutants in Water; Keith, L.H., Ed; Ann Arbor Sci.: Ann Arbor,
MI; 1976.
3. Richard J.J.; Junk, G.A. Journal AWWA 1977, 69, 62.
4. Budde, W.L.; Eichelberger, J.W. Organic Analyses Using Gas Chromatography-
Mass Spectrometry; Ann Arbor Science: Ann Arbor, MI; 1978.
5. Glaser, J.A.; et al. Environmental Science and Technology 1981, 15, 1426.
6. Methods for the Determination of Organic Compounds in Finished Drinking
Water and Raw Source Water; U.S. Environmental Protection Agency. Office
of Research and Development. Environmental Monitoring and Support
Laboratory. ORD Publication Offices of Center for Environmental Research
Information: Cincinnati, OH 1986.
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION
LIMITS (MDL) FOR 1,2-DIBROMOETHANE (EDB) AND
1.2-DIBROMO-3-CHLOROPROPANE (DBCP)
Analyte
Retention Time, Minutes
Column A Column B MDL (jig/L)
EDB
DBCP
9.5
17.3
8.9
15.0
0.01
0.01
Column A: Durawax-DX 3
Column B: DB-1
TABLE 2.
SINGLE LABORATORY ACCURACY AND PRECISION
FOR EDB AND DBCP IN TAP WATER
Analyte
EDB
DBCP
Number
of
Samples
7
7
7
7
7
7
Spike
Concentration
(M9A)
0.03
0.24
50.0
0.03
0.24
50.0
Average
Recovery
(%)
114
98
95
90
102
94
Relative
Standard
Deviation
(%)
9.5
11.8
4.7
11.4
8.3
4.8
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TABLE 3.
ACCURACY AND PRECISION AT 2.0
OVER A 4-WEEK STUDY PERIOD
Analyte
EDB
DBCP
Matrix1
RW-A
GW
GW-A
TW
TW-A
RW-A
GW
GW-A
TW
TW-A
Number
of Samples
16
15
16
16
16
16
16
16
16
16
Average
Accuracy
(% Recovery)
104
101
96
93
93
105
105
101
95
94
Relative
Std. Dev.
(%)
4.7
2.5
4.7
6.3
6.1
8.2
6.2
8.4
10.1
6.9
RW-A = Organic-free reagent water at pH 2
GW = Ground water, ambient pH
GW-A = Ground water at pH 2
TW = Tap water, ambient pH
TW-A = Tap water at pH 2
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FIGURE 1.
SAMPLE CHROMATOGRAM FOR EXTRACT OF WATER SPIKED
AT 0.114 M9/L WITH EDB AND DBCP
COLUMN: Fused silica capillary
LIQUID PHASE: Durawax-DX3
FILM THICKNESS: 0.25 urn
COLUMN DIMENSIONS: 30 M x 0.317
10
I I I 111 I I I
i i
2 4 • • 10 11 14 It It 20 22 24 2t
TIME (MIN)
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METHOD 8011
1,2-DIBROMOETHANE AND l,2-DIBROMO-3-CHLOROPROPANE
BY MICROEXTRACTION AND GAS CHROMATOGRAPHY
7 2 Check
ins t rutnent
performance
7 4 1 Add
NaCl to
samples
743 Add
hexane and
ex t ract
sample
744 Put
part of
extract in
vial
745 Save
remainder of
extract for
possible
reanalysis
7 5 Analyze
by GC
7 6 Determine
sample
volume
7 7 Calculate
concent rations
Stop
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METHOD 8015A
NONHALOGENATED VOLATILE ORGANICS BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
I.I Method 8015 is used to determine the concentration of various
nonhalogenated volatile organic compounds. The following compounds can be
determined by this method:
Appropriate Technique
Direct
Compound Name CAS No.a Purge-and-Trap Injection
Di ethyl ether
Ethanol
Methyl ethyl ketone (MEK)
Methyl isobutyl ketone (MIBK)
60-29-7
64-17-5
78-93-3
108-10-1
b
i
PP
PP
b
b
b
b
a Chemical Abstract Services Registry Number.
b Adequate response using this technique
i Inappropriate technique for this analyte
pp Poor purging efficiency, resulting in high EQLs
2.0 SUMMARY OF METHOD
2.1 Method 8015 provides gas chromatographic conditions for the detection
of certain nonhalogenated volatile organic compounds. Samples may be introduced
into the GC using direct injection or purge-and-trap (Method 5030). Ground water
samples must be analyzed by Method 5030. A temperature program is used in the
gas chromatograph to separate the organic compounds. Detection is achieved by
a flame ionization detector (FID).
2.2 The method provides an optional gas chromatographic column that may
be helpful in resolving the analytes from co-eluting non-target compounds and for
analyte confirmation.
3.0 INTERFERENCES
3.1 Refer to Method 5030 and 8000.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly chlorofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A trip blank prepared from
organic-free reagent water and carried through sampling and subsequent storage
and handling can serve as a check on such contamination.
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4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas Chromatograph - Analytical system complete with gas
chromatograph suitable for on-column injections or purge-and-trap sample
introduction and all required accessories, including detectors, column
supplies, recorder, gases, and syringes. A data system for measuring peak
heights and/or peak areas is recommended.
4.1.2 Columns
4.1.2.1 Column 1 - 8 ft x 0.1 in. ID stainless steel or
glass column packed with 1% SP-1000 on Carbopack-B 60/80 mesh or
equivalent.
4.1.2.2 Column 2 - 6 ft x 0.1 in. ID stainless steel or
glass column packed with n-octane on Porasil-C 100/120 mesh
(Durapak) or equivalent.
4.1.3 Detector - Flame ionization (FID).
4.2 Sample introduction apparatus - Refer to Method 5030 for the
appropriate equipment for sample introduction purposes.
4.3 Syringes - A 5 ml Luerlok glass hypodermic and a 5 ml, gas-tight with
shutoff valve.
4.4 Volumetric flasks, Class A - Appropriate sizes with ground glass
stoppers.
4.5 Microsyringes - 10 and 25 /zL with a 0.006 in. ID needle (Hamilton
702N or equivalent) and a 100 /LtL.
4.6 Analytical balance - 0.0001 g.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH. Pesticide quality or equivalent. Store away from
other solvents.
5.4 Stock standards - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standards in
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methanol using assayed liquids.
5.4.1 Place about 9.8 ml of methanol in a 10 ml tared, ground-glass
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 minutes or until all alcohol wetted surfaces have dried. Weigh
the flask to the nearest 0.0001 g.
5.4.2 Using a 100 /iL syringe, immediately add two or more drops of
assayed reference material to the flask; then reweigh. The liquid must
fall directly into the alcohol without contacting the neck of the flask.
5.4.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.4.4 Transfer the stock standard solution into a bottle with a
Teflon lined screw-cap. Store, with minimal headspace, at -10°C to -20°C
and protect from light.
5.4.5 Standards must be replaced after 6 months, or sooner if
comparison with check standards indicates a problem.
5.5 Secondary dilution standards - Using stock standard solutions, pre-
pare in methanol secondary dilution standards, as needed, that contain the
compounds of interest, either singly or mixed together. The secondary dilution
standards should be prepared at concentrations such that the aqueous calibration
standards prepared in Section 5.5 will bracket the working range of the
analytical system. Secondary dilution standards should be stored with minimal
headspace for volatiles and should be checked frequently for signs of degradation
or evaporation, especially just prior to preparing calibration standards from
them.
5.6 Calibration standards - Calibration standards at a minimum of five
concentrations are prepared in water from the secondary dilution of the stock
standards. One of the concentrations should be at a concentration near, but
above, the method detection limit. The remaining concentrations should
correspond to the expected range of concentrations found in real samples or
should define the working range of the GC. Each standard should contain each
analyte for detection by this method (e.g. some or all of the compounds listed
in Section 1.1 may be included). In order to prepare accurate aqueous standard
solutions, the following precautions must be observed:
5.6.1 Do not inject more than 20 jiiL of alcoholic standards into
100 ml of water.
5.6.2 Use a 25 /xL Hamilton 702N microsyringe or equivalent
(variations in needle geometry will adversely affect the ability to
deliver reproducible volumes of methanolic standards into water).
5.6.3 Rapidly inject the alcoholic standard into the filled
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volumetric flask. Remove the needle as fast as possible after injection.
5.6.4 Mix aqueous standards by inverting the flask three times only.
5.6.5 Fill the sample syringe from the standard solution contained
in the expanded area of the flask (do not use any solution contained in
the neck of the flask).
5.6.6 Never use pipets to dilute or transfer samples or aqueous
standards.
5.6.7 Aqueous standards are not stable and should be discarded after
1 hour, unless properly sealed and stored. The aqueous standards can be
stored up to 24 hours, if held in sealed vials with zero headspace.
5.7 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.7.1 Prepare calibration standards at a minimum of five
concentrations for each parameter of interest as described in Section 5.6.
5.7.2 Prepare a spiking solution containing each of the internal
standards using the procedures described in Sections 5.4 and 5.5. It is
recommended that the secondary dilution standard be prepared at a
concentration of 15 ng/juL of each internal standard compound. The
addition of 10 fj,L of this standard to 5.0 ml of sample or calibration
standard would be equivalent to 30
5.7.3 Analyze each calibration standard according to Section 7.0,
adding 10 /j,l of internal standard spiking solution directly to the
syringe.
5.8 Surrogate standards - The analyst should monitor both the performance
of the analytical system and the effectiveness of the method in dealing with each
sample matrix by spiking each sample, standard, and water blank with one or two
surrogate compounds recommended to encompass the range of temperature program
used in this method. From stock standard solutions prepared as in Section 5.4,
add a volume to give 750 jug of each surrogate to 45 ml of water contained in a
50 ml volumetric flask, mix, and dilute to volume for a concentration of
15 ng//iL. Add 10 /xL of this surrogate spiking solution directly into the 5 ml
syringe with every sample and reference standard analyzed. If the internal
standard calibration procedure is used, the surrogate compounds may be added
directly to the internal standard spiking solution (Section 5.7.2).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Section 4.1.
8015A - 4 Revision 1
July 1992
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7.0 PROCEDURE
7.1 Volatile compounds are introduced into the gas chromatograph either
by direct injection or purge-and-trap (Method 5030). Method 5030 may be used
directly on ground water samples or low-concentration contaminated soils and
sediments. For high-concentration soils or sediments, methanolic extraction, as
described in Method 5030, may be necessary prior to purge-and-trap analysis.
Method 5030 also provides guidance on the analysis of aqueous miscible and non-
aqueous miscible liquid wastes (see Section 7.4.1.1).
7.2 Chromatographic conditions (Recommended)
7.2.1 Column 1
Carrier gas (Helium) flow rate: 40 mL/min
Temperature program:
Initial temperature: 45°C, hold for 3 minutes
Program: 45°C to 220°C at 8°C/min
Final temperature: 220°C, hold for 15 minutes.
7.2.2 Column 2
Carrier gas (Helium) flow rate: 40 mL/min
Temperature program:
Initial temperature: 50°C, hold for 3 minutes
Program: 50°C to 170°C at 6°C/min
Final temperature: 170°C, hold for 4 minutes.
7.3 Calibration - Refer to Method 8000 for proper calibration techniques.
7.3.1 Calibration must take place using the same sample introduction
method that will be used to analyze actual samples (see Section 7.4.1).
7.3.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.4 Gas Chromatographic analysis
7.4.1 Introduce volatile compounds into the gas chromatograph using
either Method 5030 (purge-and-trap method) or the direct injection method.
If the internal standard calibration technique is used, add 10 jiiL of
internal standard to the sample prior to purging.
7.4.1.1 Direct injection - In very limited applications
(e.g. aqueous process wastes), direct injection of the sample into
the GC system with a 10 juL syringe may be appropriate. One such
application is for verification of the alcohol content of an aqueous
sample prior to determining if the sample is ignitable (Methods 1010
or 1020). In this case, it is suggested that direct injection be
used. The detection limit is very high (approximately 10,000 M9/L);
therefore, it is only permitted when concentrations in excess of
10,000 jug/L are expected or for water-soluble compounds that do not
purge. The system must be calibrated by direct injection (bypassing
the purge-and-trap device).
8015A - 5 Revision 1
July 1992
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Non-aqueous miscible wastes may also be analyzed by direct
injection if the concentration of target analytes in the sample
falls within the calibration range. If dilution of the sample is
necessary, follow the guidance for High Concentration samples in
Method 5030, Section 7.3.3.2.
7.4.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.*
7.4.3 Record the sample volume purged or injected and the resulting
peak sizes (in area units or peak heights).
7.4.4 Calculation of concentration is covered in Method 8000.
7.4.5 If analytical interferences are suspected, or for the purpose
of confirmation, analysis using the second GC column is recommended.
7.4.6 If the response for a peak is off-scale, prepare a dilution of
the sample with water. The dilution must be performed on a second aliquot
of the sample which has been properly sealed and stored prior to use.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 8000 for gas chromatographic procedures. Quality control to ensure the
proper operation of the purge-and-trap device is covered in Method 5030.
8.2 Quality control required to validate the GC system operation is found
in Method 8000, Section 8.6.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if recovery is within limits (limits established by performing
QC procedure outlined in Method 8000, Section 8.10).
8.3.1 If recovery is not within limits, the following is required:
• Check to be sure that there are no errors in calculations,
surrogate solutions, and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Re-extract and re-analyze the sample if none of the above are
a problem or flag the data as "estimated concentration".
9.0 METHOD PERFORMANCE
9.1 The accuracy and precision obtained will be determined by the sample
matrix, sample introduction technique, and calibration procedures used.
8015A - 6 Revision 1
July 1992
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9.2 Specific method performance information will be provided as it
becomes available.
10.0 REFERENCES
1. Bellar, T.A., and J.J. Lichtenberg, Determining Volatile Organics at
' ' " J. Amer. Water Works
Bellar, T.A., and J.J. Lichtenberg, Determining
Microgram-per-Liter Levels by Gas Chromatography,
Assoc., 66(12). pp. 739-744 (1974).
2. Bellar, T.A., and J.J. Lichtenberg, Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds, in Van Hall, ed., Measurement of Organic Pollutants in Water
and Wastewater, ASTM STP 686, pp. 108-129, 1979.
3. Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters: Category 11 - Purgeables and Category 12 -
Acrolein, Acrylonitrile, and Dichlorodifluoromethane, Report for EPA
Contract 68-03-2635 (in preparation).
8015A - 7 Revision 1
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METHOD 8015A
NONHALOGENATED VOLATILE ORGANICS BY GAS CHROMATOGRAPHY
Start
7 2 Set
chroma tographic
condi11ona
7 3 Calibrate
(refer to
Method 8000)
741 Introduce
sample into CC
by direct
infection or
purge-and-trap
742 Follow
Method 8000
for analysis
sequence,
etc
744 Record
volume purged
or
injected,and
peak sizes
7 4 5 Calculate
concentrations
(refer to
Method 8000)
7 4 6 Are
analytical
interferences
suspected?
7 4 7 Is peak
response off
scale?
746 Analyze
sample using
second CC
column
747 Dilute
second
aliquot of
sample
8015A - 8
Revision 1
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METHOD 8020A
AROMATIC VOLATILE ORGANICS BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8020 is used to determine the concentration of various
aromatic volatile organic compounds. The following compounds can be determined
by this method:
Appropriate Technique
Direct
Compound Name CAS No.a Purge-and-Trap Injection
Benzene
Chlorobenzene
1,2-Dichlorobenzene
1, 3 -Di chlorobenzene
1,4-Dichlorobenzene
Ethyl benzene
Toluene
Xylenes
a Chemical Abstract
b adequate response
71-43-2
108-90-7
95-50-1
541-73-1
106-46-7
100-41-4
108-88-3
Services Registry Number.
by this technique.
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
1.2 Table 1 lists the method detection limit for each target analyte in
organic-free reagent water. Table 2 lists the estimated quantitation limit (EQL)
for other matrices.
2.0 SUMMARY OF METHOD
2.1 Method 8020 provides chromatographic conditions for the detection of
aromatic volatile compounds. Samples can be introduced into the GC using direct
injection or purge-and-trap (Method 5030). Ground water samples must be
determined using Method 5030. A temperature program is used in the gas
chromatograph to separate the organic compounds. Detection is achieved by a
photo-ionization detector (PID).
2.2 If interferences are encountered, the method provides an optional gas
chromatographic column that may be helpful in resolving the analytes from the
interferences and for analyte confirmation.
3.0 INTERFERENCES
3.1 Refer to Method 5030 and 8000.
8020A - 1 Revision 1
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3.2 Samples can be contaminated by diffusion of volatile organics
(particularly chlorofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A field sample blank prepared from
organic-free reagent water and carried through sampling and subsequent storage
and handling can serve as a check on such contamination.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas Chromatograph - Analytical system complete with gas
chromatograph suitable for on-column injections or purge-and-trap sample
introduction and all required accessories, including detectors, column
supplies, recorder, gases, and syringes. A data system for measuring peak
heights and/or peak areas is recommended.
4.1.2 Columns
4.1.2.1 Column 1: 6 ft x 0.082 in ID #304 stainless steel
or glass column packed with 5% SP-1200 and 1.75% Bentone-34 on
100/120 mesh Supelcoport, or equivalent.
4.1.2.2 Column 2: 8 ft x 0.1 in ID stainless steel or
glass column packed with 5% l,2,3-Tris(2-cyanoethoxy)propane on
60/80 mesh Chromosorb W-AW, or equivalent.
4.1.3 Detector - Photoionization (PID) (h-Nu Systems, Inc. Model
PI-51-02 or equivalent).
4.2 Sample introduction apparatus - Refer to Method 5030 for the
appropriate equipment for sample introduction purposes.
4.3 Syringes - A 5 mL Luerlok glass hypodermic and a 5 mL, gas-tight with
shutoff valve.
4.4 Volumetric flask, Class A - Appropriate sizes with ground glass
stoppers.
4.5 Microsyringe - 10 and 25 p.1 with a 0.006 in ID needle (Hamilton 702N
or equivalent) and a 100 juL.
4.6 Analytical balance - 0.0001 g.
5.0 REAGENTS
5.1 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2 Methanol (CH3OH) - pesticide quality or equivalent. Store away from
other solvents.
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5.3 Stock standards - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standards in
methanol using assayed liquids. Because of the toxicity of benzene and
1,4-dichlorobenzene, primary dilutions of these materials should be prepared in
a hood.
5.3.1 Place about 9.8 mL of methanol in a 10 ml tared ground glass
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 min or until all alcohol wetted surfaces have dried. Weigh the
flask to the nearest 0.0001 g.
5.3.2 Using a 100 /zL syringe, immediately add two or more drops of
assayed reference material to the flask; then reweigh. The liquid must
fall directly into the alcohol without contacting the neck of the flask.
5.3.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.3.4 Transfer the stock standard solution into a Teflon-sealed
screw-cap bottle. Store, with minimal headspace, at 4°C and protect from
light.
5.3.5 All standards must be replaced after 6 months, or sooner if
comparison with check standards indicates a problem.
5.4 Secondary dilution standards: Using stock standard solutions,
prepare in methanol secondary dilution standards, as needed, that contain the
compounds of interest, either singly or mixed together. The secondary dilution
standards should be prepared at concentrations such that the aqueous calibration
standards prepared in Section 5.5 will bracket the working range of the
analytical system. Secondary dilution standards should be stored with minimal
headspace for volatiles and should be checked frequently for signs of degradation
or evaporation, especially just prior to preparing calibration standards from
them.
5.5 Calibration standards: Calibration standards at a minimum of five
concentrations are prepared in organic-free reagent water from the secondary
dilution of the stock standards. One of the concentrations should be at a
concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations found
in real samples or should define the working range of the GC. Each standard
should contain each analyte for detection by this method (e.g., some or all of
the compounds listed in the target analyte list may be included). In order to
prepare accurate aqueous standard solutions, the following precautions must be
observed.
8020A - 3 Revision 1
September 1994
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5.5.1 Do not inject more than 20 /j.1 of alcoholic standards into
100 ml of organic-free reagent water.
5.5.2 Use a 25 /iL Hamilton 702N microsyringe or equivalent
(variations in needle geometry will adversely affect the ability to
deliver reproducible volumes of methanolic standards into water).
5.5.3 Rapidly inject the alcoholic standard into the filled
volumetric flask. Remove the needle as fast as possible after injection.
5.5.4 Mix aqueous standards by inverting the flask three times only.
5.5.5 Fill the sample syringe from the standard solution contained
in the expanded area of the flask (do not use any solution contained in
the neck of the flask).
5.5.6 Never use pipets to dilute or transfer samples or aqueous
standards.
5.5.7 Aqueous standards are not stable and should be discarded after
1 hr, unless properly sealed and stored. The aqueous standards can be
stored up to 24 hr, if held in sealed vials with zero headspace.
5.6 Internal standards (if internal standard calibration is used): To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
Alpha,alpha,alpha-trifluorotoluene has been used successfully as an internal
standard.
5.6.1 Prepare calibration standards at a minimum of five
concentrations for each parameter of interest as described in Section 5.5.
5.6.2 Prepare a spiking solution containing each of the internal
standards using the procedures described in Sections 5.3 and 5.4. It is
recommended that the secondary dilution standard be prepared at a
concentration of 15 mg/L of each internal standard compound. The addition
of 10 IJ.L of this standard to 5.0 ml of sample or calibration standard
would be equivalent to 30 M9/L.
5.6.3 Analyze each calibration standard according to Section 7.0,
adding 10 /uL of internal standard spiking solution directly to the
syringe.
5.7 Surrogate standards: The analyst should monitor both the performance
of the analytical system and the effectiveness of the method in dealing with each
sample matrix by spiking each sample, standard, and organic-free reagent water
blank with surrogate compounds (bromochlorobenzene, bromofluorobenzene, 1,1,1-
trifluorotoluene, fluorobenzene, and difluorobenzene are recommended) which
encompass the range of the temperature program used in this method. From stock
8020A - 4 Revision 1
September 1994
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standard solutions prepared as in Section 5.3, add a volume to give 750 /ug of
each surrogate to 45 ml of organic-free reagent water contained in a 50 ml
volumetric flask, mix, and dilute to volume for a concentration of 15 ng/juL.
Add 10 jttL of this surrogate spiking solution directly into the 5 ml syringe with
every sample and reference standard analyzed. If the internal standard
calibration procedure is used, the surrogate compounds may be added directly to
the internal standard spiking solution (Section 5.6.2).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Volatile compounds are introduced into the gas chromatograph either
by direct injection or purge-and-trap (Method 5030). Method 5030 may be used
directly on ground water samples or low-concentration contaminated soils and
sediments. For medium-concentration soils or sediments, methanolic extraction,
as described in Method 5030, may be necessary prior to purge-and-trap analysis.
Method 5030 also provides guidance on the analysis of aqueous miscible and non-
aqueous miscible liquid wastes (see Section 7.4.1.1 below).
7.2 Gas chromatography conditions (Recommended):
7.2.1 Column 1:
Carrier gas (He) flow rate: 36 mL/min
For lower boiling compounds:
Initial temperature: 50°C, hold for 2 min;
Temperature program: 50°C to 90°C at 6°C/min, hold until
all compounds have eluted.
For higher boiling range of compounds:
Initial temperature: 50°C, hold for 2 min;
Temperature program: 50°C to 110°C at 3°C/min, hold until
all compounds have eluted.
Column 1 provides outstanding separations for a wide variety of
aromatic hydrocarbons. Column 1 should be used as the primary analytical
column because of its unique ability to resolve para-, meta-, and ortho-
aromatic isomers.
7.2.2 Column 2:
Carrier gas (He) flow rate: 30 mL/min
Initial temperature: 40°C, hold for 2 min;
Temperature program: 40°C to 100°C at 2°C/min, hold until
all compounds have eluted.
8020A - 5 Revision 1
September 1994
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Column 2, an extremely high polarity column, has been used for a
number of years to resolve aromatic hydrocarbons from alkanes in complex
samples. However, because resolution between some of the aromatics is not
as efficient as with Column 1, Column 2 should be used as a confirmatory
column.
7.3 Calibration: Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point on
the calibration curve.
7.3.1 Calibration must take place using the same sample introduction
method that will be used to analyze actual samples (see Section 7.4.1).
7.3.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.4 Gas chromatographic analysis:
7.4.1 Introduce volatile compounds into the gas chromatograph using
either Method 5030 (purge-and-trap method) or the direct injection method.
If the internal standard calibration technique is used, add 10 /jL of
internal standard to the sample prior to purging.
7.4.1.1 Direct injection: In very limited applications
(e.g., aqueous process wastes), direct injection of the sample into
the GC system with a 10 /nL syringe may be appropriate. The
detection limit is very high (approximately 10,000 M9/L); therefore,
it is only permitted when concentrations in excess of 10,000 /zg/L
are expected or for water soluble compounds that do not purge. The
system must be calibrated by direct injection (bypassing the purge-
and-trap device).
Non-aqueous miscible wastes may also be analyzed by direct
injection if the concentration of target analytes in the sample
falls within the calibration range. If dilution of the sample is
necessary, follow the guidance for High Concentration samples in
Method 5030, Section 7.3.3.2.
7.4.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
7.4.3 Table 1 summarizes the estimated retention times and detection
limits for a number of organic compounds analyzable using this method. An
example of the separation achieved by Column 1 is shown in Figure 1.
Figure 2 shows an example of the separation achieved using Column 2.
7.4.4 Record the sample volume purged or injected and the resulting
peak sizes (in area units or peak heights).
7.4.5 Calculation of concentration is covered in Method 8000.
8020A - 6 Revision 1
September 1994
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7.4.6 If analytical interferences are suspected, or for the purpose
of confirmation, analysis using the second GC column is recommended.
7.4.7 If the response for a peak is off scale, i.e., beyond the
calibration range of the standards, prepare a dilution of the sample with
organic-free reagent water. The dilution must be performed on a second
aliquot of the sample which has been properly sealed and stored prior to
use.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 8000 for gas chromatographic procedures. Quality control to ensure the
proper operation of the purge-and-trap device is covered in Method 5030.
8.2 Quality control required to validate the GC system operation is found
in Method 8000.
8.2.1 The quality control check sample concentrate (Method 8000)
should contain each parameter of interest at a concentration of 10 mg/L
in methanol.
8.2.2 Table 3 indicates the calibration and QC acceptance criteria
for this method. Table 4 gives method accuracy and precision as functions
of concentration for the analytes of interest. The contents of both
tables should be used to evaluate a laboratory's ability to perform and
generate acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if recovery is within limits (limits established by performing
QC procedure outlined in Method 8000).
8.3.1 If recovery is not within limits, the following is required.
• Check to be sure that there are no errors in
calculations, surrogate solutions and internal
standards. Also, check instrument performance.
• Recalculate the data and/or reanalyze the extract if any
of the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above
are a problem or flag the data as "estimated
concentration".
8020A - 7 Revision 1
September 1994
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9.0 METHOD PERFORMANCE
9.1 This method was tested by 20 laboratories using organic-free reagent
water, drinking water, surface water, and three industrial wastewaters spiked at
six concentrations over the range 2.1 - 500 jiig/L. Single operator precision,
overall precision, and method accuracy were found to be directly related to the
concentration of the parameter and essentially independent of the sample matrix.
Linear equations to describe these relationships are presented in Table 4.
9.2 The accuracy and precision obtained will be determined by the sample
matrix, sample introduction technique, and by the calibration procedure used.
9.3 The method detection limits reported in Table 1 were generated under
optimum analytical conditions by an Agency contractor (Ref. 7) as guidance, and
may not be readily achievable by all laboratories at all times.
10.0 REFERENCES
1. Bellar, T.A., and J.J. Lichtenberg, J. Amer. Water Works Assoc., 66(12),
pp. 739-744, 1974.
2. Bellar, T.A., and J.J. Lichtenberg, "Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds", in Van Hall (ed.), Measurement of Organic Pollutants in Water
and Wastewater, ASTM STP 686, pp. 108-129, 1979.
3. Dowty, B.J., S.R. Antoine, and J.L. Laseter, "Quantitative and Qualitative
Analysis of Purgeable Organics by High Resolution Gas Chromatography and
Flame lonization Detection", in Van Hall, ed., Measurement of Organic
Pollutants in Water and Wastewater. ASTM STP 686, pp. 24-35, 1979.
4. Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 11 - Purgeables and Category 12 -
Acrolein, Acrylonitrile, and Dichlorodifluoromethane. Report for EPA
Contract 68-03-2635.
5. "EPA Method Validation Study 24, Method 602 (Purgeable Aromatics)", report
for EPA Contract 68-03-2856.
6. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule", October 26, 1984.
7. Gebhart, J.E., S.V. Lucas, S.J. Naber, A.M. Berry, T.H. Danison and H.M.
Burkholder, "Validation of SW-846 Methods 8010, 8015, and 8020"; Report
for EPA Contract 68-03-1760, Work Assignment 2-15; US EPA, EMSL-
Cincinnati, 1987."
8020A - 8 Revision 1
September 1994
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
FOR AROMATIC VOLATILE ORGANICS
Compound
Benzene
Chlorobenzeneb
1,4-Dichlorobenzene
1,3-Dichlorobenzene
1,2-Dichlorobenzene
Ethyl Benzene
Toluene
Xylenes
Retention
(min)
Col. 1
3.33
9.17
16.8
18.2
25.9
8.25
5.75
time
Col. 2
2.75
8.02
1'6. 2
15.0
19.4
6.25
4.25
Method
detection
limit3
(M9/L)
0.2
0.2
0.3
0.4
0.4
0.2
0.2
a Using purge-and-trap method (Method 5030). See Sec. 9.3.
b Chlorobenzene and m-xylene may co-elute on some columns.
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQLs)
FOR VARIOUS MATRICES3
Matrix
Factor
Ground water
Low-concentration soil
Water miscible liquid waste
High-concentration soil and sludge
Non-water miscible waste
10
10
500
1250
1250
EQL = [Method detection limit (see Table 1)] X [Factor found in this
table]. For non-aqueous samples, the factor is on a wet-weight basis.
Sample EQLs are highly matrix-dependent. The EQLs determined herein are
provided for guidance and may not always be achievable.
8020A - 9
Revision 1
September 1994
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TABLE 3.
QC ACCEPTANCE CRITERIA3
Parameter
Benzene
Chlorobenzene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Di chlorobenzene
Ethyl benzene
Toluene
Range
for Q
(M9/L)
15.4-24.6
16.1-23.9
13.6-26.4
14.5-25.5
13.9-26.1
12.6-27.4
15.5-24.5
Limit
for s
(M9/L)
4.1
3.5
5.8
5.0
5.5
6.7
4.0
Range
for x
(M9/L)
10.0-27.9
12.7-25.4
10.6-27.6
12.8-25.5
11.6-25.5
10.0-28.2
11.2-27.7
Range
P> Ps
(%)
39-150
55-135
37-154
50-141
42-143
32-160
46-148
Q =
s =
x =
P, Ps =
Concentration measured in QC check sample, in
Standard deviation of four recovery measurements, in M9/L-
Average recovery for four recovery measurements, in jug/L.
Percent recovery measured.
Criteria from 40 CFR Part 136 for Method 602, using packed columns, and
were calculated assuming a check sample concentration of 20 fj.g/1. These
criteria are based directly upon the method performance data in Table 4.
Where necessary, the limits for recovery have been broadened to assure
applicability of the limits to concentrations below those used to develop
Table 1. When capillary columns are used, see Method 8021 for performance
data.
8020A - 10
Revision 1
September 1994
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TABLE 4.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION
Parameter
Benzene
Chlorobenzene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Di chlorobenzene
Ethyl benzene
Toluene
Accuracy, as
recovery, x'
(M9/L)
0.92C+0.57
0.95C+0.02
0.93C+0.52
0.96C-0.04
0.93C-0.09
0.94C+0.31
0.94C+0.65
Single analyst
precision, sr'
(M9/L)
0.09x+0.59
0.09X+0.23
0.17X-0.04
0.15x-0.10
0.15X+0.28
0.17X+0.46
0.09X+0.48
Overall
precision,
S' (M9/L)
0.21X+0.56
0.17X+0.10
0.22X+0.53
0.19X+0.09
0.20X+0.41
0.26X+0.23
0.18x+0.71
x'
Expected recovery for one or more measurements of a sample
containing concentration C, in
Expected single analyst standard deviation of measurements at an
average concentration of x, in
S'
C
x
Expected inter! aboratory standajrd deviation of measurements at an
average concentration found of x, in
True value for the concentration, in /^g/L.
Average recovery found for measurements of samples containing a
concentration of C, in M9/L.
8020A - 11
Revision 1
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Figure 1
Chromatogram of Aromatic Volatile Organics
(column 1 conditions)
Column:
Program:
Detector:
Sample:
5% SP-1200/1.75% Bentone 34
50°C-2 Minutes, 6°C/Min. to 90°C
Photoionization
0.40 jig/L Standard Mixture
8 10 12 14
RETENTION TIME (MINUTES}
16
18
20 22
8020A - 12
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Figure 2
Chromatogram of Aromatic Volatile Organics
(column 2 conditions)
Column:
Program:
Detector:
Sample:
5% l,2,3-Tris(2-Cyanoethoxy)Propane on Chromosorb-W
40°C-2 Minutes, 2°C/Min. to 100*C
Photoionization
2.0 /jg/L Standard Mixture
I
mm
12 If
(MINUTU>
24
8020A - 13
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METHOD 8020A
AROMATIC VOLATILE ORGANICS BY GAS CHROMATOGRAPHY
Start
7.1 Introduce compounds
into gas chromatograph
by direct injection or
purge-and-trap
(Method 5030)
7.2 Set gas
chromatograph
condition.
7.3 Calibrate
(refer to Method 8000)
7.4.1 Introduce
volatile compounds
into gas chromatograph
by purge-and-trap or
direct injection.
7.4.2 Follow Method
8000 for analysis
sequence, etc.
7.4.4 Record volume
purged or injected
and peak sizes.
7.4.5 Calculate
concentration
(refer to Method 8000)
7.4.6 Are
analytical
interferences
suspected?
7.4.7 Is
response for
a peak
off-scale?
7.4.6 Analyze using
second GC column.
7.4.7 Dilute second
aliquot of sample.
8020A -14
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METHOD 8021A
HALOGENATED VOLATILES BY GAS CHROMATOGRAPHY USING
PHOTOIONIZATION AND ELECTROLYTIC CONDUCTIVITY DETECTORS
IN SERIES: CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8021 is used to determine volatile organic compounds in a
variety of solid waste matrices. This method is applicable to nearly all types
of samples, regardless of water content, including ground water, aqueous sludges,
caustic liquors, acid liquors, waste solvents, oily wastes, mousses, tars,
fibrous wastes, polymeric emulsions, filter cakes, spent carbons, spent
catalysts, soils, and sediments. The following compounds can be determined by
this method:
Analyte
Appropriate Technique
CAS No.a Purge-and-Trap
Direct
Injection
Benzene
Bromobenzene
Bromochl oromethane
Bromodi chl oromethane
Bromoform
Bromomethane
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chl orodi bromomethane
Chloroethane
Chloroform
Chloromethane
2-Chlorotoluene
4-Chlorotoluene
l,2-Dibromo-3-chloropropane
1,2-Dibromoethane
Di bromomethane
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1, 4 -Di chl orobenzene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1, 1-Dichloroethene
cis-1, 2-Di chl oroethene
trans -1, 2-Di chl oroethene
71-43-2
108-86-1
74-97-5
75-27-4
75-25-2
74-83-9
104-51-8
135-98-8
98-06-6
56-23-5
108-90-7
124-48-1
75-00-3
67-66-3
74-87-3
95-49-8
106-43-4
96-12-8
106-93-4
74-95-3
95-50-1
541-73-1
106-46-7
75-71-8
75-34-3
107-06-2
75-35-4
156-59-4
156-60-5
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
PP
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
8021A - 1
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Appropriate Technique
Analyte
CAS No.a Purge-and-Trap
Direct
Injection
1 , 2-Di chl oropropane
1,3-Dichloropropane
2, 2-Di chl oropropane
1,1-Dichloropropene
cis-l,3-dichloropropene
trans-l,3-dichloropropene
Ethyl benzene
Hexachlorobutadiene
Isopropyl benzene
p-Isopropyl toluene
Methylene chloride
Naphthalene
n-Propyl benzene
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
1,2,3-Trichlorobenzene
1,2,4-Trichlorobenzene
1,1,1-Tri chl oroethane
1,1,2-Trichloroethane
Trichloroethene
Tri chl orof 1 uoromethane
1 , 2 , 3 -Tri chl oropropane
1, 2, 4-Tri methyl benzene
1,3, 5 -Tri methyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
78-87-5
142-28-9
590-20-7
563-58-6
10061-01-5
10061-02-6
100-41-4
87-68-3
98-82-8
99-87-6
75-09-2
91-20-3
103-65-1
100-42-5
630-20-6
79-34-5
127-18-4
108-88-3
87-61-6
120-82-1
71-55-6
79-00-5
79-01-6
75-69-4
96-18-4
95-63-6
108-67-8
75-01-4
95-47-6
108-38-3
106-42-3
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
a Chemical Abstract Services Registry Number.
b Adequate response by thi
pp Poor purging efficiency
s technique.
resulting in high EQLs.
is
1.2 Method detection limits (MDLs) are compound dependent and vary with
purging efficiency and concentration. The MDLs for selected analytes are
presented in Table 1. The applicable concentration range of this method
compound and instrument dependent but is approximately 0.1 to 200
Analytes that are inefficiently purged from water will not be detected when
present at low concentrations, but they can be measured with acceptable accuracy
and precision when present in sufficient amounts. Determination of some
structural isomers (i.e. xylenes) may be hampered by coelution.
8021A - 2
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1.3 The estimated quantitation limit (EQL) of Method 8021A for an
individual compound is approximately 1 //g/kg (wet weight) for soil/sediment
samples, 0.1 mg/kg (wet weight) for wastes, and 1 /zg/L for ground water (see
Table 3). EQLs will be proportionately higher for sample extracts and samples
that require dilution or reduced sample size to avoid saturation of the detector.
1.4 This method is recommended for use only by analysts experienced in
the measurement of purgeable organics at the low /Ltg/L level or by experienced
technicians under the close supervision of a qualified analyst.
1.5 The toxicity or carcinogenicity of chemicals used in this method has
not been precisely defined. Each chemical should be treated as a potential
health hazard, and exposure to these chemicals should be minimized. Each
laboratory is responsible for maintaining awareness of OSHA regulations regarding
safe handling of chemicals used in this method. Additional references to
laboratory safety are available for the information of the analyst (references
4 and 6).
1.6 The following method analytes have been tentatively classified as
known or suspected human or mammalian carcinogens: benzene, carbon tetrachloride,
1,4-dichlorobenzene, 1,2-dichloroethane, hexachloro-butadiene, 1,1,2,2-
tetrachloroethane, 1,1,2-trichloroethane, chloroform, 1,2-dibromoethane,
tetrachloroethene, trichloroethene, and vinyl chloride. Pure standard materials
and stock standard solutions of these compounds should be handled in a hood. A
NIOSH/MESA approved toxic gas respirator should be worn when the analyst handles
high concentrations of these toxic compounds.
2.0 SUMMARY OF METHOD
2.1 Method 8021 provides gas chromatographic conditions for the
detection of halogenated and aromatic volatile organic compounds. Samples can
be analyzed using direct injection or purge-and-trap (Method 5030). Ground water
samples must be analyzed using Method 5030 (where applicable). A temperature
program is used in the gas chromatograph to separate the organic compounds.
Detection is achieved by a photoionization detector (PID) and an electrolytic
conductivity detector (HECD) in series.
2.2 Tentative identifications are obtained by analyzing standards under
the same conditions used for samples and comparing resultant GC retention times.
Confirmatory information can be gained by comparing the relative response from
the two detectors. Concentrations of the identified components are measured by
relating the response produced for that compound to the response produced by a
compound that is used as an internal standard.
3.0 INTERFERENCES
3.1 Refer to Methods 5030 and 8000.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly chlorofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A trip blank prepared from
8021A - 3 Revision 1
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organic-free reagent water and carried through sampling and subsequent storage
and handling can serve as a check on such contamination.
3.3 Sulfur dioxide is a potential interferant in the analysis for vinyl
chloride.
4.0 APPARATUS AND MATERIALS
4.1 Sample introduction apparatus - Refer to Method 5030 for the
appropriate equipment for sample introduction purposes.
4.2 Gas Chromatograph - capable of temperature programming; equipped
with variable-constant differential flow controllers, subambient oven controller,
photoionization and electrolytic conductivity detectors connected with a short
piece of uncoated capillary tubing, 0.32-0.5 mm ID, and data system.
4.2.1 Column - 60 m x 0.75 mm ID VOCOL wide-bore capillary column
with 1.5 nm film thickness (Supelco Inc., or equivalent).
4.2.2 Photoionization detector (PID) (Tracer Model 703, or
equivalent).
4.2.3 Electrolytic conductivity detector (HECD) (Tracor Hall Model
700-A, or equivalent).
4.3 Syringes - 5 ml glass hypodermic with Luer-Lok tips.
4.4 Syringe valves - 2-way with Luer ends (Teflon or Kel-F).
4.5 Microsyringe - 25 juL with a 2 in. x 0.006 in. ID, 22° bevel needle
(Hamilton #702N or equivalent).
4.6 Microsyringes - 10, 100 juL.
4.7 Syringes - 0.5, 1.0, and 5 ml, gas-tight with shut-off valve.
4.8 Bottles - 15 ml, Teflon lined with screw-cap or crimp top.
4.9 Analytical balance - 0.0001 g.
4.10 Refrigerator.
4.11 Volumetric flasks, Class A - Appropriate sizes with ground glass
stoppers.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests.
Unless otherwise indicated, it is intended that all inorganic reagents shall
conform to the specifications of the Committee on Analytical Reagents of the
American Chemical Society, where such specifications are available. Other grades
8021A - 4 Revision 1
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may be used, provided it is first ascertained that the reagent is of sufficiently
high purity to permit its use without lessening the accuracy of the
determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH - Pesticide quality or equivalent, demonstrated to
be free of analytes. Store away from other solvents.
5.4 Vinyl chloride, (99.9% pure), CH2=CHC1. Vinyl chloride is available
from Ideal Gas Products, Inc., Edison, New Jersey and from Matheson, East
Rutherford, New Jersey, as well as from other sources. Certified mixtures of
vinyl chloride in nitrogen at 1.0 and 10.0 ppm (v/v) are available from several
sources.
5.5 Stock standards - Stock solutions may either be prepared from pure
standard materials or purchased as certified solutions. Prepare stock standards
in methanol using assayed liquids or gases, as appropriate. Because of the
toxicity of some of the organohalides, primary dilutions of these materials of
the toxicity should be prepared in a hood.
NOTE: If direct injection is used, the solvent system of standards must
match that of the sample. It is not necessary to prepare high
concentration aqueous mixed standards when using direct injection.
5.5.1 Place about 9.8 ml of methanol in a 10 mL tared ground glass
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 minutes until all alcohol-wetted surfaces have dried. Weigh the
flask to the nearest 0.1 mg.
5.5.2 Add the assayed reference material, as described below.
5.5.2.1 Liquids: Using a 100 jiL syringe, immediately add
two or more drops of assayed reference material to the flask; then
reweigh. The liquid must fall directly into the alcohol without
contacting the neck of the flask.
5.5.2.2 Gases: To prepare standards for any compounds
that boil below 30°C (e.g. bromomethane, chloroethane,
chloromethane, dichlorodifluoromethane, trichlorofluoromethane,
vinyl chloride), fill a 5 ml valved gas-tight syringe with the
reference standard to the 5.0 ml mark. Lower the needle to 5 mm
above the methanol meniscus. Slowly introduce the reference
standard above the surface of the liquid. The heavy gas rapidly
dissolves in the methanol. This may also be accomplished by using
a lecture bottle equipped with a Hamilton Lecture Bottle Septum
(#86600). Attach Teflon tubing to the side-arm relief valve and
direct a gentle stream of gas into the methanol meniscus.
5.5.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
8021A - 5 Revision 1
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to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.5.4 Transfer the stock standard solution into a bottle with a
Teflon lined screw-cap or crimp top. Store, with minimal headspace, at
-10°C to -20°C and protect from light.
5.5.5 Prepare fresh stock standards for gases weekly or sooner if
comparison with check standards indicates a problem. Reactive compounds
such as 2-chloroethyl vinyl ether and styrene may need to be prepared more
frequently. All other standards must be replaced after six months. Both
gas and liquid standards must be monitored closely by comparison to the
initial calibration curve and by comparison to QC check standards. It may
be necessary to replace the standards more frequently if either check
exceeds a 20% drift.
5.6 Prepare secondary dilution standards, using stock standard
solutions, in methanol, as needed, that contain the compounds of interest, either
singly or mixed together. The secondary dilution standards should be prepared
at concentrations such that the aqueous calibration standards prepared in Sec.
5.7 will bracket the working range of the analytical system. Secondary dilution
standards should be stored with minimal headspace for volatiles and should be
checked frequently for signs of degradation or evaporation, especially just prior
to preparing calibration standards from them.
5.7 Calibration standards, at a minimum of five concentration levels are
prepared in organic-free reagent water from the secondary dilution of the stock
standards. One of the concentration levels should be at a concentration near,
but above, the method detection limit. The remaining concentration levels should
correspond to the expected range of the concentrations found in real samples or
should define the working range of the GC. Standards (one or more) should
contain each analyte for detection by this method. In order to prepare accurate
aqueous standard solutions, the following precautions must be observed.
NOTE: Prepare calibration solutions for use with direct injection analyses
in water at the concentrations required.
5.7.1 Do not inject more than 20 /^L of alcoholic standards into
100 mL of water.
5.7.2 Use a 25 juL Hamilton 702N microsyringe or equivalent
(variations in needle geometry will adversely affect the ability to
deliver reproducible volumes of methanolic standards into water).
5.7.3 Rapidly inject the alcoholic standard into the filled
volumetric flask. Remove the needle as fast as possible after injection.
5.7.4 Mix aqueous standards by inverting the flask three times.
8021A - 6 Revision 1
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5.7.5 Fill the sample syringe from the standard solution contained
in the expanded area of the flask (do not use any solution contained in
the neck of the flask).
5.7.6 Never use pipets to dilute or transfer samples or aqueous
standards.
5.7.7 Aqueous standards are not stable and should be discarded after
one hour, unless properly sealed and stored. The aqueous standards can
be stored up to 12 hours, if held in sealed vials with zero headspace.
5.7.8 Optionally calibration using a certified gaseous mixture can
be accomplished daily utilizing commercially available gaseous analyte
mixture of bromomethane, chloromethane, chloroethane, vinyl chloride,
dichlorodifluoromethane and trichlorofluoromethane in nitrogen. These
mixtures of documented quality are stable for as long as six months
without refrigeration. (VOA-CYL III, RESTEK Corporation, Cat. #20194 or
equivalent).
5.8 Internal standards - Prepare a spiking solution containing
fluorobenzene and 2-bromo-l-chloropropane in methanol, using the procedures
described in Sees. 5.5 and 5.6. It is recommended that the secondary dilution
standard be prepared at a concentration of 5 mg/L of each internal standard
compound. The addition of 10 nl of such a standard to 5.0 ml of sample or
calibration standard would be equivalent to 10 M9/L.
5.9 Surrogate standards - The analyst should monitor both the
performance of the analytical system and the effectiveness of the method in
dealing with each sample matrix by spiking each sample, standard, and reagent
blank with two or more surrogate compounds. A combination of 1,4-dichlorobutane
and bromochlorobenzene is recommended to encompass the range of the temperature
program used in this method. From stock standard solutions prepared as in Sec.
5.5, add a volume to give 750 /^g of each surrogate to 45 mL of organic-free
reagent water contained in a 50 ml volumetric flask, mix, and dilute to volume
for a concentration of 15 ng//xL. Add 10 p,l of this surrogate spiking solution
directly into the 5 mL syringe with every sample and reference standard analyzed.
If the internal standard calibration procedure is used, the surrogate compounds
may be added directly to the internal standard spiking solution (Sec. 5.8).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Volatile compounds are introduced into the gas chromatograph either
by direct injection or purge-and-trap (Method 5030). Method 5030 may be used
directly on ground water samples or low-concentration contaminated soils and
sediments. For medium-concentration soils or sediments, methanolic extraction,
as described in Method 5030, may be necessary prior to purge-and-trap analysis.
8021A - 7 Revision 1
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7.2 Gas chromatography conditions (Recommended)
7.2.1 Set up the gas chromatograph system so that the
photoionization detector (PID) is in series with the electrolytic
conductivity detector (HECD).
7.2.2 Oven settings:
Carrier gas (Helium) Flow rate: 6 mL/min.
Temperature program
Initial temperature: 10°C, hold for 8 minutes at
Program: 10°C to 180°C at 4°C/"un
Final temperature: 180°C, hold until all expected
compounds have eluted.
7.2.3 The carrier gas flow is augmented with an additional 24 ml of
helium flow before entering the photoionization detector. This make-up
gas is necessary to ensure optimal response from both detectors.
7.2.4 These halogen-specific systems eliminate misidentifications
due to non-organohalides which are coextracted during the purge step. A
Tracer Hall Model 700-A detector was used to gather the single laboratory
accuracy and precision data presented in Table 2. The operating
conditions used to collect these data are:
Reactor tube: Nickel, 1/16 in OD
Reactor temperature: 810°C
Reactor base temperature: 250°C
Electrolyte: 100% n-Propyl alcohol
Electrolyte flow rate: 0.8 mL/min
Reaction gas: Hydrogen at 40 mL/min
Carrier gas plus make-up gas: Helium at 30 mL/min
7.2.5 A sample chromatogram obtained with this column is presented
in Figure 5. This column was used to develop the method performance
statements in Sec. 9.0. Estimated retention times and MDLs that can be
achieved under these conditions are given in Table 1. Other columns or
element specific detectors may be used if the requirements of Sec. 8.0 are
met.
7.3 Calibration - Refer to Method 8000 for proper calibration
techniques. Use Table 1 and especially Table 2 for guidance on selecting the
lowest point on the calibration curve.
7.3.1 Calibration must take place using the same sample introduction
method that will be used to analyze actual samples (see Sec. 7.4.1).
7.3.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.4 Gas chromatographic analysis
8021A - 8 Revision 1
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7.4.1 Introduce volatile compounds into the gas chromatograph using
either Method 5030 (purge-and-trap method) or the direct injection method
(see Sec. 7.4.1.1). If the internal standard calibration technique is
used, add 10 /zL of internal standard to the sample prior to purging.
7.4.1.1 Direct injection - In very limited applications
(e.g. aqueous process wastes) direct injection of the sample into
the GC system with a 10 /zL syringe may be appropriate. The
detection limit is very high (approximately 10,000 M9/L)> therefore,
it is only permitted where concentrations in excess of 10,000 M9A
are expected or for water-soluble compounds that do not purge. The
system must be calibrated by direct injection (bypassing the purge-
and-trap device).
7.4.2 Follow Sec. 7.6 in Method 8000 for instructions on the
analysis sequence, appropriate dilutions, establishing daily retention
time windows, and identification criteria. Include a mid-concentration
standard after each group of 10 samples in the analysis sequence.
7.4.3 Table 1 summarizes the estimated retention times on the two
detectors for a number of organic compounds analyzable using this method.
7.4.4 Record the sample volume purged or injected and the resulting
peak sizes (in area units or peak heights).
7.4.5 Calculation of concentration is covered in Method 8000.
7.4.6 If analytical interferences are suspected, or for the purpose
of confirmation, analysis using a second GC column is recommended.
7.4.7 If the response for a peak is off-scale, i.e., beyond the
calibration range of the standards, prepare a dilution of the sample with
organic-free reagent water. The dilution must be performed on a second
aliquot of the sample which has been properly sealed and stored prior to
use.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 8000 for gas chromatographic procedures. Quality control to ensure the
proper operation of the purge-and-trap device is covered in Method 5030.
8.2 Quality control required to validate the GC system operation is
found in Method 8000.
8.2.1 The quality control reference sample (Method 8000) should
contain each parameter of interest at a concentration of 10 mg/L in
methanol.
8.2.2 Table 2 gives method accuracy and precision as functions of
concentration for the analytes of interest.
8021A - 9 Revision 1
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8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if recovery is within limits (limits established by performing
QC procedure outlined in Method 8000).
8.3.1 If recovery is not within limits, the following is required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any
of the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above
are a problem or flag the data as "estimated
concentration".
9.0 METHOD PERFORMANCE
9.1 Method detection limits for these analytes have been calculated from
data collected by spiking organic-free reagent water at 0.1 /ng/L. These data
are presented in Table 1.
9.2 This method was tested in a single laboratory using organic-free
reagent water spiked at 10 jug/L. Single laboratory precision and accuracy data
for each detector are presented for the method analytes in Table 2.
10.0 REFERENCES
1. Volatile Organic Compounds in Water by Purqe-and-Trap Capillary Column Gas
Chromatography with Photoionization and Electrolytic Conductivity
Detectors in Series, Method 502.2. Rev. 2.0 (1989); Methods for the
Determination of Organic Compounds in Drinking Water", Environmental
Monitoring Systems Laboratory, Cincinnati, OH, EPA/600/4-88/039, December,
1988
2. The Determination of Halogenated Chemicals in Water by the Purge and Trap
Method, Method 502.1; Environmental Protection Agency, Environmental
Monitoring and Support Laboratory: Cincinnati, Ohio 45268, September,
1986.
3. Volatile Aromatic and Unsaturated Organic Compounds in Water by Purge and
Trap Gas Chromatography, Method 503.1; Environmental Protection Agency,
Environmental Monitoring and Support Laboratory: Cincinnati, Ohio,
September, 1986.
4. Glaser, J.A.; Forest, D.L.; McKee, G.D.; Quave, S.A.; Budde, W.L. "Trace
Analyses for Wastewaters"; Environ. Sci. Technol. 1981, 15, 1426.
8021A - 10 Revision 1
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5. Bellar, T.A.; Lichtenberg, J.J. The Determination of Synthetic Organic
Compounds in Water by Purge and Sequential Trapping Capillary Column Gas
Chromatography; U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory: Cincinnati, Ohio, 45268.
8021A - 11 Revision 1
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TABLE 1.
CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS (MDL) FOR
VOLATILE ORGANIC COMPOUNDS ON PHOTOIONIZATION DETECTION (PID) AND
HALL ELECTROLYTIC CONDUCTIVITY DETECTOR (HECD) DETECTORS
Analyte
Di chl orod i f 1 uoromethane
Chloromethane
Vinyl Chloride
Bromomethane
Chloroethane
Tri chl orof 1 uoromethane
1,1-Dichloroethene
Methylene Chloride
trans-l,2-Dichloroethene
1,1-Dichl oroethane
2,2-Dichloropropane
cis-l,2-Di chloroethane
Chloroform
Bromochl oromethane
1,1,1 -Tri chl oroethane
1 , 1-Dichl oropropene
Carbon Tetrachloride
Benzene
1,2-Dichloroethane
Trichloroethene
1,2-Dichloropropane
Bromodi chl oromethane
Dibromomethane
Toluene
1,1, 2 -Tri chl oroethane
Tetrachl oroethene
1,3-Dichloropropane
Di bromochl oromethane
1,2-Dibromoethane
Chlorobenzene
Ethyl benzene
1,1,1 , 2-Tetrachl oroethane
m-Xylene
p-Xylene
o-Xylene
Styrene
Isopropyl benzene
Bromoform
1,1,2 , 2-Tetrachl oroethane
1,2,3-Trichloropropane
PID
Ret. Time8
minute
_b
-
9.88
-
-
-
16.14
-
19.30
-
-
23.11
-
-
-
25.21
-
26.10
-
27.99
-
-
-
31.95
-
33.88
-
-
-
36.56
36.72
-
36.98
36.98
38.39
38.57
39.58
-
-
-
HECD
Ret. Time
minute
8.47
9.47
9.93
11.95
12.37
13.49
16.18
18.39
19.33
20.99
22.88
23.14
23.64
24.16
24.77
25.24
25.47
-
26.27
28.02
28.66
29.43
29.59
-
33.21
33.90
34.00
34.73
35.34
36.59
-
36.80
-
-
-
-
-
39.75
40.35
40.81
PID
MDL
M9/L
0.02
ND°
0.05
0.02
0.02
0.009
0.02
0.01
0.05
0.003
0.005
0.01
0.01
0.02
0.01
0.05
HECD
MDL
M9/L
0.05
0.03
0.04
1.1
0.1
0.03
0.07
0.02
0.06
0.07
0.05
0.01
0.02
0.01
0.03
0.02
0.01
0.03
0.01
0.006
0.02
2.2
ND
0.04
0.03
0.03
0.8
0.01
0.005
1.6
0.01
0.4
8021A - 12
Revision 1
September 1994
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TABLE 1.
(Continued)
Analyte
PID
Ret. Time8
minute
HECD
Ret. Time
minute
PID
MDL
HECD
MDL
n-Propylbenzene 40.87
Bromobenzene 40.99
1,3,5-Trimethylbenzene 41.41
2-Chlorotoluene 41.41
4-Chlorotoluene 41.60
tert-Butylbenzene 42.92
1,2,4-Trimethylbenzene 42.71
sec-Butyl benzene 43.31
p-Isopropyltoluene 43.81
1,3-Dichlorobenzene 44.08
1,4-Dichlorobenzene 44.43
n-Butylbenzene 45.20
1,2-Dichlorobenzene 45.71
l,2-Dibromo-3-Chloropropane
1,2,4-Trichlorobenzene 51.43
Hexachlorobutadiene 51.92
Naphthalene 52.38
1,2,3-Trichlorobenzene 53.34
Internal Standards
Fluorobenzene 26.84
2-Bromo-1-chloropropane
41.03
41.45
41.63
44.11
44.47
45.74
48.57
51.46
51.96
53.37
33.08
0.004
0.006
0.004
ND
0.02
0.06
0.05
0.02
0.01
0.02
0.007
0.02
0.05
0.02
0.06
0.06
ND
0.03
0.01
0.01
0.02
0.01
0.02
3.0
0.03
0.02
0.03
Retention times determined on 60 m x 0.75 mm ID VOCOL capillary column.
Program: Hold at 10°C for 8 minutes, then program at 4°C/min to 180°C, and
hold until all expected compounds have eluted.
b Dash (-) indicates detector does not respond.
c ND = Not determined.
8021A - 13
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TABLE 2.
SINGLE LABORATORY ACCURACY AND PRECISION DATA
FOR VOLATILE ORGANIC COMPOUNDS IN WATERd
Photoionization
Detector
Analyte
Benzene
Bromobenzene
Bromochl oromethane
Bromodi chl oromethane
Bromoform
Bromomethane
n-Butylbenzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chl oromethane
2-Chlorotoluene
4-Chlorotoluene
1 ,2-Dibromo-3-chloropropane
Di bromochl oromethane
1,2-Dibromoethane
Dibromomethane
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-1,2 Dichloroethene
tr ans-1, 2-Di chl oroethene
1 ,2-Dichloropropane
1,3-Dichloropropane
2,2-Dichloropropane
1 , 1 -Di chl oropropene
Ethyl benzene
Hexachlorobutadiene
Isopropyl benzene
p- 1 sopropyl toluene
Recovery,8
%
99
99
-
-
-
-
100
97
98
-
100
-
-
-
NDC
101
-
-
-
-
102
104
103
-
-
-
100
ND
93
-
-
-
103
101
99
98
98
Standard
Deviation
of Recovery
1.2
1.7
-
-
-
-
4.4
2.6
2.3
-
1.0
-
-
-
ND
1.0
-
-
-
-
2.1
1.7
2.2
-
-
-
2.4
ND
3.7
-
-
-
3.6
1.4
9.5
0.9
2.4
Hall Electrolytic
Conductivity Detector
Standard
Recovery,8 Deviation
% of Recovery
_b
97
96
97
106
97
-
-
-
92
103
96
98
96
97
97
86
102
97
109
100
106
98
89
100
100
103
105
99
103
100
105
103
-
98
-
-
.
2.7
3.0
2.9
5.5
3.7
-
-
-
3.3
3.7
3.8
2.5
8.9
2.6
3.1
9.9
3.3
2.7
7.4
1.5
4.3
2.3
5.9
5.7
3.8
2.9
3.5
3.7
3.8
3.4
3.6
3.4
-
8.3
-
-
8021A - 14
Revision 1
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TABLE 2.
(Continued)
Analyte
Photoionization
Detector
Recovery,
Standard
Deviation
of Recovery
Hall Electrolytic
Conductivity Detector
Recovery,
Standard
Deviation
of Recovery
Methyl ene chloride
Naphthalene
n-Propyl benzene
Styrene
1,1, 1 ,2-Tetrachloroethane
1,1,2,2-Tetrachloroethane
Tetrachl oroethene
Toluene
1,2,3-Trichlorobenzene
1,2,4-Trichlorobenzene
1,1, 1 -Tri chl oroethane
1 , 1 , 2 -Tri chl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
1 , 2 , 4 -Tri methyl benzene
1 , 3 , 5 -Tri methyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
-
102
103
104
-
-
101
99
106
104
-
-
100
-
-
99
101
109
99
100
99
-
6.3
2.0
1.4
-
-
1.8
0.8
1.9
2.2
-
-
0.78
-
-
1.2
1.4
5.4
0.8
1.4
0.9
97
-
-
-
99
99
97
-
98
102
104
109
96
96
99
-
-
95
-
-
-
2.8
-
-
-
2.3
6.8
2.4
-
3.1
2.1
3.4
6.2
3.5
3.4
2.3
-
-
5.6
-
-
Recoveries and standard deviations were determined from seven samples and spiked at
10 jug/L of each analyte. Recoveries were determined by internal standard method. Internal
standards were: Fluorobenzene for PID, 2-Bromo-l-chloropropane for HECD.
b Detector does not respond.
c ND = Not determined.
This method was
reference 5).
tested in a single laboratory using water spiked at 10 M9/L (see
8021A - 15
Revision 1
September 1994
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TABLE 3.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQL)
FOR VARIOUS MATRICES8
Matrix Factor
Ground water 10
Low-concentration soil 10
Water miscible liquid waste 500
High-concentration soil and sludge 1250
Non-water miscible waste 1250
EQL = [Method detection limit (see Table 1)] X [Factor found in
this table]. For non-aqueous samples, the factor is on a wet-
weight basis. Sample EQLs are highly matrix-dependent. The EQLs
listed herein are provided for guidance and may not always be
achievable.
8021A - 16 Revision 1
September 1994
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FIGURE 1.
PURGING DEVICE
BUT 114 IN 0.0
>— 14 MM 00
INLET 1(4 IN. O 0
EXIT 1M IN. 0.0
10 MM GLASS FRTT
SAMPLE INLET
2-WAY SYRINGE VALVE
tr CM 20 GAUGE SYRINGE NEEOU
6 MM 0 0 RUMER SEPTUM
MUTT 1/4 IN 00
vie IN o o
y~ STAINLESS STEEL
13X
MCXECUCAR SIEVE
PUflQE GAS FILTER
PUAGEOAS
8021A - 17
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FIGURE 2.
TRAP PACKINGS AND CONSTRUCTION TO INCLUDE DESORB CAPABILITY
PACKING DETAIL
~t- 3 MM GLASS *OX
CONSTRUCTION OCTAJC
CM SlUO
'S CM THA* 3C
— • CM J««. OV-1
8021A - 18
Revision 1
September 1994
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FIGURE 3.
PURGE-AND-TRAP SYSTEM - PURGE MODE
CAflttEPGAS
PLOW CONTROL
pressure
REGULATOR
uomo tfuecnoN PORTS
COLUMN OVEN
*
13X MOLECULAR
S46VE FILTER
I
CONFIRMATORY COLUMN
TO 0€TKTOW
ANALYTICAL COLUMN
OPTIONAL 4JSORT COLUMN
S€L£CnON VALVf
». /- TRAP INLET
TRAP
22"C
PURGING
D€V>C€
NOTE.
ALL UNCS BCTWCEN TRAP
ANO QC SHOULD K HCATEO
8021A - 19
Revision 1
September 1994
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FIGURE 4.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - DESORB MODE
CARRIER GAS
FLOW CONTROL
PRESSURE
REGULATOR
PURGE GAS
FLOW CONTROL
13X MOLECULAR
SIEVE FILTER
UOUIO INJECTION PORTS
r— COLUMN OVEN
JUUV-
CONFIRMATORY COLUMN
TO DETECTOR
ANALYTICAL COLUMN
OPTIONAL *PORT COLUMN
SELECTION VALVE
TRAP INLET
TRAP
200 *C
i PURGING
1 DEVICE
NOTE.
ALL UNES BETWEEN TRAP
AND GC SHOULD BE HEATED
TO«TC.
8021A - 20
Revision 1
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FIGURE 5.
GAS CHROMATOGRAM OF VOLATILE ORGAN ICS
COLUMNI 60 METED x 0.73 MM I.D. VOCOL CAPILLARV
AMI TMAT VOC'l WITH NAU. • flO tH SCAIES
58 ST f= R S3 8CS 8 SSSS538
S^GS £ 8 5 iSSS
JL_JL_L
x .z x
J
ILJ
LJOllL.
*- JK I
UN d>-K I
8021A - 21
Revision 1
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METHOD 8021A
HALOGENATED VOLATILES BY GAS CHROMATOGRAPHY USING PHOTOIONIZATION
AND ELECTROLYTIC CONDUCTIVITY DETECTORS IN SERIES:
CAPILLARY COLUMN TECHNIQUE
( Start j
7.2 Set
chromatographic
conditions.
7.3 Refer to
Method 8000 for
calibration techniques.
7.4.1 Introduce
sample into GC using
direct injection or
purge-and-trap.
7.4.4 Record
sample volume
introduced into GC
and peak sizes.
7.4.5 Refer
to Method 8000 for
calculations.
7.4.6 Are
analytical
interferences
suspected?
7.4.7 Is peak
response off
scale?
Reanalyze sample
using second GC
column.
Dilute and reanalyze
second aliquot of
sample.
8021A - 22
Revision 1
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METHOD 8030A
ACROLEIN AND ACRYLONITRILE BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8030 is used to determine the concentration of the following
volatile organic compounds:
Compound Name CAS No.a
Acrolein (Propenal) 107-02-8
Acrylonitrile 107-13-1
a Chemical Abstract Services Registry Number.
1.2 Table 1 lists chromatographic conditions and method detection limits
for acrolein and acrylonitrile in organic-free reagent water. Table 2 lists the
estimated quantitation limit (EQL) for other matrices.
2.0 SUMMARY OF METHOD
2.1 Method 8030 provides gas chromatographic conditions for the detection
of the target analytes. Samples can be analyzed using direct injection or purge-
and-trap (Method 5030). Tenax should be used as the trap packing material.
Ground water samples must be analyzed using Method 5030. A temperature program
is used in the gas chromatograph to separate the organic compounds. Detection
is achieved by a flame ionization detector (FID).
2.2 The method provides an optional gas chromatographic column that may
be helpful in resolving the analytes from co-eluting non-target compounds and for
analyte confirmation.
3.0 INTERFERENCES
3.1 Refer to Methods 5030 and 8000.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly chlorofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A trip blank prepared from
organic-free reagent water and carried through sampling and subsequent storage
and handling can serve as a check on such contamination.
8030A - 1 Revision 1
July 1992
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4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas Chromatograph - Analytical system complete with gas
chromatograph suitable for on-column injections or purge-and-trap sample
introduction and all required accessories, including detectors, column
supplies, recorder, gases, and syringes. A data system for measuring peak
height and/or peak area is recommended.
4.1.2 Columns
4.1.2.1 Column 1 - 10 ft x 2 mm ID stainless steel or
glass packed with Porapak-QS (80/100 mesh) or equivalent.
4.1.2.2 Column 2 - 6 ft x 0.1 in. ID stainless steel or
glass packed with Chromosorb 101 (60/80 mesh) or equivalent.
4.1.3 Detector - Flame ionization (FID).
4.2 Sample introduction apparatus - Refer to Method 5030 for the
appropriate equipment for sample introduction purposes.
4.3 Syringes - A 5 ml Luer-lok glass hypodermic and a 5 mL, gas-tight
with shutoff valve.
4.4 Volumetric flasks, Class A - Appropriate sizes with ground glass
stoppers.
4.5 Microsyringes - 10 and 25 p,L with a 0.006 in. ID needle
(Hamilton 702N, or equivalent) and a 100 /iL.
4.6 Analytical balance - 0.0001 g.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water: All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Hydrochloric acid, HC1 - 1:1 (v/v).
5.4 Sodium hydroxide, NaOH - ION solution. Dissolve 40 g NaOH in
organic-free reagent water and dilute to 100 ml.
5.5 Stock standards - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standards in
8030A - 2 Revision 1
July 1992
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organic-free reagent water using assayed liquids. Because acrolein and
acrylonitrile are lachrymators, primary dilutions of these compounds should be
prepared in a hood.
5.5.1 Place about 9.8 ml of organic-free reagent water in a 10
mL tared ground-glass stoppered volumetric flask. For acrolein standards
the water must be adjusted to pH 4-5 using hydrochloric acid (1:1 v/v) or
sodium hydroxide (ION), if necessary. Weigh the flask to the nearest
0.0001 g.
5.5.2 Using a 100 /itL syringe, immediately add two or more drops
of assayed reference material to the flask, then reweigh. The liquid must
fall directly into the water without contacting the neck of the flask.
5.5.3 Reweigh, dilute to volume, stopper, and then mix by
inverting the flask several times. Calculate the concentration in
milligrams per liter (mg/L) from the net gain in weight. When compound
purity is assayed to be 96% or greater, the weight may be used without
correction to calculate the concentration of the stock standard.
Commercially prepared stock standards may be used at any concentration, if
they are certified by the manufacturer or by an independent source.
5.5.4 Transfer the stock standard solution into a bottle with
a Teflon lined screw-cap. Store, with minimal headspace, at 4°C and
protect from light.
5.5.5 Prepare fresh standards daily.
5.6 Secondary dilution standards - Prepare secondary dilution standards
as needed, in organic-free reagent water, from the stock standard solutions. The
secondary dilution standards must contain the compounds of interest, either
singly or mixed together. The secondary dilution standards should be prepared
at concentrations such that the aqueous calibration standards prepared in Section
5.7 will bracket the working range of the analytical system. Secondary dilution
standards should be stored with minimal headspace, and should be checked
frequently for signs of degradation or evaporation, especially just prior to
preparing calibration standards from them.
5.7 Calibration standards - Prepare calibration standards in organic-free
reagent water from the secondary dilution standards at a minimum of five
concentrations. One of the concentrations should be at a concentration near, but
above, the method detection limit. The remaining concentrations should
correspond to the expected range of concentrations found in real samples, or
should define the working range of the GC. Each standard should contain each
analyte for detection by this method. In order to prepare accurate aqueous
standard solutions, the following precautions must be observed.
5.7.1 Use a 25 jitL Hamilton 702N microsyringe, or equivalent,
(variations in needle geometry will adversely affect the ability to
deliver reproducible volumes of standards into water).
5.7.2 Never use pipets to dilute or transfer samples or aqueous
standards.
8030A - 3 Revision 1
July 1992
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5.7.3 Standards must be prepared daily.
5.8 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.8.1 Prepare calibration standards at a minimum of five
concentrations for each parameter of interest, as described in Section
5.7.
5.8.2 Prepare a spiking solution containing each of the internal
standards, using the procedures described in Sections 5.5 and 5.6. It is
recommended that the secondary dilution standard be prepared at a
concentration of 15 mg/L of each internal standard compound. The addition
of 10 /iL of this standard to 5.0 ml of sample or calibration standard
would be equivalent to 30
5.8.3 Analyze each calibration standard according to Section
7.0, adding 10 nl of internal standard spiking solution directly to the
syringe.
5.9 Surrogate standards - The analyst should monitor both the performance
of the analytical system and the effectiveness of the method in dealing with each
sample matrix by spiking each sample, standard, and organic-free reagent water
blank with one or two surrogate compounds (e.g. compounds similar in analytical
behavior to the analytes of interest but which are not expected to be present in
the sample) recommended to encompass the range of the temperature program used
in this method. From stock standard solutions prepared as in Section 5.5, add
a volume to give 750 M9 of each surrogate to 45 ml of organic-free reagent water
contained in a 50 mL volumetric flask, mix, and dilute to volume for a
concentration of 15 ng/juL. Add 10 juL of this surrogate spiking solution
directly into the 5 ml syringe with every sample and reference standard analyzed.
If the internal standard calibration procedure is used, the surrogate compounds
may be added directly to the internal standard spiking solution (Section 5.8.2).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Volatile compounds are introduced into the gas chromatograph either
by direct injection or heated purge-and-trap (Method 5030). Method 5030 may be
used directly on ground water samples or low-concentration contaminated soils and
sediments. For high-concentration soils or sediments, methanolic extraction, as
described in Method 5030, may be necessary prior to purge-and-trap analysis.
8030A - 4 Revision 1
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7.2 Gas chromatographlc conditions (Recommended)
7.2.1 Column 1:
Helium flow rate = 30 mL/min
Temperature program:
Initial temperature = 110°C, hold for 1.5 minutes
Program = 110°C to 150°C, heating as
rapidly as possible
Final temperature = 150°C, hold for 20 minutes.
7.2.2 Column 2:
Helium flow rate = 40 mL/min
Temperature program:
Initial temperature = 80°C, hold for 4 minutes
Program = 80°C to 120°C at 50°C/"rin
Final temperature = 120°C, hold for 12 minutes.
7.3 Calibration - Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point on
the calibration curve.
7.3.1 Calibration must take place using the same sample
introduction method that will be used to analyze actual samples (see
Section 7.4.1).
7.3.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.4 Gas chromatographic analysis
7.4.1 Introduce volatile compounds into the gas chromatograph
using either Method 5030 (heated purge-and-trap method using Tenax as the
trap packing material) or the direct injection method. If the internal
standard calibration technique is used, add 10 juL of the internal standard
to the sample prior to purging.
7.4.1.1 Direct injection - In very limited applications
(e.g. aqueous process wastes), direct injection of the sample into
the GC system with a 10 pi syringe may be appropriate. The
detection limit is very high (approximately 10,000 /ug/L); therefore,
it is only permitted when concentrations in excess of 10,000 ng/L
are expected or for water-soluble compounds that do not purge. The
system must be calibrated by direct injection (bypassing the purge-
and-trap device).
7.4.2 Follow Method 8000 for instructions on the analysis
sequence, appropriate dilutions, establishing daily retention time
windows, and identification criteria. Include a mid-concentration
standard after each group of 10 samples in the analysis sequence.
7.4.3 Table 1 summarizes the estimated retention times and
detection limits for a number of organic compounds analyzable using this
8030A - 5 Revision 1
July 1992
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method. Figure 1 illustrates the chromatographic separation of acrolein
and of acrylonitrile using Column 1.
7.4.4 Record the sample volume purged or injected and the
resulting peak sizes (in area units or peak heights).
7.4.5 Calculation of concentration is covered in Method 8000.
7.4.6 If analytical interferences are suspected, or for the
purpose of confirmation, analysis using the second GC column is
recommended.
7.4.7 If the response for a peak is off-scale, prepare a
dilution of the sample with organic-free reagent water. The dilution must
be performed on a second aliquot of the sample which has been properly
sealed and stored prior to use.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures and
Method 8000 for gas chromatographic procedures. Quality control to ensure the
proper operation of the purge-and-trap device is covered in Method 5030.
8.2 Procedures to check the GC system operation are found in Method 8000,
Section 8.6.
8.2.1 The quality control check sample concentrate (Method 8000,
Section 8.6) should contain each parameter of interest at a concentration
of 25 mg/L in water.
8.2.2 Table 3 indicates the calibration and QC acceptance
criteria for this method. Table 4 gives single laboratory accuracy and
precision for the analytes of interest. The contents of both Tables
should be used to evaluate a laboratory's ability to perform and generate
acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if recovery is within limits (limits established by performing
QC procedure outlined in Method 8000, Section 8.10).
8.3.1 If recovery is not within limits, the following is
required.
• Check to be sure that there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of the
above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are a
problem or flag the data as "estimated concentration".
8030A - 6 Revision 1
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9.0 METHOD PERFORMANCE
9.1 In a single laboratory, the average recoveries and standard
deviations presented in Table 4 were obtained using Method 5030. Seven replicate
samples were analyzed at each spike concentration.
9.2 The accuracy and precision obtained will be determined by the sample
matrix, sample introduction technique, and by the calibration procedure used.
10.0 REFERENCES
1. Bellar, T.A. and J.J. Lichtenberg, J. Amer. Water Works Assoc., 66(12).
pp. 739-744, 1974.
2. Bellar, T.A. and J.J. Lichtenberg, "Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds," in Van Hall, ed., Measurement of Organic Pollutants in Water
and Wastewater, ASTM STP 686, pp. 108-129, 1979.
3. Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters, Category 11: Purgeables and Category 12:
Acrolein, Acrylonitrile, and Dichlorodifluoromethane, Report for EPA
Contract 68-03-2635 (in preparation).
4. Going, J., et al., Environmental Monitoring Near Industrial Sites -
Acrylonitrile, Office of Toxic Substances, U.S. EPA, Washington, DC, EPA
560/6-79-003, 1979.
5. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
6. Kerns, E.H., et al. "Determination of Acrolein and Acrylonitrile in Water
by Heated Purge and Trap Technique," U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, Ohio 45268,
1980.
7. "Evaluation of Method 603," Final Report for EPA Contract 68-03-1760 (in
preparation).
8030A - 7 Revision 1
July 1992
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
Retention time (min) Method detection
Compound Col. 1 Col. 2 limit8 (/xg/L)
Acrolein
Acrylonitrile
10.6
12.7
8.2
9.8
0.7
0.5
Based on using purge-and-trap, Method 5030.
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION
LIMITS (EQLs) FOR VARIOUS MATRICES8
Matrix Factor6
Ground water 10
Low-concentration soil 10
Water miscible liquid waste 500
High-concentration soil and sludge 1250
Non-water miscible waste 1250
Sample EQLs are highly matrix dependent. The EQLs listed herein
are provided for guidance and may not always be achievable.
EQL = [Method detection limit (Table 1)] X [Factor (Table 2)].
For non-aqueous samples, the factor is on a wet-weight basis.
8030A - 8 Revision 1
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TABLE 3.
CALIBRATION AND QC ACCEPTANCE CRITERIA3
Analyte
Acrolein
Acrylonitrile
Range
for Q
(M9/L)
45.9 - 54.1
41.2 - 58.8
Limit
for S
(M9/L)
4.6
9.9
Range
for x
(M9/L)
42.9 - 60.1
33.1 - 69.9
Range
P> Ps
(%)
88-118
71-135
Q = Concentration measured in QC check sample, in M9/L-
S = Standard deviation of four recovery measurements, in /zg/L.
R = Average recovery for four recovery measurements, in jig/L.
P, P = Percent recovery measured.
Criteria from 40 CFR Part 136 for Method 603
assuming a QC check sample concentration of 50
and were calculated
TABLE 4.
SINGLE LABORATORY ACCURACY AND PRECISION
Parameter
Acrolein
Acrylonitrile
AW
POTW
Spike
cone.
(M9/L)
5.0
50.0
5.0
50.0
5.0
100.0
5.0
50.0
20.0
100.0
10.0
100.0
ASTM Type
Average
recovery
(M9/L)
5.2
51.4
4.0
44.4
0.1
9.3
4.2
51.4
20.1
101.3
9.1
104.0
II water.
Prechlorination secon
Standard
deviation
(M9/L)
0.2
0.7
0.2
0.8
0.1
1.1
0.2
1.5
0.8
1.5
0.8
3.2
dary effluent
Average
percent
recovery
104
103
80
89
2
9
84
103
100
101
91
104
from a munici
Sampl e
matrix
AW
AW
POTW
POTW
IW
IW
AW
AW
POTW
POTW
IW
IW
pal sewage
IW
treatment plant.
Industrial wastewater containing an
reactant.
unidentified acrolein
8030A - 9
Revision 1
July 1992
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Figure 1
Gas Chromatogram of Acrolein and Acrylonitrile
Column: Por«pafc.Qs
110«C lot is mm
««tod to 1SO«C
Ootoctor: FUm«
<* «0 78 90 10.5 120 US ISO
METCNTlONTlMi. MIN.
8030A - 10
Revision 1
July 1992
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METHOD 8030A
ACROLEIN AND ACRYLONITRILE BY GAS CHROMATOGRAPHY
Start
7 1 Int reduce
compounds into gas
chromatograph by
direct injection or
purge-and- trap
(Method 5030)
7 2 Set gas
chromatograph
condition.
7 3 Calibrate
(refer to Method
8000)
7 4 1 Int r oduce
volatile compounds
into gas
chromatograph by
purge-and-trap or
direct injection
742 Folio* Method
8000 for analysis
sequence, etc
7 4 4 Record volume
purged or injected
and peak sizes
7 4 5 Calculate
concent ration
(refer to Method
8000)
7 4 6 Analyze us ing
second GC column
7 4 7 Dilute second
aliquot of sample
8030A - 11
Revision 1
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METHOD 8031
ACRYLONITRUE BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8031 is used to determine the concentration of acrylonitrile
in water. This method may also be applicable to other matrices. The following
compound can be determined by this method:
Compound Name CAS No.'
Acrylonitrile 107-13-1
a Chemical Abstract Services Registry Number.
1.2 The estimated quantitation limit of Method 8031 for determining the
concentration of acrylonitrile in water is approximately 10 p.g/1.
1.3 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 A measured sample volume is micro-extracted with methyl tert-butyl
ether. The extract is separated by gas chromatography and measured with a
Nitrogen/Phosphorus detector.
3.0 INTERFERENCES
3.1 Method interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware that leads to discrete
artifacts and/or elevated baselines in gas chromatograms. All of these materials
must be routinely demonstrated to be free from interferences under the conditions
of the analysis by running laboratory reagent blanks.
3.2 Samples can be contaminated by diffusion of volatile organics around
the septum seal into the sample during handling and storage. A field blank
should be prepared from organic-free reagent water and carried through the
sampling and sample handling protocol to serve as a check on such contamination.
3.3 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
8031 - 1 Revision 0
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sample syringe must be rinsed out between samples with solvent. Whenever an
unusually concentrated sample is encountered, it should be followed by the
analysis of solvent to check for cross contamination.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph system
4.1.1 Gas chromatograph, analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detector, analytical columns, recorder, gases, and
syringes. A data system for measuring peak heights and/or peak areas is
recommended.
4.1.2 Column: Porapak Q - 6 ft., 80/10 Mesh, glass column, or
equivalent.
4.1.3 Nitrogen/Phosphorus detector.
4.2 Materials
4.2.1 Grab sample bottles - 40 mL VOA bottles.
4.2.2 Mixing bottles - 90 ml bottle with a Teflon lined cap.
4.2.3 Syringes - 10 /uL and 50 juL.
4.2.4 Volumetric flask (Class A) - 100 ml.
4.2.5 Graduated cylinder - 50 ml.
4.2.6 Pipet (Class A) - 5, 15, and 50 ml.
4.2.7 Vials - 10 ml.
4.3 Preparation
4.3.1 Prepare all materials to be used as described in Chapter 4 for
volatile organics.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
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5.2 General
5.2.1 Methanol, CH3OH - Pesticide quality, or equivalent.
5.2.2 Organic-free reagent water. All references to water in this
method refer to organic-free reagent water, as defined in Chapter One.
5.2.3 Methyl tert-butyl ether, CH3Ot-C4H9 - Pesticide quality, or
equivalent.
5.2.4 Acrylonitrile, H2C:CHCN, 98%.
5.3 Stock standard solution
5.3.1 Stock standard solutions - Can be prepared from pure standard
materials or can be purchased as certified solutions. Prepare stock
standards in organic-free reagent water using assayed liquids.
5.3.2 The stock standard solution may be prepared by volume or by
weight. Stock solutions must be replaced after one year, or sooner if
comparison with the check standards indicates a problem.
CAUTION: Acrylonitrile is toxic. Standard preparation should be
performed in a laboratory fume hood.
5.3.2.1 To prepare the stock standard solution by volume:
inject 10 /uL of acrylonitrile (98%) into a 100 mL volumetric flask
with a syringe. Make up to volume with methanol.
5.3.2.2 To prepare the stock standard solution by weight:
Place about 9.8 ml of organic-free reagent water into a 10 ml
volumetric flask before weighing the flask and stopper. Weigh the
flask and record the weight to the nearest 0.0001 g. Add two drops
of pure acrylonitrile, using a 50 juL syringe, to the flask. The
liquid must fall directly into the water, without contacting the
inside wall of the flask. Stopper the flask and then reweigh.
Dilute to volume with organic-free reagent water. Calculate the
concentration from the net gain in weight.
5.4 Working standard solutions
5.4.1 Prepare a minimum of 5 working standard solutions that cover
the range of analyte concentrations expected in the samples. Working
standards of 20, 40, 60, 80, and 100 M9/L may be prepared by injecting 10,
20, 30, 40, and 50 /uL of the stock standard solution prepared in Sec.
5.3.2.1 into 5 separate 90 ml mixing bottles containing 40 ml of organic-
free reagent water.
5.4.2 Inject 15 ml of methyl tert-butyl ether into each mixing
bottle, shake vigorously, and let stand 5 minutes, or until layers have
separated.
8031 - 3 Revision 0
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5.4.3 Remove 5 ml of top layer by pipet, and place in a 10 mL vial.
5.4.4 Keep all standard solutions below 4°C until used.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Sample Extraction
7.1.1 Pour 40 mL of the sample into a 90 mL mixing bottle. Pipet 15
mL of Methyl tert-butyl ether into the mixing bottle. Shake vigorously
for about 2 min. and let stand for about 5 min. Remove about 5 mL of the
top layer and store in a 10 mL vial.
7.2 Chromatographic Conditions (Recommended)
Carrier Gas (He) flow rate: 35 mL/min.
Column Temperature: 180° C, Isothermal
Injection port temperature: 250° C
Detector temperature: 250° C
Detector Current (DC): 18 volts
Gases: Hydrogen, 3 mL/min; Air, 290 mL/min.
7.3 Calibration of GC
7.3.1 On a daily basis, inject 3 pi of methyl tert-butyl ether
directly into the GC to flush the system. Also purge the system with
methyl tert-butyl ether injections between injections of standards and
samples.
7.3.2 Inject 3 jiL of a sample blank (organic-free reagent water
carried through the sample storage procedures and extracted with methyl
tert-butyl ether).
7.3.3 Inject 3 /xL of at least five standard solutions: one should
be near the detection limit; one should be near, but below, the expected
concentrations of the analyte; one should be near, but above, the expected
concentrations of the analyte. The range of standard solution
concentrations used should not exceed the working range of the GC system.
7.3.4 Prepare a calibration curve using the peak areas of the
standards (retention time of acrylonitrile under the conditions of Sec.
7.2 is approximately 2.3 minutes). If the calibration curve deviates
significantly from a straight line, prepare a new calibration curve with
the existing standards, or, prepare new standards and a new calibration
curve. See Method 8000, Sec. 7.4.2, for additional guidance on
calibration by the external standard method.
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7.4 Sample Analysis
7.4.1 Inject 3 juL of the sample extract, using the same
chromatographic conditions used to prepare the standard curve. Calculate
the concentration of acrylonitrile in the extract, using the area of the
peak, against the calibration curve prepared in Sec. 7.3.4.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
8.2 Prior to preparation of stock solutions, methanol and methyl
tert-butyl ether reagents should be analyzed gas chromatographically under the
conditions described in Sec. 7.2, to determine possible interferences with the
acrylonitrile peak. If the solvent blanks show contamination, a different batch
of solvents should be used.
9.0 METHOD PERFORMANCE
9.1 Method 8031 was tested in a single laboratory over a period of days.
Duplicate samples and one spiked sample were run for each calculation. The GC
was calibrated daily. Results are presented in Table 1.
10.0 REFERENCES
1. K.L. Anderson, "The Determination of Trace Amounts of Acrylonitrile in
Water by Specific Nitrogen Detector Gas Chromatograph", American Cynamid
Report No. WI-88-13, 1988.
8031 - 5 Revision 0
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TABLE 1
SINGLE LABORATORY METHOD PERFORMANCE
CONCENTRATION
SAMPLE SPIKE (jxg/l.) % RECOVERY
A 60 100
B 60 105
C 40 86
D 40 100
E 40 88
F 60 94
Average 96
8031 - 6 Revision 0
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METHOD 8031
ACRYLONITRILE BY GAS CHROMATOGRAPHY
C
Start
7.1 .1 Extract 40 mL
of sample with methyl
t-butyl ether in 90 mL
bottle.
1
r
7.2 Set
Chromatographic
conditions.
>
r
7.3.1 Flush GC
system with 30 uL
methyl t-butyl ether.
>
f
7.3.2 Analyze 3 uL
of sample blank.
^
1
7.3.3 - 7.3.4 Establish
calibration curve with
at least 5 stds.
>
r
7.4 Sample Analysis
^
r
Stop
8031 - 7
Revision 0
September 1994
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METHOD 8032
ACRYLAMIDE BY GAS CHROMATOGRAPHV
1.0 SCOPE AND APPLICATION
1.1 Method 8032 is used to determine trace amounts of acrylamide monomer
in aqueous matrices. This method may be applicable to other matrices. The
following compound can be determined by this method:
Compound Name CAS No.a
Acrylamide 79-06-01
a Chemical Abstract Services Registry Number.
1.2 The method detection limit (MDL) in clean water is 0.032 M9/L-
1.3 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Method 8032 is based on bromination of the acrylamide double bond.
The reaction product (2,3-dibromopropionamide) is extracted from the reaction
mixture with ethyl acetate, after salting out with sodium sulfate. The extract
is cleaned up using a Florisil column, and analyzed by gas chromatography with
electron capture detection (GC/ECD).
2.2 Compound identification should be supported by at least one
additional qualitative technique. Analysis using a second gas chromatographic
column or gas chromatography/mass spectrometry may be used for compound
confirmation.
3.0 INTERFERENCES
3.1 No interference is observed from sea water or in the presence of 8.0%
of ammonium ions derived from ammonium bromide. Impurities from potassium
bromide are removed by the Florisil clean up procedure.
8032 - 1 Revision 0
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4.0 APPARATUS AND MATERIALS
4.1 Gas chromatographic System
4.1.1 Gas chromatograph suitable for on-column injections with all
required accessories, including detector, analytical columns, recorder,
gases, and syringes. A data system for measuring peak heights and/or peak
areas is recommended.
4.1.2 Column: 2 m x 3 mm glass column, 5% FFAP (free fatty acid
polyester) on 60-80 mesh acid washed Chromosorb W, or equivalent.
4.1.3 Detector: electron capture detector.
4.2 Kuderna-Danish (K-D) apparatus.
4.2.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.2.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.2.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.2.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Separatory funnel - 150 ml.
4.4 Volumetric flask (Class A) - 100 ml, with ground glass stopper;
25 ml, amber, with ground glass stopper.
4.5 Syringe - 5 mL.
4.6 Microsyringes - 5 jiL, 100 jiL.
4.7 Pipets (Class A).
4.8 Glass column (30 cm x 2 cm).
4.9 Mechanical shaker.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
8032 - 2 Revision 0
September 1994
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such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Solvents
5.3.1 Ethyl acetate, C2H5C02C2H5. Pesticide quality, or equivalent.
5.3.2 Diethyl ether, C2H5OC2H5. Pesticide quality, or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl alcohol preservative must be
added to each liter of ether.
5.3.3 Methanol, CH3OH. Pesticide quality, or equivalent.
5.3.4 Benzene, C6H6. Pesticide quality, or equivalent.
5.3.5 Acetone, CH3COCH3. Pesticide quality, or equivalent.
5.4 Saturated bromine water. Prepare by shaking organic-free reagent
water with bromine and allowing to stand for 1 hour, in the dark, at 4°C. Use
the aqueous phase.
5.5 Sodium sulfate (anhydrous, granular), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.6 Sodium thiosulfate, Na2S203, 1 M aqueous solution.
5.7 Potassium bromide, KBr, prepared for infrared analysis.
5.8 Concentrated hydrobromic acid, HBr, specific gravity 1.48.
5.9 Acrylamide monomer, H2C:CHCONH2, electrophoresis reagent grade,
minimum 95% purity.
5.10 Dimethyl phthalate, C6H4(COOCH3)2, 99.0% purity.
5.11 Florisil (60/100 mesh): Prepare Florisil by activating at 130°C for
at least 16 hours. Alternatively, store Florisil in an oven at 130°C. Before
use, cool the Florisil in a desiccator. Pack 5 g of the Florisil, suspended in
benzene, in a glass column (Sec. 4.8).
5.12 Stock standard solutions
5.12.1 Prepare a stock standard solution of acrylamide monomer
as specified in Sec. 5.12.1.1. When compound purity is assayed to be 96%
8032 - 3 Revision 0
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or greater, the weight can be used without correction to calculate the
concentration of the stock standard. Commercially prepared standards can
be used at any concentration if they are certified by the manufacturer or
by an independent source.
5.12.1.1 Dissolve 105.3 mg of acrylamide monomer in
organic-free reagent water in a 100 mL volumetric flask, and dilute
to the mark with organic-free reagent water. Dilute the solution of
acrylamide monomer so as to obtain standard solutions containing
0.1 - 10 mg/L of acrylamide monomer.
5.13 Calibration standards
5.13.1 Dilute the acrylamide stock solution with organic-free
reagent water to produce standard solutions containing 0.1-5 mg/L of
acrylamide. Prior to injection the calibration standards are reacted and
extracted in the same manner as environmental samples (Sec. 7).
5.14 Internal standards
5.14.1 The suggested internal standard is dimethyl phthalate.
Prepare a solution containing 100 mg/L of dimethyl phthalate in ethyl
acetate. The concentration of dimethyl phthalate in the sample extracts
and calibration standards should be 4 mg/L.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Bromination
7.1.1 Pipet 50 mL of sample into a 100 mL glass stoppered flask.
Dissolve 7.5 g of potassium bromide into the sample, with stirring.
7.1.2 Adjust the pH of the solution with concentrated hydrobromic
acid until the pH is between 1 and 3.
7.1.3 Wrap the flask with aluminum foil in order to exclude light.
Add 2.5 mL of saturated bromine water, with stirring. Store the flask and
contents in the dark, at 0°C, for at least 1 hour.
7.1.4 After reacting the solution for at least an hour, decompose
the excess of bromine by adding 1 M sodium thiosulfate solution, dropwise,
until the color of the solution is discharged.
7.1.5 Add 15 g of sodium sulfate, using a magnetic stirrer to effect
vigorous stirring.
8032 - 4 Revision 0
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7.2 Extraction
7.2.1 Transfer the solution into a 150 mL separatory funnel. Rinse
the reaction flask three times with 1 ml aliquots of organic-free reagent
water. Transfer the rinsings into the separatory funnel.
7.2.2 Extract the aqueous solution with two 10 mL portions of ethyl
acetate for 2 min each, using a mechanical shaker (240 strokes per min).
Dry the organic phase with 1 g of sodium sulfate.
7.2.3 Transfer the organic phase into a 25 mL amber volumetric
flask. Rinse the sodium sulfate with three 1.5 ml portions of ethyl
acetate and combine the rinsings with the organic phase.
7.2.4 Add exactly 100 jug of dimethyl phthalate to the flask and make
the solution up to the 25 ml mark with ethyl acetate. Inject 5 /xL
portions of this solution into the gas chromatograph.
7.3 Florisil cleanup: Whenever interferences are observed, the samples
should be cleaned up as follows.
7.3.1 Transfer the dried extract into a Kuderna-Danish evaporator
with 15 ml of benzene. Evaporate the solvent at 70°C under reduced
pressure, and concentrate the solution to about 3 mL.
7.3.2 Add 50 mL of benzene and subject the solution to Florisil
column chromatography at a flow rate of 3 mL/min. Elute the column first
with 50 mL of diethyl ether/benzene (1:4) at a flow rate of 5 mL/min, and
then with 25 mL of acetone/benzene (2:1) at a flow rate of 2 mL/min.
Discard all of the first eluate and the initial 9 mL portion of the second
eluate, and use the remainder for the determination, using dimethyl
phthalate (4 mg/L) as an internal standard.
NOTE: Benzene is toxic, and should be only be used under a
ventilated laboratory hood.
7.4 Gas chromatographic conditions:
Nitrogen carrier gas flow rate: 40 mL/min
Column temperature: 165°C.
Injector temperature: 180°C
Detector temperature: 185°C.
Injection volume: 5 ^L
7.5 Calibration:
7.5.1 Inject 5 juL of a sample blank (organic-free reagent water
carried through all sample storage, handling, bromination and extraction
procedures).
7.5.2 Prepare standard solutions of acrylamide as described in Sec.
5.13.1. Brominate and extract each standard solution as described in
Sees. 7.1 and 7.2.
8032 - 5 Revision 0
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7.5.2.1 Inject 5 p,i of each of a minimum of five standard
solutions: one should be near the detection limit; one should be
near, but below, the expected concentrations of the analyte; one
should be near, but above, the expected concentrations of the
analyte.
7.5.2.2 Prepare a calibration curve using the peak areas
of the standards. If the calibration curve deviates significantly
from a straight line, prepare a new calibration curve with the
existing standards, or, prepare new standards and a new calibration
curve. See Method 8000, Sec. 7.4.3, for additional guidance on
calibration by the internal standard method.
7.5.2.3 Calculate the response factor for each standard
according to Equation 1.
(Ps) (Mis)
RF = - Equation 1
(Pis) (MA)
RF = Response factor
Ps = Peak height of acryl amide
Mis = Amount of internal standard injected (ng)
Pjs = Peak height of internal standard
MA = Amount of acryl amide injected (ng)
7.5.3 Calculate the mean response factor according to Equation 2.
RF = - Equation 2
RF = Mean response factor
RF = Response factors from standard analyses
(calculated in Equation 1)
n = Number of analyses
7.6 Gas chromatographic analysis:
7.6.1 Inject 5 /iL portions of each sample (containing 4 mg/L
internal standard) into the gas chromatograph. An example GC/ECD
chromatogram is shown in Figure 1.
7.6.2 The concentration of acrylamide monomer in the sample is given
by Equation 3.
(PA) (MJ
[A] = - — - Equation 3
(Pis) (RF) (V() (V.)
[A] = Concentration of acrylamide monomer in sample (mg/L)
8032 - 6 Revision 0
September 1994
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PA = Peak height of acryl amide monomer
Mis = Amount of internal standard injected (ng)
Vs = Total volume of sample (mL)
P^ = Peak height of internal standard
RF = Mean response factor from Equation 2
Vi = Injection volume
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
9.0 METHOD PERFORMANCE
9.1 The following performance data have been generated under the
conditions described in this method:
9.1.1 The calibration curve for Method 8032 is linear over the range
0-5 jug/L of acryl amide monomer.
9.1.2 The limit of detection for an aqueous solution is 0.032 M9/L-
9.1.3 The yields of the brominated compound are 85.2 + 3.3% and 83.3
+ 0.9%, at fortification concentrations of 1.0 and 5.0 M9/U respectively.
9.2 Table 1 provides the recoveries of acrylamide monomer from river
water, sewage effluent, and sea water.
9.3 The recovery of the bromination product as a function of the amount
of potassium bromide and hydrobromic acid added to the sample is shown in
Figure 2.
9.4 The effect of the reaction time on the recovery of the bromination
product is shown in Figure 3. The yield was constant when the reaction time was
more than 1 hour.
9.5 Figure 4 shows the recovery of the bromination product as a function
of the initial pH from 1 to 7.35. The yield was constant within this pH range.
The use of conventional buffer solutions, such as sodium acetate - acetic acid
solution or phosphate solution, caused a significant decrease in yield.
10.0 REFERENCES
1. Hashimoto, A., "Improved Method for the Determination of Acrylamide
Monomer in Water by Means of Gas-Liquid Chromatography with an Electron-
capture Detector," Analyst, 101:932-938, 1976.
8032 - 7 Revision 0
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Q.
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Figure 1
I
A
B
I 10 12 14
Timt/min
16
Typical gas chromatograms of the bromination product obtained from aqueous
acrylamide monomer solution:
A.
B.
BL,
Peaks:
1.
I.
4-7.
Untreated
With Florisil cleanup
Chromatogram of blank, concentrated five-fold before gas chromatographic
analysis.
2,3-Dibromopropionamide
Dimethyl phthalate
Impurities from potassium bromide
Sample size = 100 ml; acrylamide monomer = 0.1 jug
8032 - 9
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Figure 2
0 5 10 IS 20 23
Amount of KBr/g ptr SO ml
L^_^^^^^BO^^«X>a^^^B^^^^B«^a«^^^U>
0 2 « 6 8 10
Amount of HBr/ml ptr SO ml
Effect of (A) potassium bromide and (B) hydrobromic acid on the yield of
bromination. Sample size = 50 ml; acrylamide monomer = 0.25 /ng
8032 - 10
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Figure 3
24
Effect of reaction time on the bromination. Reaction conditions:
50 ml of sample;
0.25 /xg of acrylamide monomer;
7.5 g of potassium bromide;
2.5 ml of saturated bromine water
Extraction conditions:
15 g of sodium sulfate;
extraction at pH 2;
solvent = 10 ml of ethyl acetate (X2)
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Figure 4
100
0 O 0
so
012345671
PM
Effect of initial pH on the bromination. Reaction and extraction conditions as
in Figure 3. The pH was adjusted to below 3 with concentrated hydrobromic acid,
and to 4-5 with dilute hydrobromic acid. Reaction at pH 6 was in distilled
water. pH 7.35 was achieved by careful addition of dilute sodium hydroxide
solution. The broken line shows the result obtained by the use of sodium acetate
- acetic acid buffer solution.
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METHOD 8032
ACRYLAMIDE BY GAS CHROMATOGRAPHY
Start
T
7.1 Bromination
\
7.1 .1 Dissolve 7.5 g KBr into
50 mL sample in flask.
1
7.1 .2 Adjust soln. pH with
concentrated HBr to between
1 and 3.
1
7. 1.3 Wrap soln. flask with
aluminum. Add 2.5 ml satd.
bromine water, stir, store at
0 C for 1 hr
I
7.1 4Add1 M sodium
thiosulfate dropwise to flask to
decompose excess bromine.
1
7.1. 5 Add 15 g sodium
sulfate, and stir
*
7.2 Extraction
1
7.2.1 Transfer flask soln. to
sap. funnel along with rinses.
1
7.2.2 Extract soln. twice w/ethyl
acetate. Dry organic phase
using sodium sulfate.
t
7.2.3 Transfer organic phase
and rinses into amber
glass flask.
1
7.2.4 Add 1 00 ug dimethyl
phthalate to flask, dilute to
mark. Inject 5 uL into GC.
1
7.3 Florisil Cleanup
1
7.3.1 Transfer dried extract to
K-D assembly w/benzene.
Concentrate to 3 ml at 70 C
under reduced pressure.
I
7.3.2 Add 50 ml benzene to
solution. Pass soln. through
Florisil column. Elutewith
diethyl ether/benzene, then
acetone/benzene. Collect
the second eluton train (less
initial 9 ml) for analysis.
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METHOD 8032
continued
7.4 GC Conditions
7.5 Calibration
I
7.5.1 Inject 5 uL sample blank.
7.5.2 Brominate and extract std.
solns. similar to the samples
.1 Inject 5 uL of each of the
minimum 5 stds
.2 Plot peak are vs. [}
.3 Calculate response factor
(RF) for each [ ].
7.5.3 Calculate mean RF from
eqn.2^
7.6 GC Analysis
7.6.1 Inject 5 uL sample containing
internal std. into GC.
7.6.2 Calculate acrylamide monomer
concentration in sample using
eqn. 3.
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METHOD 8040A
PHENOLS BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8040 is used to determine the concentration of various
phenolic compounds. The following compounds can be determined by this method:
Compound Name
Appropriate Technique
CAS No.8 3510 3520 3540 3550 3580
2-sec-Butyl-4,6-dinitrophenol
(DNBP, Dinoseb)
4-Chloro-3-methylphenol
2-Chlorophenol
Cresols (methyl phenols)
2-Cyclohexyl-4,6-dinitrophenol
2,4-Dichlorophenol
2,6-Dichlorophenol
2,4-Dimethylphenol
2,4-Dinitrophenol
2-Methyl-4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
Tetrachlorophenols
Trichlorophenols
2,4,6-Trichlorophenol
88-85-7 X ND ND ND X
59-50-7 X X X X X
95-57-8 X X X X X
1319-77-3 X ND ND ND X
131-89-5 X ND ND ND LR
120-83-2 X X X X X
87-65-0 X ND ND ND X
105-67-9 X X X X X
51-28-5 X X X X X
534-52-1 X X X X X
88-75-5 X X X X X
100-02-7 X X X X X
87-86-5 X X X X X
108-95-2 DC(28) X X X X
25167-83-3 X ND ND ND X
25167-82-2 X X X X X
88-06-2 X X X X X
a Chemical Abstract Services Registry Number.
DC = Unfavorable distribution coefficient (number in parenthesis is percent
recovery).
LR = Low response.
ND = Not determined.
X = Greater than 70 percent recovery by this technique.
1.2 Table 1 lists the method detection limit for the target analytes in
water. Table 2 lists the estimated quantitation limit (EQL) for all matrices.
2.0 SUMMARY OF METHOD
2.1 Method 8040 provides gas chromatographic conditions for the detection
of phenolic compounds. Prior to analysis, samples must be extracted using
appropriate techniques (see Chapter Two for guidance). Both neat and diluted
organic liquids (Method 3580, Waste Dilution) may be analyzed by direct
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injection. A 2 to 5 /iL sample is injected into a gas chromatograph using the
solvent flush technique, and compounds in the GC effluent are detected by a flame
ionization detector (FID).
2.2 Method 8040 also provides for the preparation of pentafluorobenzyl-
bromide (PFB) derivatives, with additional cleanup procedures for electron
capture gas chromatography. This is to lower the detection limits of some
phenols and to aid the analyst in the elimination of interferences.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All of these materials must be demonstrated to be free
from interferences, under the conditions of the analysis, by analyzing reagent
blanks. Specific selection of reagents and purification of solvents by
distillation in all-glass systems may be required.
3.3 Interferences coextracted from samples will vary considerably from
source to source, depending upon the waste being sampled. Although general
cleanup techniques are recommended as part of this method, unique samples may
require additional cleanup.
3.4 The decomposition of some analytes under basic extraction conditions
has been demonstrated. Specifically, phenols may react to form tannates. These
reactions increase with increasing pH, and are decreased by the shorter reaction
times available in Method 3510.
3.5 The flame ionization detector (FID) is very susceptible to false
positives caused by the presence of hydrocarbons commonly found in samples from
waste sites. The problem may be minimized by applying acid-base cleanup (Method
3650) and/or alumina column chromatography (Method 3611) prior to GC/FID analysis
or using the derivatization technique and analyzing by GC/electron capture
detector. Initial site investigation should always be performed utilizing GC/MS
analysis to characterize the site and determine the feasibility of utilizing
Method 8040 with a GC/FID.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas Chromatograph - Analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detectors, column supplies, recorder, gases, and
syringes. A data system for measuring peak areas and/or peak heights is
recommended.
4.1.2 Columns
4.1.2.1 Column for underivatized phenols - 1.8 m x 2.0 mm
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ID glass column packed with 1% SP-1240DA on Supelcoport 80/100 mesh,
or equivalent.
4.1.2.2 Column for derivatized phenols - 1.8 m x 2 mm ID
glass column packed with 5% OV-17 on Chromosorb W-AW-DMCS 80/100
mesh, or equivalent.
4.1.3 Detectors - Flame ionization (FID) and electron capture (ECD).
4.2 Reaction vial - 20 ml, with Teflon lined screw-cap or crimp top.
4.3 Volumetric flask, Class A - Appropriate sizes with ground-glass
stoppers.
4.4 Kuderna-Danish (K-D) apparatus
4.4.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025 or
equivalent). Ground-glass stopper is used to prevent evaporation of
extracts.
4.4.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps or
equivalent.
4.4.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.4.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.4.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.5 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.6 Water bath - Heated, with concentric ring cover, capable of
temperature control (+ 5°C). The bath should be used in a hood.
4.7 Microsyringe - 10 juL-
4.8 Syringe - 5 ml.
4.9 Balance - analytical, 0.0001 g.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
8040A - 3 Revision 1
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5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Hexane, CH3(CH2)4CH3 - Pesticide quality or equivalent.
5.4 2-Propanol, (CH3)2CHOH - Pesticide quality or equivalent.
5.5 Toluene, C6H5CH3 - Pesticide quality or equivalent.
5.6 Derivatization reagent - Add 1 ml pentafluorobenzyl bromide and 1 g
18-crown-6-ether to a 50 ml volumetric flask and dilute to volume with
2-propanol. Prepare fresh weekly. This operation should be carried out in a
hood. Store at 4°C and protect from light.
5.6.1 Pentafluorobenzyl bromide (alpha-Bromopentafluorotoluene),
C6F5CH2Br. 97% minimum purity.
NOTE: This chemical is a lachrymator.
5.6.2 18-crown-6-ether (1,4,7,10,13,16-Hexaoxacyclooctadecane) -
98% minimum purity.
NOTE: This chemical is highly toxic.
5.7 Potassium carbonate (Powdered), K2C03.
5.8 Stock standard solutions
5.8.1 Prepare stock standard solution at a concentration of
1000 mg/L by dissolving 0.0100 g of assayed reference material in
2-propanol and diluting to volume in a 10 ml volumetric flask. Larger
volumes can be used at the convenience of the analyst. When compound
purity is assayed to be 96% or greater, the weight can be used without
correction to calculate the concentration of the stock standard.
Commercially prepared stock standards can be used at any concentration if
they are certified by the manufacturer or by an independent source.
5.8.2 Transfer the stock standard solutions into bottles with Teflon
lined screw-caps or crimp tops. Store at 4°C and protect from light.
Stock standards should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards
from them.
5.8.3 Stock standard solutions must be replaced after one year, or
sooner if comparison with check standards indicates a problem.
5.9 Calibration standards - Prepare calibration standards at a minimum
of five concentrations through dilution of the stock standards with 2-propanol.
One of the concentrations should be at a concentration near, but above, the
method detection limit. The remaining concentrations should correspond to the
expected range of concentrations found in real samples or should define the
working range of the GC. Calibration solutions must be replaced after six
months, or sooner, if comparison with check standards indicates a problem.
8040A - 4 Revision 1
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5.10 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.10.1 Prepare calibration standards at a minimum of five
concentrations for each analyte as described in Section 5.9.
5.10.2 To each calibration standard, add a known constant
amount of one or more internal standards, and dilute to volume with 2-
propanol.
5.10.3 Analyze each calibration standard according to Section
7.0.
5.11 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (if necessary), and analytical system and the
effectiveness of the method in dealing with each sample matrix by spiking each
sample, standard, and organic-free reagent water blank with phenolic surrogates
(e.g. 2-fluorophenol and 2,4,6-tribromophenol) recommended to encompass the range
of the temperature program used in this method. Method 3500 details instructions
on the preparation of acid surrogates. Deuterated analogs of analytes should not
be used as surrogates for gas chromatographic analysis due to coelution problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1. Extracts must be stored under refrigeration and analyzed within 40
days of extraction.
7.0 PROCEDURE
7.1 Extraction
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a pH of
less than or equal to 2 with methylene chloride, using either Method 3510
or 3520. Solid samples are extracted using either Method 3540 or 3550,
and non-aqueous samples using Method 3580. Extracts obtained from
application of either Method 3540 or 3550 should undergo Acid-Base
Partition Cleanup, using Method 3650.
7.1.2 Prior to gas chromatographic analysis, the extraction solvent
must be exchanged to 2-propanol. The exchange is performed as follows:
7.1.2.1 Following concentration of the extract to 1 mL
using the macro-Snyder column, allow the apparatus to cool and drain
for at least 10 minutes.
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7.1.2.2 Remove the micro-Snyder column and rinse its lower
joint into the concentrator tube with a minimum amount of 2-
propanol. Adjust the extract volume to 1.0 ml. Stopper the
concentrator tube and store refrigerated at 4°C if further
processing will not be performed immediately. If the extract will
be stored longer than two days, it should be transferred to a vial
with a Teflon lined screw-cap or crimp top. If the extract requires
no further derivatization or cleanup, proceed with gas
chromatographic analysis.
7.2 Gas chromatographic conditions (Recommended)
7.2.1 Column for underivatized phenols -
Carrier gas (N2) flow rate: 30 mL/min
Initial temperature: 80°C
Temperature program: 80°C to 150°C at 8°C/min
Final Temperature: 150°C, hold until all compounds have
eluted.
7.2.2 Column for derivatized phenols -
Carrier gas (5% methane/95% argon)
flow rate: 30 mL/min
Initial temperature: 200°C
Temperature program: isothermal, hold until all
compounds have eluted.
7.3 Calibration - Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point on
the calibration curve.
7.3.1 The procedure for internal or external calibration may be used
for the underivatized phenols. Refer to Method 8000 for a description of
each of these procedures. If derivatization of the phenols is required,
the method of external calibration should be used by injecting five or
more concentrations of calibration standards that have also undergone
derivatization and cleanup prior to instrument calibration.
7.4 Gas chromatographic analysis
7.4.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 /nL of internal standard to the sample prior to
injection.
7.4.2 Phenols are to be determined on a gas chromatograph equipped
with a flame ionization detector according to the conditions listed for
the 1% SP-1240DA column (Section 7.2.1). Table 1 summarizes estimated
retention times and sensitivities that should be achieved by this method
for clean water samples. Estimated quantitation limits for other
matrices are list in Table 2.
7.4.3 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
8040A - 6 Revision 1
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identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
7.4.4 An example of a GC/FID chromatogram for certain phenols is
shown in Figure 1. Other packed or capillary (open-tubular) columns,
chromatographic conditions, or detectors may be used if the requirements
of Section 8.2 are met.
7.4.5 Record the sample volume injected and the resulting peak sizes
(in area units or peak heights).
7.4.6 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes. See Method 8000 for calculation equations.
7.4.7 If peak detection using the SP-1240DA column with the flame
ionization detector is prevented by interferences, PFB derivatives of the
phenols should be analyzed on a gas chromatograph equipped with an
electron capture detector according to the conditions listed for the 5%
OV-17 column (Section 7.2.2). The derivatization and cleanup procedure
is outlined in Sections 7.5 through 7.6. Table 3 summarizes estimated
retention times for derivatives of some phenols using the conditions of
this method.
7.4.8 Figure 2 shows a GC/ECO chromatogram of PFB derivatives of
certain phenols.
7.4.9 Record the sample volume injected and the resulting peak sizes
(in area units or peak heights).
7.4.10 Determine the identity and quantity of each component
peak in the sample chromatogram which corresponds to the compounds used
for calibration purposes. The method of external calibration should be
used (see Method 8000 for guidance). The concentration of the individual
compounds in the sample is calculated as follows:
Concentration
where:
A = Mass of underivatized phenol represented by area of peak
in sample chromatogram, determined from calibration
curve (see Method 8000), ng.
Vt = Total amount of column eluate or combined fractions from
which V, was taken, juL.
B = Total volume of hexane added in Section 7.5.5, ml.
D = Total volume of 2-propanol extract prior to
derivatization, ml.
8040A - 7 Revision 1
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V, = Volume injected, nl.
X = Volume of water extracted, mL, or weight of nonaqueous
sample extracted, g, from Section 7.1. Either the dry
or wet weight of the nonaqueous sample may be used,
depending upon the specific application of the data.
C * Volume of hexane sample solution added to cleanup column
(Method 3630), mL.
E = Volume of 2-propanol extract carried through
derivatization in Section 7.5.1, ml.
7.5 Derivatization - If interferences prevent measurement of peak area
during analysis of the extract by flame ionization gas chromatography, the
phenols must be derivatized and analyzed by electron capture gas chromatography.
7.5.1 Pipet a 1.0 ml aliquot of the 2-propanol stock standard
solution or of the sample extract into a glass reaction vial. Add 1.0 ml
derivatization reagent (Section 5.3). This amount of reagent is
sufficient to derivatize a solution whose total phenolic content does not
exceed 300 mg/L.
7.5.2 Add approximately 0.003 g of potassium carbonate to the
solution and shake gently.
7.5.3 Cap the mixture and heat it for 4 hours at 80°C in a hot water
bath.
7.5.4 Remove the solution from the hot water bath and allow it to
cool.
7.5.5 Add 10 ml hexane to the reaction vial and shake vigorously for
1 minute. Add 3.0 mL organic-free reagent water to the reaction vial and
shake for 2 minutes.
7.5.6 Decant the organic layer into a concentrator tube and cap with
a glass stopper. Proceed with cleanup procedure.
7.6 Cleanup
7.6.1 Cleanup of the derivatized extracts takes place using Method
3630 (Silica Gel Cleanup), in which specific instructions for cleanup of
the derivatized phenols appear.
7.6.2 Following column cleanup, analyze the samples using GC/ECD, as
described starting in Section 7.4.7.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
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the extraction method used. If extract cleanup was performed, follow the QC in
Method 3600 and in the specific cleanup method.
8.2 Procedures to check the GC system operation are found in Method 8000,
Section 8.6.
8.2.1 The quality control check sample concentrate (Method 8000,
Section 8.6) should contain each analyte of interest at a concentration
of 100 mg/L in 2-propanol.
8.2.2 Table 4 indicates the calibration and QC acceptance criteria
for this method. Table 5 gives method accuracy and precision as
functions of concentration for the analytes. The contents of both tables
should be used to evaluate a laboratory's ability to perform and generate
acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000, Section 8.10).
8.3.1 If recovery is not within limits, the following is required.
• Check to be sure that there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are a
problem or flag the data as "estimated concentration."
9.0 METHOD PERFORMANCE
9.1 The method was tested by 20 laboratories using organic-free reagent
water, drinking water, surface water, and three industrial wastewaters spiked at
six concentrations over the range 12 to 450 /zg/L. Single operator precision,
overall precision, and method accuracy were found to be directly related to the
concentration of the analyte and essentially independent of the sample matrix.
Linear equations to describe these relationships for a flame ionization detector
are presented in Table 5.
9.2 The accuracy and precision obtained will be affected by the sample
matrix, sample-preparation technique, and calibration procedures used.
10.0 REFERENCES
1. Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 3 - Chlorinated Hydrocarbons and
Category 8 - Phenols. Report for EPA Contract 68-03-2625 (in
preparation).
8040A - 9 Revision 1
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U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for
the Analysis of Pollutants Under the Clean Water Act; Final Rule and
Interim Final Rule and Proposed Rule," October 26, 1984.
"Determination of Phenols in Industrial and Municipal Wastewaters,"
Report for EPA Contract 68-03-2625 (in preparation).
"EPA Method Validation Study Test Method 604 (Phenols)," Report for EPA
Contract 68-03-2625 (in preparation).
Kawahara, F.K. "Microdetermination of Derivatives of Phenols and
Mercaptans by Means of Electron Capture Gas Chromatography," Analytical
Chemistry, 40, 1009, 1968.
Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
8040A - 10 Revision 1
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TABLE 1.
FLAME IONIZATION GAS CHROMATOGRAPHY OF PHENOLS8
Method
Retention time Detection
Analyte (minutes) limit
2-sec-Butyl-4,6-dinitrophenol (DNBP)
4-Chloro-3-methylphenol 7.50 0.36
2-Chlorophenol 1.70 0.31
Cresols (methyl phenols)
2-Cyclohexyl-4,6-dinitrophenol
2,4-Dichlorophenol 4.30 0.39
2,6-Dichlorophenol
2,4-Dimethylphenol 4.03 0.32
2,4-Dinitrophenol 10.00 13.0
2-Methyl-4,6-dinitrophenol 10.24 16.0
2-Nitrophenol 2.00 0.45
4-Nitrophenol 24.25 2.8
Pentachlorophenol 12.42 7.4
Phenol 3.01 0.14
Tetrachlorophenols
Trichlorophenols
2,4,6-Trichlorophenol 6.05 0.64
8 - 1% SP-1240DA on Supelcoport 80/100 mesh column.
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION
LIMITS (EQLs) FOR VARIOUS MATRICES8
Matrix Factor6
Ground water 10
Low-concentration soil by sonication with GPC cleanup 670
High-concentration soil and sludges by sonication 10,000
Non-water miscible waste 100,000
a Sample EQLs are highly matrix-dependent. The EQLs listed herein are provided
'for guidance and may not always be achievable.
b EQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For non-
aqueous samples, the factor is on a wet-weight basis.
8040A - 11 Revision 1
July 1992
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TABLE 3.
ELECTRON CAPTURE GAS CHROMATOGRAPHY OF PFB DERIVATIVES8
Parent compound
4-Ch1oro-2-methyl phenol
2-Chlorophenol
2,4-Dichlorophenol
2,4-Dimethylphenol
2,4-Dinitrophenol
2-Methyl -4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
Retention
time
(min)
4.8
3.3
5.8
2.9
46.9
36.6
9.1
14.0
28.8
1.8
7.0
Method
detection
limit (M9/L)
1.8
0.58
0.68
0.63
0.77
0.70
0.59
2.2
0.58
- 5% OV-17 on Chromosorb W-AW-DMCS 80/100 mesh column.
8040A - 12 Revision 1
July 1992
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TABLE 4.
QC ACCEPTANCE CRITERIA8
Analyte
4-Chloro-3-methyl phenol
2-Chlorophenol
2,4-Dichlorophenol
2,4-Dimethylphenol
4, 6-Dinitro-2-methyl phenol
2,4-Dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
Test
cone.
(M9/L)
100
100
100
100
100
100
100
100
100
100
100
Limit
for s
(M9/L)
16.6
27.0
25.1
33.3
25.0
36.0
22.5
19.0
32.4
14.1
16.6
Range
for x
(M9/L)
56.7-113.4
54.1-110.2
59.7-103.3
50.4-100.0
42.4-123.6
31.7-125.1
56.6-103.8
22.7-100.0
56.7-113.5
32.4-100.0
60.8-110.4
Recovery
Range
(%)
99-122
38-126
44-119
24-118
30-136
12-145
43-117
13-110
36-134
23-108
53-119
s
x
a
= Standard deviation of four recovery measurements, in
= Average recovery for four recovery measurements, in jug/L.
Criteria from 40 CFR Part 136 for Method 604. These criteria are based
directly upon the method performance data in Table 5. Where necessary, the
limits for recovery have been broadened to assure applicability of the
limits to concentrations below those used to develop Table 5.
8040A - 13
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TABLE 5.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Analyte
4-Chloro-3-methyl phenol
2-Chlorophenol
2,4-Dichlorophenol
2, 4-Dimethyl phenol
4, 6-Dinitro-2-methyl phenol
2,4-Dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
Accuracy, as
recovery, x'
(M9/L)
0.87C-1.97
0.83C-0.84
0.81C+0.48
0.62C-1.64
0.84C-1.01
0.80C-1.58
0.81C-0.76
0.46C+0.18
0.83C+2.07
0.43C+0.11
0.86C-0.40
Single analyst
precision, s '
(M9/D
O.llx-0.21
O.lSx+0.20
0.17X-0.02
O.SOx-0.89
0.15X+1.25
0.27X-1.15
O.lBx+0.44
0.17X+2.43
0.22X-0.58
0.20X-0.88
O.lOx+0.53
Overal 1
precision,
S' (ng/D
0.16X+1.41
0.21X+0.75
O.lSx+0.62
0.25x+0.48
0.19X+5.85
0.29X+4.51
0.14x+3.84
0.19X+4.79
0.23X+0.57
O.Ux+0.77
0.13x+2.40
X'
S'
C
x
Expected recovery for one or more measurements of a sample
containing a concentration of C, in /xg/L.
Expected single analyst standard deviation of measurements at an
average concentration of x, in M9/L-
Expected interlaboratory standard deviation of measurements at an
average concentration found of x, in M9/L*
True value for the concentration, in /xg/L.
Average recovery found for measurements of samples containing a
concentration of C, in M9/L.
8From 40 CFR Part 136 for Method 604.
8040A - 14
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July 1992
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Figure 1
Gas Chromatogram of Phenols
Column: 1% SM2400A on Soptieooort
Program: 80°C 0 Minute* 8°/Minuta to 1SO°C
Dttactor: Flama lonisation
8 12 16 20
RETENTION TIME (MINUTES)
24
21
8040A - 15
Revision 1
July 1992
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Figure 2
Gas Chromatogram of PFB Derivatives of Phenols
Column: 5% OV-17 on OtremoMrb W-AW
200°C
Electron Cteturt
A_
12 16 20 24
RETENTION TIME (MINUTES)
21 32
8040A - 16
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July 1992
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METHOD 8040A
PHENOLS BY GAS CHROMATOGRAPHY
7 1 1 Choose
appropriate
ex t raction
method (refer
to Chapter 2)
7 1 2
Exchange
OKt raction
solvent to
2 -propanol
7 2 Set gas
chromatography
conditions
7 3 Refer to
Method 8000
for proper
cal ibra to. on
techniques
3 1 la N.
atization x. Yea
phenols j— — — +
quired' /
731 Inject at
least 5
concentrations
of ca 1 ibra tion
s tandar ds
7 4 Perform
CC analysis
(see Method
8000)
7 1 analyze
using GC/FID
8040A - 17
Revision 1
July 1992
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METHOD 8040A
(Continued)
7 5 Prepare
derivatives
749 Record
sample volume
injected and
peak sizes
7 k Cleanup
using Method
3630
7 4 10
Identitify and
quantitate each
component peak
7 4 7 Analyze
PFB
deriva tives
using CC/ECD
7 4 10
Calculate
concentration
Stop
8040A - 18
Revision 1
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METHOD 8060
PHTHALATE ESTERS
1.0 SCOPE AND APPLICATION
1.1 Method 8060 Is used to determine the concentration of various
phthalate esters. Table 1 indicates compounds that may be determined by this
method and lists the method detection limit for each compound in reagent
water. Table 2 lists the practical quantisation limit (PQL) for other
matrices.
2.0 SUMMARY OF METHOD
2.1 Method 8060 provides gas chromatographic conditions for the
detection of ppb levels of phthalate esters. Prior to use of this method,
appropriate sample extraction techniques must be used. Both neat and diluted
organic liquids (Method 3580, Waste Dilution) may be analyzed by direct
injection. A 2- to 5-uL aliquot of the extract is injected into a gas
chromatograph (GC) using the solvent flush technique, and compounds in the GC
effluent are detected by an electron capture detector (ECD) or a flame
ionization detector (FID). Ground water samples should be determined by ECD.
2.2 The method provides a second gas chromatographic column that may be
helpful in resolving the analytes from interferences that may occur and for
analyte confirmation.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Phthalate esters contaminate many types of products commonly found
in the laboratory. The analyst must demonstrate that no phthalate residues
contaminate the sample or solvent extract under the conditions of analysis.
Plastics, in particular, must be avoided because phthalates are commonly used
as plasticizers and are easily extracted from plastic materials. Serious
phthalate contamination may result at any time if consistent quality control
is not practiced.
3.3 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing
misinterpretation of gas chromatograms. All these materials must be
demonstrated to be free from interferences, under the conditions of the
analysis, by analyzing method blanks. Specific selection of reagents and
purification of solvents by distillation in all-glass systems may be required.
3.4 Interferences coextracted from samples will vary considerably from
source to source, depending upon the waste being sampled. Although general
cleanup techniques are recommended as part of this method, unique samples may
require additional cleanup.
8060 - 1
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Date September 1986
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TABLE 1. RETENTION TIME AND DETECTION LIMIT INFORMATION FOR PHTHALATE ESTERS
Retention time (mln) Method detection
limit (ug/L)
Compound Col. la Col. 2b ECD FID
Benzyl butyl phthalate
B1 s (2-ethyl hexyl ) phthal ate
Di-n-butyl phthalate
D1 ethyl phthalate
Dimethyl phthalate
Di-n-octyl phthalate
*6.94
*8.92
8.65
2.82
2.03
*16.2
**5.11
**10.5
3.50
1.27
0.95
**8.0
0.34
2.0
0.36
0.49
0.29
3.0
15
20
14
31
19
31
aColumn 1: Supelcoport 100/120 mesh coated with 1.5% SP-2250/1.95% SP-
2401 packed in a 180-cm x 4-mm I.D. glass column with carrier gas at 60
mL/min flow rate. Column temperature is 180*C, except where * indicates
220*C. Under these conditions the retention time of Aldrin is 5.49 min
at 180*C and 1.84 min at 220*C.
bColumn 2: Supelcoport 100/120 mesh with 3% OV-1 in a 180-cm x 4-mm I.D.
glass column with carrier gas at 60 mL/min flow rate. Column temperature
is 200*C, except where ** indicates 220*C. Under these conditions the
retention time of Aldrin is 3.18 min at 200*C and 1.46 min at 220*C.
TABLE 2. DETERMINATION OF PRACTICAL QUANTITATION LIMITS (PQL) FOR VARIOUS
MATRICESa
Matrix Factorb
Ground water 10
Low-level soil by sonication with GPC cleanup 670
High-level soil and sludges by sonication 10,000
Non-water miscible waste 100,000
aSample PQLs are highly matrix-dependent. The PQLs listed herein are
provided for guidance and may not always be achievable.
bPQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For non-
aqueous samples, the factor is on a wet-weight basis.
8060 - 2
Revision
Date September 1986
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4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph;
4.1.1 Gas chromatograph: Analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detectors, column supplies, recorder, gases, and
syringes. A data system for measuring peak areas and/or peak heights is
recommended.
4.1.2 Columns:
4.1.2.1 Column 1: 1.8-m x 4-mm I.D. glass column packed with
1.5% SP-2250/1.95% SP-2401 on Supelcoport 100/120 mesh or
equivalent.
4.1.2.2 Column 2: 1.8-m x 4-mm I.D. glass column packed with
3% OV-1 on Supelcoport 100/120 mesh or equivalent.
4.1.3 Detectors: Flame ionization (FID) or electron capture (ECD).
4.2 Volumetric flask; 10-, 50-, and 100-mL, ground-glass stopper.
4.3 Kuderna-Danish (K-D) apparatus:
4.3.1 Concentrator tube: 10-mL, graduated (Kontes K-570050-1025 or
equivalent). Ground-glass stopper is used to prevent evaporation of
extracts
4.3.2 Evaporation flask: 500-mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs.
4.3.3 Snyder column: Three-ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column: Two-ball micro (Kontes K-569001-0219 or
equivalent).
4.4 Boiling chips; Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.5 Water bath; Heated, with concentric ring cover, capable of
temperature control (+5*C). The bath should be used in a hood.
4.6 Mlcrosyringe; 10-uL.
4.7 Syringe; 5-mL.
4.8 Vials; Glass, 2- and 20-mL capacity with Teflon-lined screw cap.
8060 - 3
Revision
Date September 1986
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5.0 REAGENTS
5.1 Solvents: Hexane, acetone, isooctane (2,2,4-trimethylpentane)
(pesticide quality or equivalent).
5.2 Stock standard solutions;
5.2.1 Prepare stock standard solutions at a concentration of 1.00
ug/uL by dissolving 0.0100 g of assayed reference material in isooctane
and diluting to volume in a 10-mL volumetric flask. Larger volumes can
be used at the convenience of the analyst. When compound purity is
assayed to be 96% or greater, the weight can be used without correction
to calculate the concentration of the stock standard. Commercially
prepared stock standards can be used at any concentration if they are
certified by the manufacturer or by an independent source.
5.2.2 Transfer the stock standard solutions into Teflon-sealed
screw-cap bottles. Store at 4*C and protect from light. Stock standards
should be checked frequently for signs of degradation or evaporation,
especially just prior to preparing calibration standards from them.
5.2.3 Stock standard solutions must be replaced after one year, or
sooner if comparison with check standards indicates a problem.
5.3 Calibration standards: Calibration standards at a minimum of five
concentrationlevelsshouldbe prepared through dilution of the stock
standards with isooctane. One of the concentration levels should be at a
concentration near, but above, the method detection limit. The remaining
concentration levels should correspond to the expected range of concentrations
found in real samples or should define the working range of the GC.
Calibration solutions must be replaced after six months, or sooner if
comparison with check standards indicates a problem.
5.4 Internal standards (if internal standard calibration is used); To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.4.1 Prepare calibration standards at a minimum of five
concentration levels for each analyte of interest as described in
Paragraph 5.3.
5.4.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with isooctane.
5.4.3 Analyze each calibration standard according to Section 7.0.
5.5 Surrogate standards; The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system and the effec-
tiveness of the method in dealing with each sample matrix by spiking each
8060 - 4
Revision 0
Date September 1986
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sample, standard, and reagent water blank with one or two surrogates (e.g.,
phthalates that are not expected to be In the sample) recommended to encompass
the range of the temperature program used in this method. Method 3500,
Section 5.3.1.1, details instructions on the preparation of base/neutral
surrogates. Deuterated analogs of analytes should not be used as surrogates
for gas chromatographic analysis due to coelution problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1. Extracts must be stored under refrigeration and analyzed within
40 days of extraction.
7.0 PROCEDURE
7.1 Extraction:
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a
neutral, or as is, pH with methylene chloride, using either Method 3510
or 3520. Solid samples are extracted using either Method 3540 or 3550.
7.1.2 Prior to gas chromatographic analysis, the extraction solvent
must be exchanged to hexane. The exchange is performed during the K-D
procedures listed in all of the extraction methods. The exchange is
performed as follows.
7.1.2.1 Following K-D of the methylene chloride extract to
1 mL using the macro-Snyder column, allow the apparatus to cool and
drain for at least 10 min.
7.1.2.2 Momentarily remove the Snyder column, add 50 mL of
hexane, a new boiling chip, and reattach the macro-Snyder column.
Concentrate the extract using 1 mL of hexane to prewet the Snyder
column. Place the K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete concentration in 5-10 min. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 1 mL, remove the K-D apparatus and allow it to drain and
cool for at least 10 min. The extract will be handled differently
at this point, depending on whether or not cleanup is needed. If
cleanup is not required, proceed to Paragraph 7.1.2.3. If cleanup
is needed, proceed to Paragraph 7.1.2.4.
7.1.2.3 If cleanup of the extract is not required, remove the
Snyder column and rinse the flask and its lower joint into the
concentrator tube with 1-2 mL of hexane. A 5-mL syringe is
recommended for this operation. Adjust the extract volume to
8060 - 5
Revision 0
Date September 1986
-------
10.0 ml. Stopper the concentrator tube and store refrigerated at
4*C if further processing will not be performed immediately. If the
extract will be stored longer than two days, it should be
transferred to a Teflon-sealed screw-cap vial. Proceed with gas
chromatographic analysis.
7.1.2.4 If cleanup of the extract is required, remove the
Snyder column and rinse the flask and its lower joint into the
concentrator tube with a minimum amount of hexane. A 5-mL syringe
is recommended for this operation. Add a clean boiling chip to the
concentrator tube and attach a two-ball micro-Snyder column. Prewet
the column by adding about 0.5 mL of hexane to the top. Place the
micro-K-D apparatus on the water bath (80*C) so that the
concentrator tube is partially immersed 1n the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete concentration in 5-10 min. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 0.5 ml, remove the K-D apparatus and allow it to drain and
cool for at least 10 min.
7.1.2.5 Remove the micro-Snyder column and rinse the flask and
its lower joint into the concentrator tube with 0.2 ml of hexane.
Adjust the extract volume to 2.0 ml and proceed with either Method
3610 or 3620.
7.2 Gas chromatpgraphy conditions (Recommended); The analysis for
phthalate esters may Be conducted using either a flame ionization or an
electron capture detector. The ECD may, however, provide substantially better
sensitivity.
7.2.1 Column 1: Set 5% methane/95% argon carrier gas flow at 60
mL/min flow rate. Set column temperature at 180*C isothermal.
7.2.2 Column 2: Set 5% methane/95% argon carrier gas flow at 60
mL/min flow rate. Set column temperature at 200*C isothermal.
7.3 Calibration; Refer to Method 8000 for proper calibration
techniques"! Use Table 1 and especially Table 2 for guidance on selecting the
lowest point on the calibration curve.
7.3.1 The procedure for internal or external
used. Refer to Method 8000 for a description
procedures.
calibration may be
of each of these
7.3.2 If cleanup is performed on the samples, the analyst should
process a series of standards through the cleanup procedure and then
analyze the samples by GC, This will confirm elution patterns and the
absence of interferents from the reagents.
8060 - 6
Revision 0
Date September 1986
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7.4 Gas chromatographic analysis;
7.4.1 Refer to Method 8000. If the Internal standard calibration
technique 1s used, add 10 uL of Internal standard to the sample prior to
injection.
7.4.2 Follow Section 7.6 1n Method 8000 for Instructions on the
analysis sequence, appropriate dilutions, establishing dally retention
time windows, and Identification criteria. Include a mid-level standard
after each group of 10 samples 1n the analysis sequence.
7.4.3 Examples of GC/ECD chromatograms for phthalate esters are
shown in Figures 1 and 2.
7.4.4 Record the sample volume Injected and the resulting peak
sizes (in area units or peak heights).
7.4.5 Using either the internal or external calibration procedure
(Method 8000), determine the Identity and quantity of each analyte peak
1n the sample chromatogram. See Section 7.8 of Method 8000 for
calculation equations.
7.4.6 If peak detection and Identification are prevented due to
interferences, the hexane extract may undergo cleanup using either Method
3610 or 3620.
7.5 Cleanup;
7.5.1 Proceed with either Method 3610 or 3620, using the 2-mL
hexane extracts obtained from Paragraph 7.1.2.5.
7.5.2 Following cleanup, the extracts should be analyzed by GC, as
described in the previous paragraphs and in Method 8000.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction 1s covered 1n Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the
QC in Method 3600 and 1n the specific cleanup method.
8.2 Procedures to check the GC system operation are found in Method
8000, Section 8.6.
8.2.1 The quality control check sample concentrate (Method 8000,
Section 8.6) should contain each analyte of interest at the following
concentrations in acetone: butyl benzyl phthalate, 10 ug/mL; bis(2-
ethylhexyl) phthalate, 50 ug/mL; di-n-octyl phthalate, 50 ug/mL; and any
other phthalate, 25 ug/mL.
8060 - 7
Revision
Date September 1986
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Column: 1.5%SP-2250+
1.95% SP-2401 on Supticoport
Ttmperaturt: 180°C
Dttictor: Electron Capture
j t
J t
0 2 4 6 8 10 12
RETENTION TIME (MINUTES)
Figure 1. Gas chromatogram of phthalates (example 1).
8060 - 8
Revision 0
Date September 1986
-------
Column: 1.5% SP-2250+
1.95% SP-2401 on Supelcoport
Temperature: 18(X>C
Detector: Electron Capture
4 8 12 16
RETENTION TIME (MINUTES)
18
Figure 2. Gas chromatogram of phthalates (example 2).
8060 - 9
Revision 0
Date September 1986
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8.2.2 Table 3 indicates the calibration and QC acceptance criteria
for this method. Table 4 gives method accuracy and precision as
functions of concentration for the analytes of interest. The contents of
both Tables should be used to evaluate a laboratory's ability to perform
and generate acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000, Section 8.10).
8.3.1 If recovery is not within limits, the following is required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are
a problem or flag the data as "estimated concentration."
9.0 METHOD PERFORMANCE
9.1 The method was tested by 16 laboratories using reagent water,
drinking water, surface water, and three industrial wastewaters spiked at six
concentrations over the range 0.7 to 106 ug/L. Single operator precision,
overall precision, and method accuracy were found to be directly related to
the concentration of the analyte and essentially independent of the sample
matrix. Linear equations to describe these relationships for a flame
ionization detector are presented in Table 4.
9.2 The accuracy and precision obtained will be determined by the sample
matrix, sample-preparation technique, and calibration procedures used.
10.0 REFERENCES
1. Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 1 - Phthalates. Report for EPA Contract
68-03-2606 (in preparation).
2. "Determination of Phthalates in Industrial and Municipal Wastewaters,"
EPA-600/4-81-063, U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory, Cincinnati, Ohio 45268, October 1981.
3. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
8060 - 10
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Date September 1986
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4. "EPA Method Validation Study 16, Method 606 (Phthalate Esters)," Report
for EPA Contract 68-03-2606 (in preparation).
5. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim Final
Rule and Proposed Rule," October 26, 1984.
6. Provost, L.P. and R.S. Elder, "Interpretation of Percent Recovery Data,"
American Laboratory, 15, pp. 58-63, 1983.
8060 - 11
Revision
Date September 1986
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TABLE 3. QC ACCEPTANCE CRITERIA3
Parameter
Bi s (2-ethy 1 hexy 1 ) phthal ate
Butyl benzyl phthal ate
Di-n-butyl phthal ate
Di ethyl phthal ate
Dimethyl phthal ate
Di-n-octyl phthal ate
Test
cone.
(ug/L)
50
10
25
25
25
50
Limit
for s
(ug/L)
38.4
4.2
8.9
9.0
9.5
13.4
Range
for 7
(ug/L)
1.2-55.9
5.7-11.0
10.3-29.6
1.9-33.4
1.3-35.5
D-50.0
Range
P, PS
(%)
D-158
30-136
23-136
D-149
D-156
D-114
s = Standard deviation of four recovery measurements, in ug/L.
7 = Average recovery for four recovery measurements, in ug/L.
P, Ps = Percent recovery measured.
D = Detected; result must be greater than zero.
Criteria from 40 CFR Part 136 for Method 606. These criteria are based
directly upon the method performance data in Table 4. Where necessary, the
limits for recovery have been broadened to assure applicability of the limits
to concentrations below those used to develop Table 4.
8060 - 12
Revision
Date September 1986
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TABLE 4. METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION3
Parameter
Bis(2-ethylhexyl) phthalate
Butyl benzyl phthalate
Di-n-butyl phthalate
Di ethyl phthalate
Dimethyl phthalate
Di-n-octyl phthalate
Accuracy, as
recovery, x1
(ug/L)
0.53C+2.02
0.82C+0.13
0.79C+0.17
0.70C+0.13
0.73C+0.17
0.35C-0.71
Single analyst
precision, sr'
(ug/L)
0.807-2.56
0.267+0.04
0.237+0.20
0.277+0.05
0.267+0.14
0.387+0.71
Overall
precision,
S1 (ug/L)
0.737-0.17
0.257+0.07
0.297+0.06
0.457+0.11
0.447+0.31
0.627+0.34
x1 = Expected recovery for one or more measurements of a sample
containing a concentration of C, in ug/L.
sr' = Expected single analyst standard deviation of measurements at an
average concentration of 7, in ug/L.
S1 = Expected interlaboratory standard deviation of measurements at an
average concentration found of 7, in ug/L.
C = True value for the concentration, in ug/L.
7 = Average recovery found for measurements of samples containing a
concentration of C, in ug/L.
Criteria from 40 CFR Part 136 for Method 606.
8060 - 13
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METHOD 8O60
PHTHALATE ESTERS
( Start J
7.1.
(see
ll
—> Choose
appropriate
extraction
procedure
Chapter 2)
o
7.4
Perform GC
analysis (see
Method 8000)
7.1.21
Exchange
extract-
ion solvent to
hexane
during micro
K-O procedures
7.2
Set gas
chromatography
conditions
Is IdentlfIce-^S.
tlon C detection
Cleanup
using Method
361O or 362O)
7
1 Refer to
Method BOOO
for proper
calibration
techniques
7.3.2
of
Process
a aeries
standards
through cleanup
procedure:
analyze by GC
8060 - 14
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Date September 1986
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METHOD 8061
PHTHALATE ESTERS BY CAPILLARY GAS CHROMATOGRAPHY
WITH ELECTRON CAPTURE DETECTION (GC/ECD)
1.0 SCOPE AND APPLICATION
1.1 Method 8061 is used to determine the identities and concentrations
of various phthalate esters in liquid, solid and sludge matrices. The following
compounds can be determined by this method:
Compound Name CAS No."
Benzyl benzoate (I.S.) 120-51-4
Bis(2-ethylhexyl) phthalate 117-81-7
Butyl benzyl phthalate 85-68-7
Di-n-butyl phthalate 84-74-2
Diethyl phthalate 84-66-2
Dimethyl phthalate 131-11-3
Di-n-octyl phthalate 117-84-0
a Chemical Abstract Services Registry Number.
1.2 Table 1 lists the method detection limits (MDL) for the target
analytes in a water matrix. The MDLs for the components of a specific sample may
differ from those listed in Table 1 because MDLs depend on the nature of
interferences in the sample matrix. Table 2 lists the estimated quantisation
limits (EQL) for other matrices.
1.3 When this method is used to analyze for any or all of the target
analytes, compound identification should be supported by at least one additional
qualitative technique. This method describes conditions for parallel column,
dual electron capture detector analysis which fulfills the above requirement.
Retention time information obtained on two megabore fused-silica open tubular
columns is given in Table 1. Alternatively, gas chromatography/mass spectrometry
could be used for compound confirmation.
1.4 The following compounds, bis(2-n-butoxyethyl) phthalate, bis(2-
ethoxyethyl) phthalate, bis(2-methoxyethyl) phthalate, bis(4-methyl-2-pentyl)
phthalate, diamyl phthalate, dicyclohexyl phthalate, dihexyl phthalate,
diisobutyl phthalate, dinonyl phthalate, and hexyl 2-ethylhexyl phthalate can
also be analyzed by this method and may be used as surrogates.
1.5 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatographs and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
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2.0 SUMMARY OF METHOD
2.1 A measured volume or weight of sample (approximately 1 liter for
liquids, 10 to 30 grams for solids and sludges) is extracted by using the
appropriate sample extraction technique specified in Methods 3510, 3540, 3541,
and 3550. Method 3520 is not recommended for the extraction of aqueous samples
because the longer chain esters (dihexyl phthalate, bis(2-ethylhexyl) phthalate,
di-n-octyl phthalate, and dinonyl phthalate) tend to adsorb to the glassware and
consequently, their extraction recoveries are <40 percent. Aqueous samples are
extracted at a pH of 5 to 7, with methylene chloride, in a separatory funnel
(Method 3510). Alternatively, particulate-free aqueous samples could be filtered
through membrane disks that contain C18-bonded silica. The phthalate esters are
retained by the silica and, later eluted with acetonitrile. Solid samples are
extracted with hexane/acetone (1:1) or methylene chloride/acetone (1:1) in a
Soxhlet extractor (Methods 3540/3541) or with an ultrasonic extractor (Method
3550). After cleanup, the extract is analyzed by gas chromatography with
electron capture detection (GC/ECD).
2.2 The sensitivity of Method 8061 usually depends on the level of
interferences rather than on instrumental limitations. If interferences prevent
detection of the analytes, cleanup of the sample extracts is necessary. Either
Method 3610 or 3620 alone or followed by Method 3660, Sulfur Cleanup, may be used
to eliminate interferences in the analysis. Method 3640, Gel Permeation Cleanup,
is applicable for samples that contain high amounts of lipids and waxes.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Interferences coextracted from the samples will vary considerably
from waste to waste. While general cleanup techniques are referenced or provided
as part of this method, unique samples may require additional cleanup approaches
to achieve desired sensitivities for the target analytes.
3.3 Glassware must be scrupulously clean. All glassware require
treatment in a muffle furnace at 400 °C for 2 to 4 hrs, or thorough rinsing with
pesticide-grade solvent, prior to use. Refer to Chapter 4, Sec. 4.1.4, for
further details regarding the cleaning of glassware. Volumetric glassware should
not be heated in a muffle furnace.
If Soxhlet extractors are baked in the muffle furnace, care must be taken
to ensure that they are dry (breakage may result if any water is left in the
side-arm). Thorough rinsing with hot tap water, followed by deionized water and
acetone is not an adequate decontamination procedure. Even after a Soxhlet
extractor was refluxed with acetone for three days, with daily solvent changes,
the concentrations of bis(2-ethylhexyl) phthalate were as high as 500 ng per
washing. Storage of glassware in the laboratory introduces contamination, even
if the glassware is wrapped in aluminum foil. Therefore, any glassware used in
Method 8061 should be cleaned immediately prior to use.
3.4 Florisil and alumina may be contaminated with phthalate esters and,
therefore, use of these materials in sample cleanup should be employed
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cautiously. If these materials are used, they must be obtained packaged in glass
(plastic packaging will contribute to contamination with phthalate esters).
Washing of these materials prior to use with the solvent(s) used for elution
during extract cleanup was found helpful, however, heating at 320 °C for Florisil
and 210 CC for alumina is recommended. Phthalate esters were detected in
Florisil cartridge method blanks at concentrations ranging from 10 to 460 ng,
with 5 phthalate esters in the 105 to 460 ng range. Complete removal of the
phthalate esters from Florisil cartridges does not seem possible, and it is
therefore desirable to keep the steps involved in sample preparation to a
minimum.
3.5 Paper thimbles and filter paper must be exhaustively washed with the
solvent that will be used in the sample extraction. Soxhlet extraction of paper
thimbles and filter paper for 12 hrs with fresh solvent should be repeated for
a minimum of three times. Method blanks should be obtained before any of the
precleaned thimbles or filter papers are used. Storage of precleaned thimbles
and filter paper in precleaned glass jars covered with aluminum foil is
recommended.
3.6 Glass wool used in any step of sample preparation should be a
specially treated pyrex wool, pesticide grade, and must be baked at 400°C for
4 hrs. immediately prior to use.
3.7 Sodium sulfate must be obtained packaged in glass (plastic packaging
will contribute to contamination with phthalate esters), and must be purified by
heating at 400 °C for 4 hrs. in a shallow tray, or by precleaning with methylene
chloride (Sec. 5.3). To avoid recontamination, the precleaned material must be
stored in glass-stoppered glass bottles, or glass bottles covered with precleaned
aluminum foil. The storage period should not exceed two weeks. To minimize
contamination, extracts should be dried directly in the glassware in which they
are collected by adding small amounts of precleaned sodium sulfate until an
excess of free flowing material is noted.
3.8 The presence of elemental sulfur will result in large peaks which
often mask the region of the compounds eluting before dicyclohexyl phthalate
(Compound No. 14) in the gas chromatograms shown in Figure 1. Method 3660 is
suggested for removal of sulfur.
3.9 Waxes and lipids can be removed by Gel Permeation Chromatography
(Method 3640). Extracts containing high concentrations of lipids are viscous,
and may even solidify at room temperature.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatography
4.1.1 Gas chromatograph, analytical system complete with gas
chromatograph suitable for on-column and split/splitless injections and
all required accessories, including detector, analytical columns,
recorder, gases, and syringes. A data system for measuring peak heights
and/or peak areas is recommended.
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4.1.1.1 Eight inch injection tee (Supelco, Inc., Catalog
No. 2-3665, or equivalent) or glass Y splitter for megabore columns
(J&W Scientific, -"press-fit", Catalog No. 705-0733, or equivalent).
4.1.2 Columns
4.1.2.1 Column 1, 30 m x 0.53 mm ID, 5% phenyl/95% methyl
silicone fused-silica open tubular column (DB-5, J&W Scientific, or
equivalent), 1.5 /xm film thickness.
4.1.2.2 Column 2, 30 m x 0.53 mm ID, 14% cyanopropyl
phenyl silicone fused-silica open tubular column (DB-1701, J&W
Scientific, or equivalent), 1,0 ptm film thickness.
4.1.3 Detector - Dual electron capture detector (ECD)
4.2 Glassware, see Methods 3510, 3540, 3541, 3550, 3610, 3620, 3640, and
3660 for specifications.
4.3 Kuderna-Danish (K-D) apparatus.
4.3.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.3.2 Evaporation flask - 500 ml (Kontes K-570001-500 or equiva-
lent). Attach to concentrator tube with springs, clamps, or equivalent.
4.3.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.3.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.3.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.4 Boiling chips, approximately 10/40 mesh. Heat to 400 °C for 30 min,
or Soxhlet-extract with methylene chloride prior to use.
4.5 Water bath, heated, with concentric ring cover, capable of
temperature control (+ 2°C).
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is
first ascertained that the reagent is of sufficiently high purity to permit its
use without lessening the accuracy of the determination.
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5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400 "C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride. If the sodium sulfate is precleaned with methylene chloride,
a method blank must be analyzed, demonstrating that there is no interference from
the sodium sulfate.
5.4 Solvents:
5.4.1 Hexane, C6H14 - Pesticide quality, or equivalent.
5.4.2 Methylene chloride, CH2C12 - Pesticide quality, or equivalent.
5.4.3 Acetone, CH3COCH3 - Pesticide quality, or equivalent.
5.4.4 Acetonitrile, CH3CN - HPLC grade.
5.4.5 Methanol, CH3OH - HPLC grade.
5.4.6 Diethyl Ether, C2H5OC2H5 - Pesticide quality, or equivalent.
Must be free of peroxides, as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl alcohol preservative must be
added to each liter of ether.
5.5 Stock standard solutions:
5.5.1 Prepare stock standard solutions at a concentration of
1000 mg/L by dissolving 0.0100 g of assayed reference material in hexane,
and diluting to volume in a 10 ml volumetric flask. When compound purity
is assayed to be 96 percent or greater, the weight can be used without
correction to calculate the concentration of the stock standard.
Commercially prepared stock standard solutions can be used at any
concentration if they are certified by the manufacturer or by an
independent source.
5.5.2 Transfer the stock standard solutions into glass vials with
Teflon lined screw-caps or crimp tops. Store at 4 °C and protect from
light. Stock standard solutions should be checked periodically by gas
chromatography for signs of degradation or evaporation, especially just
prior to preparation of calibration standards.
5.5.3 Stock standard solutions must be replaced after 6 months, or
sooner if comparison with check standards indicates a problem.
5.6 Calibration standards: Calibration standards are prepared at a
minimum of five concentrations for each parameter of interest through dilution
of the stock standard solutions with hexane. One of the concentrations should
be at a concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations found
in real samples, or should define the working range of the GC. Calibration
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solutions must be replaced after 1 to 2 months, or sooner if comparison with
calibration verification standards indicates a problem.
5.7 Internal standards (if internal standard calibration is used): To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Benzyl benzoate has been tested and
found appropriate for Method 8061.
5.7.1 Prepare a spiking solution of benzyl benzoate in hexane at
5000 mg/L. Addition of 10 juL of this solution to 1 ml of sample extract
is recommended. The spiking concentration of the internal standard should
be kept constant for all samples and calibration standards. Store the
internal standard spiking solution at 4 °C in glass vials with Teflon
lined screw-caps or crimp tops. Standard solutions should be replaced
when ongoing QC (Sec. 8) indicates a problem.
5.8 Surrogate standards: The analyst should monitor the performance of
the extraction, cleanup (when used), analytical system, and the effectiveness of
the method in dealing with each sample matrix by spiking each sample, standard,
and blank with surrogate compounds. Three surrogates may be used for Method 8061
in addition to those listed in Sec. 1.4: diphenyl phthalate, diphenyl
isophthalate, and dibenzyl phthalate. However, the compounds listed in Sec. 1.4
are recommended.
5.8.1 Prepare a surrogate standard spiking solution, in acetone,
which contains 50 ng//xL of each compound. Addition of 500 juL of this
solution to 1 L of water or 30 g solid sample is equivalent to 25 fj.g/1 of
water or 830 jug/kg of solid sample. The spiking concentration of the
surrogate standards may be adjusted accordingly, if the final volume of
extract is reduced below 2 ml for water samples or 10 ml for solid
samples. Store the surrogate spiking solution at 4 °C in glass vials with
Teflon lined screw-caps or crimp tops. The solution must be replaced
after 6 months, or sooner if ongoing QC (Sec. 8) indicates problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Extraction:
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a pH of
5 to 7 with methylene chloride in a separatory funnel (Method 3510).
Method 3520 is not recommended for the extraction of aqueous samples
because the longer chain esters (dihexyl phthalate bis(2-ethylhexyl)
phthalate, di-n-octyl phthalate, and dinonyl phthalate) tend to adsorb to
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the glassware and consequently, their extraction recoveries are
<40 percent. Solid samples are extracted with hexane/acetone (1:1) or
methylene chloride/acetone (1:1) in a Soxhlet extractor (Methods
3540/3541) or with an ultrasonic extractor (Method 3550). Immediately
prior to extraction, spike 500 /uL of the surrogate standard spiking
solution (concentration = 50 ng//iL) into 1 L aqueous sample or 30 g solid
sample.
7.1.2 Extraction of particulate-free aqueous samples using
C18-extraction disks (optional):
7.1.2.1 Disk preconditioning: Place the C18-extraction disk
into the filtration apparatus and prewash the disk with 10 to 20 ml
of acetonitrile. Apply vacuum to pull the solvent through the disk.
Maintain vacuum to pull air through for 5 min. Follow with 10 ml of
methanol. Apply vacuum and pull most of the methanol through the
disk. Release vacuum before the disk gets dry. Follow with 10 ml
organic-free reagent water. Apply vacuum and pull most of the water
through the disk. Release the vacuum before the disk gets dry.
7.1.2.2 Sample preconcentration: Add 2.5 ml of methanol to
the 500 ml aqueous sample in order to get reproducible results.
Pour the sample into the filtration apparatus. Adjust vacuum so
that it takes approximately 20 min to process the entire sample.
After all of the sample has passed through the membrane disk, pull
air through the disk for 5 to 10 min. to remove any residual water.
7.1.2.3 Sample elution: Break the vacuum and place the tip
of the filter base into the test tube that is contained inside the
suction flask. Add 10 ml of acetonitrile to the graduated funnel,
making sure to rinse the walls of the graduated funnel with the
solvent. Apply vacuum to pass the acetonitrile through the membrane
disk.
7.1.2.4 Extract concentration (if necessary): Concentrate
the extract to 2 mL or less, using either the micro Snyder column
technique (Sec. 7.1.2.4.1) or nitrogen blowdown technique (Sec.
7.1.2.4.2).
7.1.2.4.1 Micro Snyder Column Technique
7.1.2.4.1.1 Add one or two clean boiling chips to
the concentrator tube and attach a two ball micro Snyder
column. Prewet the column by adding about 0.5 mL of
acetonitrile to the top of the column. Place the K-D
apparatus in a hot water bath (15-20°C above the boiling
point of the solvent) so that the concentrator tube is
partially immersed in the hot water and the entire lower
rounded surface of the flask is bathed with hot vapor.
Adjust the vertical position of the apparatus and the
water temperature, as required, to complete the
concentration in 5-10 minutes. At the proper rate of
distillation the balls of .the column will actively
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chatter, but the chambers will not flood. When the
apparent volume of liquid reaches 0.5 ml, remove the K-D
apparatus from the water bath and allow it to drain and
cool for at least 10 minutes. Remove the Snyder column
and rinse the flask and its lower joints with about
0.2 ml of solvent and add to the concentrator tube.
Adjust the final volume to 1.0-2.0 ml with solvent.
7.1.2.4.2 Nitrogen Slowdown Technique
7.1.2.4.2.1 Place the concentrator tube in a warm
water bath (approximately 35 °C) and evaporate the
solvent volume to the required level using a gentle
stream of clean, dry nitrogen (filtered through a column
of activated carbon).
CAUTION: Do not use plasticized tubing between
the carbon trap and the sample.
7.1.2.4.2.2 The internal wall of the tube must be
rinsed down several times with acetonitrile during the
operation. During evaporation, the solvent level in the
tube must be positioned to prevent water from condensing
into the sample (i.e., the solvent level should be below
the level of the water bath). Under normal operating
conditions, the extract should not be allowed to become
dry.
7.2 Solvent Exchange: Prior to Florisil cleanup or gas chromatographic
analysis, the methylene chloride and methylene chloride/acetone extracts obtained
in Sec. 7.1.1 must be exchanged to hexane, as described in Sees. 7.2.1 through
7.2.3. Exchange is not required for the acetonitrile extracts obtained in
Sec. 7.1.2.4.
7.2.1 Add one or two clean boiling chips to the flask and attach a
three ball Snyder column. Concentrate the extract as described in Sec.
7.1.2.4.1, using 1 ml of methylene chloride to prewet the column, and
completing the concentration in 10-20 minutes. When the apparent volume
of liquid reaches 1-2 ml, remove the K-D apparatus from the water bath and
allow it to drain and cool for at least 10 minutes.
7.2.2 Momentarily remove the Snyder column, add 50 ml of hexane, a
new boiling chip, and attach the macro Snyder column. Concentrate the
extract as described in Sec. 7.1.2.4.1, using 1 ml of hexane to prewet the
Snyder column, raising the temperature of the water bath, if necessary, to
maintain proper distillation, and completing the concentration in 10-20
minutes. When the apparent volume of liquid reaches 1-2 ml, remove the
K-D apparatus and allow it to drain and cool for at least 10 min.
7.2.3 Remove the Snyder column and rinse the flask and its lower
joint into the concentrator tube with 1 to 2 ml hexane. A 5 ml syringe is
recommended for this operation. Adjust the extract volume to 2 ml for
water samples, using either the micro Snyder column technique (Sec.
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7.1.2.4.1) or nitrogen blowdown technique (Sec. 7.1.2.4.2), or 10 mL for
solid samples. Stopper the concentrator tube and store at 4 °C if further
processing will be performed immediately. If the extract will be stored
for two days or longer, it should be transferred to a glass vial with a
Teflon lined screw-cap or crimp top. Proceed with the gas chromatographic
analysis.
7.3 Cleanup/Fractionation:
7.3.1 Cleanup may not be necessary for extracts from a relatively
clean sample matrix. If polychlorinated biphenyls (PCBs) and
organochlorine pesticides are known to be present in the sample, use the
procedure outlined in Methods 3610 or 3620. When using column cleanup,
collect Fraction 1 by eluting with 140 ml (Method 3610) or 100 ml
(Method 3620) of 20-percent diethyl ether in hexane. Note that, under
these conditions, bis(2-methoxyethyl) phthalate, bis(2-ethoxyethyl)
phthalate, and bis(2-n-butoxyethyl) phthalate are not recovered from the
Florisil column. The elution patterns and compound recoveries are given
in Table 3.
7.3.2 Methods 3610 and 3620 also describe procedures for sample
cleanup using Alumina and Florisil Cartridges. With this method,
bis(2-methoxyethyl) phthalate, bis(2-ethoxyethyl) phthalate, and
bis(2-n-butoxyethyl) phthalate are recovered quantitatively.
7.4 Gas chromatographic conditions (recommended):
7.4.1 Column 1 and Column 2 (Sec. 4.1.2):
Carrier gas (He) = 6 mL/min.
Injector temperature = 250 "C.
Detector temperature = 320 °C.
Column temperature:
Initial temperature = 150 °C, hold for 0.5 min.
Temperature program = 150 °C to 220 °C at 5 °C/min.,
followed by 220 °C to 275 °C at 3
"C/min.
Final temperature = 275 °C hold for 13 min.
7.4.2 Table 1 gives the retention times and MDLs that can be
achieved by this method for the 16 phthalate esters. An example of the
separations achieved with the DB-5 and DB-1701 fused-silica open tubular
columns is shown in Figure 1.
7.5 Calibration:
7.5.1 Refer to Method 8000 for proper calibration techniques. Use
Tables 1 and 2 for guidance on selecting the lowest point on the
calibration curve.
7.5.2 The procedure for internal or external calibration may be
used. Refer to Method 8000 for the description of each of these
procedures.
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7.6 Gas chromatographic analysis:
7.6.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 juL of internal standard solution at 5000 mg/L
to the sample prior to injection.
7.6.2 Follow Method 8000 for instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria.
7.6.3 Record the sample volume injected and the resulting peak
areas.
7.6.4 Using either the internal or the external calibration
procedure (Method 8000), determine the identity and the quantity of each
component peak in the sample chromatogram which corresponds to the
compounds used for calibration purposes.
7.6.5 If the response of a peak exceeds the working range of the
system, dilute the extract and reanalyze.
7.6.6 Identify compounds in the sample by comparing the retention
times of the peaks in the sample chromatogram with those of the peaks in
standard chromatograms. The retention time window used to make
identifications is based upon measurements of actual retention time
variations over the course of 10 consecutive injections. Three times the
standard deviation of the retention time can be used to calculate a
suggested window size.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the QC
specified in Method 3600 and in the specific cleanup method.
8.2 Quality control required to evaluate the GC system operation is found
in Method 8000.
8.2.1 The quality control check sample concentrate (Method 8000)
should contain the test compounds at 5 to 10 ng/juL.
8.3 Calculate the recoveries of the surrogate compounds for all samples,
method blanks, and method spikes. Determine if the recoveries are within limits
established by performing QC procedures outlined in Method 8000.
8.3.1 If the recoveries are not within limits, the following are
required:
8.3.1.1 Make sure there are no errors in calculations,
surrogate solutions and internal standards. Also check instrument
performance.
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8.3.1.2 Recalculate the data and/or reanalyze the extract
if any of the above checks reveal a problem.
8.3.1.3 Reextract and reanalyze the sample if none of the
above are a problem, or flag the data as "estimated concentration."
8.4 An internal standard peak area check must be performed on all
samples. The internal standard must be evaluated for acceptance by determining
whether the measured area for the internal standard deviates by more than 30
percent from the average area for the internal standard in the calibration
standards. When the internal standard peak area is outside that limit, all
samples that fall outside the QC criteria must be reanalyzed.
8.5 GC/MS confirmation: Any compounds confirmed by two columns may also
be confirmed by GC/MS if the concentration is sufficient for detection by GC/MS
as determined by the laboratory-generated detection limits.
8.5.1 The GC/MS would normally require a minimum concentration of 10
ng//iL in the final extract for each single-component compound.
8.5.2 The sample extract and associated blank should be analyzed by
GC/MS as per Sec. 7.0 of Method 8270. Normally, analysis of a blank is
not required for confirmation analysis, however, analysis for phthalates
is a special case because of the possibility for sample contamination
through septum punctures, etc.
8.5.3 A reference standard of the compound must also be analyzed by
GC/MS. The concentration of the reference standard must be at a
concentration that would demonstrate the ability to confirm the phthalate
esters identified by GC/ECD.
8.6 Include a mid-concentration calibration standard after each group of
20 samples in the analysis sequence. The response factors for the
mid-concentration calibration must be within + 15 percent of the average values
for the multiconcentration calibration.
8.7 Demonstrate through the analyses of standards that the Florisil
fractionation scheme is reproducible. When using the fractionation schemes given
in Methods 3610 or 3620, batch-to-batch variations in the composition of the
alumina or Florisil material may cause variations in the recoveries of the
phthalate esters.
9.0 METHOD PERFORMANCE
9.1 The MDL is defined in Chapter One. The MDL concentrations listed in
Table 1 were obtained using organic-free reagent water. Details on how to
determine MDLs are given in Chapter One. The MDL actually achieved in a given
analysis will vary, as it is dependent on instrument sensitivity and matrix
effects.
9.2 This method has been tested in a single laboratory by using different
types of aqueous samples and solid samples which were fortified with the test
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compounds at two concentrations. Single-operator precision, overall precision,
and method accuracy were found to be related to the concentration of the
compounds and the type of matrix. Results of the single-laboratory method
evaluation are presented in Tables 4 and 5.
9.3 The accuracy and precision obtained is determined by the sample
matrix, sample preparation technique, cleanup techniques, and calibration
procedures used.
10.0 REFERENCES
1. Glazer, J.A.; Foerst, G.D.; McKee, G.D.; Quave, S.A., and Budde, W.L.,
"Trace Analyses for Wastewaters," Environ. Sci. and Technol. 15: 1426,
1981.
2. Lopez-Avila, V., Baldin, E., Benedicto, J., Milanes, J., and Beckert,
W.F., "Application of Open-Tubular Columns to SW-846 GC Methods", EMSL-Las
Vegas, 1990.
3. Beckert, W.F. and Lopez-Avila, V., "Evaluation of SW-846 Method 8060 for
Phthalate Esters", Proceedings of Fifth Annual Testing and Quality
Assurance Symposium, USEPA, 1989.
8061 - 12
Revision 0
September 1994
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TABLE 2.
ESTIMATED QUANTITATION LIMITS (EQL) FOR VARIOUS MATRICES8
Matrix Factor
Groundwater 10
Low-concentration soil by ultrasonic extraction 670
with GPC cleanup
High-concentration soil and sludges by ultrasonic 10,000
extraction
Non-water miscible waste 100,000
EQL = [Method detection limit (see Table 1)] X [Factor found in this
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Sample EQLs are highly matrix-dependent. The EQLs determined herein are
provided for guidance and may not always be achievable.
8061 - 15 Revision 0
September 1994
-------
TABLE 3.
AVERAGE RECOVERIES OF METHOD 8061 COMPOUNDS USING METHODS 3610 AND 3620
Compound
Dimethyl phthalate
Diethyl phthalate
Diisobutyl phthalate
Di-n-butyl phthalate
Bis(4-methyl-2-pentyl) phthalate
Bis(2-methoxyethyl) phthalate
Diamyl phthalate
Bis(2-ethoxyethyl) phthalate
Hexyl 2-ethylhexyl phthalate
Dihexyl phthalate
Benzyl butyl phthalate
Bis(2-n-butoxyethyl) phthalate
Bis(2-ethylhexyl) phthalate
Dicyclohexyl phthalate
Di-n-octyl phthalate
Dinonyl phthalate
Alumina
column8
64.5
62.5
77. 0
76.5
89.5
70.5
75.0
67.0
90.5
73.0
87.0
62.5
91.0
84.5
108
71.0
Florisil
column8
40.0
57.0
80.0
85.0
84.5
0
81.5
0
105
74.5
90.0
0
82.0
83.5
115
72.5
Alumina
cartridge13
101
103
104
108
103
64.1°
103
111
101
108
103
108
97.6
97.5
112
97.3
Florisil
cartridge*1
89.4
97.3
91.8
102
105
78. 3e
94.5
93.6
96.0
96.8
98.6
91.5
97.5
90.5
97.1
105
8 2 determinations; alumina and Florisil chromatography performed according
to Methods 3610 and 3620, respectively.
b 2 determinations, using 1 g alumina cartridges; Fraction 1 was eluted with
5 ml of 20-percent acetone in hexane. 40 ^9 of each component was spiked
per cartridge.
c 36.8 percent was recovered by elution with an additional 5 ml of
20-percent acetone in hexane.
d 2 determinations, using 1 g Florisil cartridges; Fraction 1 was eluted
with 5 mL of 10-percent acetone in hexane. 40 jug of each component was
spiked per cartridge.
e 14.4 percent was recovered by elution with an additional 5 ml of
10-percent acetone in hexane.
8061 - 16
Revision 0
September 1994
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Figure 1
DB-S
30 m x 0.53 mm 10
1.5-jimRm
1
6
UJ
IS
IS
11 12 SU-1 SU-2 SU-3
14
16
SU-2 SU-3
12 SU-1 15 T T 16
13
DB-1701
30 m x 0.53 mm ID
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10
11
r
^
LI
uu
L_
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10
20
TIME (min)
30
40
GC/ECD chromatograms of a composite phthalate esters standard (concentration
10 ng/jiL per compound) analyzed on a DB-5 and a DB-1701 fused-silica open
tubular column. Temperature program: 150°C (0.5 min hold) to 220°C at
5°C/min, then to 275°C (13 min hold) at 3°C/nrin.
8061 - 19
Revision 0
September 1994
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METHOD 8061
PHTHALATE ESTERS BY CAPILLARY GAS CHROMATOGRAPHY
WITH ELECTRON CAPTURE DETECTION (GC/ECD)
Start
7 1 Extraction
7 1 1 Refer to Chapter 2 for
guidance on choosing
an extraction procedure
Recommendations given
7 1 2 Determine spike sample
recovery and detection limit
for each new sample matrix
and a given extraction
procedure.
7 1 3 Aqueous sample extraction
with C18 disks
1 Precondition disks using
solvent tram
2 Concentrate sample
analytes on disk.
3 Elute sample analytes
with acetonitrile.
4 Concentrate extract'
1 Micro-Snyder Column
Technique
2 Nitrogen Slowdown
Technique
1 Evaporate solvent to
desired level
2 Rinse tube walls
frequently and avoid
evaporating to dryness
7 2 Solvent Exchange to Hexane [
7 2 1 Evaporate extract volume to
1 -2 ml using K-D assembly
722 Add hexane to K-D assembly
and evaporate to 1 -2 ml
723 Rinse K-D components and
adjust volume to desired level
7 3 Cleanup/Fracdonation
73.1 Cleanup may not be
necessary for extracts with
clean sample matrices
Fraction collection and
methods outlined for other
compd groups of interest.
732 Flonsil Cartridge Cleanup
1 Check each lot of Flonsil
cartridges for analyte
recovery by eluOng and
analyzing a composite std
2 Wash and adjust solvent
Mow through cartridges
3 Place culture tubes or 5 ml
vol flasks for eluate
collection
4 Transfer appropriate extract
volume on cartridge
5 Elute the cartridges and
dilute to mark on flask
Transfer eluate to glass
vials for concentration
733 Collect 2 fractions if PCBs
and organochlonne pesticides
are known to be present
7 4 Gas Chromatograph
7 4 1 Set GC operating parameters
742 Table 1 and Figure 1 show
MDLs and analyte retention
times
8061 - 20
Revision 0
September 1994
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METHOD 8061
(CONTINUED)
7 5 Calibration
1
75.1 See Method 8000 tor
calibration technique
7.5 2 Refer to Method 8000 tor
internal/external std
procedure.
' 6 GC Analysis
7 6 1 Refer to Method 8000
762 Follow Section 7 6 in
Method 8000 tor
instructions on analysis
sequence, dilutions.
retention time windows.
and identification criteria.
7 6.3 Record injection volume
and sample peak areas
764 Identify and quantify each
component peak using the
internal or external std
procedure
765 Dilute extracts which
show analyte levels
outside of the calibration
range.
1
766 Identify compounds in the
sample by comparing
retention times in the
sample and the standard
chromatograms
\
I
8061 - 21
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METHOD 8070
NITROSAMINES BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 This method covers the determination of certain nitrosamines. The
following compounds can be determined by this method:
Appropriate Technique
Compound Name CAS No.a 3510 3520 3540 3550 3580
N-Nitrosodimethylamine 62-75-9 X X X X
N-Nitrosodiphenylamine 86-30-6 X X X X
N-Nitrosodi-n-propylamine 621-64-7 X X X X
a Chemical Abstract Services Registry Number.
X Greater than 70 percent recovery by this preparation technique.
X
X
X
1.2 This is a gas chromatographic (GC) method applicable to the
determination of the parameters listed above in municipal and industrial
discharges. When this method is used to analyze unfamiliar samples for any or
all of the compounds above, compound identifications should be supported by at
least one additional qualitative technique. This method describes analytical
conditions for a second gas chromatographic column that can be used to confirm
measurements made with the primary column. Method 8270 provides gas
chromatograph/mass spectrometer (GC/MS) conditions appropriate for the
qualitative and quantitative confirmation of results for
N-nitrosodi-n-propylamine. In order to confirm the presence of
N-nitrosodiphenylamine, the cleanup procedure specified in Section 7.3.3 or 7.3.4
must be used. In order to confirm the presence of N-nitrosodimethylamine by
GC/MS, chromatographic column 1 of this method must be substituted for the column
recommended in Method 8270. Confirmation of these parameters using GC-high
resolution mass spectrometry or a Thermal Energy Analyzer is also recommended
practice.
1.3 The method detection limit (MDL) for each parameter is listed in
Table 1. The MDL for a specific wastewater may differ from those listed,
depending upon the nature of interferences in the sample matrix. Table 2 lists
the Estimated Quantitation Limits (EQLs) for various matrices.
1.4 The toxicity or carcinogenicity of each reagent used in this method
has not been precisely defined. However, each chemical compound should be
treated as a potential health hazard. From this viewpoint, exposure to these
chemicals must be reduced to the lowest possible concentration by whatever means
available. The laboratory is responsible for maintaining a current awareness
file of OSHA regulations regarding the safe handling of the chemicals specified
in this method. A reference file of material data handling sheets should also
8070 - 1 Revision 0
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be made available to all personnel involved in the chemical analysis.
1.5 These nitrosamines are known carcinogens. Therefore, utmost care
must be exercised in the handling of these materials. Nitrosamine reference
standards and standard solutions should be handled and prepared in a ventilated
glove box within a properly ventilated room.
1.6 N-Nitrosodiphenylamine is reported to undergo transnitrosation
reactions. Care must be exercised in the heating or concentrating of solutions
containing this compound in the presence of reactive amines.
2.0 SUMMARY OF METHOD
2.1 A measured volume of aqueous sample, approximately one liter, is
solvent extracted with methylene chloride using a separatory funnel. The
methylene chloride extract is washed with dilute HC1 to remove free amines,
dried, and concentrated to a volume of 10 ml or less. Gas chromatographic
conditions are described which permit the separation and measurement of the
compounds in the extract after it has been exchanged to methanol.
2.2 Method 8070 provides gas chromatographic conditions for the detection
of ppb concentrations of nitrosamines. Prior to use of this method, appropriate
sample extraction techniques must be used. Both neat and diluted organic liquids
(Method 3580, Waste Dilution) may be analyzed by direct injection. A 2 to 5 /xL
aliquot of the extract is injected into a gas chromatograph (GC) using the
solvent flush technique, and compounds in the GC effluent are detected by a
nitrogen-phosphorus detector (NPD) or a Thermal Energy Analyzer and the reductive
Hall detector.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Matrix interferences may be caused by contaminants that are
coextracted from the sample. The extent of matrix interferences will vary
considerably from source to source, depending upon the nature and diversity of
the industrial complex or municipality being sampled. The cleanup procedures
(Methods 3610 or 3620) can be used to overcome many of these interferences, but
unique samples may require additional cleanup approaches to achieve the MDL
listed in Table 1.
3.3 Nitrosamines contaminate many types of products commonly found in the
laboratory. The analyst must demonstrate that no nitrosamine residues
contaminate the sample or solvent extract under the conditions of analysis.
Plastics, in particular, must be avoided because nitrosamines are commonly used
as plasticizers and are easily extracted from plastic materials. Serious
nitrosamine contamination may result at any time if consistent quality control
is not practiced.
3.4 The sensitive and selective Thermal Energy Analyzer and the reductive
Hall detector may be used in place of the nitrogen-phosphorus detector when
8070 - 2 Revision 0
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interferences are encountered. The Thermal Energy Analyzer offers the highest
selectivity of the non-mass spectrometric detectors.
3.5 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All these materials must be demonstrated to be free from
interferences, under the conditions of the analysis, by analyzing reagent blanks.
Specific selection of reagents and purification of solvents by distillation in
all-glass systems may be required.
3.6 Interferences coextracted from samples will vary considerably from
source to source, depending upon the waste being sampled. Although general
cleanup techniques are recommended as part of this method, unique samples may
require additional cleanup.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph - An analytical system complete with temperature
programmable gas chromatograph suitable for on-column injection and all required
accessories including syringes, analytical columns, gases, detector, and strip-
chart recorder. A data system is recommended for measuring peak areas.
4.1.1 Column 1 - 1.8 m x 4 mm ID Pyrex glass, packed with Chromosorb
W AW, (80/100 mesh) coated with 10% Carbowax 20 M/2% KOH or equivalent.
This column was used to develop the method performance statements in
Section 9.0. Guidelines for the use of alternate column packings are
provided in Section 7.3.2.
4.1.2 Column 2-1.8mx4mmID Pyrex glass, packed with
Supelcoport (100/120 mesh) coated with 10% SP-2250, or equivalent.
4.1.3 Detector - Nitrogen-Phosphorus, reductive Hall or Thermal
Energy Analyzer. These detectors have proven effective in the analysis of
wastewaters for the parameters listed in the scope. A nitrogen-phosphorus
detector was used to develop the method performance statements in Section
9.0. Guidelines for the use of alternate detectors are provided in
Section 7.3.2.
4.2 Kuderna-Danish (K-D) apparatus
4.2.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025 or
equivalent). Calibration must be checked at the volumes employed in the
test. A ground glass stopper is used to prevent evaporation of extracts.
4.2.2 Evaporation flask - 500 ml (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.2.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
8070 - 3 Revision 0
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equivalent).
4.2.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Boiling chips - Approximately 10/40 mesh. Heat to 400°C for
30 minutes or Soxhlet extract with methylene chloride.
4.4 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 2°C). The bath should be used in a hood.
top.
4.5 Balance - Analytical, 0.0001 g.
4.6 Vials - 10 to 15 ml, amber glass with Teflon lined screw-cap or crimp
4.7 Volumetric flasks, Class A, Appropriate sizes with ground glass
stoppers.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all inorganic reagents shall conform to
the specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may be
used, provided it is first ascertained that the reagent is of sufficiently high
purity to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH - Pesticide quality or equivalent.
5.4 Isooctane, (CH3)3CCH2CH(CH3)2 - Pesticide quality or equivalent.
5.5 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.6 Stock standard solutions (1000 mg/L) - Stock standard solutions can
be prepared from pure standard materials or purchased as certified solutions.
5.6.1 Prepare stock standard solutions by accurately weighing
0.1000 + 0.0010 g of pure material. Dissolve the material in pesticide
quality methanol and dilute to volume in a 100 ml volumetric flask.
Larger volumes can be used at the convenience of the analyst. If compound
purity is certified at 96% or greater, the weight can be used without
correction to calculate the concentration of the stock standard.
Commercially prepared stock standards can be used at any concentration if
they are certified by the manufacturer or by an independent source.
5.6.2 Transfer the stock standard solutions into bottles with Teflon
lined screw-caps or crimp tops. Store at 4°C and protect from light.
8070 - 4 Revision 0
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Stock standard solutions should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them.
5.6.3 Stock standard solutions must be replaced after six months, or
sooner if comparison with check standards indicates a problem.
5.7 Calibration standards - A minimum of five concentrations should be
prepared through dilution of the stock standards with isooctane. One of the
concentrations should be at a concentration near, but above, the method detection
limit. The remaining concentrations should correspond to the expected range of
concentrations found in real samples or should define the working range of the
GC. Calibration solutions must be replaced after six months, or sooner if
comparison with check standards indicates a problem.
5.8 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.8.1 Prepare calibration standards at a minimum of five
concentrations for each analyte of interest, as described in Section 5.7.
5.8.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with isooctane.
5.8.3 Analyze each calibration standard according to Section 7.0.
5.9 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system and the effectiveness
of the method in dealing with each sample matrix by spiking each sample,
standard, and reagent blank with one or two surrogates (e.g. nitrosamines that
are not expected to be in the sample) recommended to encompass the range of the
temperature program used in this method. Method 3500 details instructions on the
preparation of base/neutral surrogates. Deuterated analogs of analytes should
not be used as surrogates for gas chromatographic analysis due to coelution
problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1. Extracts must be stored at 4°C and analyzed within 40 days of
extraction.
8070 - 5 Revision 0
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7.0 PROCEDURE
7.1 Extraction
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a
neutral, or as is, pH with methylene chloride, using either Method 3510 or
3520. Solid samples are extracted using either Method 3540 or 3550.
7.1.2 Prior to gas chromatographic analysis, the extraction solvent
must be exchanged to methanol. The exchange is performed during the K-D
procedures listed in all of the extraction methods. The exchange is
performed as follows.
7.1.2.1 Following K-D of the methylene chloride extract to
1 ml using the macro-Snyder column, allow the apparatus to cool and drain
for at least 10 minutes.
7.1.2.2 Momentarily remove the Snyder column, add 50 ml of
methanol, a new boiling chip, and reattach the macro-Snyder column.
Concentrate the extract using 1 ml of methanol to prewet the Snyder
column. Place the K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete concentration in 5-10 minutes. At the proper
rate of distillation the balls of the column will actively chatter,
but the chambers will not flood. When the apparent volume of liquid
reaches 1 ml, remove the K-D apparatus and allow it to drain and
cool for at least 10 minutes. The extract will be handled
differently at this point, depending on whether or not cleanup is
needed. If cleanup is not required, proceed to Section 7.1.2.3. If
cleanup is needed, proceed to Section 7.1.2.4.
7.1.2.3 If cleanup of the extract is not required, remove
the Snyder column and rinse the flask and its lower joint into the
concentrator tube with 1-2 ml of methanol. A 5 ml syringe is
recommended for this operation. Adjust the extract volume to
10.0 ml. Stopper the concentrator tube and store refrigerated at
4°C if further processing will not be performed immediately. If the
extract will be stored longer than two days, it should be
transferred to a vial with a Teflon lined screw-cap or crimp top.
Proceed with gas chromatographic analysis.
7.1.2.4 If cleanup of the extract is required, remove the
Snyder column and rinse the flask and its lower joint into the
concentrator tube with a minimum amount of methylene chloride. A 5
ml syringe is recommended for this operation. Add a clean boiling
chip to the concentrator tube and attach a two ball micro-Snyder
column. Prewet the column by adding about 0.5 ml of methylene
chloride to the top. Place the micro K-D apparatus on the water
bath (80°C) so that the concentrator tube is partially immersed in
the hot water. Adjust the vertical position of the apparatus and
the water temperature, as required, to complete concentration in 5-
8070 - 6 Revision 0
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10 minutes. At the proper rate of distillation the balls of the
column will actively chatter, but the chambers will not flood. When
the apparent volume of liquid reaches 0.5 ml, remove the K-D
apparatus and allow it to drain and cool for at least 10 minutes.
7.1.2.5 Remove the micro-Snyder column and rinse the flask
and its lower joint into the concentrator tube with 0.2 ml of
methylene chloride. Adjust the extract volume to 2.0 ml and proceed
with either Method 3610, 3620, or 3640.
7.1.3 If N-nitrosodiphenylamine is to be measured by gas
chromatography, the analyst must first use a cleanup column to eliminate
diphenylamine interference (Methods 3610 or 3620). If N-
nitrosodiphenylamine is of no interest, the analyst may proceed directly
with gas chromatographic analysis (Section 7.3).
7.2 Cleanup
7.2.1 Cleanup procedures may not be necessary for a relatively clean
sample matrix. The cleanup procedure recommended in this method has been
used for the analysis of various clean waters and industrial effluents. If
particular circumstances demand the use of an alternative cleanup
procedure, the analyst must determine the elution profile and demonstrate
that the recovery of each compound of interest is no less than 85%.
Diphenylamine, if present in the original sample extract must be separate
from the nitrosamines if N-nitrosodiphenylamine is to be determined by
this method.
7.2.2 Proceed with either Method 3610 or 3620, using the 2 ml
methylene chloride extracts obtained from Section 7.1.2.5.
7.2.3 Following cleanup, the extracts should be analyzed by GC, as
described in the previous paragraphs and in Method 8000.
7.3 Gas Chromatography
7.3.1 N-nitrosodiphenylamine completely reacts to form diphenylamine
at the normal operating temperatures of a GC injection port (200 to 250°C).
Thus, N-nitrosodiphenylamine is chromatographed and detected as
diphenylamine. Accurate determination depends on removal of diphenylamine
that may be present in the original extract prior to GC (see Section
7.1.3).
7.3.2 Table 1 summarizes the recommended operating conditions for
the gas chromatograph. This table includes retention times and MDLs that
were obtained under these conditions. Examples of the parameter
separations achieved by these columns are shown in Figures 1 and 2. Other
packed columns, chromatographic conditions, or detectors may be used if
the requirements of Section 8.2 are met. Capillary (open-tubular) columns
may also be used if the relative standard deviations of responses for
replicate injections are demonstrated to be less than 6% and the
requirements of Section 8.2 are met.
8070 - 7 Revision 0
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7.4 Calibration - Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point on
the calibration curve.
7.4.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.4.2 If cleanup is performed on the samples, the analyst should
process a series of standards through the cleanup procedure and then
analyze the samples by GC. This will confirm elution patterns and the
absence of interferents from the reagents.
7.5 Gas chromatographic analysis
7.5.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 /il_ of internal standard to the sample prior to
injection.
7.5.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration check standard after
each group of 10 samples in the analysis sequence.
7.5.3 Examples of GC/NPD chromatograms for nitrosamines are shown in
Figures 1 and 2.
7.5.4 Record the sample volume injected and the resulting peak sizes
(in area units or peak heights).
7.5.5 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each analyte peak in
the sample chromatogram. See Method 8000 for calculation equations.
7.5.6 If peak detection and identification are prevented due to
interferences, the hexane extract may undergo cleanup using either Method
3610 or 3620.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the QC
in Method 3600 and in the specific cleanup method.
8.2 Procedures to check the GC system operation are found in Method 8000,
Section 8.6.
8.2.1 The quality control (QC) reference sample concentrate (Method
8000, Section 8.6) should contain each analyte of interest at 20 mg/L.
8.2.2 Table 3 indicates the calibration and QC acceptance criteria
for this method. Table 4 gives method accuracy and precision as functions
8070 - 8 Revision 0
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of concentration for the analytes of interest. The contents of both
Tables should be used to evaluate a laboratory's ability to perform and
generate acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000, Section 8.10).
8.3.1 If recovery is not within limits, the following is required.
• Check to be sure that there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are a
problem or flag the data as "estimated concentration.
9.0 METHOD PERFORMANCE
9.1 This method has been tested for linearity of recovery from spiked
organic-free reagent water and has been demonstrated to be applicable for the
concentration range from 4 x MDL to 1000 x MDL.
9.2 In a single laboratory (Southwest Research Institute), using spiked
wastewater samples, the average recoveries presented in Table 2 were obtained.
Each spiked sample was analyzed in triplicate on three separate occasions. The
standard deviation of the percent recovery is also included in Table 2.
10.0 REFERENCES
1. Fed. Regist. 1984, 49, 43234; October 26.
2. "Determination of Nitrosamines in Industrial and Municipal Wastewaters";
Report for EPA Contract 68-03-2606, in preparation.
3. Burgess, E.M.; Lavanish, J.M. "Photochemical Decomposition of N-
nitrosamines"; Tetrahedron Letters 1964, 1221.
4. Methods for Chemical Analysis of Water and Wastes; U.S. Environmental
Protection Agency. Office of Research and Development. Environmental
Monitoring and Support Laboratory. ORD Publication Offices of Center for
Environmental Research Information: Cincinnati, OH, 1979; EPA-600/4-79-
020.
5. "Method Detection Limit and Analytical Curve Studies EPA Methods 606, 607,
608"; U.S. Environmental Protection Agency. Environmental Monitoring and
Support Laboratory, Cincinnati, OH, special letter report for EPA Contract
68-03-2606.
8070 - 9 Revision 0
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
Analyte
Retention Time
(minutes)
Column 1 Column 2
Method
Detection Limit
(M9/L)
N-Nitrosodimethylamine
N-Ni trosodi -n-propyl ami ne
N-Ni trosodi phenyl ami nea
4.1
12.1
12. 8b
0.88
4.2
6.4C
0.15
0.46
0.81
Column 1 conditions:
Carrier gas (He) flow rate:
Column temperature:
Column 2 conditions:
Carrier gas (He) flow rate:
Column temperature:
40 mL/min
Isothermal,
indicated.
40 mL/min
Isothermal,
indicated.
at 110°C, except as otherwise
at 120°C, except as otherwise
a Measured as diphenylamine.
b Determined isothermally at 220°C.
c Determined isothermally at 210°C.
TABLE 2.
SINGLE OPERATOR ACCURACY AND PRECISION
Average Standard Spike
Percent Deviation Range
Number
of Matrix
Analyte
Types
N-Nitrosodimethylamine
N-Ni trosodi phenyl ami ne
N-Ni trosodi -n-propyl ami ne
Recovery % (M9/L)
32
79
61
3.7
7.1
4.1
0.8
1.2
9.0
Analyses
29
29
29
5
5
5
8070 - 10
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TABLE 3.
QC ACCEPTANCE CRITERIA
Test Limit Range Recovery
Cone. for s for X Range
Analyte (ng/L)
N-Nitrosodimethyl amine 20 3.4 4.6-20.0 13-109
N-Nitrosodiphenylamine 20 6.1 2.1-24.5 D-139
N-Nitrosodi-n-propylamine 20 5.7 11.5-26.8 45-146
s = Standard deviation for four recovery measurements, in yg/L.
X = Average recovery for four recovery measurements, in jig/L.
D = Detected, result must be greater than zero.
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TABLE 4.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION
Analyte
N-Nitrosodimethylamine
N-Ni trosodi phenyl ami ne
N-Nitroso-n-propylamine
Accuracy, as
recovery, X'
(ng/D
0.37C+0.06
0.64C+0.52
0.96C-0.07
Single
analyst
precision,
s/ (ng/L)
0.25X-0.04
0.36X-1.53
0.15X+0.13
Overall
precision,
S' (ng/L)
0.25X+0.11
0.46X-0.47
0.21X+0.15
X' = Expected recovery for one or more measurements of a sample
containing a concentration of C, in |ig/L.
sr' = Expected single analyst standard deviation of measurements at an
average concentration found of )(, in (ig/L.
C = True value for the concentration, in ng/L.
X = Average recovery found for measurements of samples containing a
concentration of C, in jig/L.
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FIGURE 1.
GAS CHROMATOGRAM OF NITROSAMINES
Column: 10% Ctrbowt* 20M + 2%
KOH on Chromosorb W-AW
Tfmpertturf. 110°
Detector: Phosphorus/Nitrogtn
6 3 10 12 14
on tim*. minutt*
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FIGURE 2.
GAS CHROMATOGRAM OF N-NITROSOOIPHENYLAMINE AS OIPHENYLAMINE
Column: 10% CerboweM 20M + 2% KOH on
Chromosorb W-AW
Tempertture: 220°C.
Detector: Phosphorus/Nitrogen
I
2 4 6 8 tO 12 14 16 19
Retention time, minutet
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METHOD 8070
NITROSAMINES BY GAS CHROMATOGRAPHY
711 Choo«e
apprpnale
••traction
procedure
712 Perform
solvent exchange
using methanol
7124 Perform
micro-K-0 procedure
uting methylene
chloride perform
Method 3610 or
3620. proceed «ith
CC analysis
)(••
0
7 1 2 3 Adjuit
• •tract. volun* and
procavd with
•nalyaxa or *tor*
in «ppropnat«
mannvr
713 Parform
coluan cleanup
uitng M.thod 3610
or 3620
7 3 2 Refer to
Table 1 for
operating
conditions for the
CC
7 4 Refer to Method
8000 for proper
calibration
technique*
7 S 1 Refer to
Method 8000 for
guidance on CC
analysis
7 S 4/7 S S Record
sample volume
injected and
resul ting peak
sue/perform
appropr la t«
calcula tions (refer
to Method 8000)
Stop
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METHOD 8080A
ORGANOCHLORINE PESTICIDES AND POLYCHLORINATED BIPHENYLS
BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8080 is used to determine the concentration of various
organochlorine pesticides and polychlorinated biphenyls (PCBs). The following
compounds can be determined by this method:
Compound Name
CAS No.'
Aldrin
a-BHC
0-BHC
5-BHC
7-BHC (Lindane)
Chlordane (technical)
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Heptachlor
Heptachlor epoxide
4,4'-Methoxychlor
Toxaphene
Aroclor-1016
Aroclor-1221
Aroclor-1232
Aroclor-1242
Aroclor-1248
Aroclor-1254
Aroclor-1260
309-00-2
319-84-6
319-85-7
319-86-8
58-89-9
12789-03-6
72-54-8
72-55-9
50-29-3
60-57-1
959-98-8
33212-65-9
1031-07-8
72-20-8
7421-93-4
76-44-8
1024-57-3
72-43-5
8001-35-2
12674-11-2
1104-28-2
11141-16-5
53469-21-9
12672-29-6
11097-69-1
11096-82-5
a Chemical Abstract Services Registry Number.
1.2 Table 1 lists the method detection limit for each compound in
organic-free reagent water. Table 2 lists the estimated quantitation limit (EQL)
for other matrices.
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2.0 SUMMARY OF METHOD
2.1 Method 8080 provides gas chromatographic conditions for the detection
of ppb concentrations of certain organochlorine pesticides and PCBs. Prior to
the use of this method, appropriate sample extraction techniques must be used.
Both neat and diluted organic liquids (Method 3580, Waste Dilution) may be
analyzed by direct injection. A 2 to 5 jitl sample is injected into a gas
chromatograph (GC) using the solvent flush technique, and compounds in the GC
effluent are detected by an electron capture detector (ECD) or an electrolytic
conductivity detector (HECD).
2.2 The sensitivity of Method 8080 usually depends on the concentration
of interferences rather than on instrumental limitations. If interferences
prevent detection of the analytes, Method 8080 may also be performed on samples
that have undergone cleanup. Method 3620, Florisil Column Cleanup, by itself or
followed by Method 3660, Sulfur Cleanup, may be used to eliminate interferences
in the analysis.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Interferences by phthalate esters can pose a major problem in
pesticide determinations when using the electron capture detector. These
compounds generally appear in the chromatogram as large late-eluting peaks,
especially in the 15% and 50% fractions from the Florisil cleanup. Common
flexible plastics contain varying amounts of phthalates. These phthalates are
easily extracted or leached from such materials during laboratory operations.
Cross contamination of clean glassware routinely occurs when plastics are handled
during extraction steps, especially when solvent-wetted surfaces are handled.
Interferences from phthalates can best be minimized by avoiding contact with any
plastic materials. Exhaustive cleanup of reagents and glassware may be required
to eliminate background phthalate contamination. The contamination from
phthalate esters can be completely eliminated with a microcoulometric or
electrolytic conductivity detector.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas Chromatograph: Analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detectors, column supplies, recorder, gases, and
syringes. A data system for measuring peak heights and/or peak areas is
recommended.
4.1.2 Columns
4.1.2.1 Column 1: Supelcoport (100/120 mesh) coated with
1.5% SP-2250/1.95% SP-2401 packed in a 1.8 m x 4 mm ID glass column
or equivalent.
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4.1.2.2 Column 2:, Supelcoport (100/120 mesh) coated with
3% OV-1 inal.8mx4mmID glass column or equivalent.
4.1.3 Detectors: Electron capture (ECD) or electrolytic
conductivity detector (HECD).
4.2 Kuderna-Danish (K-D) apparatus:
4.2.1 Concentrator tube: 10 mL, graduated (Kontes K-570050-1025 or
equivalent). A ground-glass stopper is used to prevent evaporation of
extracts.
4.2.2 Evaporation flask: 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.2.3 Snyder column: Three ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Snyder column: Two ball micro (Kontes K-569001-0219 or
equivalent).
4.2.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Boiling chips: Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.4 Water bath: Heated, with concentric ring cover, capable of
temperature control (±5°C). The bath should be used in a hood.
4.5 Volumetric flasks, Class A: sizes as appropriate with ground-glass
stoppers.
4.6 Microsyringe: 10 nl.
4.7 Syringe: 5 ml.
4.8 Vials: Glass, 2, 10, and 20 ml capacity with Teflon-lined screw caps
or crimp tops.
4.9 Balances: Analytical, 0.0001 g and Top loading, 0.01 g.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
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5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Solvents
5.3.1 Hexane, C6H14 - Pesticide quality or equivalent.
5.3.2 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.3.3 Toluene, C6H5CH3 - Pesticide quality or equivalent.
5.3.4 Isooctane, (CH3)3CCH2CH(CH3)2 - Pesticide quality or equivalent.
5.4 Stock standard solutions:
5.4.1 Prepare stock standard solutions at a concentration of
1000 mg/L by dissolving 0.0100 g of assayed reference material in
isooctane and diluting to volume in a 10 ml volumetric flask. A small
volume of toluene may be necessary to put some pesticides in solution.
Larger volumes can be used at the convenience of the analyst. When
compound purity is assayed to be 96% or greater, the weight can be used
without correction to calculate the concentration of the stock standard.
Commercially prepared stock standards can be used at any concentration if
they are certified by the manufacturer or by an independent source.
5.4.2 Transfer the stock standard solutions into vials with Teflon-
lined screw caps or crimp tops. Store at 4°C and protect from light.
Stock standards should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards from
them.
5.4.3 Stock standard solutions must be replaced after one year, or
sooner if comparison with check standards indicates a problem.
5.5 Calibration standards: Calibration standards at a minimum of five
concentrations for each parameter of interest are prepared through dilution of
the stock standards with isooctane. One of the concentrations should be at a
concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations found
in real samples or should define the working range of the GC. Calibration
solutions must be replaced after six months, or sooner, if comparison with check
standards indicates a problem.
5.6 Internal standards (if internal standard calibration is used): To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.6.1 Prepare calibration standards at a minimum of five
concentrations for each analyte of interest as described in Sec. 5.5.
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5.6.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with isooctane.
5.6.3 Analyze each calibration standard according to Sec. 7.0.
5.7 Surrogate standards: The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system and the effectiveness
of the method in dealing with each sample matrix by spiking each sample,
standard, and organic-free reagent water blank with pesticide surrogates.
Because GC/ECD data are much more subject to interference than GC/MS, a secondary
surrogate is to be used when sample interference is apparent. Two surrogate
standards (tetrachloro-m-xylene (TCMX) and decachlorobiphenyl) are added to each
sample; however, only one need be calculated for recovery. Proceed with
corrective action when both surrogates are out of limits for a sample (Sec. 8.3).
Method 3500 indicates the proper procedure for preparing these surrogates.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes, Sec.
4.1. Extracts must be stored under refrigeration and analyzed within 40 days of
extraction.
7.0 PROCEDURE
7.1 Extraction:
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a
neutral, or as is, pH with methylene chloride, using either Method 3510 or
3520. Solid samples are extracted using Method 3540, 3541, or 3550.
7.1.2 Prior to gas chromatographic analysis, the extraction solvent
must be exchanged to hexane. The exchange is performed during the K-D
procedures listed in all of the extraction methods. The exchange is
performed as follows.
7.1.2.1 Following K-D of the methylene chloride extract to
1 mL using the macro-Snyder column, allow the apparatus to cool and
drain for at least 10 min.
7.1.2.2 Increase the temperature of the hot water bath to
about 90°C. Momentarily remove the Snyder column, add 50 mL of
hexane, a new boiling chip, and reattach the macro-Snyder column.
Concentrate the extract using 1 mL of hexane to prewet the Snyder
column. Place the K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete concentration in 5-10 min. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
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reaches 1 ml, remove the K-D apparatus and allow it to drain and
cool for at least 10 min.
7.1.2.3 Remove the Snyder column and rinse the flask and
its lower joint into the concentrator tube with 1-2 ml of hexane.
A 5 ml syringe is recommended for this operation. Adjust the
extract volume to 10.0 ml. Stopper the concentrator tube and store
refrigerated at 4°C, if further processing will not be performed
immediately. If the extract will be stored longer than two days, it
should be transferred to a vial with a Teflon-lined screw cap or
crimp top. Proceed with gas chromatographic analysis if further
cleanup is not required.
7.2 Gas chromatography conditions (Recommended):
7.2.1 Column 1:
Carrier gas (5% methane/95% argon) flow rate: 60 mL/min
Column temperature: 200°C isothermal
When analyzing for the low molecular weight PCBs (PCB 1221-PCB
1248), it is advisable to set the oven temperature to 160°C.
7.2.2 Column 2:
Carrier gas (5% methane/95% argon) flow rate: 60 mL/min
Column temperature: 200°C isothermal
When analyzing for the low molecular weight PCBs (PCB 1221-PCB
1248), it is advisable to set the oven temperature to 140°C.
7.2.3 When analyzing for most or all of the analytes in this method,
adjust the oven temperature and column gas flow to provide sufficient
resolution for accurate quantitation of the analytes. This will normally
result in a retention time of 10 to 12 minutes for 4,4'-DDT, depending on
the packed column used.
7.3 Calibration: Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point on
the calibration curve.
7.3.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.3.2 Because of the low concentration of pesticide standards
injected on a GC/ECD, column adsorption may be a problem when the GC has
not been used for a day. Therefore, the GC column should be primed or
deactivated by injecting a PCB or pesticide standard mixture approximately
20 times more concentrated than the mid-concentration standard. Inject
this prior to beginning initial or daily calibration.
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7.4 Gas chromatographic analysis:
7.4.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 /xL of internal standard to the sample prior to
injection.
7.4.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
NOTE: A 72 hour sequence is not required with this method.
7.4.3 Examples of GC/ECD chromatograms for various pesticides and
PCBs are shown in Figures 1 through 5.
7.4.4 Prime the column as per Sec. 7.3.2.
7.4.5 DDT and endrin are easily degraded in the injection port if
the injection port or front of the column is dirty. This is the result of
buildup of high boiling residue from sample injection. Check for
degradation problems by injecting a mid-concentration standard containing
only 4,4'-DDT and endrin. Look for the degradation products of 4,4'-DDT
(4,4'-DDE and 4,4'-ODD) and endrin (endrin ketone and endrin aldehyde).
If degradation of either DDT or endrin exceeds 20%, take corrective action
before proceeding with calibration, by following the GC system maintenance
outlined in of Method 8000. Calculate percent breakdown as follows:
Total DDT degradation peak area (DDE + ODD)
% breakdown = x 100
for 4,4'-DDT Total DDT peak area (DDT + DDE + ODD)
Total endrin degradation peak area
(endrin aldehyde + endrin ketone)
% breakdown = x 100
for Endrin Total endrin peak area (endrin +
endrin aldehyde + endrin ketone)
7.4.6 Record the sample volume injected and the resulting peak sizes
(in area units or peak heights).
7.4.7 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes.
7.4.8 If peak detection and identification are prevented due to
interferences, the hexane extract may need to undergo cleanup using Method
3620. The resultant extract(s) may be analyzed by GC directly or may
undergo further cleanup to remove sulfur using Method 3660.
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'0
7.5 Cleanup:
7.5.1 Proceed with Method 3620, followed by, if necessary, Method
3660, using the 10 ml hexane extracts obtained from Sec. 7.1.2.3.
7.5.2 Following cleanup, the extracts should be analyzed by GC, as
described in the previous sections and in Method 8000.
7.5.3 If only PCBs are to be measured in a sample, the sulfuric
acid/permanganate cleanup (Method 3665), followed by Silica Cleanup
(Method 3630) or Florisil Cleanup (Method 3620), is recommended.
7.6 Calculations (excerpted from U.S. FDA, RAM):
7.6.1 Calculation of Certain Residues: Residues which are mixtures
of two or more components present problems in measurement. When they are
found together, e.g., toxaphene and DDT, the problem of quantitation
becomes even more difficult. In the following sections suggestions are
offered for handling toxaphene, chlordane, PCB, DDT, and BHC. A 10%
DC-200 stationary phase column was used to obtain the chromatograms in
Figures 6-9.
7.6.2 Toxaphene: Quantitative calculation of toxaphene or Strobane
is difficult, but reasonable accuracy can be obtained. To calculate
toxaphene on GC/ECD: (a) adjust sample size so that toxaphene major peaks
are 10-30% full-scale deflection (FSD); (b) inject a toxaphene standard
that is estimated to be within +10 ng of the sample; (c) construct the
baseline of standard toxaphene between its extremities; and (d) construct
the baseline under the sample, using the distances of the peak troughs to
baseline on the standard as a guide (Figures 7, 8, and 9). This procedure
is made difficult by the fact that the relative heights and widths of the
peaks in the sample will probably not be identical to the standard. A
toxaphene standard that has been passed through a Florisil column will
show a shorter retention time for peak X and an enlargement of peak Y.
7.6.3 Toxaphene and DDT: If DDT is present, it will superimpose
itself on toxaphene peak V. To determine the approximate baseline of the
DDT, draw a line connecting the trough of peaks U and V with the trough of
peaks W and X and construct another line parallel to this line which will
just cut the top of peak W (Figure 61). This procedure was tested with
ratios of standard toxaphene-DDT mixtures from 1:10 to 2:1 and the results
of added and calculated DDT and toxaphene by the "parallel lines" method
of baseline construction were within 10% of the actual values in all
cases.
7.6.3.1 A series of toxaphene residues have been
calculated using total peak area for comparison to the standard and
also using area of the last four peaks only in both sample and
standard. The agreement between the results obtained by the two
methods justifies the use of the latter method for calculating
toxaphene in a sample where the early eluting portion of the
toxaphene chromatogram is interfered with by other substances.
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7.6.3.2 The baseline for methoxychlor superimposed on
toxaphene (Figure 8b) was constructed by overlaying the samples on
a toxaphene standard of approximately the same concentration (Figure
8a) and viewing the charts against a lighted background.
7.6.4 Chlordane is a technical mixture of at least 11 major
components and 30 or more minor ones. Gas chromatography-mass
spectrometry and nuclear magnetic resonance analytical techniques have
been applied to the elucidation of the chemical structures of the many
chlordane constituents. Figure 9a is a chromatogram of standard chlor-
dane. Peaks E and F are responses to trans- and cis-chlordane, respec-
tively. These are the two major components of technical chlordane, but
the exact percentage of each in the technical material is not completely
defined and is not consistent from batch to batch. Other labelled peaks
in Figure 9a are thought to represent: A, monochlorinated adduct of
pentachlorocyclopentadiene with cyclopentadiene; B, coelution of
heptachlor and a-chlordene; C, coelution of /3-chlordene and 7-chlordene;
D, a chlordane analog; G, coelution of cis-nonachlor and "Compound K," a
chlordane isomer. The right "shoulder" of peak F is caused by trans-
nonachlor.
7.6.4.1 The GC pattern of a chlordane residue may differ
considerably from that of the technical standard. Depending on the
sample substrate and its history, residues of chlordane can consist
of almost any combination of constituents from the technical
chlordane, plant and/or animal metabolites, and products of
degradation caused by exposure to environmental factors such as
water and sunlight. Only limited information is available on which
residue GC patterns are likely to occur in which samples types, and
even this information may not be applicable to a situation where the
route of exposure is unusual. For example, fish exposed to a recent
spill of technical chlordane will contain a residue drastically
different from a fish whose chlordane residue was accumulated by
ingestion of smaller fish or of vegetation, which in turn had
accumulated residues because chlordane was in the water from
agricultural runoff.
7.6.4.2 Because of this inability to predict a chlordane
residue GC pattern, it is not possible to prescribe a single method
for the quantitation of chlordane residues. The analyst must judge
whether or not the residue's GC pattern is sufficiently similar to
that of a technical chlordane reference material to use the latter
as a reference standard for quantitation.
7.6.4.3 When the chlordane residue does not resemble
technical chlordane, but instead consists primarily of individual,
identifiable peaks, quantitate each peak separately against the
appropriate reference materials and report the individual residues.
(Reference materials are available for at least 11 chlordane
constituents, metabolites or degradation products which may occur in
the residue.)
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7.6.4.4 When the GC pattern of the residue resembles that
of technical chlordane, quantitate chlordane residues by comparing
the total area of the chlordane chromatogram from peaks A through F
(Figure 9a) in the sample versus the same part of the standard
chromatogram. Peak G may be obscured in a sample by the presence of
other pesticides. If G is not obscured, include it in the
measurement for both standard and sample. If the heptachlor epoxide
peak is relatively small, include it as part of the total chlordane
area for calculation of the residue. If heptachlor and/or
heptachlor epoxide are much out of proportion as in Figure 6j,
calculate these separately and subtract their areas from total area
to give a corrected chlordane area. (Note that octachlor epoxide,
a metabolite of chlordane, can easily be mistaken for heptachlor
epoxide on a nonpolar GC column.)
7.6.4.5 To measure the total area of the chlordane
chromatogram, proceed as in Sec. 7.6.2 on toxaphene. Inject an
amount of technical chlordane standard which will produce a
chromatogram in which peaks E and F are approximately the same size
as those in the sample chromatograms. Construct the baseline
beneath the standard from the beginning of peak A to the end of peak
F as shown in Figure 9a. Use the distance from the trough between
peaks E and F to the baseline in the chromatogram of the standard to
construct the baseline in the chromatogram of the sample. Figure 9b
shows how the presence of toxaphene causes the baseline under
chlordane to take an upward angle. When the size of peaks E and F
in standard and sample chromatograms are the same, the distance from
the trough of the peaks to the baselines should be the same.
Measurement of chlordane area should be done by total peak area if
possible.
NOTE: A comparison has been made of the total peak area
integration method and the addition of peak heights
method for several samples containing chlordane. The
peak heights A, B, C, D, E, and F were measured in
millimeters from peak maximum of each to the baseline
constructed under the total chlordane area and were then
added together. These results obtained by the two
techniques are too close to ignore this method of "peak
height addition" as a means of calculating chlordane.
The technique has inherent difficulties because not all
the peaks are symmetrical and not all are present in the
same ratio in standard and in sample. This method does
offer a means of calculating results if no means of
measuring total area is practical.
7.6.5 Polychlorinated biphenyls (PCBs): Quantitation of residues of
PCB involves problems similar to those encountered in the quantitation of
toxaphene, Strobane, and chlordane. In each case, the chemical is made up
of numerous compounds. So the chromatograms are multi-peak. Also in each
case, the chromatogram of the residue may not match that of the standard.
7.6.5.1 Mixtures of PCBs of various chlorine contents were
sold for many years in the U.S. by the Monsanto Co. under the
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tradename Aroclor (1200 series and 1016). Though these Aroclors are
no longer marketed, the PCBs remain in the environment and are
sometimes found as residues in foods, especially fish.
7.6.5.2 PCB residues are quantitated by comparison to one
or more of the Aroclor materials, depending on the chromatographic
pattern of the residue. A choice must be made as to which Aroclor
or mixture of Aroclors will produce a chromatogram most similar to
that of the residue. This may also involve a judgment about what
proportion of the different Aroclors to combine to produce the
appropriate reference material.
7.6.5.3 Quantitate PCB residues by comparing total area or
height of residue peaks to total area of height of peaks from
appropriate Aroclor(s) reference materials. Measure total area or
height response from common baseline under all peaks. Use only
those peaks from the sample that can be attributed to
chlorobiphenyls. These peaks must also be present in the
chromatogram of the reference materials. Mixtures of Aroclors may
be required to provide the best match of GC patterns of sample and
reference.
7.6.6 DDT: DDT found in samples often consists of both o,p'- and
p,p'-DDT. Residues of DDE and ODD are also frequently present. Each
isorner of DDT and its metabolites should be quantitated using the pure
standard of that compound and reported as such.
7.6.7 Hexachlorocyclohexane (BHC, from the former name, benzene
hexachloride): Technical grade BHC is a cream-colored amorphous solid
with a very characteristic musty odor; it consists of a mixture of six
chemically distinct isomers and one or more heptachloro-cyclohexanes and
octachloro-cyclohexanes.
7.6.7.1 Commercial BHC preparations may show a wide
variance in the percentage of individual isomers present. The
elimination rate of the isomers fed to rats was 3 weeks for the a-,
7-, and 5-isomers and 14 weeks for the /3-isomer. Thus it may be
possible to have any combination of the various isomers in different
food commodities. BHC found in dairy products usually has a large
percentage of /3-isomer.
7.6.7.2 Individual isomers (a, 0, 7, and 6) were injected
into gas chromatographs equipped with flame ionization,
microcoulometric, and electron capture detectors. Response for the
four isomers is very nearly the same whether flame ionization or
microcoulometric GLC is used. The a-, 7-, and
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8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the QC
in Method 3600 and in the specific cleanup method.
8.2 Quality control required to evaluate the GC system operation is found
in Method 8000.
8.2.1 The quality control check sample concentrate (Method 8000)
should contain each single-component parameter of interest at the
following concentrations in acetone or other water miscible solvent:
4,4'-DDD, 10 mg/L; 4,4'-DDT, 10 mg/L; endosulfan II, 10 mg/L; endosulfan
sulfate, 10 mg/L; endrin, 10 mg/L; and any other single-component
pesticide, 2 mg/L. If this method is only to be used to analyze for PCBs,
chlordane, or toxaphene, the QC check sample concentrate should contain
the most representative multi-component parameter at a concentration of 50
mg/L in acetone.
8.2.2 Table 3 indicates the QC acceptance criteria for this method.
Table 4 gives method accuracy and precision as functions of concentration
for the analytes of interest. The contents of both Tables should be used
to evaluate a laboratory's ability to perform and generate acceptable data
by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000).
8.3.1 If recovery is not within limits, the following is required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any
of the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above
are a problem or flag the data as "estimated
concentration".
8.4 GC/MS confirmation: Any compounds confirmed by two columns may also
be confirmed by GC/MS if the concentration is sufficient for detection by GC/MS
as determined by the laboratory generated detection limits.
8.4.1 The GC/MS would normally require a minimum concentration of 10
ng/jLtL in the final extract, for each single-component compound.
8.4.2 The pesticide extract and associated blank should be analyzed
by GC/MS as per Sec. 7.0 of Method 8270.
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8.4.3 The confirmation may be from the GC/MS analysis of the
base/neutral-acid extractables extracts (sample and blank). However, if
the compounds are not detected in the base/neutral-acid extract even
though the concentration is high enough, a GC/MS analysis of the pesticide
extract should be performed.
8.4.4 A reference standard of the compound must also be analyzed by
GC/MS. The concentration of the reference standard must be at a level
that would demonstrate the ability to confirm the pesticides/PCBs
identified by GC/ECD.
9.0 METHOD PERFORMANCE
9.1 The method was tested by 20 laboratories using organic-free reagent
water, drinking water, surface water, and three industrial wastewaters spiked at
six concentrations. Concentrations used in the study ranged from 0.5 to 30 |ug/L
for single-component pesticides and from 8.5 to 400 jug/L for multi-component
parameters. Single operator precision, overall precision, and method accuracy
were found to be directly related to the concentration of the parameter and
essentially independent of the sample matrix. Linear equations to describe these
relationships for an electron capture detector are presented in Table 4.
9.2 The accuracy and precision obtained will be determined by the sample
matrix, sample-preparation technique, optional cleanup techniques, and
calibration procedures used.
10.0 REFERENCES
1. U.S. EPA, "Development and Application of Test Procedures for Specific
Organic Toxic Substances in Wastewaters, Category 10: Pesticides and
PCBs," Report for EPA Contract 68-03-2605.
2. U.S. EPA, "Interim Methods for the Sampling and Analysis of Priority
Pollutants in Sediments and Fish Tissue," Environmental Monitoring and
Support Laboratory, Cincinnati, OH 45268, October 1980.
3. Pressley, T.A., and J.E. Longbottom, "The Determination of Organohalide
Pesticides and PCBs in Industrial and Municipal Wastewater: Method 617,"
U.S. EPA/EMSL, Cincinnati, OH, EPA-600/4-84-006, 1982.
4. "Determination of Pesticides and PCB's in Industrial and Municipal
Wastewaters, U.S. Environmental Protection Agency," Environmental
Monitoring and Support Laboratory, Cincinnati, OH 45268, EPA-600/4-82-023,
June 1982.
5. Goerlitz, D.F. and L.M. Law, Bulletin for Environmental Contamination and
Toxicology, 6, 9, 1971.
6. Burke, J.A., "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
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7. Webb, R.G. and A.C. McCall, "Quantitative PCB Standards for Electron
Capture Gas Chromatography," Journal of Chromatographic Science, JJ_, 366,
1973.
8. Millar, J.D., R.E. Thomas and H.J. Schattenberg, "EPA Method Study 18,
Method 608: Organochlorine Pesticides and PCBs," U.S. EPA/EMSL, Research
Triangle Park, NC, EPA-600/4-84-061, 1984.
9. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
11. U.S. Food and Drug Administration, Pesticide Analytical Manual, Vol. 1,
June 1979.
12. Sawyer, L.D., JAOAC, 56, 1015-1023 (1973), 61 272-281 (1978), 61 282-291
(1978).
13. Stewart, J. "EPA Verification Experiment for Validation of the SOXTEC* PCB
Extraction Procedure"; Oak Ridge National Laboratory, Oak Ridge, TN,
37831-6138; October 1988.
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TABLE 1.
GAS CHROMATOGRAPHY OF PESTICIDES AND PCBsa
Analyte
Aldrin
a-BHC
0-BHC
5-BHC
7-BHC (Lindane)
Chlordane (technical)
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Heptachlor
Heptachlor epoxide
Methoxychlor
Toxaphene
PCB-1016
PCB-1221
PCB-1232
PCB-1242
PCB-1248
PCB-1254
PCB-1260
Retention
Col. 1
2.40
1.35
1.90
2.15
1.70
e
7.83
5.13
9.40
5.45
4.50
8.00
14.22
6.55
11.82
2.00
3.50
18.20
e
e
e
e
e
e
e
e
time (min)
Col. 2
4.10
1.82
1.97
2.20
2.13
e
9.08
7.15
11.75
7.23
6.20
8.28
10.70
8.10
9.30
3.35
5.00
26.60
e
e
e
e
e
e
e
e
Method
Detection
limit (Mg/L)
0.004
0.003
0.006
0.009
0.004
0.014
0.011
0.004
0.012
0.002
0.014
0.004
0.066
0.006
0.023
0.003
0.083
0.176
0.24
nd
nd
nd
0.065
nd
nd
nd
aU.S. EPA. Method 617. Organochlorine Pesticides and PCBs.
Monitoring and Support Laboratory, Cincinnati, Ohio 45268.
e = Multiple peak response.
nd = not determined.
Environmental
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TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQLs) FOR VARIOUS MATRICES8
Matrix Factor
Ground water 10
Low-concentration soil by sonication with GPC cleanup 670
High-concentration soil and sludges by sonication 10,000
Non-water miscible waste 100,000
EQL = [Method detection limit (see Table 1)] X [Factor found in this
table]. For non-aqueous samples, the factor is on a wet-weight
basis. Sample EQLs are highly matrix-dependent. The EQLs listed
herein are provided for guidance and may not always be achievable.
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TABLE 3.
QC ACCEPTANCE CRITERIA8
Analyte
Aldrin
a-BHC
0-BHC
6-BHC
7-BHC
Chlordane
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan Sulfate
Endrin
Heptachlor
Heptachlor epoxide
Toxaphene
PCB-1016
PCB-1221
PCB-1232
PCB-1242
PCB-1248
PCB-1254
PCB-1260
Test
cone.
(M9/L)
2.0
2.0
2.0
2.0
2.0
50
10
2.0
10
2.0
2.0
10
10
10
2.0
2.0
50
50
50
50
50
50
50
50
Limit
for s
(M9/L)
0.42
0.48
0.64
0.72
0.46
10.0
2.8
0.55
3.6
0.76
0.49
6.1
2.7
3.7
0.40
0.41
12.7
10.0
24.4
17.9
12.2
15.9
13.8
10.4
Range
for x
(M9/L)
1.08-2.24
0.98-2.44
0.78-2.60
1.01-2.37
0.86-2.32
27.6-54.3
4.8-12.6
1.08-2.60
4.6-13.7
1.15-2.49
1.14-2.82
2.2-17.1
3.8-13.2
5.1-12.6
0.86-2.00
1.13-2.63
27.8-55.6
30.5-51.5
22.1-75.2
14.0-98.5
24.8-69.6
29.0-70.2
22.2-57.9
18.7-54.9
Range
P> PS
(%)
42-122
37-134
17-147
19-140
32-127
45-119
31-141
30-145
25-160
36-146
45-153
D-202
26-144
30-147
34-111
37-142
41-126
50-114
15-178
10-215
39-150
38-158
29-131
8-127
s = Standard deviation of four recovery measurements, in /xg/L.
x = Average recovery for four recovery measurements, in M9/L.
P, Ps = Percent recovery measured.
D
Detected; result must be greater than zero.
"Criteria from 40 CFR Part 136 for Method 608. These criteria are based directly
upon the method performance data in Table 4. Where necessary, the limits for
recovery have been broadened to assure applicability of the limits to
concentrations below those used to develop Table 4.
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TABLE 4.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Analyte
Aldrin
a-BHC
0-BHC
-------
Figure 1
Gas Chromatogram of Pesticides
Column: 1.5% SP-2250+
1.95% SP-2401 on Supelcoport
Temperature: 200°C
Detector: Electron Capture
I 12
WTlNT1ON TIME (MINUTES)
II
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Figure 2
Gas Chromatogram of Chlordane
Column: 1.5% SP-2250+
1.95% SP-2401 on Supelcoport
Temperature: 200°C
Detector: Electron Capture
4 I 12
RCTENTION TlMf (MINUTES)
16
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Figure 3
Gas Chromatogram of Toxaphene
Column: 1.5% SP-2250+
1.95% SP-2401 on Supelcoport
Temperature: 200°C
Detector: Electron Capture
f 10 14 II
NfTtNTION TIM! (MINUTIS)
22
26
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Figure 4
Gas Chromatogram of Aroclor 1254
Column: 1.5% SP-2250+
1.95% SP-2401 on Supelcoport
Temperature: 200°C
Detector: Electron Capture
• 10 14
MITtNTlON TIMI (MINUTES)
It
22
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Figure 5
Gas Chromatogram of Aroclor 1260
Column: 1.5% SP-2250+
1.95% SP-2401 on Supelcoport
Temperature: 200°C
Detector: Electron Capture
10 U II
MITINTION TIMI (MINUTU)
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Figure 6
1 a
J..L
Fig.6--Baseline construction for some typical gas chromotagraphic peaks.
a: symmetrical separated flat baseline; b and c: overlapp flat baseline;
d: separated (pen does not return to baseline between peaks); e: separated
sloping baseline; f: separated (pen goes below baseline between peaks);
g: a- and 7-BHC sloping baseline; h: a-,0- and 7-BHC sloping baseline;
i: chlordane flat baseline; j: heptachlor and heptachlor epoxide super-
imposed on chlordane; k: chair-shaped peaks, unsymmetrical peak;
1: p,p'-DDT superimposed on toxaphene.
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Figure 7
Fig.- 7a -- Baseline construction for multiple residues with standard
toxaphene.
Fig.- 7b -- Baseline construction for multiple residues with toxaphene,
DDE and o,p'-, and p,p'-DDT
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Figure 8
Fig.- 8a -- Baseline construction for multiple residues: standard toxaphene.
Fig.- 8b --
Baseline construction for multiple residues: rice bran with BHC,
toxaphene, DDT, and methoxychlor.
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Figure 9
Fig.- 9a -- Baseline construction for multiple residues: standard chlordane.
Fig.- 9b -- Baseline construction for multiple residues: rice bran with
chlordane, toxaphene, and DDT.
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METHOD 8080A
ORGANOCHLORINE PESTICIDES AND POLYCHLORINATED BIPHENYLS
BY GAS CHROMATOGRAPHY
Start
7.1.1 Choose
appropriate extraction
procedure.
7.3.2 Prime or
deactivate the GC
column prior to
daily calibration.
7.1.2 Exchange
extraction solvent
to hexane.
7.2 Set
chromatographic
conditions.
7.3 Refer to
Method 8000 for
proper calibration
techniques.
7.4 Perform
GC analysis.
7.4.8
is peak
detection and
identification
prevented?
7.6.1 Do
residues have
two or more
components?
7.5.1 Cleanup
using Method 3620
or 3660 if necessary.
7.6 Calculate
concentrations.
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METHOD 8081
ORGANOCHLORINE PESTICIDES AND PCBs AS AROCLORS BY GAS
CHROMATOGRAPHY: CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8081 is used to determine the concentrations of various
organochlorine pesticides and polychlorinated biphenyls (PCBs) as Aroclors, in
extracts from solid and liquid matrices. Open-tubular, capillary columns were
employed with electron capture detectors (ECD) or electrolytic conductivity
detectors (ELCD). When compared to the packed columns, these fused-silica, open-
tubular columns offer improved resolution, better selectivity, increased
sensitivity, and faster analysis. The list below is annotated to show whether a
single- or dual-column analysis system was used to identify each target analyte.
Compound Name
CAS Registry No.
Aldrina-b
Aroclor-1016a'b
Aroclor-1221a'b
Aroclor-1232a'b
Aroclor-1242a'b
Aroclor-1248a-b
Aroclor-1254a-b
Aroclor-1260a'b
a-BHCa'b
j8-BHCa'b
7-BHC (Lindane)a'b
5-BHCa-b
Chlorobenzilateb
a-Chlordaneb
7-Chlordanea'b
DBCPb
4,4'-DDDa-b
4,4'-DDEa'b
4,4'-DDTa'b
Diallateb
Dieldrina'b
Endosulfan Ia-b
Endosulfan IIa-b
Endosulfan sulfatea'b
Endrina'b
Endrin aldehydea'b
Endrin ketoneb
309-00-2
12674-11-2
1104-28-2
11141-16-5
53469-21-9
12672-29-6
11097-69-1
11096-82-5
319-84-6
319-85-7
58-89-9
319-86-8
510-15-6
5103-71-9
5103-74-2
96-12-8
72-54-8
72-55-9
50-29-3
2303-16-4
60-57-1
959-98-8
33213-65-9
1031-07-8
72-20-8
7421-93-4
53494-70-5
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Compound Name CAS Registry No.
Heptachlora'b 76-44-8
Heptachlor epoxidea-b 1024-57-3
Hexachlorobenzeneb 118-74-1
Hexachlorocyclopentadieneb 77-47-4
Isodrin6 465-73-6
Keponeb 143-50-0
Methoxychlora-b 72-43-5
Toxaphenea'b 8001-35-2
Single-column analysis
Dual-column analysis
1.2 The analyst must select columns, detectors and calibration procedures
most appropriate for the specific analytes of interest in a study. Matrix-
specific performance data must be established and the stability of the analytical
system and instrument calibration must be established for each analytical matrix
(e.g., hexane solutions from sample extractions, diluted oil samples, etc.).
1.3 Although performance data are presented for many of the listed
chemicals, it is unlikely that all of them could be determined in a single
analysis. This limitation results because the chemical and chromatographic
behavior of many of these chemicals can result in co-elution. Several
cleanup/fractionation schemes are provided in this method and in Method 3600.
Any chemical is a potential method interference when it is not a target analyte.
1.4 Several multi-component mixtures (i.e., Aroclors and Toxaphene) are
listed as target compounds. When samples contain more than one multi-component
analyte, a higher level of analyst expertise is required to attain acceptable
levels of qualitative and quantitative analysis. The same is true of multi-
component analytes that have been subjected to environmental degradation or
degradation by treatment technologies. These result in "weathered" Aroclors (or
any other multi-component mixtures) that may have significant differences in peak
patterns than those of standards. In these cases, individual congener analyses
may be preferred over total mixture analyses.
1.5 Compound identification based on single column analysis should be
confirmed on a second column, or should be supported by at least one other
qualitative technique. This method describes analytical conditions for a second
gas chromatographic column that can be used to confirm the measurements made with
the primary column. GC/MS Method 8270 is also recommended as a confirmation
technique if sensitivity permits (Sec. 8).
1.6 This method describes a dual column option. The option allows a
hardware configuration of two analytical columns joined to a single injection
port. The option allows one injection to be used for dual column analysis.
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Analysts are cautioned that the dual column option may not be appropriate when
the instrument is subject to mechanical stress, many samples are to be run in a
short period, or when contaminated samples are analyzed.
1.7 This method is restricted to use by or under the supervision of
analysts experienced in the use of a gas chromatograph (GC) and in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
1.8 Extracts suitable for analysis by this method may also be analyzed
for organophosphorus pesticides (Method 8141). Some extracts may also be
suitable for triazine herbicide analysis, if low recoveries (normally samples
taken for triazine analysis must be preserved) are not a problem.
1.9 The following compounds may also be determined using this method:
Compound Name
CAS Registry No.
Alachlora'b
Captafolb
Captanb
Chloronebb
Chloropropylateb
Chlorothalonilb
DCPAb
Dichloneb
Dicofolb
Etridiazoleb
Halowax-1000b
Halowax-1001b
Halowax-1013b
Halowax-1014b
Halowax-1051b
Halowax-1099b
Mirexb
Nitrofenb
PCNBb
Perthaneb
Propachlorb
Strobaneb
tra/?s-Nonachlorb
trans-Permethrinb
Trifluralinb
15972-60-8
2425-06-1
133-06-2
2675-77-6
99516-95-7
1897-45-6
1861-32-1
117-80-6
115-32-2
2593-15-9
58718-66-4
58718-67-5
12616-35-2
12616-36-3
2234-13-1
39450-05-0
2385-85-5
1836-75-5
82-68-8
72-56-0
1918-16-17
8001-50-1
39765-80-5
51877-74-8
1582-09-8
Single-column analysis
Dual-column analysis
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2.0 SUMMARY OF METHOD
2.1 A measured volume or weight of sample (approximately 1 L for 1 iquids,
2 g to 30 g for solids) is extracted using the appropriate sample extraction
technique. Liquid samples are extracted at neutral pH with methylene chloride
using either a separatory funnel (Method 3510) or a continuous liquid-liquid
extractor (Method 3520). Solid samples are extracted with hexane-acetone (1:1)
or methylene chloride-acetone (1:1) using either Soxhlet extraction (Method
3540), Automated Soxhlet (Method 3541), or Ultrasonic Extraction (Method 3550).
A variety of cleanup steps may be applied to the extract, depending on (1) the
nature of the coextracted matrix interferences and (2) the target analytes.
After cleanup, the extract is analyzed by injecting a 1-juL sample into a gas
chromatograph with a narrow- or wide-bore fused silica capillary column and
electron capture detector (GC/ECD) or an electrolytic conductivity detector
(GC/ELCD).
3.0 INTERFERENCES
3.1 Refer to Methods 3500 (Sec. 3, in particular), 3600, and 8000.
3.2 Sources of interference in this method can be grouped into three
broad categories: contaminated solvents, reagents or sample processing hardware;
contaminated GC carrier gas, parts, column surfaces or detector surfaces; and the
presence of coeluting compounds in the sample matrix to which the ECD will
respond. Interferences coextracted from the samples will vary considerably from
waste to waste. While general cleanup techniques are referenced or provided as
part of this method, unique samples may require additional cleanup approaches to
achieve desired degrees of discrimination and quantitation.
3.3 Interferences by phthalate esters introduced during sample
preparation can pose a major problem in pesticide determinations. These
materials may be removed prior to analysis using Gel Permeation Cleanup -
pesticide option (Method 3640) or as Fraction III of the silica gel cleanup
procedure (Method 3630). Common flexible plastics contain varying amounts of
phthalate esters which are easily extracted or leached from such materials during
laboratory operations. Cross-contamination of clean glassware routinely occurs
when plastics are handled during extraction steps, especially when solvent-wetted
surfaces are handled. Interferences from phthalate esters can best be minimized
by avoiding contact with any plastic materials and checking all solvents and
reagents for phthalate contamination. Exhaustive cleanup of solvents, reagents
and glassware may be required to eliminate background phthalate ester
contamination.
3.4 Glassware must be scrupulously cleaned. Clean all glassware as soon
as possible after use by rinsing with the last solvent used. This should be
followed by detergent washing with hot water, and rinses with tap water and
organic-free reagent water. Drain the glassware and dry in an oven at 130°C for
several hours or rinse with methanol and drain. Store dry glassware in a clean
environment.
3.5 The presence of elemental sulfur will result in broad peaks that
interfere with the detection of early-eluting organochlorine pesticides. Sulfur
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contamination should be expected with sediment samples. Method 3660 is suggested
for removal of sulfur. Since the recovery of Endrin aldehyde (using the TBA
procedure) is drastically reduced, this compound must be determined prior to
sulfur cleanup.
3.6 Waxes, lipids, and other high molecular weight co-extractables can
be removed by Gel-Permeation Cleanup (Method 3640).
3.7 It may be difficult to quantitate Aroclor patterns and single
component pesticides together. Some pesticides can be removed by sulfuric
acid/permanganate cleanup (Method 3665) and silica fractionation (Method 3630).
Guidance on the identification of PCBs is given in Sec. 7.
3.8 The following target analytes coelute using single column analysis:
DB 608 Trifluralin/Dial late isomers
PCNP/Dichlone/Isodrin
DDD/Endosulfan II
DB 1701 Captan/Chlorobenzilate
Captafol/Mirex
DDD/Endosulfan II
Methoxychlor/Endosulfan sulfate
3.8.1 Other halogenated pesticides or industrial chemicals may
interfere with the analysis of pesticides. Certain co-eluting
organophosphorus pesticides are eliminated by the Gel Permeation
Chromatography cleanup - pesticide option (Method 3640). Co-eluting
chlorophenols are eliminated by Silica gel (Method 3630), Florisil (Method
3620), or Alumina (Method 3610) cleanup.
3.9 The following compounds coelute using the dual column analysis. Two
temperature programs are provided for the same pair of columns as option 1 and
option 2 for dual column analysis. In general, the DB-5 column resolves fewer
compounds that the DB-1701:
3.9.1 DB-5/DB-1701, thin film, slow ramp: See Sec. 7 and Table 6.
DB-5 trans-Permethrin/Heptachlor epoxide
Endosulfan I/a-Chlordane
Perthane/Endrin
Endosulfan II/Chloropropylate/Chiorobenzi1 ate
4,4'-DDT/Endosulfan sulfate
Methoxychlor/Di cofol
Perthane/Endrin and Chiorobenzilate/Endosulfan II/Chloropropylate
will also co-elute on DB-5 after moderate deterioration in column
performance.
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DB-1701 Chlorothalonil/B-BHC
(5-BHC/DCPA/trans-Permethrin
or-Chlordane/trans-Nonachlor
Captan/Dieldrin
Chiorobenzi1 ate/Chioropropylate
Chlorothalonil/B-BHC and or-Chlordane/trans-Nonachlor will co-elute
on the DB-1701 column after moderate deterioration in column performance.
Nitrofen, Dichlone, Carbophenothion, Dichloran and Kepone were
removed from the composite mixture because of extensive peak tailing on
both columns. Simazine and Atrazine give poor responses on the ECD
detector. Triazine compounds should be analyzed using Method 8141 (NPD
option).
3.9.2 DB-5/DB-1701, thick film, fast ramp: See Sec. 7 and Table 7.
DB-5 Diallate/a-BHC
Perthane/Endosulfan II
Chiorobenzi1 ate/Chioropropylate
Endrin/Nitrofen
4,4'-DDT/Endosulfan sulfate
Methoxychlor/Di colfol
DB-1701 or-Chl ordane/trans-Nonachl or (partially resolved)
4,4'-DDD/Endosulfan II (partially resolved)
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph: an analytical system complete with gas
chromatograph suitable for on-column and split-splitless injection and all
required accessories including syringes, analytical columns, gases, electron
capture detectors (ECD), and recorder/integrator or data system.
The columns listed in this section were used to develop the method
performance data. Their specification is not intended to prevent laboratories
from using columns that are developed after promulgation of the method.
Laboratories may use other capillary columns if they document method performance
data (e.g. chromatographic resolution, analyte breakdown, and MDLs) equal to or
better than those provided with the method.
4.1.1 Single-column Analysis:
4.1.1.1 Narrow-bore columns:
4.1.1.1.1 Column 1 - 30 m x 0.25 or 0.32 mm internal
diameter (ID) fused silica capillary column chemically bonded
with SE-54 (DB 5 or equivalent), 1 /im film thickness.
4.1.1.1.2 Column 2 - 30 m x 0.25 mm ID fused silica
capillary column chemically bonded with 35 percent phenyl
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methylpolysiloxane (DB 608, SPB 608, or equivalent), 25 /urn
coating thickness, 1 /zm film thickness.
4.1.1.1.3 Narrow bore columns should be installed in
split/splitless (Grob-type) injectors.
4.1.1.2 Wide-bore columns
4.1.1.2.1 Column 1 - 30 m x 0.53 mm ID fused silica
capillary column chemically bonded with 35 percent phenyl
methylpolysiloxane (DB 608, SPB 608, RTx-35, or equivalent),
0.5 urn or 0.83 jum film thickness.
4.1.1.2.2 Column 2 - 30 m x 0.53 mm ID fused silica
capillary column chemically bonded with 50 percent phenyl
methylpolysiloxane (DB 1701, or equivalent), 1.0 jum film
thickness.
4.1.1.2.3 Column 3 - 30 m x 0.53 mm ID fused silica
capillary column chemically bonded with SE-54 (DB 5, SPB 5,
RTx5, or equivalent), 1.5 /xm film thickness.
4.1.1.2.4 Wide-bore columns should be installed in 1/4
inch injectors, with deactivated liners designed specifically
for use with these columns.
4.1.2 Dual Column Analysis:
4.1.2.1 Column pair 1:
4.1.2.1.1 J&W Scientific press-fit Y-shaped glass 3-
way union splitter (J&W Scientific, Catalog no. 705-0733) or
Restek Y-shaped fused-silica connector (Restek, Catalog no.
20405), or equivalent.
4.1.2.1.2 30 m x 0.53 m ID DB-5 (J&W Scientific),
1.5 /Lim film thickness, or equivalent.
4.1.2.1.3 30 m x 0.53 mm ID DB-1701 (J&W Scientific),
1.0 jum film thickness, or equivalent.
4.1.2.2 Column pair 2:
4.1.2.2.1 Splitter 2 - Supelco 8 in. glass injection
tee, deactivated (Supelco, Catalog no. 2-3665M), or
equivalent.
4.1.2.2.2 30 m x 0.53 m ID DB-5 (J&W Scientific),
0.83 /Ltm film thickness, or equivalent.
4.1.2.2.3 30 m x 0.53 mm ID DB-1701 (J&W Scientific),
1.0 jum film thickness, or equivalent.
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4.1.3 Column rinsing kit: Bonded-phase column rinse kit (J&W
Scientific, Catalog no. 430-3000 or equivalent).
4.2 Glassware (see Methods 3510, 3520, 3540, 3541, 3550, 3630, 3640,
3660, and 3665 for specifications).
4.3 Kuderna-Danish (K-D) apparatus. See extraction methods for specifics.
5.0 REAGENTS
5.1 Reagent or pesticide grade chemicals shall be used in all tests.
Unless otherwise indicated, it is intended that all reagents shall conform to
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
NOTE: Store the standard solutions (stock, composite, calibration,
internal, and surrogate) at 4°C in Teflon-sealed containers in the
dark. When a lot of standards is prepared, it is recommended that
aliquots of that lot be stored in individual small vials. All stock
standard solutions must be replaced after one year or sooner if
routine QC (Sec. 8) indicates a problem. All other standard
solutions must be replaced after six months or sooner if routine QC
(Sec. 8) indicates a problem.
5.2 Solvents and reagents: As appropriate for Method 3510, 3520, 3540,
3541, 3550, 3630, 3640, 3660, or 3665: n-hexane, diethyl ether, methylene
chloride, acetone, ethyl acetate, and isooctane (2,2,4-trimethylpentane). All
solvents should be pesticide quality or equivalent, and each lot of solvent
should be determined to be phthalate free. Solvents must be exchanged to hexane
or isooctane prior to analysis.
5.2.1 Organic-free reagent water: All references to water in this
method refer to organic-free reagent water as defined in Chapter One.
5.3 Stock standard solutions (1000 mg/L): Can be prepared from pure
standard materials or can be purchased as certified solutions.
5.3.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure compound. Dissolve the compound in isooctane or hexane
and dilute to volume in a 10-mL volumetric flask. If compound purity is
96 percent or greater, the weight can be used without correction to
calculate the concentration of the stock standard solution. Commercially
prepared stock standard solutions can be used at any concentration if they
are certified by the manufacturer or by an independent source.
5.3.2 6-BHC, Dieldrin, and some other standards may not be
adequately soluble in isooctane. A small amount of acetone or toluene
should be used to dissolve these compounds during the preparation of the
stock standard solutions.
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5.4 Composite stock standard: Can be prepared from individual stock
solutions. For composite stock standards containing less than 25 components,
take exactly 1 ml of each individual stock solution at 1000 mg/L, add solvent,
and mix the solutions in a 25-mL volumetric flask. For example, for a composite
containing 20 individual standards, the resulting concentration of each component
in the mixture, after the volume is adjusted to 25 ml, will be 1 mg/25 ml. This
composite solution can be further diluted to obtain the desired concentrations.
For composite stock standards containing more than 25 components, use volumetric
flasks of the appropriate volume (e.g., 50 ml, 100 ml).
5.5 Calibration standards should be prepared at a minimum of five
concentrations by dilution of the composite stock standard with isooctane or
hexane. The concentrations should correspond to the expected range of
concentrations found in real samples and should bracket the linear range of the
detector.
5.5.1 Although all single component analytes can be resolved on a
new 35 percent phenyl methyl silicone column (e.g., DB-608), two
calibration mixtures should be prepared for the single component analytes
of this method.
5.5.2 This procedure is established to (1) minimize potential
resolution and quantitation problems on confirmation columns or on older
35 percent phenyl methyl silicone (e.g. DB-608) columns and (2) allow
determination of Endrin and DDT breakdown for method QC (Sec. 8).
5.5.3 Separate calibration standards are required for each multi-
component target analyte, with the exception of Aroclors 1016 and 1260,
which can be run as a mixture.
5.6 Internal standard (optional):
5.6.1 Pentachloronitrobenzene is suggested as an internal standard
for the single column analysis, when it is not considered to be a target
analyte. l-Bromo-2-nitrobenzene is a suggested option. Prepare the
standard to complement the concentrations found in Sec. 5.5.
5.6.2 Make a solution of 1000 mg/L of l-bromo-2-nitrobenzene for
dual-column analysis. Dilute it to 500 ng/^L for spiking, then use a
spiking volume of 10 /xL/mL of extract.
5.7 Surrogate standards: The performance of the method should be
monitored using surrogate compounds. Surrogate standards are added to all
samples, method blanks, matrix spikes, and calibration standards.
5.7.1 For the single column analysis, use decachlorobiphenyl as the
primary surrogate. However, if recovery is low, or late-eluting compounds
interfere with decachlorobiphenyl, then tetrachloro-m-xylene should be
evaluated as a surrogate. Proceed with corrective action when both
surrogates are out of limits for a sample (Sec. 8.2). Method 3500, Sec.
5, indicates the proper procedure for preparing these surrogates.
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5.7.2 For the dual column analysis make a solution of 1000 mg/L of
4-chloro-3-nitrobenzotrifluoride and dilute to 500 ng//iL. Use a spiking
volume of 100 pi for a 1 L aqueous sample. Store the spiking solutions
at 4°C in Teflon-sealed containers in the dark.
6.0 SAMPLE COLLECTION, PRESERVATION AND HANDLING
6.1 See Chapter 4, Organic Analytes, Sec. 4.
6.2 Extracts must be stored under refrigeration in the dark and analyzed
within 40 days of extraction.
7.0 PROCEDURE
7.1 Extraction:
7.1.1 Refer to Chapter Two and Method 3500 for guidance in choosing
the appropriate extraction procedure. In general, water samples are
extracted at a neutral pH with methylene chloride as a solvent using a
separatory funnel (Method 3510) or a continuous liquid-liquid extractor
(Method 3520). Extract solid samples with hexane-acetone (1:1) using one
of the Soxhlet extraction (Method 3540 or 3541) or ultrasonic extraction
(Method 3550) procedures.
NOTE: Hexane/acetone (1:1) may be more effective as an extraction
solvent for organochlorine pesticides and PCBs in some
environmental and waste matrices than is methylene
chloride/acetone (1:1). Use of hexane/acetone generally
reduces the amount of co-extracted interferences and improves
signal/noise.
7.1.2 Spiked samples are used to verify the applicability of the
chosen extraction technique to each new sample type. Each sample type
must be spiked with the compounds of interest to determine the percent
recovery and the limit of detection for that sample (Sec. 5). See Method
8000 for guidance on demonstration of initial method proficiency as well
as guidance on matrix spikes for routine sample analysis.
7.2 Cleanup/Fractionation:
7.2.1 Cleanup procedures may not be necessary for a relatively clean
sample matrix, but most extracts from environmental and waste samples will
require additional preparation before analysis. The specific cleanup
procedure used will depend on the nature of the sample to be analyzed and
the data quality objectives for the measurements. General guidance for
sample extract cleanup is provided in this section and in Method 3600.
7.2.1.1 If a sample is of biological origin, or contains
high molecular weight materials, the use of GPC cleanup/pesticide
option (Method 3640) is recommended. Frequently, one of the
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adsorption chromatographic cleanups may also be required following
the GPC cleanup.
7.2.1.2 If only PCBs are to be measured in a sample, the
sulfuric acid/permanganate cleanup (Method 3665) is recommended.
Additional cleanup/fractionation by Alumina Cleanup (Method 3610),
Silica-Gel Cleanup (Method 3630), or Florisil Cleanup (Method 3620),
may be necessary.
7.2.1.3 If both PCBs and pesticides are to be measured in
the sample, isolation of the PCB fraction by Silica Cleanup (Method
3630) is recommended.
7.2.1.4 If only pesticides are to be measured, cleanup by
Method 3620 or Method 3630 is recommended.
7.2.1.5 Elemental sulfur, which may appear in certain
sediments and industrial wastes, interferes with the electron
capture gas chromatography of certain pesticides. Sulfur should be
removed by the technique described in Method 3660, Sulfur Cleanup.
7.3 GC Conditions: This method allows the analyst to choose between
a single column or a dual column configuration in the injector port. Either
wide- or narrow-bore columns may be used. Identifications based on retention
times from a single column must be confirmed on a second column or with an
alternative qualitative technique.
7.3.1 Single Column Analysis:
7.3.1.1 This capillary GC/ECD method allows the analyst
the option of using 0.25-0.32 mm ID capillary columns (narrow-bore)
or 0.53 mm ID capillary columns (wide-bore). Performance data are
provided for both options. Figures 1-6 provide example
chromatograms.
7.3.1.2 The use of narrow-bore columns is recommended when
the analyst requires greater chromatographic resolution. Use of
narrow-bore columns is suitable for relatively clean samples or for
extracts that have been prepared with one or more of the clean-up
options referenced in the method. Wide-bore columns (0.53 mm) are
suitable for more complex environmental and waste matrices.
7.3.1.3 For the single column method of analysis, using
wide-bore capillary columns, Table 1 lists average retention times
and method detection limits (MDLs) for the target analytes in water
and soil matrices. For the single column method of analysis, using
narrow-bore capillary columns, Table 2 lists average retention times
and method detection limits (MDLs) for the target analytes in water
and soil matrices. The MDLs for the components of a specific sample
may differ from those listed in Tables 1 and 2 because they are
dependent upon the nature of interferences in the sample matrix.
Table 3 lists the Estimated Quantitation Limits (EQLs) for other
matrices. Table 4 lists the GC operating conditions for the single
column method of analysis.
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7.3.2 Dual Column Analysis:
7.3.2.1 The dual-column/dual-detector approach involves
the use of two 30 m x 0.53 mm ID fused-silica open-tubular columns
of different polarities, thus different selectivities towards the
target compounds. The columns are connected to an injection tee and
ECD detectors. Retention times for the organochlorine analytes on
dual columns are in Table 5. The GC operating conditions for the
compounds in Table 5 are in Table 6. Multicomponent mixtures of
Toxaphene and Strobane were analyzed separately (Figures 7 and 8)
using the GC operating conditions found in Table 7. Seven Aroclor
mixtures and six Halowax mixtures were analyzed under the conditions
outlined in Table 7 (Figures 9 through 21). Figure 22 is a sample
chromatogram for a mixture of organochlorine pesticides. The
retention times of the individual components detected in these
mixtures are given in Tables 8 and 9.
7.3.2.1.1 Operating conditions for a more heavily
loaded DB-5/DB-1701 pair are given in Table 7. This column
pair was used for the detection of multi component
organochlorine compounds.
7.3.2.1.2 Operating conditions for a DB-5/DB-1701
column pair with thinner films, a different type of splitter,
and a slower temperature programming rate are provided in
Table 6. These conditions gave better peak shapes for
compounds such as Nitrofen and Dicofol. Table 5 lists the
retention times for the compounds detected on this column
pair.
7.4 Calibration:
7.4.1 Prepare calibration standards using the procedures in Sec. 5.
Refer to Method 8000 (Sec. 7) for proper calibration techniques for both
initial calibration and calibration verification. The procedure for
either internal or external calibration may be used, however, in most
cases external standard calibration is used with Method 8081. This is
because of the sensitivity of the electron capture detector and the
probability of the internal standard being affected by interferences.
Because several of the pesticides may co-elute on any single column,
analysts should use two calibration mixtures (see Sec. 3.8). The specific
mixture should be selected to minimize the problem of peak overlap.
NOTE: Because of the sensitivity of the electron capture detector,
the injection port and column should always be cleaned prior
to performing the initial calibration.
7.4.1.1 Method 8081 has many multi-component target
analytes. For this reason, the target analytes chosen for
calibration should be limited to those specified in the project
plan. For instance, some sites may require analysis for the
organochlorine pesticides only or the PCBs only. Toxaphene and/or
technical Chlordane may not be specified at certain sites. In
addition, where PCBs are specified in the project plan, a mixture of
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Aroclors 1016 and 1260 will suffice for the initial calibration of
all Aroclors, since they include all congeners present in the
different regulated Aroclors. A mid-point calibration standard of
all Aroclors (for Aroclor pattern recognition) must be included with
the initial calibration so that the analyst is familiar with each
Aroclor pattern and retention times on each column.
7.4.1.2 For calibration verification (each 12 hr shift)
all target analytes required in the project plan must be injected
with the following exception for the Aroclors. For sites that
require PCB analysis, include only the Aroclors that are expected to
be found at the site. If PCBs are required, but it is unknown which
Aroclors may be present, the mid-concentration Aroclors 1016/1260
mixture only, may be injected. However, if specific Aroclors are
found at the site during the initial screening, it is required that
the samples containing Aroclors be reinjected with the proper mid-
concentration Aroclor standards.
7.4.2 Because of the low concentration of pesticide standards
injected on a GC/ECD, column adsorption may be a problem when the GC has
not been used for a day or more. Therefore, the GC column should be
primed or deactivated by injecting a PCB or pesticide standard mixture
approximately 20 times more concentrated than the mid-concentration
standard. Inject this standard mixture prior to beginning the initial
calibration or calibration verification.
CAUTION: Several analytes, including Aldrin, may be observed in
the injection just following this system priming.
Always run an acceptable blank prior to running any
standards or samples.
7.4.3 Retention time windows:
7.4.3.1 Before establishing the retention time windows,
make sure the gas chromatographic system is within optimum operating
conditions. The width of the retention time window should be based
upon actual retention times of standards measured over the course of
72 hours. See Method 8000 for details.
7.4.3.2 Retention time windows shall be defined as plus or
minus three times the standard deviation of the absolute retention
times for each standard. However, the experience of the analyst
should weigh heavily in the interpretation of the chromatograms.
For multicomponent standards (i.e., PCBs), the analyst should use
the retention time window but should primarily rely on pattern
recognition. Sec. 7.5.4 provides guidance on the establishment of
absolute retention time windows.
7.4.3.3 Certain analytes, particularly Kepone, are subject
to changes in retention times. Dry Kepone standards prepared in
hexane or isooctane can produce gaussian peaks. However, Kepone
extracted from samples or standards exposed to water or methanol may
produce peaks with broad tails that elute later than the standard
(0-1 minute). This shift is presumably the result of the formation
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of a hemi-acetal from the ketone functionality. Method 8270 is
recommended for Kepone.
7.5 Gas chromatographic analysis:
7.5.1 Set up the GC system using the conditions described in Tables
4, 6, or 7. An initial oven temperature at or below 140-150°C is required
to resolve the four BHC isomers. A final temperature of 240-270°C is
required to elute decachlorobiphenyl. Use of injector pressure
programming will improve the chromatography of late eluting peaks.
7.5.2 Verify calibration each 12 hour shift by injecting calibration
verification standards prior to conducting any analyses. See Sec. 7.4.1.2
for special guidance on calibration verification of PCBs. A calibration
standard must also be injected at intervals of not less than once every
twenty samples (after every 10 samples is recommended to minimize the
number of samples requiring re-injection when QC limits are exceeded) and
at the end of the analysis sequence. The calibration factor for each
analyte to be quantitated must not exceed a ±15 percent difference when
compared to the initial calibration curve. When this criterion is
exceeded, inspect the gas chromatographic system to determine the cause
and perform whatever maintenance is necessary before verifying calibration
and proceeding with sample analysis. If routine maintenance does not
return the instrument performance to meet the QC requirements (Sec. 8.2)
based on the last initial calibration, then a new initial calibration must
be performed.
7.5.2.1 Analysts should use high and low concentrations of
mixtures of single-component analytes and multi-component analytes
for calibration verification.
7.5.3 Sample injection may continue for as long as the calibration
verification standards and standards interspersed with the samples meet
instrument QC requirements. It is recommended that standards be analyzed
after every 10 (required after every 20 samples), and at the end of a set.
The sequence ends when the set of samples has been injected or when
qualitative and/or quantitative QC criteria are exceeded.
7.5.3.1 Each sample analysis must be bracketed with an
acceptable initial calibration, calibration verification standard(s)
(each 12 hr shift), or calibration standards interspersed within the
samples. All samples that were injected after the standard that
last met the QC criteria must be reinjected.
7.5.3.2 Although analysis of a single mid-concentration
standard (standard mixture or multi-component analyte) will satisfy
the minimum requirements, analysts are urged to use different
calibration verification standards during organochlorine
pesticide/PCB analyses. Also, multi-level standards (mixtures or
multi-component analytes) are highly recommended to ensure that
detector response remains stable for all analytes over the
calibration range.
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7.5.4 Establish absolute retention time windows for each analyte.
Use the absolute retention time for each analyte from standards analyzed
during that 12 hour shift as the midpoint of the window. The daily
retention time window equals the midpoint + three times the standard
deviations.
7.5.4.1 Tentative identification of an analyte occurs when
a peak from a sample extract falls within the daily retention time
window.
7.5.4.2 Validation of gas chromatographic system
qualitative performance: Use the calibration standards analyzed
during the sequence to evaluate retention time stability. If any of
the standards fall outside their daily retention time windows, the
system is out of control. Determine the cause of the problem and
correct it.
7.5.5 Record the volume injected to the nearest 0.05 juL and the
resulting peak size in area units. Using either the internal or the
external calibration procedure (Method 8000), determine the identity and
the quantity of each component peak in the sample chromatogram which
corresponds to the compounds used for calibration purposes.
7.5.5.1 If the responses exceed the calibration range of
the system, dilute the extract and reanalyze. Peak height
measurements are recommended over peak area integration when
overlapping peaks cause errors in area integration.
7.5.5.2 If partially overlapping or coeluting peaks are
found, change columns or try GC/MS quantitation, see Sec. 8 and
Method 8270.
7.5.5.3 If the peak response is less than 2.5 times the
baseline noise level, the validity of the quantitative result may be
questionable. The analyst should consult with the source of the
sample to determine whether further concentration of the sample is
warranted.
7.5.6 Identification of mixtures (i.e. PCBs and Toxaphene) is based
on the characteristic "fingerprint" retention time and shape of the
indicator peak(s); and quantitation is based on the area under the
characteristic peaks as compared to the area under the corresponding
calibration peak(s) of the same retention time and shape generated using
either internal or external calibration procedures.
7.5.7 Quantitation of the target compounds is based on: 1) a
reproducible response of the ECD or ELCD within the calibration range; and
2) a direct proportionality between the magnitude of response of the
detector to peaks in the sample extract and the calibration standards.
Proper quantitation requires the appropriate selection of a baseline from
which the area or height of the characteristic peak(s) can be determined.
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7.5.8 If compound identification or quantitation is precluded due to
interference (e.g., broad, rounded peaks or ill-defined baselines are
present) cleanup of the extract or replacement of the capillary column or
detector is warranted. Rerun the sample on another instrument to
determine if the problem results from analytical hardware or the sample
matrix. Refer to Method 3600 for the procedures to be followed in sample
cleanup.
7.6 Quantitation of Multiple Component Analytes:
7.6.1 Multi-component analytes present problems in measurement.
Suggestions are offered in the following sections for handing Toxaphene,
Chlordane, PCBs, DDT, and BHC.
7.6.2 Toxaphene: Toxaphene is manufactured by the chlorination of
camphenes, whereas Strobane results from the chlorination of a mixture of
camphenes and pinenes. Quantitative calculation of Toxaphene or Strobane
is difficult, but reasonable accuracy can be obtained. To calculate
Toxaphene on GC/ECD: (a) adjust the sample size so that the major
Toxaphene peaks are 10-70% of full-scale deflection (FSD); (b) inject a
Toxaphene standard that is estimated to be within +10 ng of the sample;
(c) quantitate using the five major peaks or the total area of the
Toxaphene pattern.
7.6.2.1 To measure total area, construct the baseline of
standard Toxaphene between its extremities; and construct the
baseline under the sample, using the distances of the peak troughs
to baseline on the standard as a guide. This procedure is made
difficult by the fact that the relative heights and widths of the
peaks in the sample will probably not be identical to the standard.
7.6.2.2 A series of Toxaphene residues have been
calculated using the total peak area for comparison to the standard
and also using the area of the last four peaks only, in both sample
and standard. The agreement between the results obtained by the two
methods justifies the use of the latter method for calculating
Toxaphene in a sample where the early eluting portion of the
Toxaphene chromatogram shows interferences from other substances
such as DDT.
7.6.3 Chlordane is a technical mixture of at least 11 major
components and 30 or more minor components. Trans- and c/s-Chlordane (a
and 7, respectively), are the two major components of technical Chlordane.
However, the exact percentage of each in the technical material is not
completely defined, and is not consistent from batch to batch.
7.6.3.1 The GC pattern of a Chlordane residue may differ
considerably from that of the technical standard. Depending on the
sample substrate and its history, residues of Chlordane can consist
of almost any combination of: constituents from the technical
Chlordane, plant and/or animal metabolites, and products of
degradation caused by exposure to environmental factors such as
water and sunlight.
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7.6.3.2 Whenever possible, when a Chlordane residue does
not resemble technical Chlordane, the analyst should quantitate the
peaks of ff-Chlordane, 7-Chlordane, and Heptachlor separately against
the appropriate reference materials, and report the individual
residues.
7.6.3.3 When the GC pattern of the residue resembles that
of technical Chlordane, the analyst may quantitate Chlordane
residues by comparing the total area of the Chlordane chromatogram
using the five major peaks or the total area. If the Heptachlor
epoxide peak is relatively small, include it as part of the total
Chlordane area for calculation of the residue. If Heptachlor and/or
Heptachlor epoxide are much out of proportion, calculate these
separately and subtract their areas from the total area to give a
corrected Chlordane area. (Note that octachloro epoxide, a
metabolite of Chlordane, can easily be mistaken for Heptachlor
epoxide on a nonpolar GC column.)
7.6.3.4 To measure the total area of the Chlordane
chromatogram, inject an amount of technical Chlordane standard which
will produce a chromatogram in which the major peaks are
approximately the same size as those in the sample chromatograms.
7.6.4 Polychlorinated biphenyls (PCBs): Quantitation of residues of
PCBs involves problems similar to those encountered in the quantitation of
Toxaphene, Strobane, and Chlordane. In each case, the material is made up
of numerous compounds which generate multi-peak chromatograms. Also, in
each case, the chromatogram of the residue may not .match that of the
standard.
7.6.4.1 Mixtures of PCBs of various chlorine contents were
sold for many years in the U.S. by the Monsanto Co. under the trade
name Aroclor (1200 series and 1016). Although these Aroclors are no
longer marketed, the PCBs remain in the environment and are
sometimes found as residues in foods, especially fish. The Aroclors
most commonly found in the environment are 1242, 1254, and 1260.
7.6.4.2 PCB residues are generally quantitated by
comparison to the most similar Aroclor standard. A choice must be
made as to which Aroclor is most similar to that of the residue and
whether that standard is truly representative of the PCBs in the
sample.
7.6.4.3 PCB Quantitation option #1- Quantitate the PCB
residues by comparing the total area of the chlorinated biphenyl
peaks to the total area of peaks from the appropriate Aroclor
reference material. Measure the total area or height response from
the common baseline under all the peaks. Use only those peaks from
the sample that can be attributed to chlorobiphenyls. These peaks
must also be present in the chromatogram of the reference materials.
Option #1 should not be used if there are interference peaks within
the Aroclor pattern, especially if they overlap PCB congeners.
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7.6.4.4 PCB Quantitation option #2- Quantitate the PCB
residues by comparing the responses of 3 to 5 major peaks in each
appropriate Aroclor standard with the peaks obtained from the
chlorinated biphenyls in the sample extract. The amount of Aroclor
is calculated using an individual response factor for each of the
major peaks. The results of the 3 to 5 determinations are averaged.
Major peaks are defined as those peaks in the Aroclor standards that
are at least 25% of the height of the largest Aroclor peak. Late-
eluting Aroclor peaks are generally the most stable in the
environment.
7.6.4.5 When samples appear to contain weathered PCBs,
treated PCBs, or mixtures of Aroclors, the use of Aroclor standards
is not appropriate. Several diagnostic peaks useful for identifying
non-Aroclor PCBs are given in Table 10. Analysts should examine
chromatograms containing these peaks carefully, as these samples may
contain PCBs. PCB concentrations may be estimated from specific
congeners by adding the concentration of the congener peaks listed
in Table 11. The congeners are analyzed as single components. This
approach will provide reasonable accuracy for Aroclors 1016, 1232,
1242 and 1248 but will underestimate the concentrations of Aroclors
1254, 1260 and 1221. It is highly recommended that heavily
weathered, treated, or mixed Aroclors be analyzed using GC/MS if
concentration permits.
7.6.5 Hexachlorocyclohexane (BHC, from the former name, benzene
hexachloride): Technical grade BHC is a cream-colored amorphous solid
with a very characteristic musty odor; it consists of a mixture of six
chemically distinct isomers and one or more heptachlorocyclohexanes and
octachlorocyclohexanes. Commercial BHC preparations may show a wide
variance in the percentage of individual isomers present. Quantitate
each isomer (a, |8, 7, and 5) separately against a standard of the
respective pure isomer.
7.6.6 DDT: Technical DDT consists primarily of a mixture of 4,4'-
DDT (approximately 75%) and 2,4'-DDT (approximately 25%). As DDT
weathers, 4,4'-DDE, 2,4'-DDE, 4,4'-DDD, and 2,4'-DDD are formed. Since
the 4,4'-isomers of DDT, DDE, and ODD predominate in the environment,
these are the isomers normally regulated by US EPA and should be
quantitated against standards of the respective pure isomer.
7.7 Suggested chromatography maintenance: Corrective measures may require
any one or more of the following remedial actions.
7.7.1 Splitter connections: For dual columns which are connected
using a press-fit Y-shaped glass splitter or a Y-shaped fused-silica
connector (J&W Scientific or Restek), clean and deactivate the splitter
port insert or replace with a cleaned and deactivated splitter. Break off
the first few inches (up to one foot) of the injection port side of the
column. Remove the columns and solvent backflush according to the
manufacturer's instructions. If these procedures fail to eliminate the
degradation problem, it may be necessary to deactivate the metal injector
body and/or replace the columns.
8081 - 18 Revision 0
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7.7.1.1 GC injector ports can be of critical concern,
especially in the analysis of DDT and Endrin. Injectors that are
contaminated, chemically active, or too hot can cause the
degradation ("breakdown") of the analytes. Endrin and DDT breakdown
to Endrin aldehyde, Endrin ketone, ODD, or DDE. When such breakdown
is observed, clean and deactivate the injector port, break off at
least 0.5 M of the column and remount it. Check the injector
temperature and lower it to 205°C, if required. Endrin and DDT
breakdown is less of a problem when ambient on-column injectors are
used.
7.7.2 Metal injector body: Turn off the oven and remove the
analytical columns when the oven has cooled. Remove the glass injection
port insert (instruments with on-column injection). Lower the injection
port temperature to room temperature. Inspect the injection port and
remove any noticeable foreign material.
7.7.2.1 Place a beaker beneath the injector port inside
the oven. Using a wash bottle, serially rinse the entire inside of
the injector port with acetone and then toluene; catch the rinsate
in the beaker.
7.7.2.2 Prepare a solution of a deactivating agent (Sylon-
CT or equivalent) following manufacturer's directions. After all
metal surfaces inside the injector body have been thoroughly coated
with the deactivation solution, rinse the injector body with
toluene, methanol, acetone, then hexane. Reassemble the injector
and replace the columns.
7.7.3 Column rinsing: The column should be rinsed with several
column volumes of an appropriate solvent. Both polar and nonpolar
solvents are recommended. Depending on the nature of the sample residues
expected, the first rinse might be water, followed by methanol and
acetone; methylene chloride is a good final rinse and in some cases may be
the only solvent required. The column should then be filled with
methylene chloride and allowed to stand flooded overnight to allow
materials within the stationary phase to migrate into the solvent. The
column is then flushed with fresh methylene chloride, drained, and dried
at room temperature with a stream of ultrapure nitrogen.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control (QC) procedures
including matrix spikes, duplicates and blanks. Quality control to validate
sample extraction is covered in Method 3500 and in the extraction method
utilized. If an extract cleanup was performed, follow the QC in Method 3600 and
in the specific cleanup method.
8.2 Quality control requirements for the GC system, including calibration
and corrective actions, are found in Method 8000. The following steps are
recommended as additional method QC.
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8.2.1 The QC Reference Sample concentrate (Method 3500) should
contain the organochlorine pesticides at 10 mg/L for water samples. If
this method is to be used for analysis of Aroclors, Chlordane, or
Toxaphene only, the QC Reference Sample should contain the most
representative multi-component mixture at a concentration of 50 mg/L in
acetone. The frequency of analysis of the QC reference sample analysis is
equivalent to a minimum of 1 per 20 samples or 1 per batch if less than 20
samples. If the recovery of any compound found in the QC reference sample
is less than 80 percent or greater than 120 percent of the certified
value, the laboratory performance is judged to be out of control, and the
problem must be corrected. A new set of calibration standards should be
prepared and analyzed.
8.2.2 Calculate surrogate standard recovery on all samples, blanks,
and spikes. Determine if the recovery is within limits (limits
established by performing QC procedures outlined in Method 8000).
If recovery is not within limits, the following are required:
8.2.2.1 Confirm that there are no errors in calculations,
surrogate solutions and internal standards. Also, check instrument
performance.
8.2.2.2 Examine chromatograms for interfering peaks and
for integrated areas.
8.2.2.3 Recalculate the data and/or reanalyze the extract
if any of the above checks reveal a problem.
8.2.2.4 Reextract and reanalyze the sample if none of the
above are a problem or flag the data as "estimated concentration."
8.2.3 Include a calibration standard after each group of 20 samples
(it is recommended that a calibration standard be included after every 10
samples to minimize the number of repeat injections) in the analysis
sequence as a calibration check. The response factors for the calibration
should be within 15 percent of the initial calibration. When this
continuing calibration is out of this acceptance window, the laboratory
should stop analyses and take corrective action.
8.2.4 Whenever quantitation is accomplished using an internal
standard, internal standards must be evaluated for acceptance. The
measured area of the internal standard must be no more than 50 percent
different from the average area calculated during calibration. When the
internal standard peak area is outside the limit, all samples that fall
outside the QC criteria must be reanalyzed.
8.3 DDT and Endrin are easily degraded in the injection port. Breakdown
occurs when the injection port liner is contaminated high boiling residue from
sample injection or when the injector contains metal fittings. Check for
degradation problems by injecting a standard containing only 4,4'-DDT and Endrin.
Presence of 4,4'-DDE, 4,4'-ODD, Endrin ketone or Endrin indicates breakdown. If
degradation of either DDT or Endrin exceeds 15%, take corrective action before
proceeding with calibration.
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8.3.1 Calculate percent breakdown as follows:
% breakdown Total DDT degradation peak area (DDE + ODD)
for 4, 4' -DDT = - x 100
peak areas (DDT + DDE + ODD)
Total endrin degradation peak area
% breakdown (Endrin aldehyde + Endrin ketone)
for Endrin = - x 100
peak areas (Endrin + aldehyde + ketone)
8.3.2 The breakdown of DDT and Endrin should be measured before
samples are analyzed and at the beginning of each 12 hour shift. Injector
maintenance and recalibration should be completed if the breakdown is
greater than 15% for either compound (Sec. 8.2.3).
8.4 GC/MS confirmation may be used for single column analysis. In
addition, any compounds confirmed by two columns should also be confirmed by
GC/MS if the concentration is sufficient for detection by GC/MS.
8.4.1 Full -scan GC/MS will normally require a minimum concentration
near 10 ng/jLtL in the final extract for each single-component compound.
Ion trap or selected ion monitoring will normally require a minimum
concentration near 1 ng/jul_.
8.4.2 The GC/MS must be calibrated for the specific target
pesticides when it is used for quantitative analysis.
8.4.3 GC/MS may not be used for single column confirmation when
concentrations are below 1
8.4.4 GC/MS confirmation should be accomplished by analyzing the
same extract used for GC/ECD analysis and the associated blank.
8.4.5 Use of the base/neutral -acid extract and associated blank may
be used if the surrogates and internal standards do not interfere and it
is demonstrated that the analyte is stable during acid/base partitioning.
However, if the compounds are not detected in the base/neutral -acid
extract even though the concentrations are high enough, a GC/MS analysis
of the pesticide extract should be performed.
8.4.6 A QC reference sample of the compound must also be analyzed by
GC/MS. The concentration of the QC reference standard must demonstrate
the ability to confirm the pesticides/Aroclors identified by GC/ECD.
8.5 Whenever silica gel (Method 3630) or Florisil (Method 3620) cleanup
is used, the analyst must demonstrate that the fractionation scheme is
reproducible. Batch to batch variation in the composition of the silica gel
material or overloading the column may cause a change in the distribution
patterns of the organochlorine pesticides and PCBs. When compounds are found in
two fractions, add the concentrations in the fractions, and corrections for any
additional dilution.
8081 - 21 Revision 0
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9.0 METHOD PERFORMANCE
9.1 The MDL is defined in Chapter One. The MDL concentrations listed in
Tables 1 and 2 were obtained using organic-free reagent water and sandy loam
soil.
9.2 The chromatographic separations in this method have been tested in
a single laboratory by using clean hexane and liquid and solid waste extracts
that were spiked with the test compounds at three concentrations. Single-
operator precision, overall precision, and method accuracy were found to be
related to the concentration of the compound and the type of matrix.
9.3 This method has been applied in a variety of commercial laboratories
for environmental and waste matrices. Performance data were obtained for a
limited number of target analytes spiked into sewage sludge and dichloroethene
still bottoms at high concentration levels. These data are provided in Tables
12 and 13.
9.4 The accuracy and precision obtainable with this method depend on the
sample matrix, sample preparation technique, optional cleanup techniques, and
calibration procedures used.
9.5 Single laboratory accuracy data were obtained for organochlorine
pesticides in a clay soil. The spiking concentration was 500 M9/kg. The
spiking solution was mixed into the soil and then immediately transferred to the
extraction device and immersed in the extraction solvent. The spiked sample was
then extracted by Method 3541 (Automated Soxhlet). The data represent a single
determination. Analysis was by capillary column gas chromatography/electron
capture detector following Method 8081 for the organochlorine pesticides. These
data are listed in Table 14 and were taken from Reference 14.
9.6 Single laboratory recovery data were obtained for PCBs in clay and
soil. Oak Ridge National Laboratory spiked Aroclors 1254 and 1260 at
concentrations of 5 and 50 ppm into portions of clay and soil samples and
extracted these spiked samples using the procedure outlined in Method 3541.
Multiple extractions using two different extractors were performed. The extracts
were analyzed by Method 8081. The data are listed in Table 15 and were taken
from Reference 15.
9.7 Multi-laboratory accuracy and precision data were obtained for PCBs
in soil. Eight laboratories spiked Aroclors 1254 and 1260 into three portions
of 10 g of Fuller's Earth on three non-consecutive days, followed by immediate
extraction using Method 3541. Six of the laboratories spiked each Aroclor at 5
and 50 mg/kg and two laboratories spiked each Aroclor at 50 and 500 mg/kg. All
extracts were analyzed by Oak Ridge National Laboratory, Oak Ridge, TN, using
Method 8081. These data are listed in Table 16 and were taken from Reference 13.
8081 - 22 Revision 0
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10.0 REFERENCES
1. Lopez-Avila, V.; Baldin, E.; Benedicto, J; Milanes, J.; Beckert. W. F.
Application of Open-Tubular Columns to SW-846 GC Methods"; final report to
the U.S. Environmental Protection Agency on Contract 68-03-3511; Mid-
Pacific Environmental Laboratory, Mountain View, CA, 1990.
2. Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 10 - Pesticides and PCB Report for
the U.S. Environmental Protection Agency on Contract 68-03-2606.
3. Goerlitz, D.F.; Law, L.M. "Removal of Elemental Sulfur Interferences from
Sediment Extracts for Pesticide Analysis"; Bull. Environ. Contam. Toxicol.
1971, 6, 9.
4. Ahnoff, M.; Josefsson, B. "Cleanup Procedures for PCB Analysis on River
Water Extracts"; Bull. Environ. Contam. Toxicol. 1975, 13, 159.
5. Jensen, S.; Renberg, L.; Reutergardth, L. "Residue Analysis of Sediment
and Sewage Sludge for Organochlorines in the Presence of Elemental
Sulfur"; Anal. Chem. 1977, 49, 316-318.
6. Wise, R.H.; Bishop, D.F.; Williams, R.T.; Austern, B.M. "Gel Permeation
Chromatography in the GC/MS Analysis of Organics in Sludges"; U.S.
Environmental Research Laboratory. Cincinnati, OH 45268.
7. Pionke, H.B.; Chesters, G.; Armstrong, D.E. "Extraction of Chlorinated
Hydrocarbon Insecticides from Soil"; Agron. J. 1968, 60, 289.
8. Burke, J.A.; Mills, P.A.; Bostwick, D.C. "Experiments with Evaporation of
Solutions of Chlorinated Pesticides"; J. Assoc. Off. Anal. Chem. 1966, 49,
999.
9. Glazer, J.A., et al. "Trace Analyses for Wastewaters"; Environ. Sci. and
Technol. 1981, 15, 1426.
10. Marsden, P.O., "Performance Data for SW-846 Methods 8270, 8081, and 8141,"
EMSL-LV, EPA/600/4-90/015.
11. Marsden, P.O., "Analysis of PCBs", EMSL-LV, EPA/600/8-90/004
12. Erickson, M. Analytical Chemistry of PCBs, Butterworth Publishers, Ann
Arbor Science Book (1986).
13. Stewart, J. "EPA Verification Experiment for Validation of the SOXTEC" PCB
Extraction Procedure"; Oak Ridge National Laboratory, Oak Ridge, TN,
37831-6138; October 1988.
14. Lopez-Avila, V. (Beckert, W., Project Officer), "Development of a Soxtec
Extraction Procedure for Extracting Organic Compounds from Soils and
Sediments", EPA 600/X-91/140, US EPA, Environmental Monitoring Systems
Laboratory-Las Vegas, October 1991.
8081 - 23 Revision 0
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15. Stewart, J.H.; Bayne, C.K.; Holmes, R.L.; Rogers, W.F.; and Maskarinec,
M.P., "Evaluation of a Rapid .Quantitative Organic Extraction System for
Determining the Concentration of PCB in Soils", Proceedings of the USEPA
Symposium on Waste Testing and Quality Assurance. Oak Ridge National
Laboratory, Oak Ridge, TN 37831-6131; July 11-15, 1988.
8081 - 24 Revision 0
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TABLE 1
GAS CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION
LIMITS FOR THE ORGANOCHLORINE PESTICIDES AND PCBs AS AROCLORS
USING WIDE-BORE CAPILLARY COLUMNS
SINGLE COLUMN METHOD OF ANALYSIS
Compound
Aldrin
a-BHC
6-BHC
£-BHC
y-BHC (Lindane)
a-Chlordane
7-Chlordane
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan Sulfate
Endrin
Endrin aldehyde
Heptachlor
Heptachlor epoxide
Methoxychlor
Toxaphene
Aroclor-1016
Aroclor-1221
Aroclor-1232
Aroclor-1242
Aroclor-1248
Aroclor-1254
Aroclor-1260
Retention
DB 608b
11.84
8.14
9.86
11.20
9.52
15.24
14.63
18.43
16.34
19.48
16.41
15.25
18.45
20.21
17.80
19.72
10.66
13.97
22.80
MR
MR
MR
MR
MR
MR
MR
MR
Water = Organic-free reagent
Time (min)
DB 1701b
12.50
9.46
13.58
14.39
10.84
16.48
16.20
19.56
16.76
20.10
17.32
15.96
19.72
22.36
18.06
21.18
11.56
15.03
22.34
MR
MR
MR
MR
MR
MR
MR
MR
water.
MDLa Water
(M9/L)
0.034
0.035
0.023
0.024
0.025
0.008
0.037
0.050
0.058
0.081
0.044
0.030
0.040
0.035
0.039
0.050
0.040
0.032
0.086
NA
0.054
NA
NA
NA
NA
NA
0.90
MDLa Soil
(M9/kg)
2.2
1.9
3.3
1.1
2.0
1.5
4.2
2.5
3.6
NA
2.1
2.4
3.6
3.6
1.6
2.0
2.1
5.7
NA
57.0
NA
NA
NA
NA
NA
70.0
Soil = Sandy loam soil .
MR = Multiple
NA = Data not
peak responses.
available.
MDL is the method detection limit. MDL was determined from the
analysis of seven replicate aliquots of each matrix processed
through the entire analytical method (extraction, silica gel
cleanup, and GC/ECD analysis). MDL = t(n-l, 0.99) x SD, where t(n-
1, 0.99) is the Student's t value appropriate for a 99% confidence
interval and a standard deviation with n-1 degrees of freedom, and
SD is the standard deviation of the seven replicate measurements.
See Table 4 for GC operating conditions.
8081 - 25
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TABLE 2
GAS CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION
LIMITS FOR THE ORGANOCHLORINE PESTICIDES AND PCBs AS AROCLORS
USING NARROW-BORE CAPILLARY COLUMNS
SINGLE COLUMN METHOD OF ANALYSIS
Compound
Retention Time (min)
DB 608b DB 5b
MDLa Water MDL8 Soil
(M9/L) (M9/kg)
Aldrin
<7-BHC
6-BHC
-------
TABLE 2
(Continued)
MDL is the method detection limit. MDL was determined from the
analysis of seven replicate aliquots of each matrix processed
through the entire analytical method (extraction, cleanup, and
GC/ECD analysis). MDL = t(n-l, 0.99) x SD, where t(n-l, 0.99) is
the Student's t value appropriate for a 99% confidence interval and
a standard deviation with n-1 degrees of freedom, and SD is the
standard deviation of the seven replicate measurements.
30 m x 0.25 mm ID DB-608 1 /urn film thickness, see Table 4 for GC
operating conditions.
30 m x 0.25 mm ID DB-5 1 jum film thickness, see Table 4 for GC
operating conditions.
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TABLE 3
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQLs) FOR VARIOUS MATRICES8
Matrix Factor
Ground water 10
Low-concentration soil by sonication with GPC cleanup 670
High-concentration soil and sludges by sonication 10,000
Non-water miscible waste 100,000
EQL = [Method detection limit for water (see Table 1 or Table 2) wide-
bore or narrow-bore options] x [Factor found in this table]. For
nonaqueous samples, the factor is on a wet-weight basis. Sample EQLs
are highly matrix-dependent. The EQLs to be determined herein are
provided for guidance and may not always be achievable.
8081 - 28 Revision 0
September 1994
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TABLE 4
GC OPERATING CONDITIONS FOR ORGANOCHLORINE COMPOUNDS
SINGLE COLUMN ANALYSIS
Narrow-bore columns:
Narrow-bore Column 1 - 30 m x 0.25 or 0.32 mm internal diameter (ID) fused
silica capillary column chemically bonded with SE-54 (DB-5 or equivalent), 1
/im film thickness.
Carrier gas (He)
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
16 psi
225°C
300°C
100°C, hold 2 minutes
100°C to 160°C at 15°C/min, followed
by 160°C to 270°C at 5°C/nnr)
270°C
Narrow-bore Column 2 - 30 m x 0.25 mm ID fused silica capillary column
chemically bonded with 35 percent phenyl methylpolysiloxane (DB-608, SPB-608,
or equivalent), 25 jitm coating thickness, 1 /urn film thickness
Carrier gas (N2)
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
20 psi
225°C
300°C
160°C,
160°C
290°C,
hold 2 minutes
to 290°C at 5°C/min
hold 1 min
Wide-bore columns:
Wide-bore Column 1 - 30 m x 0.53 mm ID fused silica capillary column
chemically bonded with 35 percent phenyl methylpolysiloxane (DB-608, SPB-608,
RTx-35, or equivalent), 0.5 /j,m or 0.83 ,um film thickness.
Wide-bore Column 2 - 30 m x 0.53 mm ID fused silica capillary column
chemically bonded with 50 percent phenyl methylpolysiloxane (DB-1701, or
equivalent), 1.0 urn film thickness.
Carrier gas (He)
Makeup gas
argon/methane (P-5 or P-10) or N2
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
5-7 mL/minute
30 mL/min
250°C
290°C
150°C, hold 0.5 minute
150°C to 270°C at 5°C/min
270°C, hold 10 min
(continued)
8081 - 29
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September 1994
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TABLE 4 (Continued)
GC OPERATING CONDITIONS FOR ORGANOCHLORINE COMPOUNDS
SINGLE COLUMN ANALYSIS
Wide-bore Columns (continued)
Wide-bore Column 3 - 30 m x 0.53 mm ID fused silica capillary column
chemically bonded with SE-54 (DB-5, SPB-5, RTx-5, or equivalent), 1.5
thickness.
Carrier gas (He) 6 mL/minute
Makeup gas
argon/methane (P-5 or P-10) or N2 30 mL/min
Injector temperature 205°C
Detector temperature 290°C
Initial temperature 140°C, hold 2 min
Temperature program 140°C to 240°C at 10°C/min,
hold 5 minutes at 240°C,
240°C to 265°C at 5°C/min
Final temperature 265°C, hold 18 min
8081 - 30 Revision 0
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TABLE 5
RETENTION TIMES OF THE ORGANOCHLORINE PESTICIDES'
DUAL COLUMN METHOD OF ANALYSIS
Compound
DBCP
Hexachl orocycl opentadi ene
Etridiazole
Chloroneb
Hexachl orobenzene
Dial late
Propachlor
Trifluralin
a-BHC
PCNB
7-BHC
Heptachlor
Aldrin
Alachlor
Chlorothalonil
Alachlor
/3-BHC
Isodrin
DC PA
5-BHC
Heptachlor epoxide
Endosulfan-I
•y-Chlordane
a-Chlordane
trans-Nonachlor
4,4'-DDE
Dieldrin
Captan
Perthane
Endrin
Chloropropylate
Chi orobenzi late
Nitrofen
4,4'-DDD
Endosulfan II
4,4'-DDT
Endrin aldehyde
Mi rex
Endosulfan sulfate
CAS No.
96-12-8
77-47-4
2593-15-9
2675-77-6
118-74-1
2303-16-4
1918-16-17
1582-09-8
319-84-6
82-68-8
58-89-9
76-44-8
309-00-2
15972-60-8
1897-45-6
15972-60-8
319-85-7
465-73-6
1861-32-1
319-86-8
1024-57-3
959-98-8
5103-74-2
5103-71-9
39765-80-5
72-55-9
60-57-1
133-06-2
72-56-0
72-20-8
99516-95-7
510-15-6
1836-75-5
72-54-8
33213-65-9
50-29-3
7421-93-4
2385-85-5
1031-07-8
DB-5
RT(min)
2.14
4.49
6.38
7.46
12.79
12.35
9.96
11.87
12.35
14.47
14.14
18.34
20.37
18.58
15.81
18.58
13.80
22.08
21.38
15.49
22.83
25.00
24.29
25.25
25.58
26.80
26.60
23.29
28.45
27.86
28.92
28.92
27.86
29.32
28.45
31.62
29.63
37.15
31.62
DB-1701
RT(min)
2.84
4.88
8.42
10.60
14.58
15.07
15.43
16.26
17.42
18.20
20.00
21.16
22.78
24.18
24.42
24.18
25.04
25.29
26.11
26.37
27.31
28.88
29.32
29.82
30.01
30.40
31.20
31.47
32.18
32.44
34.14
34.42
34.42
35.32
35.51
36.30
38.08
38.79
40.05
continued
8081 - 31
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Compound
Methoxychlor
Captafol
Endrin ketone
trans-Permethrin
Kepone
Dicofol
Dichlone
a,a'-Dibromo-m-xylene
2-Bromobiphenyl
TABLE 5
(Continued)
CAS No.
72-43-5
2425-06-1
53494-70-5
51877-74-8
143-50-0
115-32-2
117-80-6
DB-5
RT(min)
35.33
32.65
33.79
41.50
31.10
35.33
15.17
9.17
8.54
DB-1701
RT(min)
40.31
41.42
42.26
45.81
b
b
b
11.51
12.49
3The GC operating conditions were as follows: 30-m x 0.53-mm ID DB-5
(0.83-jum film thickness) and 30-m x 0.53-mm ID DB-1701 (1.0-/im film
thickness) connected to an 8-in injection tee (Supelco Inc.). Temperature
program: 140°C (2-min hold) to 270°C (1-min hold) at 2.8°C/min; injector
temperature 250°C; detector temperature 320°C; helium carrier gas 6 mL/min;
nitrogen makeup gas 20 mL/min.
detected at 2 ng per injection.
8081 - 32 Revision 0
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Column 1:
TABLE 6
GC OPERATING CONDITIONS FOR ORGANOCHLORINE PESTICIDES
FOR DUAL COLUMN METHOD OF ANALYSIS
LOW TEMPERATURE, THIN FILM
Type: DB-1701 (J&W) or equivalent
Dimensions: 30 m x 0.53 mm ID
Film Thickness (jiim): 1.0
Column 2:
Type: DB-5 (J&W) or equivalent
Dimensions: 30 m x 0.53 mm ID
Film Thickness (fj,m): 0.83
Carrier gas flowrate (mL/min): 6 (Helium)
Makeup gas flowrate (mL/min): 20 (Nitrogen)
Temperature program: 140°C (2 min hold) to 270°C (1 min hold) at 2.8°C/min
Injector temperature: 250°C
Detector temperature: 320°C
Injection volume: 2 p.1
Solvent: Hexane
Type of injector: Flash vaporization
Detector type: Dual ECD
Range: 10
Attenuation: 64 (DB-1701J/32 (DB-5)
Type of splitter: Supelco 8 in injection tee
8081 - 33 Revision 0
September 1994
-------
Column 1:
TABLE 7
GC OPERATING CONDITIONS FOR ORGANOCHLORINE PESTICIDES
FOR THE DUAL COLUMN METHOD OF ANALYSIS
HIGH TEMPERATURE, THICK FILM
Type: DB-1701 (J&W) or equivalent
Dimensions: 30 m x 0.53 mm ID
Film Thickness: 1.0 p,m
Column 2:
Type: DB-5 (J&W) or equivalent
Dimensions: 30 m x 0.53 mm ID
Film Thickness: 1.5 urn
Carrier gas flowrate (mL/min): 6 (Helium)
Makeup gas flowrate (mL/min): 20 (Nitrogen)
Temperature program: 150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min
then to 275°C (10 min hold) at 4°C/min.
Injector temperature: 250°C
Detector temperature: 320°C
Injection volume: 2 til
Solvent: Hexane
Type of injector: Flash vaporization
Detector type: Dual ECD
Range: 10
Attenuation: 64 (DB-170IJ/64 (DB-5)
Type of splitter: J&W Scientific press-fit Y-shaped inlet splitter
8081 - 34 Revision 0
September 1994
-------
TABLE 8 SUMMARY OF RETENTION TIMES (MIN) OF AROCLORS
ON THE DB-5 COLUMN8
DUAL SYSTEM OF ANALYSIS
Peak
No.b
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
Aroclor
1016
8.41
8.77
8.98
9.71
10.49
10.58
10.90
11.23
11.88
11.99
12.27
12.66
12.98
13.18
13.61
13.80
13.96
14.48
14.63
14.99
15.35
16.01
16.27
Aroclor
1221
5.85
7.63
8.43
8.77
8.99
10.50
10.59
11.24
12.29
12.68
12.99
Aroclor
1232
5.85
7.64
8.43
8.78
9.00
10.50
10.59
10.91
11.24
11.90
12.00
12.29
12.69
13.00
13.19
13.63
13.82
13.97
14.50
14.64
15.02
15.36
16.14
16.29
17.04
17.22
17.46
18.41
18.58
18.83
19.33
20.03
21.18
Aroclor
1242
7.57
8.37
8.73
8.94
9.66
10.44
10.53
10.86
11.18
11.84
11.95
12.24
12.64
12.95
13.14
13.58
13.77
13.93
14.46
14.60
14.98
15.32
15.96
16.08
16.26
17.19
17.43
17.92
18.16
18.37
18.56
18.80
19.30
19.97
20.46
20.85
21.14
22.08
Aroclor
1248
8.95
10.45
10.85
11.18
11.85
12.24
12.64
12.95
13.15
13.58
13.77
13.93
14.45
14.60
14.97
15.31
16.08
16.24
16.99
17.19
17.43
17.69
17.91
18.14
18.36
18.55
18.78
19.29
19.92
20.45
20.83
21.12
21.36
22.05
Aroclor
1254
13.59
13.78
13.90
14.46
14.98
15.32
16.10
16.25
16.53
16.96
17.19
17.44
17.69
17.91
18.14
18.36
18.55
18.78
19.29
19.48
19.81
19.92
20.28
20.57
20.83
20.98
21.38
21.78
22.04
22.38
22.74
22.96
23.23
23.75
Aroclor
1260
13.59
16.26
16.97
17.21
18.37
18.68
18.79
19.29
19.48
19.80
20.28
20.57
20.83
21.38
21.78
22.03
22.37
22.73
22.95
23.23
23.42
23.73
Pesticide elating at same
retention time
Chlorothalonil (11.18)
Captan (16.21)
gamma-Chlordane (16.95)
4,4'-DDE (18.38)
Dieldrin (18.59)
Chloropropylate (19.91)
Endosulfan II (19.91)
Kepone (20.99)
4,4'-DDT (21.75)
Endosulfan sulfate (21.75)
Captafol (22.71)
Endrin ketone (23.73)
"The GC operating conditions are given in Table 7.
(continued)
8081 - 35
Revision 0
September 1994
-------
TABLE 8 CONTINUED
Peak
No.
56
57
58
59
60
61
62
63
64
65
66
67
68
69
Aroclor Aroclor Aroclor Aroctor Aroclor Aroclor
1016 1221 1232 1242 1248 1254
23.99
24.27
24.61
24.93
26.22
Aroclor Pesticide e luting at same
1260 retention time
23.97
24.16
Methoxychlor (24.29)
Dicofol (24.29)
24.45
24.62
24.91
25.44
26.19 Mirex (26.19)
26.52
26.75
27.41
28.07
28.35
29.00
"The GC operating conditions are given in Table 7.
'"These are sequentially numbered from elution order and are not isomer numbers
8081 - 36
Revision 0
September 1994
-------
TABLE 9 SUMMARY OF RETENTION TIMES (MIN) OF AROCLORS
ON THE DB-1701 COLUMN8
DUAL SYSTEM OF ANALYSIS
Peak
No.b
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
Aroclor
1016
6.33
6.78
6.96
7.64
8.23
8.62
8.88
9.05
9.46
9.77
10.27
10.64
11.01
11.09
11.98
12.39
12.92
12.99
13.14
13.49
13.58
Aroclor
1221
4.45
5.38
5.78
5.86
6.34
6.78
6.96
8.23
8.63
9.06
9.79
10.29
10.65
Aroctor
1232
4.45
5.86
6.34
6.79
6.96
8.23
8.63
8.89
9.06
9.47
9.78
10.29
10.66
11.02
11.10
11.99
12.39
12.77
13.00
13.16
13.49
13.61
14.08
14.30
14.49
15.38
15.65
15.78
16.13
16.77
17.13
Aroclor
1242
6.28
6.72
6.90
7.59
8.15
8.57
8.83
8.99
9.40
9.71
10.21
10.59
10.96
11.02
11.94
12.33
12.71
12.94
13.09
13.44
13.54
13.67
14.03
14.26
14.46
15.33
15.62
15.74
16.10
16.73
17.09
17.46
17.69
18.48
19.13
Aroclor
1248
6.91
8.16
8.83
8.99
9.41
9.71
10.21
10.59
10.95
11.03
11.93
12.33
12.69
12.93
13.09
13.44
13.54
14.03
14.24
14.39
14.46
15.10
15.32
15.62
15.74
16.10
16.74
17.07
17.44
17.69
18.19
18.49
19.13
Aroclor
1254
10.95
11.93
12.33
13.10
13.24
13.51
13.68
14.03
14.24
14.36
14.56
15.10
15.32
15.61
15.74
16.08
16.34
16.44
16.55
16.77
17.07
17.29
17.43
17.68
18.17
18.42
18.59
18.86
19.10
19.42
Aroclor Pesticide e luting at same
1260 retention time
Trifluralin (6.96)
13.52
14.02
14.25
14.56
Chlordane (15.32)
16.61 4,4'-DDE (15.67)
15.79
16.19
16.34
16.45
16.77 Perthane (16.71)
17.08
17.31
17.43
17.68
18.18
18.40
18.86
19.09 Endosulfan II (19.05)
19.43
"The GC operating conditions are given in Table 7.
(continued)
8081 - 37
Revision 0
September 1994
-------
TABLE 9 CONTINUED
Peak
No.
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
Aroclor Aroclor Aroclor Aroclor Aroclor Aroclor
1016 1221 1232 1242 1248 1254
19.55
20.20
20.34
20.57 20.55
20.62
20.88
21.53
21.83
23.31
Aroclor Pesticide e luting at same
1260 retention time
19.59 4,4'-DDT (19.54)
20.21
20.43
20.66 Endrin aldehyde (20.69)
20.87
21.03
21.53
21.81
23.27
23.85
24.11
24.46
24.59
24.87
25.85
27.05
27.72
'The GC operating conditions are given in Table 7.
""These are sequentially numbered from elution order and are not isomer numbers
8081 - 38
Revision 0
September 1994
-------
TABLE 10
PEAKS DIAGNOSTIC OF PCBs OBSERVED IN 0.53 mm ID COLUMN
SINGLE COLUMN ANALYSIS
Peak RT on RT on Elution
No.c DB 608° DB 1701a Aroclorb Order
I479047661221Before TCmX
II 7.15 6.96 1221, 1232, 1248 Before a-BHC
III 7.89 7.65 1061, 1221, 1232, 1242, Before a-BHC
IV 9.38 9.00 1016, 1232, 1242, 1248, just after a-BHC on
DB-1701;just before
7-BHC on DB-608
V 10.69 10.54 1016, 1232, 1242. 1248 a-BHC and
heptachlor on DB-1701;
just after heptachlor
on DB-608
VI 14.24 14.12 1248, 1254 T-BHC and heptachlor
epoxide on DB-1701;
heptachlor epoxide and
7-Chlordane on DB-608
VII 14.81 14.77 1254 Heptachlor epoxide and
7-Chlordane on
DB-1701; a- and
7-Chlordane on DB-608
VIII 16.71 16.38 1254 DDE and Dieldrin on
DB-1701; Dieldrin and
Endrin on DB-608
IX 19.27 18.95 1254, 1260 Endosulfan II on
DB-1701; DDT on DB-608
Continued
8081 - 39 Revision 0
September 1994
-------
TABLE 10 (Continued)
PEAKS DIAGNOSTIC OF PCBs OBSERVED IN 0.53 mm ID COLUMN
SINGLE COLUMN ANALYSIS
Peak RT on RT on Elution
No. DB 608a DB 1701" Aroclorb Order
X 21.22 21.23 1260 Endrin aldehyde and
Endosulfan sulfate on
DB-1701; Endosulfan
sulfate and
Methoxychlor on
on DB-608
XI 22.89 22.46 1260 Just before endrin
ketone on DB-1701;
after endrin ketone on
DB-608
a Temperature program: T( = 150°C, hold 30 seconds; increase temperature at
5°C/minutes to 275°C.
b Underlined Aroclor indicates the largest peak in the pattern.
0 These are sequentially numbered from elution order and are not isomer
numbers
8081 - 40 Revision 0
September 1994
-------
TABLE 11 SPECIFIC PCB CONGENERS IN AROCLORS
Congener
IUPAC number
Aroclor
1016 1221 1232 1242 1248 1254 1260
Biphenyl
2CB
23DCB
34DCB
244'TCB
22'35'TCB
23'44'TCB
233'4'6PCB
23'44'5PCB
22'44'55'HCB
22'344'5'HCB
22'344'55'HpCB
22'33'44'5HpCB
..
1
5
12
28*
44
66*
110
118*
153
138
180
170
X
XXX
XXX
X X
X X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
*apparent co-elution of two major peaks:
28 with 31 (2,4',5 trichloro)
66 with 95 (2,2',3,5',6 pentachloro)
118 with 149 (2,2',3,4',5',6 hexachloro)
8081 - 41
Revision 0
September 1994
-------
TABLE 12 ANALYTE RECOVERY FROM SEWAGE SLUDGE
Compound
Sonication
Soxhlet
Hexachloroethane
2-Chloronapthalene
4-Bromodiphenyl ether
a-BHC
7-BHC
Heptachlor
Aldrin
j8-BHC
(J-BHC
Heptachlor epoxide
Endosulfan I
7-Chlordane
a-Chlordane
DDE
Dieldrin
Endrin
Endosulfan II
DDT
Endrin aldehyde
ODD
Tetrachl oro-m-xyl ene
Decachl orobi phenyl
%Recovery
80
50
118
88
55
60
92
351
51
54
52
50
49
52
89
56
52
57
45
57
71
26
%RSD
7
56
14
25
9
13
33
71
11
11
11
9
8
11
19
10
10
10
6
11
19
23
%Recovery
79
67
nd
265
155
469
875
150
57
70
70
65
66
74
327
92
88
95
42
99
82
28
%RSD
1
8
18
29
294
734
260
2
3
4
1
0
1
7
15
11
17
10
8
1
48
Concentration spiked in the sample: 500-1000 ng/g
Three replicates/sample
Extraction solvent, Method 3540 - methylene chloride
Extraction solvent, Method 3550 - methylene chloride/acetone (1:1)
Cleanup - Method 3640
GC column - DB-608, 30M X 0.53 mm ID
8081 - 42
Revision 0
September 1994
-------
TABLE 13 ANALYTE RECOVERY FROM DCE STILL BOTTOMS
Compound
Sonication
Soxhlet
Hexachloroethane
2-Chloronapthalene
4-Bromodiphenyl ether
a-BHC
/3-BHC
Heptachlor
Aldrin
l8-BHC
-------
TABLE 14
SINGLE LABORATORY ACCURACY DATA FOR THE EXTRACTION OF
ORGANOCHLORINE PESTICIDES FROM SPIKED CLAY SOIL BY METHOD 3541
(AUTOMATED SOXHLET)"
Compound Name Spike Level % Recovery
DB-5 DB-1701
a-BHC
/3-BHC
Heptachlor
Aldrin
Heptachlor epoxide
trans-Chlordane
Endosulfan I
Dieldrin
Endrin
Endosulfan II
4,4'-DDT
Mi rex
500
500
500
500
500
500
500
500
500
500
500
500
89
86
94
b
97
94
92
b
111
104
b
108
94
b
95
92
97
95
92
113
104
104
b
102
a The operating conditions for the automated Soxhlet were as follows:
immersion time 45 min; extraction time 45 min; the sample size was 10 g
clay soil, extraction solvent, 1:1 acetone/hexane. No equilibration time
following spiking.
b Not able to determine because of interference.
Data taken from Reference 14.
8081 - 44 Revision 0
September 1994
-------
TABLE 15
SINGLE LABORATORY RECOVERY DATA FOR EXTRACTION OF
PCBS FROM CLAY AND SOIL BY METHOD 3541a (AUTOMATED SOXHLET)
Matrix Compound Spike Level
(ppm)
Clay Aroclor-1254 5
Clay Aroclor-1254 50
Clay Aroclor-1260 5
Clay Aroclor-1260 50
Soil Aroclor-1254 5
Soil Aroclor-1254 50
Trial
1
2
3
4
5
6
1
2
3
4
5
6
1
2
3
4
5
6
1
2
3
4
5
6
1
2
3
4
5
1
2
3
4
5
6
Percent
Recovery6
87.0
92.7
93.8
98.6
79.4
28.3
65.3
72.6
97.2
79.6
49.8
59.1
87.3
74.6
60.8
93.8
96.9
113.1
73.5
70.1
92.4
88.9
90.2
67.3
69.7
89.1
91.8
83.2
62.5
84.0
77.5
91.8
66.5
82.3
61.6
(continued)
8081 - 45
Revision 0
September 1994
-------
TABLE 15
(continued)
Matrix Compound Spike Level
(ppm)
Soil Aroclor-1260 5
Soil Aroclor-1260 50
Trial
1
2
3
4
5
6
7
1
2
3
4
5
6
Percent
Recovery15
83.9
82.8
81.6
96.2
93.7
93.8
97.5
76.9
69.4
92.6
81.6
83.1
76.0
a The operating conditions for the automated Soxhlet were as follows:
immersion time 60 min; reflux time 60 min.
b Multiple results from two different extractors.
Data from Reference 15.
8081 - 46
Revision 0
September 1994
-------
TABLE 16. MULTI-LABORATORY PRECISION AND ACCURACY DATA
FOR THE EXTRACTION OF PCBS FROM SPIKED SOIL
BY METHOD 3541 (AUTOMATED SOXHLET)
Laboratory
Lab 1
Lab 2
Lab 3
Lab 4
Lab 5
Lab 6
Lab 7
Lab 8
All
Laboratories
Num
Average
St Dev
Num
Average
St Dev
Num
Average
St Dev
Num
Average
St Dev
Num
Average
St Dev
Num
Average
St Dev
Num
Average
St Dev
Num
Average
St Dev
Num
Average
St Dev
PCB Percent Recovery
Aroclor
1254
PCB Level
5
3.0
101.2
34.9
3.0
72.8
10.8
6.0
112.6
18.2
2 0
i4o!g
4.3
3.0
100.1
17.9
3.0
65.0
16.0
20.0
98.8
28.7
50
3.0
74.0
41.8
6.0
56.5
7.0
3.0
63.3
8.3
6.0
144.3
30.4
3.0
97.1
8.7
3.0
127.7
15.5
3.0
123.4
14.6
3.0
38.3
21.9
30.0
92.5
42.9
500
6.0
66.9
15.4
3.0
80.1
5.1
9.0
71.3
14.1
1260
PCB Level
5
3.0
83.9
7.4
3.0
70.6
2.5
6.0
100.3
13.3
3.0
138.7
15.5
3.0
82.1
7.9
3.0
92.8
36.5
21.0
95.5
25.3
50
3.0
78.5
7.4
6.0
70.1
14.5
3.0
57.2
5.6
6.0
84.8
3.8
3.0
79.5
3.1
4.0
105.9
7.9
3.0
94.1
5.2
3.0
51.9
12.8
31.0
78.6
18.0
500
6.0
74.5
10.3
3.0
77.0
9.4
9.0
75.3
9.5
All
Levels
12.0
84.4
26.0
24.0
67.0
13.3
12.0
66.0
9.1
24.0
110.5
28.5
12.0
83.5
10.3
12.0
125.4
18.4
12.0
99.9
19.0
12.0
62.0
29.1
120.0
87.6
29.7
Data from Reference 13.
8081 - 47
Revision 0
September 1994
-------
FIGURE 1.
GAS CHROMATOGRAM OF THE MIXED ORGANOCHLORINE PESTICIDE STANDARD
Start Time : 0.00 min End Time : 33.00 *?n
Scale Factor: 0 Plot Offset: 20 mV
lo« Point : 20.00
Plot Scilt: 400 M
High Point : 420.00 «V
Response [mV]
o~
iD
78-
NJ
GJ
ill i 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 I
LL
•= -4.68
:-7.99
L.67
-9.93
-10.78
11.05
-11.81
-13.65
-14.34
4l§gS
-14.92
-16.32
17.17
-17.63
an—18.56
-23.18
-23.80
•26.23
-=^28.64
Column:
Temperature program:
-0.95
-8.60
-30.19
30 m x 0.25 mm ID, DB-5
100°C (hold 2 minutes) to 160°C at 15°C/min, then at
5°C/min to 270°C; carrier He at 16 psi.
8081 - 48
Revision 0
September 1994
-------
FIGURE 2.
GAS CHROMATOGRAM OF INDIVIDUAL ORGANOCHLORINE PESTICIDE STANDARD MIX A
Start Time : 0.00 mm End lime : 33.00 nun
Scale Factor: 0 Plot Offset: 20 mv
lav Point : 20.00 mV Kign Point : 270.00 mv
Plot Scale: 250 mv
Response [mV]
i i i i i i
T, . , ,?
N)
O
O
I I I I I I I
o"
m
no"
D
r-t-
o'
3
H
ro
-
LJ
O"
'-7.93
1.60
-14.27
-17.08
20.22
0.77
22.68
-23.73
--28.52
-8.54
•-9.86
-12
.33
10.98
13.58
-17.54
18.47
-19.78
-19.24
-21.13
-23.03
-30.05
Column:
Temperature program:
30 m x 0.25 mm ID, DB-5
100°C (hold 2 minutes) to 160°C at 15°C/min, then at
5°C/min to 270°C; carrier He at 16 psi.
8081 - 49
Revision 0
September 1994
-------
FIGURE 3
GAS CHROMATOGRAM OF INDIVIDUAL ORGANOCHLORINE PESTICIDE STANDARD MIX B
Start rime : 0.00 mm
Scale Factor: 0
End Time : 33.00 mm
Plot Offset: 20 mv
Low Point : 20.00 mv
Plot Scale: 250 mv
High Point • 270 20 mv
Response [mV]
o
i i I i i
a 01
o a
I i i i i I
K) fO
o cn
o a
i i i I i i i i I i i
Q'
fl>
H
i'
fD
Ul
O"
2.74
-6.97
4—9.60
--10.71
-14.27
^-15.24
-<-i g nf.
22.00
-11.73
-14.84
-16.23
—17.08
-17.63
-18.31
19.54
-20.19
21.03
--22.68
-9.95
-8.5-
-30.04
Column:
Temperature program:
30 m x 0.25 mm ID, DB-5
100°C (hold 2 minutes) to 160°C at 15°C/min» then at
5°C/min to 270°C; carrier He at 16 psi.
8081 - 50
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FIGURE 4.
GAS CHROMATOGRAM OF THE TOXAPHENE STANDARD
Star; Tim : 0.00 mm End Tine : 33.00 nin Low Point : 20.00 mv High Point : 80.00
Scale Factor: 0 Plot Offset: 20 mV Plot Sealt: 60 UN
Response [mV]
I I I I I I I I II I I I I I I I I II ll I I ll I I II I I I I I I I I II I I I I I I I Ml I I I I I I I I II
01—
ID
rti
it"
3
T8-
Uf
24.32
Column:
Temperature program:
30 m x 0.25 mm ID, DB-5
100°C (hold 2 minutes) to 160°C at 15°C/min, then at
5°C/min to 270°C; carrier He at 16 psi.
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FIGURE 5.
GAS CHROMATOGRAM OF THE AROCLOR-1016 STANDARD
Start Time : 0.00 mm End Time : 33.00 mm Low Point : 20.00 mV High Point : 120.00 mV
Scale Factor: 0 Plot Offset: 20 mV Plot Scale: 100 mV
Response [mV]
NJ 4* 0) DO O
O O O O O
I I I I I I I I I I I I I I I I I I I I I I I II I I I I I I I I I I I II I I I I I I I I I I I 1
Uf
-1.81
-12.95
-1.03
Column:
Temperature program:
30 m x 0.25 mm ID DB-5 fused silica capillary.
100°C (hold 2 minutes) to 160°C at 15°C/niin, then at
5°C/min to 270°C; carrier He at 16 psi.
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FIGURE 6.
GAS CHROMATOGRAM OF THE TECHNICAL CHLORDANE STANDARD
Start Time : 0.00 mm
Scale Factor: 0
End Time : 33.00 mm Lou Point : 20.00 mv High Point : 220.00 mv
Plot Offset: 20 mv Plot Scale: 200 mv
Response [rnV]
o
o
J L
to
o
1
J I I
Lfl—
I'D
nT
o -*.
3 <-»
H
3'
o
-
.59
-12.92
13.60
-0.97
Column:
Temperature program:
30 m x 0.25 mm ID DB-5 fused silica capillary.
100°C (hold 2 minutes) to 160°C at 15°C/min, then at
5°C/min to 270°C; carrier He at 16 psi.
8081 - 53
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DB-1701
Li
OB-5
•T
u-
FIGURE 7. GC/ECD chromatogram of Toxaphene analyzed on a DB-5/DB-1701 fused-
silica open-tubular column pair. The GC operating conditions were
as follows: 30 m x 0.53 mm ID DB-5 (1.5-/nm film thickness) and 30
m x 0.53 mm ID DB-1701 (1.0-jum film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
8081 - 54
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0>
o-
DB-1701
o
u) IM
Ju
Or
r^
•s
DB-5
~—A.
FIGURE 8. GC/ECD chromatogram of Strobane analyzed on a DB-5/DB-1701 fused-
silica open-tubular column pair. The GC operating conditions were
as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and 30
m x 0.53 mm ID DB-1701 (1.0-/zm film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/nnn then to 275°C
(10 min hold) at 40C/nnn.
8081 - 55
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\r
0
in
rv
rxi
JL
m
-------
I
bill
DB-1701
o-
f-0
•CO
A/.
U. ^
ULL
DB-5
(• 1*1
• I-
o •
FIGURE 10. GC/ECD chromatogram of Aroclor 1221 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-,um film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-^m film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
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DB-1701
DB-5
FIGURE 11. GC/ECD chromatogram of Aroclor 1232 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-Mm film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
8081 - 58
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r-
O-
"'•?:•« 'A
r*M W T .1
\JL_jw-_UJ
DB-1701
I-J
LL
DB-5
FIGURE 12. GC/ECD chromatogram of Aroclor 1242 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-^m film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
8081 - 59
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Ill
0
(u
in
in
—in
-in
r«
ODOU 0)
tn <
Jj
DB-1701
O m
(X ft
DB-S
,
to if,
o- —
M in
* S-'
FIGURE 13. GC/ECD chromatogram of Aroclor 1248 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/nm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-pi film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
8081 - 60
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ID
r-
DB-1701
UO
0-
•o
li
DB-5
uuj UiO —
0 O>l u bi 1*1
k>
«
r«
FIGURE 14. GC/ECD chromatogram of Aroclor 1254 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/Lim film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
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OB-1701
11 " .***,'! ..
: S
DB-5
FIGURE 15. GC/ECD chromatogram of Aroclor 1260 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jum film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
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DB-1701
r- -
© -
a>
o
KI in
n 9
r- m
01 r>i
• o-
rsi r-
DB-5
fO K»TT 0-
*» <3 T 0)
01 O ® (M ID
1 . O T ® O
® •" "• ^™» «•» ^
.AN-fA.
FIGURE 16. GC/ECD chromatogram of Halowax 1000 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/im film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
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DB-1701
FIGURE 17. GC/ECD chromatogram of Halowax 1001 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-^m film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jum film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
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aryo-
•09
DB-1701
tr»
«
•o
*>
»
DB-5
« 9 f-
CD <•«(-»
v>««in mm • in •
»«*• in M • . .
*v» •»• n . -e « »
• *» • • «o ra IN P»
**ir» » m
FIGURE 18. GC/ECD chromatogram of Halowax 1099 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
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DB-1701
0 »
Hi —
0
T 9
a o-
I- «
•o r-
DB-5
«
r1
\J
FIGURE 19. GC/ECD chromatogram of Halowax 1013 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-^m film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-Mm film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/irnn then to 275°C
(10 min hold) at 4°C/min.
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OB-1701
i
DB-5
FIGURE 20. GC/ECD chromatogram of Halowax 1014 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/zm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
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DB-1701
f ". s
v » • n* « M .
M ««t - » •
*» «* It •% • - -
t 3
DB-5
i
•t • * " *
•lL*-«» *• - fk
Elf*'-? • - •»
J^Jili
its
FIGURE 21. GC/ECD chromatogram of Halowax 1051 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/im film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/nnn then to 275°C
(10 min hold) at 4°C/min.
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DB-5
I
i I
i
,
1
aw
I
DB-1701
12 ] « SU IS «
JL_
10 11 12 IS
14
n
32
IS 3* 4
4)
20.
FIGURE 22. GC/ECD chromatogram of the organochlorine pesticides analyzed on a
DB-5/DB-1701 fused-silica open-tubular column pair. The GC
operating conditions were as follows: 30 m x 0.53 mm ID DB-5 (0.83-
nm film thickness) and 30 m x 0.53 mm ID DB-1701 (1.0-/um film
thickness) connected to an 8 in injection tee (Supelco Inc.).
Temperature program: 140°C (2 min hold) to 270°C (1 min hold) at
2.8°C/min.
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METHOD 8081
ORGANOCHLORINE PESTICIDES AND PCBs AS AROCLORS BY GAS
CHROMATOGRAPHY: CAPILLARY COLUMN TECHNIQUE
7.1.1 Choose
appropriate extraction
technique (see Chapter 2)
i
7.1.2 Add specified
matrix sp*e to sample.
1
7.2 Routine cleanup/
fracttonaaon.
7.3 Set chrornalographic
i
7.4 Refer to Method 8000
tor proper calfcratton
techniques.
7.4.2 Prime or deactivate QC
column prior to caNbratton.
7.5 Perform GO analysis (see
MettwdSOOO)
7.5.8
Any sample
peak Inter-
ferences?
7.5.8 Additional
(see Section 7.2)
7.8CataJtetionof
toxaphene, chterdane, PCBs,
DDT. and BHC done here.
8081 - 70
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LIST OF TABLES
Table 1 Gas chromatographic retention times and method detection limits for
the Organochlorine Pesticides and PCBs as Aroclors using wide-bore
capillary columns, single column analysis
Table 2 Gas chromatographic retention times and method detection limits for
the Organochlorine pesticides and PCBs as Aroclors using narrow-bore
capillary columns, single column analysis
Table 3 Estimated quantitation limits (EQL) for various matrices
Table 4 GC Operating conditions for Organochlorine compounds, single column
analysis
Table 5 Retention times of the Organochlorine pesticides, dual column method
of analysis
Table 6 GC operating conditions for Organochlorine pesticides, dual column
method of analysis, low temperature, thin film
Table 7 GC operating conditions for Organochlorine pesticides, dual column
method of analysis, high temperature, thick film
Table 8 Summary of retention times (min) of Aroclors on the DB 5 column,
dual system of analysis
Table 9 Summary of retention times (min) of Aroclors on the DB 1701 column,
dual system of analysis
Table 10 Peaks diagnostic of PCBs observed in 0.53 mm ID column, single
column system of analysis
Table 11 Specific Congeners in Aroclors
Table 12 Recovery from Sewage Sludge
Table 13 Recovery DCE still bottoms
Table 14 Single Laboratory Accuracy Data for the Extraction of Organochlorine
Pesticides from Spiked Clay Soil by Method 3541 (Automated Soxhlet)
Table 15 Single Laboratory Recovery Data for Extraction of PCBs from Clay and
Soil by Method 3541 (Automated Soxhlet)
Table 16 Multi-laboratory Precision and Accuracy Data for the Extraction of
PCBs from Spiked Soil by Method 3541 (Automated Soxhlet)
8081 - 71
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LIST OF FIGURES
Figure 1. GC of the Mixed Organochlorine Pesticide Standard. The GC operating
conditions were as follows: 30 m x 0.25 mm ID DB-5 column.
Temperature program: 100°C (hold 2 minutes) to 160°C at 15°C/min,
then at 5°C/min to 270°C; carrier He at 16 psi.
Figure 2. GC of Individual Organochlorine Pesticide Standard Mix A. The GC
operating conditions were as follows: 30 m x 0.25 mm ID DB-5
column. Temperature program: 100°C (hold 2 minutes) to 160°C at
15°C/nrin> then at 5°C/mir\ to 270°C; carrier He at 16 psi.
Figure 3. GC of Individual Organochlorine Pesticide Standard Mix B. The GC
operating conditions were as follows: 30 m x 0.25 mm ID DB-5
column. Temperature program: 100°C (hold 2 minutes) to 160°C at
15°C/min> then at 5°C/min to 270°C; carrier He at 16 psi.
Figure 4. GC of the Toxaphene Standard. The GC operating conditions were as
follows: 30 m x 0.25 mm ID DB-5 column. Temperature program:
100°C (hold 2 minutes) to 160°C at 15°C/min, then at 5°C/min to 270°C;
carrier He at 16 psi.
Figure 5. GC of the Aroclor-1016 Standard. The GC operating conditions were
as follows: 30 m x 0.25 mm ID DB-5 fused silica capillary column.
Temperature program: 100°C (hold 2 minutes) to 160°C at 15°C/min,
then at 5°C/min to 270°C; carrier He at 16 psi.
Figure 6. GC of the Technical Chlordane Standard. The GC operating conditions
were as follows: 30 m x 0.25 mm ID DB-5 fused silica capillary
column. Temperature program: 100°C (hold 2 minutes) to 160°C at
15°C/niin, then at 5°C/min to 270°C; carrier He at 16 psi.
Figure 7. GC/ECD chromatogram of Toxaphene analyzed on a DB-5/DB-1701 fused-
silica open-tubular column pair. The GC operating conditions were
as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and 30
m x 0.53 mm ID DB-1701 (1.0-jum film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
Figure 8. GC/ECD chromatogram of Strobane analyzed on a DB-5/DB-1701 fused-
silica open-tubular column pair. The GC operating conditions were
as follows: 30 m x 0.53 mm ID DB-5 (1.5-/um film thickness) and 30
m x 0.53 mm ID DB-1701 (1.0-/um film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
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Figure 9. GC/ECD chromatogram of Aroclor 1016 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/im film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/zm film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
Figure 10. GC/ECD chromatogram of Aroclor 1221 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/Ltm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/xni film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
i
Figure 11. GC/ECD chromatogram of Aroclor 1232 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/xm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-pi film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
Figure 12. GC/ECD chromatogram of Aroclor 1242 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/im film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jun film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
Figure 13. GC/ECD chromatogram of Aroclor 1248 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min-
Figure 14. GC/ECD chromatogram of Aroclor 1254 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
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Figure 15. GC/ECD chromatogram of Aroclor 1260 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-|um film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/xm film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
Figure 16. GC/ECD chromatogram of Halowax 1000 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-Mm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/niin then to 275°C
(10 min hold) at 4°C/nrin.
Figure 17. GC/ECD chromatogram of, Halowax 1001 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-Mtn film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
Figure 18. GC/ECD chromatogram of Halowax 1099 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-/im film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
Figure 19. GC/ECD chromatogram of Halowax 1013 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-/im film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-^tm film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
Figure 20. GC/ECD chromatogram of Halowax 1014 analyzed on a DB-5/DB-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jum film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jum film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/miri then to 275°C
(10 min hold) at 4°C/min.
8081 - 74
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Figure 21. GC/ECD chromatogram of Halowax 1051 analyzed on a DB-5/D8-1701
fused-silica open-tubular column pair. The GC operating conditions
were as follows: 30 m x 0.53 mm ID DB-5 (1.5-jttm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-^m film thickness) connected to a J&W
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
Figure 22. GC/ECD chromatogram of the organochlorine pesticides analyzed on a
DB-5/DB-1701 fused-silica open-tubular column pair. The GC
operating conditions were as follows: 30 m x 0.53 mm ID DB-5 (0.83-
jum film thickness) and 30 m x 0.53 mm ID DB-1701 (1.0-jum film
thickness) connected to an 8 in injection tee (Supelco Inc.).
Temperature program: 140°C (2 min hold) to 270°C (1 min hold) at
2.8°C/min.
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METHOD 8090
NITROAROMATICS AND CYCLIC KETONES
1.0 SCOPE AND APPLICATION
1.1 Method 8090 is used to determine the concentration of various
nitroaromatic and cyclic ketone compounds. Table 1 indicates compounds that
may be determined by this method and lists the method detection limit for each
compound in reagent water. Table 2 lists the practical quantisation limit
(PQL) for other matrices.
2.0 SUMMARY OF METHOD
2.1 Method 8090 provides gas chromatographic conditions for the
detection of ppb levels of nitroaromatic and cyclic ketone compounds. Prior
to use of this method, appropriate sample extraction techniques must be used.
Both neat and diluted organic liquids (Method 3580, Waste Dilution) may be
analyzed by direct injection. A 2- to 5-uL aliquot of the extract is injected
into a gas chromatograph (GC) using the solvent flush technique, and compounds
in the GC effluent are detected by an electron capture detector (ECD) or a
flame ionization detector (FID). The dinitrotoluenes are determined using
ECD, whereas the other compounds amenable to this method are determined using
FID.
2.2 If interferences prevent proper detection of the analytes, the
method may also be performed on extracts that have undergone cleanup.
3.0 INTERFERENCES
3.1 Refer to Method 3500, 3600, and 8000.
3.2 Solvents, reagents, glassware, and other sample-processing hardware
may yield discrete artifacts and/or elevated baselines causing
misinterpretation of gas chromatograms. All of these materials must be
demonstrated to be free from interferences, under the conditions of the
analysis, by analyzing method blanks. Specific selection of reagents and
purification of solvents by distillation in all-glass systems may be required.
3.3 Interferences coextracted from samples will vary considerably from
source to source, depending upon the waste being sampled. Although general
cleanup techniques are recommended as part of this method, unique samples may
require additional cleanup.
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TABLE 1. GAS CHROMATOGRAPHY OF NITROAROMATICS AND ISOPHORONE
Retention time (m1n) Method detection
limit (ug/L)
Compound Col. la Col. 2b ECD FID
Isophorone
Nitrobenzene
2,4-D1n1trotoluene
2,6-D1n1trotoluene
Di nitrobenzene
Naphthoqulnone
4.49
3.31
5.35
3.52
5.72
4.31
6.54
4.75
15.7
13.7
0.02
0.01
5.7
3.6
-
-
aColumn 1: Gas-Chrom Q (80/100 mesh) coated with 1.95% QF-1/1.5% OV-17
packed 1n a 1.2-m x 2-mm or 4-mm I.D. glass column. A 2-mrn I.D. column and
nitrogen gas at 44 mL/m1n flow rate were used when determining Isophorone and
nitrobenzene by GC/FID. The column temperature was held Isothermal at 85*C.
A 4-mm I.D. column and 10% methane/90% argon carrier gas at 44 mL/m1n flow
rate were used when determining the dlnltrotoluenes by GC/ECD. The column
temperature was held Isothermal at 145*C.
bColumn 2: Gas-Chrom Q (80/100 mesh) coated with 3% OV-101 packed 1n a 3.0-
m x 2-mm or 4-mm I.D. glass column. A 2-mm I.D. column and nitrogen carrier
gas at 44 mL/m1n flow rate were used when determining Isophorone and
nitrobenzene by GC/FID. The column temperature was held Isothermal at 100*C.
A 4-mm I.D. column and 10% methane/90% argon carrier gas at 44 ml/mln flow
rate were used to determine the dlnltrotoluenes by GC/ECD. The column
temperature was held Isothermal at 150*C.
TABLE 2. DETERMINATION OF PRACTICAL QUANTITATION LIMITS (PQL) FOR VARIOUS
MATRICES3
Matrix Factorb
Ground water 10
Low-level soil by sonlcation with GPC cleanup 670
High-level soil and sludges by sonlcatlon 10,000
Non-water mlsdble waste 100,000
aSample PQLs are highly matrix-dependent. The PQLs listed herein are
provided for guidance and may not always be achievable.
Multiply the Method Detection Limits 1n Table 1 by the Factor to
determine the PQL for each analyte In the matrix to be analyzed.
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4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph;
4.1.1 Gas chromatograph: Analytical system complete with gas
chromatograph suitable for on-column Injections and all required
accessories, including detectors, column supplies, recorder, gases, and
syringes. A data system for measuring peak areas and/or peak heights is
recommended.
4.1.2 Columns:
4.1.2.1 Column 1: 1.2-m x 2- or 4-mrn I.D. glass column packed
with 1.95% QF-1/1.5% OV-17 on Gas-Chrom Q (80/100 mesh) or
equivalent.
4.1.2.2 Column 2: 3.0-m x 2- or 4-mm I.D. glass column packed
with 3% OV-101 on Gas-Chrom Q (80/100 mesh) or equivalent.
4.1.3 Detectors: Flame ionization (FID) or electron capture (ECD).
4.2 Kuderna-Danish (K-D) apparatus;
4.2.1 Concentrator tube: 10-mL, graduated (Kontes K-570050-1025 or
equivalent). Ground-glass stopper is used to prevent evaporation of
extracts
4.2.2 Evaporation flask: 500-mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs. *
4.2.3 Snyder column: Three-ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Snyder column: Two-ball micro (Kontes K-569001-0219 or
equivalent).
4.3 Boiling chips; Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.4 Water bath; Heated, with concentric ring cover, capable of
temperature control (+5*C). The bath should be used in a hood.
4.5 Volumetric flasks; 10-, 50-, and 100-mL, ground-glass stopper.
4.6 Microsyringe; 10-uL.
4.7 Syringe; 5-mL.
4.8 Vials; Glass, 2-, 10-, and 20-mL capacity with Teflon-lined screw
cap.
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5.0 REAGENTS
5.1 Solvents; hexane, acetone (pesticide quality or equivalent.)
5.2 Stock standard solutions;
5.2.1 Prepare stock standard solutions at a concentration of 1.00
ug/uL by dissolving 0.0100 g of assayed reference material 1n hexane and
diluting to volume 1n a 10-mL volumetric flask. Larger volumes can be
used at the convenience of the analyst. When compound purity 1s assayed
to be 96% or greater, the weight can be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards can be used at any concentration 1f they are certified by
the manufacturer or by an Independent source.
5.2.2 Transfer the stock standard solutions Into Teflon-sealed
screw-cap bottles. Store at 4*C and protect from light. Stock standards
should be checked frequently for signs of degradation or evaporation,
especially just prior to preparing calibration standards from them.
5.2.3 Stock standard solutions must be replaced after one year, or
sooner 1f comparison with check standards Indicates a problem.
5.3 Calibration standards; Calibration standards at a minimum of five
concentration levels are prepared through dilution of the stock standards with
hexane. One of the concentration levels should be at a concentration near,
but above, the method detection limit. The remaining concentration levels
should correspond to the expected range of concentrations found 1n real
samples or should define the working range of the GC. Calibration solutions
must be replaced after six months, or sooner 1f comparison with a check
standard indicates a problem.
5.4 Internal standards (if internal standard calibration is used); To
use this approach, the analyst must select one or more Internal standards that
are similar In analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the Internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.4.1 Prepare calibration standards at a minimum of five
concentration levels for each parameter of interest as described in
Paragraph 5.3.
5.4.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with hexane.
5.4.3 Analyze each calibration standard according to Section 7.0.
5.5 Surrogate standards; The analyst should monitor the performance of
the extraction, cleanup(when used), and analytical system and the effec-
tiveness of the method in dealing with each sample matrix by spiking each
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sample, standard, and reagent water blank with one or two surrogates (e.g., 2-
fluoroblphenyl) recommended to encompass the range of the temperature program
used 1n this method. Method 3500, Section 5.3.1.1, details Instructions on
the preparation of base/neutral surrogates. Deuterated analogs of analytes
should not be used as surrogates for gas chromatographic analysis due to
coelutlon problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the Introductory material to this chapter, Organic Analytes,
Section 4.1. Extracts must be stored under refrigeration and analyzed within
40 days of extraction.
7.0 PROCEDURE
7.1 Extraction;
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a pH
between 5 to 9 with methylene chloride, using either Method 3510 or 3520.
Solid samples are extracted using either Method 3540 or 3550.
7.1.2 Prior to gas chromatographic analysis, the extraction solvent
must be exchanged to hexane. The exchange is performed during the K-D
procedures listed in all of the extraction methods. The exchange may be
performed in one of two ways, depending on the data requirements. If the
detection limits cited in Table 1 must be achieved, the exchange should
be performed as described starting in Section 7.1.4. If these detection
limits are not necessary, solvent exchange is performed as outlined in
Section 7.1.3.
7.1.3 Solvent exchange when detection limits In Table 1 are not
required:
7.1.3.1 Following K-D of the methylene chloride extract to
1 mL using the macro-Snyder column, allow the apparatus to cool and
drain for at least 10 min.
7.1.3.2 Momentarily remove the Snyder column, add 50 mL of
hexane, a new boiling chip, and reattach the macro-Snyder column.
Concentrate the extract using 1 mL of hexane to prewet the Snyder
column. Place the K-D apparatus on the water bath so that the
concentrator tube is partially Immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete concentration in 5-10 min. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 1 mL, remove the K-D apparatus and allow it to drain and
cool for at least 10 min. The extract will be handled differently
8090 - 5
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Date September 1986
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at this point, depending on whether or not cleanup is needed. If
cleanup is not required, proceed to Paragraph 7.1.3.3. If cleanup
is needed, proceed to Paragraph 7.1.3.4.
7.1.3.3 If cleanup of the extract is not required, remove the
Snyder column and rinse the flask and its lower joint into the
concentrator tube with 1-2 mL of hexane. A 5-mL syringe is
recommended for this operation. Adjust the extract volume to
10.0 ml. Stopper the concentrator tube and store refrigerated at
4*C if further processing will not be performed immediately. If the
extract will be stored longer than two days, it should be
transferred to a Teflon-sealed screw-cap vial. Proceed with gas
chromatographic analysis.
7.1.3.4 If cleanup of the extract is required, remove the
Snyder column and rinse the flask and its lower joint into the
concentrator tube with a minimum amount of hexane. A 5-mL syringe
is recommended for this operation. Add a clean boiling chip to the
concentrator tube and attach a two-ball micro-Snyder column. Prewet
the column by adding about 0.5 ml of hexane to the top. Place the
micro-K-D apparatus on the water bath (80*C) so that the concen-
trator tube is partially immersed in the hot water. Adjust the
vertical position of the apparatus and the water temperature, as
required, to complete concentration in 5-10 min. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 0.5 ml, remove the K-D apparatus and allow it to drain and
cool for at least 10 min.
7.1.3.5 Remove the micro-Snyder column and rinse the flask and
its lower joint into the concentrator tube with 0.2 ml of hexane.
Adjust the extract volume to 2.0 ml and proceed with Method 3620.
7.1.4 Solvent exchange when detection limits listed 1n Table 1 must
be achieved:
7.1.4.1 Following K-D of the methylene chloride extract to
1 ml using the macro-Snyder column, allow the apparatus to cool and
drain for at least 10 min.
7.1.4.2 Remove the Snyder column and rinse the flask and its
lower joint into the concentrator tube with 1-2 ml of methylene
chloride. A 5-mL syringe is recommended for this operation. Add
1-2 mL of hexane, a clean boiling chip, and attach a two-ball micro-
Snyder column. Prewet the column by adding 0.5 mL of hexane to the
top. Place the micro-K-D apparatus on the water bath (60-65*C) so
that the concentrator tube is partially immersed in the hot water.
Adjust the vertical position of the apparatus and the water
temperature, as required, to complete concentration in 5-10 min. At
the proper rate of distillation the balls of the column will
actively chatter, but the chambers will not flood. When the
apparent volume of liquid reaches 0.5 mL, remove the K-D apparatus
and allow it to drain and cool for at least 10 min.
8090 - 6
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7.1.4.3 Remove the mlcro-Snyder column and rinse the flask and
its lower joint into the concentrator tube with a minimum amount of
hexane. The volume of the extract should be adjusted to 1.0 ml If
the extract will be analyzed without cleanup. If the extract will
require cleanup, adjust the volume to 2.0 ml with hexane. Stopper
the concentrator tube and store refrigerated at 4*C if further
processing will not be performed immediately. If the extract will
be stored longer than two days, it should be transferred to a
Teflon-sealed screw-cap vial. Proceed with either gas chromato-
graphic analysis or with cleanup, as necessary.
7.2 Gas chromatography conditions (Recommended); The determination of
dinitrotoluenesshouldbe performedusingGC/ECD. All other compounds
amenable to this method are to be analyzed by GC/FID.
7.2.1 Column 1: Set 10% methane/90% argon carrier gas flow at
44 mL/min flow rate. For a 2-mm I.D. column, set the temperature at 85*C
isothermal. For a 4-mm I.D. column, set the temperature at 145*C
isothermal.
7.2.2 Column 2: Set 10% methane/90% argon carrier gas flow at
44 mL/min flow rate. For a 2-mm I.D. column, set the temperature at
100*C isothermal. For a 4-mm I.D. column, set the temperature at 150*C
isothermal.
7.3 Calibration: Refer to Method 8000 for proper calibration
techniques^Use Table 1 and especially Table 2 for guidance on selecting the
lowest point on the calibration curve.
7.3.1 The procedure for internal or external standard calibration
may be used. Refer to Method 8000 for a description of each of these
procedures.
7.3.4 If cleanup is performed on the samples, the analyst should
process a series of standards through the cleanup procedure and then
analyze the samples by GC. This will confirm elution patterns and the
absence of interferents from the reagents.
7.4 Gas chromatographic analysis;
7.4.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 uL of internal standard to the sample prior to
injection.
7.4.2 Follow Section 7.6 in Method 8000 for instructions on the
analysis sequence, appropriate dilutions, establishing daily retention
time windows, and identification criteria. Include a mid-level standard
after each group of 10 samples in the analysis sequence when using FID
and after each group of 5 samples in the analysis sequence when using
ECD.
8090 - 7
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7.4.3 An example of a GC/FID chromatogram for nitrobenzene and
isophorone is shown in Figure 1. Figure 2 is an example of a GC/ECD
chromatogram of the dinitrotoluenes.
7.4.4 Record the sample volume injected and the resulting peak
sizes (in area units or peak heights).
7.4.5 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each analyte peak
in the sample chromatogram. See Section 7.8 of Method 8000 for
calculation equations.
7.4.6 If peak detection and identification are prevented due to
interferences, the hexane extract may undergo cleanup using Method 3620.
7.5 Cleanup;
7.5.1 Proceed with Method 3620, using the 2-mL hexane extracts
obtained from either Paragraph 7.1.3.5 or 7.1.4.3.
7.5.2 Following cleanup, the extracts should be analyzed by GC, as
described in the previous paragraphs and in Method 8000.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the
QC in Method 3600 and in the specific cleanup method.
8.2 Procedures to check the GC
8000, Section 8.6.
system operation are found in Method
8.2.1 The quality control check sample concentrate (Method 8000,
Section 8.6) should contain each parameter of interest in acetone at a
concentration of 20 ug/mL for each dinitrotoluene and 100 ug/mL for
isophorone and nitrobenzene.
8.2.2 Table 3 indicates the calibration and QC acceptance criteria
for this method. Table 4 gives method accuracy and precision as
functions of concentration for the analytes of interest. The contents of
both Tables should be used to evaluate a laboratory's ability to perform
and generate acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000, Section 8.10).
8090 - 8
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Date September 1986
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COLUMN: 1.5% OV-17 +1JS% QF-1
ON GAS CHROM Q
TEMPERATURE: 8S°C.
DETECTOR: FLAME IONIZATION
ui
5
i
o
24 6 8 10 12
RETENTION TIME-MINUTES
Figure 1. Gas chromatogram of nitrobcnzcna and itophorone.
8090 - 9
Revision p
Date September 1986
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COLUMN: 1.5% OV-17+1.95% OF-1
ON GAS CHROM Q
TEMPERATURE: 145®C.
DETECTOR: ELECTRON CAPTURE
2
1
o
-------
8.3.1 If recovery 1s not within limits, the following 1s required.
• Check to be sure there are no errors 1n calculations,
surrogate solutions and Internal standards. Also, check
Instrument performance.
• Recalculate the data and/or reanalyze the extract 1f any of
the above checks reveal a problem.
• Reextract and reanalyze the sample 1f none of the above are
a problem or flag the data as "estimated concentration."
9.0 METHOD PERFORMANCE
9.1 The method was tested by 18 laboratories using reagent water,
drinking water, surface water, and three industrial wastewaters spiked at six
concentrations over the range 1.0 to 515 ug/L. Single operator precision,
overall precision, and method accuracy were found to be directly related to
the concentration of the parameter and essentially independent of the sample
matrix. Linear equations to describe these relationships for a flame
ionizatlon detector are presented in Table 4.
9.2 The accuracy and precision obtained will be determined by the sample
matrix, sample-preparation technique, and calibration procedures used.
10.0 REFERENCES
1. "Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 4 - Nitroaromatlcs and Isophorone,'
Report for EPA Contract 68-03-2624 (in preparation).
2. "Determination of Nitroaromatics and Isophorone in Industrial and
Municipal Wastewaters," EPA-600/4-82-024, U.S. Environmental Protection
Agency, Environmental Monitoring and Support Laboratory, Cincinnati, Ohio
45268, June 1982.
3. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
4. "EPA Method Validation Study 19, Method 609 (Nitroaromatlcs and
Isophorone)," Report for EPA Contract 68-03-2624 (1n preparation).
5. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim Final
Rule and Proposed Rule," October 26, 1984.
6. Provost, L.P. and R.S. Elder, "Interpretation of Percent Recovery Data,"
American Laboratory, lj>, pp. 58-63, 1983.
8090 - 11
Revision
Date September 1986
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TABLE 3. QC ACCEPTANCE CRITERIA*
Parameter
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Isophorone
Nitrobenzene
Test
cone.
(ug/L)
20
20
100
100
Limit
for s
(ug/L)
5.1
4.8
32.3
33.3
Range
for X
(ug/L)
3.6-22.8
3.8-23.0
8.0-100.0
25.7-100.0
Range
P, PS
(%)
6-125
8-126
D-117
6-118
s = Standard deviation of four recovery measurements, 1n ug/L.
7 = Average recovery for four recovery measurements, 1n ug/L.
P, Ps = Percent recovery measured.
D = Detected; result must be greater than zero.
aCr1teria from 40 CFR Part 136 for Method 609. These criteria are based
directly upon the method performance data 1n Table 4. Where necessary, the
limits for recovery have been broadened to assure applicability of the limits
to concentrations below those used to develop Table 4.
8090 - 12
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Date September 1986
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TABLE 4. METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION
Parameter
2,4-D1n1trotoluene
2,4-D1n1trotoluene
Isophorene
Nitrobenzene
Accuracy, as
recovery, x1
(ug/L)
0.65C+0.22
0.66C+0.20
0.49C+2.93
0.60C+2.00
Single analyst
precision, sr'
(ug/L)
0.207+0.08
0.197+0.06
0.287+2.77
0.257+2.53
Overal 1
precision,
S' (ug/L)
0.377-0.07
0.367-0.00
0.467+0.31
0.377-0.78
x1 = Expected recovery for one or more measurements of a sample
containing a concentration of C, 1n ug/L.
sr' = Expected single analyst standard deviation of measurements at an
average concentration of 7, 1n ug/L.
S1 = Expected Interlaboratory standard deviation of measurements at an
average concentration found of 7, 1n ug/L.
C = True value for the concentration, 1n ug/L.
7 = Average recovery found for measurements of samples containing a
concentration of C, In ug/L.
8090 - 13
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METHOD B090
NITHOAHOMATICS AND CYCLIC KETONES
7.1.1
Choose
extraction
procedure from
Chapter 2
7.1.4
Concentrate to
1 mL using K-D
apparatus
7.1.3
1 Rinse
with hexane:
re-concentrate
to .5 ml_:
adjust to Z mL
7.1.3
Is cleanup
required?
Cleanup using
Method 362O
7.1.31 Rinse
1 flesh
Into concentra-
tor tube with
hexane: adjust
to 10 mL
0
Rinse
. with hexane:
concetrate to
.5 ml using K-D
Yes
7.1.4
Adjust volume
to
1 mL
7.1.4
Adjust volume
to Z mL
7.1.4
Cleanup using
Method 362O
8090 - 14
Revision Q
Date September 1986
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METHOD BO90
NITHOAROMATICS AND CYCLIC KETONES
(Continued)
Set GC column
Operating
conditions
7.3
Calibrate (see
Method 8000)
7.4 j
Perform
GC analysis
(see Method
aoooi
( Stop J
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Date September 1986
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METHOD 8100
POLYNUCLEAR AROMATIC HYDROCARBONS
1.0 SCOPE AND APPLICATION
1.1 Method 8100 is used to determine the concentration of certain
polynuclear aromatic hydrocarbons (PAH). Table 1 indicates compounds that may
be determined by this method.
1.2 The packed column gas chromatographic method described here cannot
adequately resolve the following four pairs of compounds: anthracene and
phenanthrene; chrysene and benzo(a)anthracene; benzo(b)fluoranthene and
benzo(k)fluoranthene; and dibenzo(a,h)anthracene and indeno(l,2,3-cd)pyrene.
The use of a capillary column instead of the packed column, also described in
this method, may adequately resolve these PAHs. However, unless the purpose
of the analysis can be served by reporting a quantitative sum for an
unresolved PAH pair, either liquid chromatography (Method 8310) or gas chroma-
tography/mass spectroscopy (Method 8270) should be used for these compounds.
2.0 SUMMARY OF METHOD
2.1 Method 8100 provides gas chromatographic conditions for the
detection of ppb levels of certain polynuclear aromatic hydrocarbons. Prior
to use of this method, appropriate sample extraction techniques must be used.
Both neat and diluted organic liquids (Method 3580, Waste Dilution) may be
analyzed by direct injection. A 2- to 5-uL aliquot of the extract 1s Injected
into a gas chromatograph (GC) using the solvent flush technique, and compounds
in the GC effluent are detected by a flame ionization detector (FID).
2.2 If interferences prevent proper detection of the analytes of
interest, the method may also be performed on extracts that have undergone
cleanup using silica gel column cleanup (Method 3630).
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing
misinterpretation of gas chromatograms. All of these materials must be
demonstrated to be free from interferences, under the conditions of the
analysis, by analyzing method blanks. Specific selection of reagents and
purification of solvents by distillation in all-glass systems may be required.
3.3 Interferences coextracted from samples will vary considerably from
source to source, depending upon the waste being sampled. Although general
cleanup techniques are recommended as part of this method, unique samples may
require additional cleanup.
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TABLE 1. GAS CHROMATOGRAPHY OF POLYNUCLEAR AROMATIC HYDROCARBONS3
Compound Retention time (min)
Acenaphthene 10.8
Acenaphthylene 10.4
Anthracene 15.9
Benzo(a)anthracene 20.6
Benzo(a)pyrene 29.4
Benzo
Benzo
Benzo
Benzo
b)fluoranthene 28.0
j)fluoranthene
kjfluoranthene 28.0
ghi)perylene 38.6
Chrysene 24.7
Dibenz(a,h)acridine
Dibenz(a,j)acridine
Dibenzo(a,h)anthracene 36.2
7H-Dibenzo(c,g)carbazole
Dibenzo(a,e)pyrene
Dibenzo(a,h)pyrene
Di benzo(a, i)pyrene
Fluoranthene 19.8
Fluorene 12.6
Indeno(l,2,3-cd)pyrene 36.2
3-Methylcholanthrene
Naphthalene 4.5
Phenanthrene 15.9
Pyrene 20.6
aResu1ts obtained using Column 1.
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4.0 APPARATUS AND MATERIALS
4.1 Gas chrornatograph;
4.1.1 Gas chromatograph: Analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detectors, column supplies, recorder, gases, and
syringes. A data system for measuring peak areas and/or peak heights is
recommended.
4.1.2 Columns:
4.1.2.1 Column 1: 1.8-m x 2-mm I.D. glass column packed with
3% OV-17 on Chromosorb W-AW-DCMS (100/120 mesh) or equivalent.
4.1.2.2 Column 2: 30-m x 0.25-mm I.D. SE-54 fused silica
capillary column.
4.1.2.3 Column 3: 30-m x 0.32-mm I.D. SE-54 fused silica
capillary column.
4.1.3 Detector: Flame ionizatlon (FID).
4.2 Volumetric flask; 10-, 50-, and 100-mL, ground-glass stopper.
4.3 Microsyrlnge: 10-uL.
5.0 REAGENTS
5.1 Solvents: Hexane, isooctane (2,2,4-trimethylpentane) (pesticide
quality or equivalent).
5.2 Stock standard solutions;
5.2.1 Prepare stock standard solutions at a concentration of 1.00
ug/uL by dissolving 0.0100 g of assayed reference material 1n isooctane
and diluting to volume 1n a 10-mL volumetric flask. Larger volumes can
be used at the convenience of the analyst. When compound purity is
assayed to be 96% or greater, the weight can be used without correction
to calculate the concentration of the stock standard. Commercially
prepared stock standards can be used at any concentration 1f they are
certified by the manufacturer or by an Independent source.
5.2.2 Transfer the stock standard solutions Into Teflon-sealed
screw-cap bottles. Store at 4*C and protect from light. Stock standards
should be checked frequently for signs of degradation or evaporation,
especially just prior to preparing calibration standards from them.
5.2.3 Stock standard solutions must be replaced after one year, or
sooner if comparison with check standards Indicates a problem.
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5.3 Calibration standards; Calibration standards at a minimum of five
concentrationlevelsshouldbe prepared through dilution of the stock
standards with isooctane. One of the concentration levels should be at a
concentration near, but above, the method detection limit. The remaining
concentration levels should correspond to the expected range of concentrations
found in real samples or should define the working range of the GC. Cali-
bration solutions must be replaced after six months, or sooner if comparison
with a check standard indicates a problem.
5.4 Internal standards (if internal standard calibration is used): To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.4.1 Prepare calibration standards at a minimum of five
concentration levels for each analyte of interest as described in
Paragraph 5.3.
5.4.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with isooctane.
5.4.3 Analyze each calibration standard according to Section 7.0.
5.5 Surrogate standards: The analyst should monitor the performance of
the extraction,cleanup(when used), and analytical system and the
effectiveness of the method in dealing with each sample matrix by spiking each
sample, standard, and reagent water blank with one or two surrogates (e.g., 2-
fluorobiphenyl and 1-fluoronaphthalene) recommended to encompass the range of
the temperature program used in this method. Method 3500, Section 5.3.1.1,
details instructions on the preparation of base/neutral surrogates.
Deuterated analogs of analytes should not be used as surrogates for gas
chromatographic analysis due to coelution problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1. Extracts must be stored under refrigeration and must be analyzed
within 40 days of extraction.
7.0 PROCEDURE
7.1 Extraction;
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a
neutral pH with methylene chloride, using either Method 3510 or 3520.
Solid samples are extracted using either Method 3540 or 3550. To achieve
maximum sensitivity with this method, the extract must be concentrated to
1 mL.
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7.2 Gas chromatography conditions (Recommended);
7.2.1 Column 1: Set nitrogen carrier gas flow at 40-mL/m1n flow
rate. Set column temperature at 100*C for 4 m1n; then program at 8*C/min
to a final hold at 280*C.
7.2.2 Column 2: Set helium carrier gas at 20-cm/sec flow rate.
Set column temperature at 35'C for 2 min; then program at 10*C/m1n to
265*C and hold for 12 m1n.
7.2.3 Column 3: Set helium carrier gas at 60 cm/sec flow rate.
Set column temperature at 35*C for 2 m1n; then program at 10*C/m1n to
265*C and hold for 3 mln.
7.3 Calibration: Refer to Method 8000 for proper calibration
techniques.
7.3.1 The procedure for Internal or external standard calibration
may be used. Refer to Method 8000 for a description of each of these
procedures.
7.3.2 If cleanup is performed on the samples, the analyst should
process a series of standards through the cleanup procedure and then
analyze the samples by GC. This will validate elutlon patterns and the
absence of interferents from the reagents.
7.4 Gas chromatographic analysis;
7.4.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 uL of internal standard to the sample prior to
injection.
7.4.2 Follow Section 7.6 in Method 8000 for instructions on the
analysis sequence, appropriate dilutions, establishing daily retention
time windows, and Identification criteria. Include a mid-level standard
after each group of 10 samples in the analysis sequence.
7.4.3 Record the sample volume Injected and the resulting peak
sizes (in area units or peak heights).
7.4.4 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes. See Section 7.8 of Method 8000 for calculation
equations.
7.4.5 If peak detection and identification are prevented due to
interferences, the extract may undergo cleanup using Method 3630.
7.5 Cleanup:
7.5.1 Proceed with Method 3630. Instructions are given in this
method for exchanging the solvent of the extract to hexane.
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7.5.2 Following cleanup, the extracts should be analyzed by GC, as
described in the previous paragraphs and in Method 8000.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the
QC in Method 3600 and in the specific cleanup method.
8.2 Procedures to check the GC system operation are found in Method
8000, Section 8.6.
8.2.1 The quality control check sample concentrate (Method 8000,
Section 8.6) should contain each analyte at the following concentrations
in acetonitrile: naphthalene, 100 ug/mL; acenaphthylene, 100 ug/mL;
acenaphthene, 100 ug/mL; fluorene, 100 ug/mL; phenanthrene, 100 ug/mL;
anthracene, 100 ug/mL; benzo(k)fluoranthene, 5 ug/mL; and any other PAH
at 10 ug/mL.
8.2.2 Table 2 indicates the calibration and QC acceptance criteria
for this method. Table 3 gives method accuracy and precision as
functions of concentration for the analytes of interest. The contents of
both Tables should be used to evaluate a laboratory's ability to perform
and generate acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000, Section 8.10).
8.3.1 If recovery is not within limits, the following procedures
are required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are
a problem or flag the data as "estimated concentration."
9.0 METHOD PERFORMANCE
9.1 The method was tested by 16 laboratories using reagent water,
drinking water, surface water, and three industrial wastewaters spiked at six
concentrations over the range 0.1 to 425 ug/L. Single operator precision,
overall precision, and method accuracy were found to be directly related to
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the concentration of the analyte and essentially independent of the sample
matrix. Linear equations to describe these relationships for a flame
ionization detector are presented in Table 3.
9.2 This method has been tested for linearity of spike recovery from
reagent water and has been demonstrated to be applicable over the
concentration range from 8 x MDL to 800 x MDL with the following exception:
benzo(ghi)perylene recovery at 80 x and 800 x MDL were low (35% and 45%,
respectively).
9.3 The accuracy and precision obtained will be determined by the sample
matrix, sample-preparation technique, and calibration procedures used.
10.0 REFERENCES
1. "Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 9 - PAHs," Report for EPA Contract 68-03-
2624 (in preparation).
2. Sauter, A.D., L.D. Betowski, T.R. Smith, V.A. Strickler, R.G. Beimer,
B.N. Colby, and J.E. Wilkinson, "Fused Silica Capillary Column GC/MS for the
Analysis of Priority Pollutants," Journal of HRC&CC 4, 366-384, 1981.
3. "Determination of Polynuclear Aromatic Hydrocarbons in Industrial and
Municipal Wastewaters," EPA-600/4-82-025, U.S. Environmental Protection
Agency, Environmental Monitoring and Support Laboratory, Cincinnati, Ohio
45268, September 1982.
4. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
5. "EPA Method Validation Study 20, Method 610 (Polynuclear Aromatic
Hydrocarbons)," Report for EPA Contract 68-03-2624 (in preparation).
6. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim Final
Rule and Proposed Rule," October 26, 1984.
7. Provost, L.P. and R.S. Elder, "Interpretation of Percent Recovery Data,"
American Laboratory, 15, pp. 58-63, 1983.
8100 - 7
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TABLE 2. QC ACCEPTANCE CRITERIA3
Parameter
Acenaphthene
Acenaphthylene
Anthracene
Benzo (a) anthracene
Benzo(a)pyrene
Benzo (b) f 1 uoranthene
Benzo(ghi)perylene
Benzo (k) f 1 uoranthene
Chrysene
Di benzo (a , h) anthracene
Fl uoranthene
Fluorene
Indeno (1,2, 3-cd) py rene
Naphthalene
Phenanthrene
Pyrene
Test
cone.
(ug/L)
100
100
100
10
10
10
10
5
10
10
10
100
10
100
100
10
Limit
for s
(ug/L)
40.3
45,1
28.7
4.0
4.0
3.1
2.3
2.5
4.2
2.0
3.0
43.0
3.0
40.7
37.7
3.4
Range
for 7
(ug/L)
D-105.7
22.1-112.1
11.2-112.3
3.1-11.6
0.2-11.0
1.8-13.8
D-10.7
D-7.0
D-17.5
0.3-10.0
2.7-11.1
D-119
1.2-10.0
21.5-100.0
8.4-133.7
1.4-12.1
Range
P, PS
(%)
D-124
D-139
D-126
12-135
D-128
6-150
D-116
D-159
D-199
D-110
14-123
D-142
D-116
D-122
D-155
D-140
s = Standard deviation of four recovery measurements, in ug/L.
7 = Average recovery for four recovery measurements, in ug/L.
P, Ps = Percent recovery measured.
D = Detected; result must be greater than zero.
Criteria from 40 CFR Part 136 for Method 610. These criteria are based
directly upon the method performance data in Table 3. Where necessary, the
limits for recovery have been broadened to assure applicability of the limits
to concentrations below those used to develop Table 3.
8100 - 8
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TABLE 3. METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION
Parameter
Acenaphthene
Acenaphthylene
Anthracene
Benzo (a) anthracene
Benzo (a)pyrene
Benzo (b) f 1 uoranthene
Benzo (ghi)perylene
Benzo (k) f 1 uoranthene
Chrysene
Di benzo (a , h) anthracene
Fl uoranthene
Fluorene
Ideno (1 , 2 , 3-cd) pyrene
Naphthalene
Phenanthrene
Pyrene
Accuracy, as
recovery, x1
(ug/L)
0.52C+0.54
0.69C-1.89
0.63C-1.26
0.73C+0.05
0.56C+0.01
0.78C+0.01
0.44C+0.30
0.59C+0.00
0.77C-0.18
0.41C-0.11
0.68C+0.07
0.56C-0.52
0.54C+0.06
0.57C-0.70
0.72C-0.95
0.69C-0.12
Single analyst
precision, sr'
(ug/L)
0.397+0.76
0.367+0.29
0.237+1.16
0.287+0.04
0.387-0.01
0.217+0.01
0.257+0.04
0.447-0.00
0.327-0.18
0.247+0.02
0.227+0.06
0.447-1.12
0.297+0.02
0.397-0.18
0.297+0.05
0.257+0.14
Overall
precision,
S1 (ug/L)
0.537+1.32
0.427+0.52
0.417+0.45
0.347+0.02
0.537-0.01
0.387-0.00
0.587+0.10
0.697+0.10
0.667-0.22
0.457+0.03
0.327+0.03
0.637-0.65
0.427+0.01
0.417+0.74
0.477-0.25
0.427-0.00
x1 = Expected recovery for one or more measurements of a sample
containing a concentration of C, in ug/L.
sr' = Expected single analyst standard deviation of measurements at an
average concentration of 7, in ug/L.
S1 = Expected interlaboratory standard deviation of measurements at an
average concentration found of 7, in ug/L.
C = True value for the concentration, in ug/L.
7 = Average recovery found for measurements of samples containing a
concentration of C, in ug/L.
8100 - 9
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METHOD 8100
POLYNUCLEAR AROMATIC HYDROCARBONS
C
7.1.1
Choose
« appro-
priate extract-
Ion procedure
(refer to
Chapter S)
7.2
Set gas
chrontatography
conditions
7.3
through cleanup
procedures
7.4
.Do GC analysis
(refer to
Method 8000)
Refer to
Method 6000
for proper
calibration
techniques
Q
7.5. 1
Do cleanup
using Method
3630
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METHOD 8110
HALOETHERS BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 This method covers the determination of certain haloethers. The
following compounds can be determined by this method:
Appropriate Technique
Compound Name CAS No.a 3510 3520 3540 3550 3580
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
4-Bromophenyl phenyl ether
4-Chlorophenyl phenyl ether
111-91-1
111-44-4
108-60-1
101-55-3
7005-72-3
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
a Chemical Abstract Services Registry Number.
X Greater than 70 percent recovery by this technique.
1.2 This is a gas chromatographic (GC) method applicable to the
determination of the compounds listed above in municipal and industrial
discharges. When this method is used to analyze unfamiliar samples for any or
all of the compounds above, compound identifications should be supported by at
least one additional qualitative technique. This method describes analytical
conditions of a second GC column that can be used to confirm measurements made
with the primary column. Method 8270 provides gas chromatograph/mass
spectrometer (GC/MS) conditions appropriate for the qualitative and quantitative
confirmation of results for all of the parameters listed above, using the extract
from this method.
1.3 The method detection limit (MDL, defined in Section 9.1) for each
parameter is listed in Table 1. The MDL for a specific wastewater may differ
from that listed, depending upon the nature of interferences in the sample
matrix.
1.4 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatography and in the interpretation
of gas chromatograms. Each analyst must demonstrate the ability to generate
acceptable results with this method using the procedure described in Section 8.2.
1.5 The toxicity or carcinogenicity of each reagent used in this method
has not been precisely defined. However, each chemical compound should be
treated as a potential health hazard. From this viewpoint, exposure to these
chemicals must be reduced to the lowest possible level by whatever means
available. The laboratory is responsible for maintaining a current awareness
file of OSHA regulations regarding the safe handling of the chemicals specified
in this method. A reference file of material data handling sheets should also
8110 - 1 Revision 0
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be made available to all personnel involved in the chemical analysis. Additional
references to laboratory safety are available and have been identified.
2.0 SUMMARY OF METHOD
2.1 A measured volume of sample, approximately one-liter, is solvent
extracted with methylene chloride using a separatory funnel. The methylene
chloride extract is dried and exchanged to hexane during concentration to a
volume of 10 ml or less. GC conditions are described which permit the separation
and measurement of the compounds in the extract using a halide specific detector.
2.2 Method 8110 provides gas chromatographic conditions for the detection
of ppb concentrations of haloethers. Prior to use of this method, appropriate
sample extraction techniques must be used. Both neat and diluted organic liquids
(Method 3580, Waste Dilution) may be analyzed by direct injection. A 2 to 5 nL
aliquot of the extract is injected into a gas chromatograph (GC) using the
solvent flush technique, and compounds in the GC effluent are detected by an
electrolytic conductivity detector (HECD).
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Matrix interferences may be caused by contaminants that are
coextracted from the sample. The extent of matrix interferences will vary
considerably from source to source, depending upon the nature and diversity of
the industrial complex or municipality being sampled. The cleanup procedures in
Section 7.3 can be used to overcome many of these interferences, but unique
samples may require additional cleanup approaches to achieve the MDL listed in
Table 1.
3.3 Dichlorobenzenes are known to coelute with haloethers under some gas
chromatographic conditions. If these materials are present in a sample, it may
be necessary to analyze the extract with two different column packings to
completely resolve all of the compounds.
3.4 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All these materials must be demonstrated to be free from
interferences under the conditions of the analysis, by analyzing reagent blanks.
Specific selection of reagents and purification of solvents by distillation in
all-glass systems may be required.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph - An analytical system complete with
temperature programmable gas chromatograph suitable for on-column
injection and all required accessories including syringes, analytical
columns, gases, detector, and strip-chart recorder. A data system is
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recommended for measuring peak areas.
4.1.2 Columns
4.1.2.1 Column 1 - 1.8 m x 2 mm ID pyrex glass, packed
with Supelcoport, (100/120 mesh) coated with 3% SP-1000 or
equivalent. This column was used to develop the method performance
statements in Section 9.0. Guidelines for the use of alternate
column packings are provided in Section 7.3.1.
4.1.2.2 Column 2 - 1.8 m x 2 mm ID pyrex glass, packed
with 2,6-diphenylene oxide polymer (Tenax-GC 60/80 mesh) or
equivalent.
4.1.3 Detector - Electrolytic conductivity or microcoulometric.
These detectors have proven effective in the analysis of wastewaters for
the parameters listed in the scope of this method. The Hall conductivity
detector (HECD) was used to develop the method performance statements in
Section 9.0. Guidelines for the use of alternate detectors are provided
in Section 7.3.1. Although less selective, an electron capture detector
(ECD) is an acceptable alternative.
4.2 Kuderna-Danish (K-D) apparatus
4.2.1 Concentrator tube - 10 mL graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.2.2 Evaporation flask - 500 ml (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.2.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Vials - Amber glass, 10 to 15 ml capacity, with Teflon lined screw-
cap or crimp top.
4.4 Boiling chips - Approximately 10/40 mesh. Heat to 400°C for
30 minutes or Soxhlet extract with methylene chloride.
4.5 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 2°C). The bath should be used in a hood.
4.6 Balance - Analytical, 0.0001 g.
4.7 Volumetric flasks, Class A - Appropriate sizes with ground glass
stoppers.
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5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all inorganic reagents shall conform to
the specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may be
used, provided it is first ascertained that the reagent is of sufficiently high
purity to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.4 Hexane, C6HU - Pesticide quality or equivalent.
5.5 Isooctane, (CH3)3CCH2CH(CH3)2 - Pesticide quality or equivalent.
5.6 Stock standard solutions (1000 mg/L) - Stock standard solutions can
be prepared from pure standard materials or purchased as certified solutions.
5.6.1 Prepare stock standard solutions by accurately weighing
0.1000 ± 0.0010 g of pure material. Dissolve the material in pesticide
quality acetone and dilute to volume in a 100 ml volumetric flask. Larger
volumes can be used at the convenience of the analyst. If compound purity
is certified at 96% or greater, the weight can be used without correction
to calculate the concentration of the stock standard. Commercially
prepared stock standards can be used at any concentration if they are
certified by the manufacturer or by an independent source.
5.6.2 Transfer the stock standard solutions into bottles with Teflon
lined screw-caps or crimp tops. Store at 4°C and protect from light.
Stock standard solutions should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them.
5.6.3 Stock standard solutions must be replaced after six months, or
sooner if comparison with check standards indicates a problem.
5.7 Calibration standards - Calibration standards at a minimum of five
concentrations should be prepared through dilution of the stock standards with
isooctane. One of the concentrations should be at a concentration near, but
above, the method detection limit. The remaining concentrations should
correspond to the expected range of concentrations found in real samples or
should define the working range of the GC. Calibration solutions must be
replaced after six months, or sooner if comparison with check standards indicates
a problem.
5.8 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
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5.8.1 Prepare calibration standards at a minimum of five
concentrations for each analyte of interest as described in Section 5.7.
5.8.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with isooctane.
5.8.3 Analyze each calibration standard according to Section 7.0.
5.9 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system and the effectiveness
of the method in dealing with each sample matrix by spiking each sample,
standard, and reagent blank with one or two surrogates (e.g. haloethers that are
not expected to be in the sample) recommended to encompass the range of the
temperature program used in this method. Method 3500 details instructions on the
preparation of base/neutral surrogates. Deuterated analogs of analytes should
not be used as surrogates for gas chromatographic analysis due to coelution
problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1. Extracts must be stored at 4°C and analyzed within 40 days of
extraction.
7.0 PROCEDURE
7.1 Extraction
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a
neutral, or as is, pH with methylene chloride, using either Method 3510 or
3520. Solid samples are extracted using either Method 3540 or 3550.
NOTE: Some of the haloethers are very volatile and significant
losses will occur in concentration steps if care is not
exercised. It is important to maintain a constant gentle
evaporation rate and not to allow the liquid volume to fall
below 1 to 2 mL before removing the K-D apparatus from the hot
water bath.
7.1.2 Prior to gas chromatographic analysis, the extraction solvent
must be exchanged to hexane. The exchange is performed during the K-D
procedures listed in all of the extraction methods. The exchange is
performed as follows.
7.1.2.1 Following K-D of the methylene chloride extract to
1 mL using the macro-Snyder column, allow the apparatus to cool and
drain for at least 10 minutes.
7.1.2.2 Momentarily remove the Snyder column, add 50 mL of
hexane, a new boiling chip, and reattach the macro-Snyder column.
Concentrate the extract using 1 mL of hexane to prewet the Snyder
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column. Place the K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete concentration in 5-10 minutes. At the proper
rate of distillation the balls of the column will actively chatter,
but the chambers will not flood. When the apparent volume of liquid
reaches 1 ml, remove the K-D apparatus and allow it to drain and
cool for at least 10 minutes. The extract will be handled
differently at this point, depending on whether or not cleanup is
needed. If cleanup is not required, proceed to Section 7.1.2.3. If
cleanup is needed, proceed to Section 7.1.2.4.
7.1.2.3 If cleanup of the extract is not required, remove
the Snyder column and rinse the flask and its lower joint into the
concentrator tube with 1-2 ml of hexane. A 5 ml syringe is
recommended for this operation. Adjust the extract volume to
10.0 mL. Stopper the concentrator tube and store refrigerated at
4°C if further processing will not be performed immediately. If the
extract will be stored longer than two days, it should be
transferred to a Teflon lined screw-cap vial. Proceed with gas
chromatographic analysis.
7.1.2.4 If cleanup of the extract is required, remove the
Snyder column and rinse the flask and its lower joint into the
concentrator tube with a minimum amount of hexane. A 5 ml syringe
is recommended for this operation. Add a clean boiling chip to the
concentrator tube and attach a two ball micro-Snyder column. Prewet
the column by adding about 0.5 ml of hexane to the top. Place the
micro-K-D apparatus on the water bath (80°C) so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete concentration in 5-10 minutes. At the proper
rate of distillation the balls of the column will actively chatter,
but the chambers will not flood. When the apparent volume of liquid
reaches 0.5 ml, remove the K-D apparatus and allow it to drain and
cool for at least 10 minutes.
7.1.2.5 Remove the micro-Snyder column and rinse the flask
and its lower joint into the concentrator tube with 0.2 mL of
hexane. Adjust the extract volume to 2.0 ml and proceed with either
Method 3610 or 3620.
7.2 Cleanup
7.2.1 Proceed with Method 3620, using the 2 ml hexane extracts
obtained from Section 7.1.2.5.
7.2.2 Following cleanup, the extracts should be analyzed by GC, as
described in the previous paragraphs and in Method 8000.
7.3 Gas Chromatography Conditions
7.3.1 Table 1 summarizes the recommended operating conditions for
the gas chromatograph. This table includes retention times and MDLs that
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were obtained under these conditions. Examples of the parameter
separations achieved by these columns are shown in Figures 1 and 2. Other
packed columns, chromatographic conditions, or detectors may be used if
the requirements of Section 8.2 are met. Capillary (open-tubular) columns
may also be used if the relative standard deviations of responses for
replicate injections are demonstrated to be less than 6% and the
requirements of Section 8.2 are met.
7.4 Calibration - Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point on
the calibration curve.
7.4.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.4.2 If cleanup is performed on the samples, the analyst should
process a series of standards through the cleanup procedure and then
analyze the samples by GC. This will confirm elution patterns and the
absence of interferents from the reagents.
7.5 Gas chromatographic analysis
7.5.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 nL of internal standard to the sample prior to
injection.
7.5.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration check standard after
each group of 10 samples in the analysis sequence.
7.5.3 Examples of GC/HECD chromatograms for haloethers are shown in
Figures 1 and 2.
7.5.4 Record the sample volume injected and the resulting peak sizes
(in area units or peak heights).
7.5.5 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each analyte peak in
the sample chromatogram. See Method 8000 for calculation equations.
7.5.6 If peak detection and identification are prevented due to
interferences, the hexane extract may undergo cleanup using either Method
3610 or 3620.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the QC
in Method 3600 and in the specific cleanup method.
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8.2 Procedures to check the GC system operation are found in Method 8000,
Section 8.6.
8.2.1 The quality control (QC) reference sample concentrate (Method
8000, Section 8.6) should contain each analyte of interest at 20 mg/L.
8.2.2 Table 1 indicates the recommended operating conditions,
retention times, and MDLs that were obtained under these conditions.
Table 2 gives method accuracy and precision for the analytes of interest.
The contents of both Tables should be used to evaluate a laboratory's
ability to perform and generate acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000, Section 8.10).
8.3.1 If recovery is not within limits, the following is required.
• Check to be sure that there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are a
problem or flag the data as "estimated concentration."
9.0 METHOD PERFORMANCE
9.1 This method has been tested for linearity of recovery from spiked
organic-free reagent water and has been demonstrated to be applicable for the
concentration range from 4 x MDL to 1000 x MDL.
9.2 In a single laboratory (Monsanto Research Center), using spiked
wastewater samples, the average recoveries presented in Table 2 were obtained.
Each spiked sample was analyzed in triplicate on three separate occasions. The
standard deviation of the percent recovery is also included in Table 2.
10.0 REFERENCES
1. Fed. Regist. 1984, 49, 43234; October 26.
2. Mills, P.A. "Variation of Florisil Activity: Simple Method for Measuring
Absorbent Capacity and Its Use in Standardizing Florisil Columns"; Journal
of the Association of Official Analytical Chemists 1968, 51_, 29.
3. Handbook of Analytical Quality Control in Water and Wastewater
Laboratories; U.S. Environmental Protection Agency. Office of Research and
Development. Environmental Monitoring and Support Laboratory. ORD
Publication Offices of Center for Environmental Research Information:
Cincinnati, OH, 1979; EPA-600/4-79-019.
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4. Methods for Chemical Analysis of Water and Wastes; U.S. Environmental
Protection Agency. Office of Research and Development. Environmental
Monitoring and Support Laboratory. ORD Publication Offices of Center for
Environmental Research Information: Cincinnati, OH, 1983; EPA-600/4-79-
020.
5. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects"; Journal of the Association of Official Analytical
Chemists 1965, 48, 1037.
6. "EPA Method Validation Study 21 Methods 611 (Haloethers)," Report for EPA
Contract 68-03-2633.
7. "Determination of Haloethers in Industrial and Municipal Wastewaters";
Report for EPA Contract 68-03-2633 (In preparation).
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
Analyte
Retention Time
(minutes)
Column 1 Column 2
Method
Detection Limit
(M9/L)
Bis(2-chloroisopropyl) ether
Bis(2-chloroethyl) ether
Bis(2-chloroethoxy)methane
4-Chlorophenyl phenyl ether
4-Bromophenyl phenyl ether
8.4
9.4
13.1
19.4
21.2
9.7
9.1
10.0
15.0
16.2
0.8
0.3
0.5
3.9
2.3
Column 1 conditions:
Carrier gas (He) flow rate:
Initial temperature:
Temperature program:
Final temperature:
40 mL/min
60°C> hold for 2 minutes
60°C to 230°C at 8°C/min
230°C, hold for 4 minutes
Under these conditions the retention time for aldrin is 22.6 minutes.
Column 2 conditions:
Carrier gas (He) flow rate:
Initial temperature:
Temperature program:
Final temperature:
40 mL/min
150°C, hold for 4 minutes
150°C to 310°C at 16°C/min
310°C
Under these conditions the retention time for aldrin is 18.4 minutes.
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TABLE 2.
SINGLE OPERATOR ACCURACY AND PRECISION
Average Standard Spike Number
Percent Deviation Range of Matrix
Analyte Recovery % (M9/L) Analyses Types
Bi s (2 -chl oroethoxyJmethane62 57313827 3
Bis(2-chloroethyl) ether 59 4.5 97 27 3
Bis(2-chloroisopropyl) ether 67 4.0 54 27 3
4-Bromophenyl phenyl ether 78 3.5 14 27 3
4-Chlorophenyl phenyl ether 73 4.5 30 27 3
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FIGURE 1.
GAS CHROMATOGRAM OF HALOETHERS
Column: 3% SP-10OO on Suptleoport
Progrtm: 60°C. -2 mmuttt 8"/mmut» to 23O°C.
Detector: Hull »/»ctrolytic conductivity
1
-c
5
i
5
2 4 6 8 to 12 14 16 18 20 22 24
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FIGURE 2.
GAS CHROMATOGRAM OF HALOETHERS
Column: Tenex GC
Progrem: 150°C.-4 minutes 16*/minute to 310*C.
Detector: Hall electrolytic conductivity
12 16 20
Retention time, minutes
24
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METHOD 8110
HALOETHERS BY GAS CHROMATOGRAPHY
Start
7124 Perform
micro-K D procedure
using hexane,
proceed with Method
3610 or 3620
711 Choose
appr oprlate
extraction
pr ocedure
712 Perform
solvent exchange
using hexane
7123 Adjust
extract volume and
proceed with
ana lysis or store
in appropriate
manne r
7 3 1 Refer to
Table 1 for
r ecommended
o pe ra 11ng
conditions for the
CC
4 Refer to Method
8000 for proper
ca 1 ibra tion
techniques
751 Refer to
Method 8000 for
guidance on CC
anal ys is
7 5 4 Record sample
volume injected and
resul ting
peak size
755 Perform
appr opr la te
calculations (refer
to Method 8000)
Stop
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METHOD 8120A
CHLORINATED HYDROCARBONS BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8120 is used to determine the concentration of certain
chlorinated hydrocarbons. The following compounds can be determined by this
method:
Appropriate Preparation Techniques
Compounds CAS No" 3510 3520 3540/ 3550 3580
3541
2-Chloronaphthalene
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachl orobenzene
Hexachl orobutadi ene
Hexachl orocycl ohexane
Hexachl orocycl opentadi ene
Hexachl oroethane
Pentachlorohexane
Tetrachl orobenzenes
1, 2, 4-Tri chl orobenzene
91-58-7
95-50-1
541-73-1
106-46-7
118-74-1
87-68-3
608-73-1
77-47-4
67-72-1
--
120-82-1
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
a Chemical Abstract Services Registry Number.
x Greater than 70 percent recovery by this technique
ND Not determined.
1.2 Table 1 indicates compounds that may be determined by this method and
lists the method detection limit for each compound in organic-free reagent water.
Table 2 lists the estimated quantitation limit (EQL) for other matrices.
2.0 SUMMARY OF METHOD
2.1 Method 8120 provides gas chromatographic conditions for the detection
of ppb concentrations of certain chlorinated hydrocarbons. Prior to use of this
method, appropriate sample extraction techniques must be used. Both neat and
diluted organic liquids (Method 3580, Waste Dilution) may be analyzed by direct
injection. A 2 to 5 juL aliquot of the extract is injected into a gas
chromatograph (GC), and compounds in the GC effluent are detected by an electron
capture detector (ECD).
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2.2 If interferences are encountered in the analysis, Method 8120 may
also be performed on extracts that have undergone cleanup using Method 3620.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All of these materials must be demonstrated to be free
from interferences, under the conditions of the analysis, by analyzing method
blanks. Specific selection of reagents and purification of solvents by
distillation in all glass systems may be required.
3.3 Interferences coextracted from samples will vary considerably from
source to source, depending upon the waste being sampled. Although general
cleanup techniques are recommended as part of this method, unique samples may
require additional cleanup.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph - Analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detectors, column supplies, recorder, gases, and
syringes. A data system for measuring peak areas and/or peak heights is
recommended.
4.1.2 Columns
4.1.2.1 Column 1 - 1.8 m x 2 mm ID glass column packed
with 1% SP-1000 on Supelcoport (100/120 mesh) or equivalent.
4.1.2.2 Column 2 - 1.8 m x 2 mm ID glass column packed
with 1.5% OV-1/2.4% OV-225 on Supelcoport (80/100 mesh) or
equivalent.
4.2
4.1.3 Detector - Electron capture (ECD).
Kuderna-Danish (K-D) apparatus
4.2.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts
4.2.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps or
equivalent.
4.2.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
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4.2.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.2.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.4 Water bath - Heated, with concentric ring cover, capable of
temperature control (+ 5°C). The bath should be used in a hood.
4.5 Volumetric flasks - 10, 50, and 100 ml, with ground glass stoppers.
4.6 Microsyringe - 10 juL.
4.7 Syringe - 5 ml.
4.8 Vials - Glass, 2, 10, and 20 ml capacity with Teflon lined screw-
caps or crimp tops.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Solvents
5.3.1 Hexane, C6H14. Pesticide quality or equivalent.
5.3.2 Acetone, CH3COCH3. Pesticide quality or equivalent.
5.3.3 Isooctane, C8H18. Pesticide quality or equivalent.
5.4 Stock standard solutions
5.4.1 Prepare stock standard solutions at a concentration of 1000
mg/L by dissolving 0.0100 g of assayed reference material in isooctane or
hexane and diluting to volume in a 10 ml volumetric flask. Larger volumes
can be used at the convenience of the analyst. When compound purity is
assayed to be 96% or greater, the weight can be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards can be used at any concentration if they are certified by
the manufacturer or by an independent source.
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5.4.2 Transfer the stock standard solutions into vials with Teflon
lined screw caps or crimp tops. Store at 4°C and protect from light.
Stock standards should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards.
5.4.3 Stock standard solutions must be replaced after one year, or
sooner if comparison with check standards indicates a problem.
5.5 Calibration standards - Calibration standards at a minimum of five
concentrations should be prepared through dilution of the stock standards with
isooctane or hexane. One of the concentrations should be at a concentration
near, but above, the method detection limit. The remaining concentrations should
correspond to the expected range of concentrations found in real samples or
should define the working range of the GC. Calibration solutions must be
replaced after six months, or sooner if comparison with check standards indicates
a problem.
5.6 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.6.1 Prepare calibration standards at a minimum of five
concentrations for each analyte of interest as described in Sec. 5.5.
5.6.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with isooctane or
hexane.
5.6.3 Analyze each calibration standard according to Sec. 7.0.
5.7 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system and the effectiveness
of the method in dealing with each sample matrix by spiking each sample,
standard, and organic-free reagent water blank with one or two surrogates (e.g.
chlorinated hydrocarbons that are not expected to be in the sample) recommended
to encompass the range of the temperature program used in this method. Method
3500 details instructions on the preparation of base/neutral surrogates.
Deuterated analogs of analytes should not be used as surrogates for gas
chromatographic analysis due to coelution problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes, Sec.
4.1.
6.2 Extracts must be stored under refrigeration and analyzed within 40
days of extraction.
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7.0 PROCEDURE
7.1 Extraction
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a
neutral, or as is, pH with methylene chloride, using either Method 3510 or
3520. Solid samples are extracted using either Methods 3540/3541 or 3550.
7.1.2 Prior to gas chromatographic analysis, the extraction solvent
must be exchanged to hexane. The exchange is performed during the K-D
procedures listed in all of the extraction methods. The exchange is
performed as follows.
7.1.2.1 Following K-D of the methylene chloride extract
to 1 ml using the macro Snyder column, allow the apparatus to cool
and drain for at least 10 minutes.
7.1.2.2 Momentarily remove the Snyder column, add 50 mi
of hexane, a new boiling chip, and reattach the macro Snyder column.
Concentrate the extract using 1 ml of hexane to prewet the Snyder
column. Place the K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete concentration in 5-10 minutes. At the proper
rate of distillation the balls of the column will actively chatter,
but the chambers will not flood. When the apparent volume of liquid
reaches 1 ml, remove the K-D apparatus and allow it to drain and
cool for at least 10 minutes. The extract will be handled
differently at this point, depending on whether or not cleanup is
needed. If cleanup is not required, proceed to Sec. 7.1.2.3. If
cleanup is needed, proceed to Sec. 7.1.2.4.
7.1.2.3 If cleanup of the extract is not required, remove
the Snyder column and rinse the flask and its lower joint into the
concentrator tube with 1-2 ml of hexane. A 5 ml syringe is
recommended for this operation. Adjust the extract volume to
10.0 ml. Stopper the concentrator tube and store refrigerated at 4°C
if further processing will not be performed immediately. If the
extract will be stored, longer than two days, it should be
transferred to a vial with a Teflon lined screw cap or crimp top.
Proceed with gas chromatographic analysis.
7.1.2.4 If cleanup of the extract is required, remove the
Snyder column and rinse the flask and its lower joint into the
concentrator tube with a minimum amount of hexane. A 5 ml syringe
is recommended for this operation. Add a clean boiling chip to the
concentrator tube and attach a two ball micro Snyder column. Prewet
the column by adding about 0.5 ml of hexane to the top. Place the
micro K-D apparatus on the water bath (80°C) so that the concentrator
tube is partially immersed in the hot water. Adjust the vertical
position of the apparatus and the water temperature, as required, to
complete concentration in 5-10 minutes. At the proper rate of
8120A - 5 Revision 1
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distillation the balls of the column will actively chatter, but the
chambers will not flood. When the apparent volume of liquid reaches
0.5 ml, remove the K-D apparatus and allow it to drain and cool for
at least 10 minutes.
7.1.2.5 Remove the micro Snyder column and rinse the flask
and its lower joint into the concentrator tube with 0.2 ml of
hexane. Adjust the extract volume to 2.0 ml and proceed with Method
3620.
7.2 Gas chromatographic conditions (Recommended)
7.2.1 Column 1
Carrier gas (5% methane/95% argon) flow rate = 25 mL/min
Column temperature = 65°C isothermal, unless otherwise specified
(see Table 1).
7.2.2 Column 2
Carrier gas (5% methane/95% argon) flow rate = 25 mL/min
Column temperature = 75°C isothermal, unless otherwise specified
(see Table 1).
7.3 Calibration - Refer to Method 8000 for proper calibration techniques.
Use Table 1 and especially Table 2 for guidance on selecting the lowest point on
the calibration curve.
7.3.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.3.2 If cleanup is performed on the samples, the analyst should
process a series of standards through the cleanup procedure and then
analyze the samples by GC. This will validate elution patterns and the
absence of interferents from the reagents.
7.4 Gas chromatographic analysis
7.4.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 /xL of internal standard to the sample prior to
injecting.
7.4.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
7.4.3 Examples of GC/ECD chromatograms for certain chlorinated
hydrocarbons are shown in Figures 1 and 2.
7.4.4 Record the sample volume injected and the resulting peak sizes
(in area units or peak heights).
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7.4.5 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes. See Method 8000 for calculation equations.
7.4.6 If peak detection and identification are prevented due to
interferences, the hexane extract may undergo cleanup using Method 3620.
7.5 Cleanup: If required, the samples may be cleaned up using the Methods
presented in Chapter 4.
7.5.1 Proceed with Method 3620 using the 2 ml hexane extracts
obtained from Sec. 7.1.2.5.
7.5.2 Following cleanup, the extracts should be analyzed by GC, as
described in the previous paragraphs and in Method 8000.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the QC
in Method 3600 and in the specific cleanup method.
8.2 Procedures to check the GC system operation are found in Method 8000.
8.2.1 The quality control check sample concentrate (Method 8000)
should contain each parameter of interest at the following concentrations
in acetone: hexachloro-substituted hydrocarbon, 10 mg/L; and any other
chlorinated hydrocarbon, 100 mg/L.
8.2.2 Table 3 indicates the calibration and QC acceptance criteria
for this method. Table 4 gives method accuracy and precision as functions
of concentration for the analytes of interest. The contents of both
Tables should be used to evaluate a laboratory's ability to perform and
generate acceptable data by this method.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000).
8.3.1 If recovery is not within limits, the following procedures are
required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also,
check instrument performance.
• Recalculate the data and/or reanalyze the extract if
any of the above checks reveal a problem.
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Reextract and reanalyze the sample if none of the above
are a problem or flag the data as "estimated
concentration".
9.0 METHOD PERFORMANCE
9.1 The method was tested by 20 laboratories using organic-free reagent
water, drinking water, surface water, and three industrial wastewaters spiked at
six concentrations over the range 1.0 to 356 M9/L. Single operator precision,
overall precision, and method accuracy were found to be directly related to the
concentration of the parameter and essentially independent of the sample matrix.
Linear equations to describe these relationships for a flame ionization detector
are presented in Table 4.
9.2 The accuracy and precision obtained will be determined by the sample
matrix, sample preparation technique, and calibration procedures used.
10.0 REFERENCES
1. "Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters. Category 3 - Chlorinated Hydrocarbons, and
Category 8 - Phenols," Report for EPA Contract 68-03-2625.
2. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
3. "EPA Method Validation Study 22, Method 612 (Chlorinated Hydrocarbons),"
Report for EPA Contract 68-03-2625.
4. "Method Performance for Hexachlorocyclopentadiene by Method 612,"
Memorandum from R. Slater, U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, Ohio 45268,
December 7, 1983.
5. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim
Final Rule and Proposed Rule," October 26, 1984.
6. "Determination of Chlorinated Hydrocarbons in Industrial and Municipal
Wastewaters," Report for EPA Contract 68-03-2625.
8120A - 8 Revision 1
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TABLE 1.
GAS CHROMATOGRAPHY OF CHLORINATED HYDROCARBONS
Compound
Retention time (min)
Col. 1 Col. 2
ND = Not determined.
3150°C column temperature,
b!65°C column temperature.
C100°C column temperature.
Method
Detection
limit (/ig/L)
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachl orobenzene
Hexachlorobutadiene
Hexachl orocycl ohexane
Hexachl orocycl opentadi ene
Hexachl oroethane
Pentachlorohexane
Tetrachl orobenzenes
1,2,4-Trichlorobenzene
2.7a
6.6
4.5
5.2
5.6"
7.7
ND
4.9
15.5
3.6"
9.3
6.8
7.6
10. lb
20.0
16.5°
8.3
22.3
0.94
1.14
1.19
1.34
0.05
0.34
0.40
0.03
0.05
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TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION
LIMITS (EQL) FOR VARIOUS MATRICES'
Matrix Factor
Ground water 10
Low-concentration soil by ultrasonic extraction with GPC cleanup 670
High-concentration soil and sludges by ultrasonic extraction 10,000
Non-water miscible waste 100,000
a EQL = [Method detection limit (see Table 1)] X [Factor found in this
table]. For non-aqueous samples, the factor is on a wet weight basis.
Sample EQLs are highly matrix dependent. The EQLs to be determined
herein are provided for guidance and may not always be achievable.
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TABLE 3.
QC ACCEPTANCE CRITERIA8
Parameter
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachl orobutadi ene
Hexachl orocycl opentadi ene
Hexachl oroethane
1,2, 4-Tri chl orobenzene
Test
cone.
Ug/L)
100
100
100
100
10
10
10
10
100
Limit Range
for s for x
(M9/L) (M9/L)
37.3 29.5-126.9
28.3 23.5-145.1
26.4 7.2-138.6
20.8 22.7-126.9
2.4 2.6-14.8
2.2 D-12.7
2.5 D-10.4
3.3 2.4-12.3
31.6 20.2-133.7
Range
P> P.
(%)
9-148
9-160
D-150
13-137
15-159
D-139
D-lll
8-139
5-149
s = Standard deviation of four recovery measurements, in jug/L.
x = Average recovery
P,PS = Percent recovery
D = Detected; result
a Criteria from 40
for four recovery
measured.
measurements, in fj.g/1.
must be greater than zero.
CFR Part 136 for
Method 612. These criteria are
based directly upon the method performance data in Table 4. Where
necessary, the limits for recovery have been broadened to assure
applicability of the limits to concentrations below those used to
develop Table 4.
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TABLE 4.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Parameter
Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi enea
Hexachloroethane
1 , 2 , 4-Tri chl orobenzene
Accuracy, as
recovery, x'
(M9/L)
0.75C+3.21
0.85C-0.70
0.72C+0.87
0.72C+2.80
0.87C-0.02
0.61C+0.03
0.47C
0.74C-0.02
0.76C+0.98
Single analyst
precision, s/
(M9/L)
0.28X-1.17
0.22X-2.95
0.21X-1.03
0.16X-0.48
0.14X+0.07
0.18X+0.08
0.24x
0.23X+0.07
0.23X-0.44
Overall
precision,
S' (M9/L)
0.38X-1.39
0.41X-3.92
0.49X-3.98
0.35X-0.57
0.36X-0.19
0.53X-0.12
0.50x
0.36X-0.00
0.40X-1.37
x'
V
S'
C
x
Expected recovery for one or more measurements of a sample
containing a concentration of C, in M9/L.
Expected single analyst standard deviation of measurements at an
average concentration of x, in
Expected interlaboratory standajrd deviation of measurements at an
average concentration found of x, in M9/L.
True value for the concentration, in M9/L-
Average recovery found for measurements of samples containing a
concentration of C, in /xg/L.
Estimates based upon the performance in a single laboratory.
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FIGURE 1
Column: 1.5% OV-1 + 1.5% OV-225 on Ga» Chrom. Q
Temperature: 7 5 ° C
Detector: Electron Capture
I
tt •
tit i i
4 • 12 16
RETENTION TIM! (MINUTIS)
20
Gas chromatagram of chlorinated hydrocarbons (high molecular weight compounds).
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FIGURE 2
Column: 1.5% OV-1 +1.5* OV-225 on Gas Chrom. Q
Temperature: 16 0 °C
Detector: Electron Capture
0 4 • 12 16
RETENTION TIME (MINUTES)
Gas chromatagram of chlorinated hydrocarbons (low molecular weight compounds)
8120A - 14
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METHOD 8120A
CHLORINATED HYDROCARBONS BY GAS CHROMATOGRAPHY
( Start J
7.1.1 Choose
appropriate
extraction
procedure (see
Chapter 2).
7.1.2 Exchange
extraction so/vent
to hexane during
K-D procedures.
7.2 Set gas
chromatography
conditions.
7.3 Refer to Method
8000 for proper
calibration
techniques.
7.3.2 Is
cleanup
necessary?
7.3.2 Process a
series of standards
through cleanup
procedure; analyze
by GC.
7.4 Perform GC
analysis (see
Method 8000).
7.4.5
Is identification
& detection
prevented by
interferences?
7.5.1 Cleanup using
Method 3620.
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METHOD 8121
CHLORINATED HYDROCARBONS BY GAS CHROMATOGRAPHY: CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8121 describes the determination of chlorinated hydrocarbons
in extracts prepared from environmental samples and RCRA wastes. It describes
wide-bore open-tubular, capillary column gas chromatography procedures using both
single column/single detector and dual-column/dual-detector approaches. The
following compounds can be determined by this method:
Compound Name CAS Registry No."
Benzal chloride98-87-3
Benzotrichloride 98-07-7
Benzyl chloride 100-44-7
2-Chloronaphthalene 91-58-7
1,2-Dichlorobenzene 95-50-1
1,3-Dichlorobenzene 541-73-1
1,4-Dichlorobenzene 106-46-1
Hexachlorobenzene 118-74-1
Hexachlorobutadiene 87-68-3
a-Hexachlorocyclohexane (a-BHC) 319-84-6
0-Hexachlorocyclohexane (/3-BHC) 319-85-7
7-Hexachlorocyclohexane (7-BHC) 58-89-9
6-Hexachlorocyclohexane (5-BHC) 319-86-8
Hexachlorocyclopentadiene 77-47-4
Hexachloroethane 67-72-1
Pentachlorobenzene 608-93-5
1,2,3,4-Tetrachlorobenzene 634-66-2
1,2,4,5-Tetrachlorobenzene 95-94-2
1,2,3,5-Tetrachlorobenzene 634-90-2
1,2,4-Trichlorobenzene 120-82-1
1,2,3-Trichlorobenzene 87-61-6
1,3,5-Trichlorobenzene 108-70-3
a Chemical Abstract Services Registry Number.
1.2 The dual-column/dual-detector approach involves the use of two
30 m x 0.53 mm ID fused-sil ica open-tubular columns of different polarities, thus
different selectivities towards the target compounds. The columns are connected
to an injection tee and two identical detectors. When compared to the packed
columns, the megabore fused-sil ica open-tubular columns offer improved
resolution, better selectivity, increased sensitivity, and faster analysis.
1.3 Table 1 lists method detection limits (MDL) for each compound in an
organic-free reagent water matrix. The MDLs for the compounds of a specific
sample may differ from those listed in Table 1 because they are dependent upon
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the nature of interferences in the sample matrix. Table 2 lists the estimated
quantitation limits (EQL) for other matrices.
1.4 Table 3 lists the compounds that have been determined by this method
and their retention times using the single column technique. Table 4 lists dual
column/dual detector retention time data. Figures 1 and 2 are chromatograms
showing the single column technique. Figure 3 shows a chromatogram of the target
analytes eluted from a pair of DB-5/DB-1701 columns and detected with electron
capture detectors (ECD) under the prescribed GC conditions listed in Table 2.
1.5 This method is restricted to use by or under the supervision of
analysts experienced in the use of a gas chromatograph and in the interpretation
of gas chromatograms.
2.0 SUMMARY OF METHOD
2.1 Method 8121 provides gas chromatographic conditions for the detection
of ppb concentrations of chlorinated hydrocarbons in water and soil or ppm
concentrations in waste samples. Prior to use of this method, appropriate sample
extraction techniques must be used for environmental samples (refer to Chapt. 2).
Both neat and diluted organic liquids (Method 3580) may be analyzed by direct
injection. Spiked samples are used to verify the applicability of the chosen
extraction technique to each new sample type. Analysis is accomplished by gas
chromatography utilizing an instrument equipped with wide bore capillary columns
and single or dual electron capture detectors.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000.
3.2 The electron capture detector responds to all electronegative
compounds. Therefore, interferences are possible by other halogenated compounds,
as well as phthalates and other oxygenated compounds, and, organonitrogen,
organosulfur and organophosphorus compounds. Second column confirmation or GC/MS
confirmation are necessary to ensure proper analyte identification unless
previous characterization of the sample source will ensure proper identification.
3.3 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
syringe used for injection must be rinsed out between samples with solvent.
Whenever an extract concentration exceeds that of the highest calibration
standard, it should be followed by the analysis of a solvent blank to check for
cross-contamination. Additional solvent blanks interspersed with the sample
extracts should be considered whenever the analysis of a solvent blank indicates
cross-contamination problems.
3.4 Phthalate esters, if present in a sample, will interfere only with
the BHC isomers because they elute in Fraction 2 of the Florisil procedure
described in Method 3620. The presence of phthalate esters can usually be
minimized by avoiding contact with any plastic materials and by following
standard decontamination procedures of reagents and glassware.
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3.5 The presence of elemental sulfur will result in large peaks, and can
often mask the region of compounds eluting after 1,2,4,5-tetrachlorobenzene. The
tetrabutylammonium (TBA)-sulfite procedure (Method 3660) works well for the
removal of elemental sulfur.
3.6 In certain cases some compounds coelute on either one or both
columns. In these cases the compounds must be reported as coeluting. The
mixture can be reanalyzed by GC/MS techniques, see Sec. 8.7 and Method 8270.
3.6.1 Using the dual column system of analysis the following
compounds coeluted:
DB-5 1,4-dichlorobenzene/benzyl chloride
l,2,3,5-tetrachlorobenzene/l,2,4,5-tetrachlorobenzene
l,2,3,4-tetrachlorobenzene/2-chloronaphthalene
DB-1701 benzyl chloride/1,2-dichlorobenzene/hexachloroethane
benzal chloride/1,2,4-trichlorobenzene/
hexachlorobutadiene
Some of the injections showed a separation of 1,2,4-trichlorobenzene
from the other two compounds, however, this is not always the case, so the
compounds are listed as coeluting.
3.7 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines causing misinterpretation
of gas chromatograms. All these materials must be demonstrated to be free from
interferences under the conditions of the analysis, by analyzing reagent blanks.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph: An analytical system complete with a gas
chromatograph suitable for on-column and split-splitless injection, and all
required accessories, including syringes, analytical columns, gases, and two
electron capture detectors. A data system for measuring peak areas, and dual
display of chromatograms is recommended. A GC equipped with a single GC column
and detector are acceptable, however, second column confirmation is obviously
more time consuming. Following are the single and dual column configurations
used for developing the retention time data presented in the method. The columns
listed in the dual column configuration may also be used for single column
analysis.
4.1.1 Single Column Analysis:
4.1.1.1 Column 1 - 30 m x 0.53 mm ID fused-silica
capillary column chemically bonded with trifluoropropyl methyl
silicone (DB-210 or equivalent).
4.1.1.2 Column 2 - 30 m x 0.53 mm ID fused-silica
capillary column chemically bonded with polyethylene glycol (DB-WAX
or equivalent).
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4.1.2 Dual Column Analysis:
4.1.2.1 Column 1 - 30 m x 0.53 mm ID fused-silica
open-tubular column, crosslinked and chemically bonded with 95
percent dimethyl and 5 percent diphenyl-polysiloxane (DB-5, RTx-5,
SPB-5, or equivalent), 0.83 ptm or 1.5 /im film thickness.
4.1.2.2 Column 2 - 30 m x 0.53 mm ID fused-silica
open-tubular column crosslinked and chemically bonded with 14
percent cyanopropylphenyl and 86 percent dimethyl-polysiloxane
(DB-1701, RTX-1701, or equivalent), 1.0 urn film thickness.
4.1.3 Splitter: If the splitter approach to dual column injection
is chosen, following are three suggested splitters. An equivalent
splitter is acceptable. See Sec. 7.5.1 for a caution on the use of
splitters.
4.1.3.1 Splitter 1 - J&W Scientific press-fit Y-shaped
glass 3-way union splitter (J&W Scientific, Catalog no. 705-0733).
4.1.3.2 Splitter 2 - Supelco 8 in. glass injection tee,
deactivated (Supelco, Catalog no. 2-3665M).
4,1.3.3 Splitter 3 - Restek Y-shaped fused-silica
connector (Restek, Catalog no. 20405).
4.1.4 Column rinsing kit (optional): Bonded-phase column rinse kit
(J&W Scientific, Catalog no. 430-3000 or equivalent).
4.1.5 Microsyringes - 100 jiL, 50 /zL, 10 nl (Hamilton 701 N or
equivalent), and 50 /iL (Blunted, Hamilton 705SNR or equivalent).
4.1.6 Balances - Analytical, 0.0001 g.
4.1.7 Volumetric flasks, Class A - 10 ml to 1000 mL.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the chemicals are of sufficiently high
purity to permit their use without affecting the accuracy of the determinations.
NOTE: Store the standard solutions (stock, composite, calibration,
internal, and surrogate) at 4°C in Teflon-sealed containers in the
dark. All standard solutions must be replaced after six months or
sooner if routine QC (Sec. 8) indicates a problem.
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5.2 Solvents
5.2.1 Hexane, C6H14 - Pesticide quality or equivalent.
5.2.2 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.2.3 Isooctane, (CH3)3CCH2CH(CH3}2 - Pesticide quality or equivalent.
5.3 Stock standard solutions (1000 mg/L): Can be prepared from pure
standard materials or can be purchased as certified solutions.
5.3.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure compound. Dissolve the compound in isooctane or hexane
and dilute to volume in a 10 ml volumetric flask. If compound purity is
96 percent or greater, the weight can be used without correction to
calculate the concentration of the stock standard solution. Commercially
prepared stock standard solutions can be used at any concentration if they
are certified by the manufacturer or by an independent source.
5.3.2 For those compounds which are not adequately soluble in hexane
or isooctane, mixtures of acetone and hexane are recommended.
5.4 Composite stock standard: Can be prepared from individual stock
solutions. For composite stock standards containing less than 25 components,
take exactly 1 mi of each individual stock solution at 1000 mg/L, add solvent,
and mix the solutions in a 25 ml volumetric flask. For example, for a composite
containing 20 individual standards, the resulting concentration of each component
in the mixture, after the volume is adjusted to 25 mL, will be 40 mg/L. This
composite solution can be further diluted to obtain the desired concentrations.
5.5 Calibration standards should be prepared at a minimum of five
concentrations by dilution of the composite stock standard with isooctane or
hexane. The concentrations should correspond to the expected range of
concentrations found in real samples and should bracket the linear range of the
detector. A suggested list of calibration solution standards is found in Table
7.
5.6 Recommended internal standard: Make a solution of 1000 mg/L of
1,3,5-tribromobenzene. (Two other internal standards, 2,5-dibromotoluene and
alpha,alpha'-dibromo-m-xylene, are suggested if matrix interferences are a
problem.) For spiking, dilute this solution to 50 ng//*L. Use a spiking volume
of 10 ij,L/mi of extract. The spiking concentration of the internal standards
should be kept constant for all samples and calibration standards. Store the
internal standard spiking solutions at 4"C in Teflon-sealed containers in the
dark.
5.7 Recommended surrogate standards: Monitor the performance of the
method using surrogate compounds. Surrogate standards are added to all samples,
method blanks, matrix spikes, and calibration standards. Make a solution of
1000 mg/L of 1,4-dichloronaphthalene and dilute it to 100 ng//iL. Use a spiking
volume of 100 ^l for a 1 L aqueous sample. If matrix interferences are a
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problem, two alternative surrogates are: alpha, 2,6-trichlorotoluene or
2,3,4,5,6-pentachlorotoluene.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes, Sec.
4.1.
6.2 Extracts must be stored at 4 °C and analyzed within 40 days of
extraction.
7.0 PROCEDURE
7.1 Extraction and Cleanup:
7.1.1 Refer to Chapter Two and Method 3500 for guidance on choosing
the appropriate extraction procedure. In general, water samples are
extracted at a neutral, or as is, pH with methylene chloride, using either
Method 3510 or 3520. Solid samples are extracted using either Methods
3540, 3541, or 3550 with methylene chloride/acetone (1:1) as the
extraction solvent.
7.1.2 If required, the samples may be cleaned up using Method 3620
(Florisil) and/or Method 3640 (Gel Permeation Chromatography). See
Chapter Two, Sec. 2.3.2 and Method 3600 for general guidance on cleanup
and method selection. Method 3660 is used for sulfur removal.
7.1.3 Prior to gas chromatographic analysis, the extraction solvent
must exchanged into hexane using the Kuderna-Danish concentration step
found in any of the extraction methods. Any methylene chloride remaining
in the extract will cause a very broad solvent peak.
7.2 Gas Chromatographic Conditions:
7.2.1 Retention time information for each of the analytes is
presented in Tables 3 and 4. The recommended GC operating conditions are
provided in Tables 5 and 6. Figures 1, 2 and 3 illustrate typical
chromatography of the method analytes for both the single column approach
and the dual column approach when operated at the conditions specified in
Tables 5 and 6.
7.3 Calibration:
7.3.1 Prepare calibration standards using the procedures in Sec.
5.0. Refer to Method 8000 for proper calibration procedures. The
procedure for internal or external calibration may be used.
7.3.2 Refer to Method 8000 for the establishment of retention time
windows.
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7.4 Gas chromatographic analysis:
7.4.1 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria.
7.4.2 Automatic injections of 1 pi are recommended. Hand injections
of no more than 2 /zL may be used if the analyst demonstrates quantitation
precision of < 10 percent relative standard deviation. The solvent flush
technique may be used if the amount of solvent is kept at a minimum. If
the internal standard calibration technique is used, add 10 jitL of the
internal standard to each ml of sample extract prior to injection.
7.4.3 Tentative identification of an analyte occurs when a peak from
a sample extract falls within the daily retention time window.
7.4.4 Validation of gas chromatographic system qualitative
performance: Use the midconcentration standards interspersed throughout
the analysis sequence (Sec. 7.3) to evaluate this criterion. If any of
the standards fall outside their daily retention time windows, the system
is out of control. Determine the cause of the problem and correct it (see
Sec. 7.5).
7.4.5 Record the volume injected to the nearest 0.05 /uL and the
resulting peak size in peak height or area units. Using either the
internal or the external calibration procedure (Method 8000), determine
the identity and the quantity of each component peak in the sample
chromatogram which corresponds to the compounds used for calibration
purposes. See Method 8000 for calculation equations.
7.4.6 If the responses exceed the linear range of the system, dilute
the extract and reanalyze. Peak height measurements are recommended over
peak area integration when overlapping peaks cause errors in area
integration.
7.4.7 If partially overlapping or coeluting peaks are found, change
columns or try a GC/MS technique (see Sec. 8.7 and Method 8270).
Interferences that prevent analyte identification and/or quantitation may
be removed by the cleanup techniques mentioned above.
7.4.8 If the peak response is less than 2.5 times the baseline noise
level, the validity of the quantitative result may be questionable. The
analyst should consult with the source of the sample to determine whether
further concentration of the sample is warranted.
7.5 Instrument Maintenance:
7.5.1 Injection of sample extracts from waste sites often leaves a
high boiling residue in: the injection port area, splitters when used, and
the injection port end of the chromatographic column. This residue
effects chromatography in many ways (i.e., peak tailing, retention time
shifts, analyte degradation, etc.) and, therefore, instrument maintenance
8121 - 7 Revision 0
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is very important. Residue buildup in a splitter may limit flow through
one leg and therefore change the split ratios. If this occurs during an
analytical run, the quantitative data may be incorrect. Proper cleanup
techniques will minimize the problem and instrument QC will indicate when
instrument maintenance is required.
7.5.2 Suggested chromatograph maintenance: Corrective measures may
require any one or more of the following remedial actions. Also see Sec.
7 in Method 8000 for additional guidance on corrective action for
capillary columns and the injection port.
7.5.2.1 Splitter connections: For dual columns which are
connected using a press-fit Y-shaped glass splitter or a Y-shaped
fused-silica connector, clean and deactivate the splitter or replace
with a cleaned and deactivated splitter. Break off the first few
inches (up to one foot) of the injection port side of the column.
Remove the columns and solvent backflush according to the
manufacturer's instructions. If these procedures fail to eliminate
the degradation problem, it may be necessary to deactivate the metal
injector body and/or replace the columns.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures. Quality control to validate sample extraction is covered in Method
3500 and in the extraction method utilized. If extract cleanup was performed,
follow the QC in Method 3600 and in the specific cleanup method.
8.2 Quality control required to evaluate the GC system operation is found
in Method 8000, Sec. 8.3.
8.3 Calculate surrogate standard recoveries for all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000, Sec. 8). If the recovery is
not within limits, the following are required:
8.3.1 Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check instrument
performance.
8.3.2 Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
8.3.3 Reextract and reanalyze the sample if none of the above are
a problem, or flag the data as "estimated concentrations".
8.4 Data from systems that automatically identify target analytes on the
basis of retention time or retention time indices should be reviewed by an
experienced analyst before they are reported.
8.5 When using the internal standard calibration technique, an internal
standard peak area check must be performed on all samples. The internal standard
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must be evaluated for acceptance by determining whether the measured area for the
internal standard deviates by more than 50 percent from the average area for the
internal standard in the calibration standards. When the internal standard peak
area is outside that limit, all samples that fall outside the QC criteria must
be reanalyzed.
8.6 Include a mid-concentration calibration standard after each group of
20 samples in the analysis sequence. The response factors for the
mid-concentration calibration must be within + 15 percent of the average values
for the multiconcentration calibration. When the response factors fall outside
that limit, all samples analyzed after that mid-concentration calibration
standard must be reanalyzed after performing instrument maintenance to correct
the usual source of the problem. If this fails to correct the problem, a new
calibration curve must be established.
8.7 GC/MS confirmation:
8.7.1 GC/MS techniques should be judiciously employed to support
qualitative identifications made with this method. Follow the GC/MS
operating requirements specified in Method 8270. Ensure that there is
sufficient concentration of the analyte(s) to be confirmed, in the extract
for GC/MS analysis.
8.7.2 When available, chemical ionization mass spectra may be
employed to aid in the qualitative identification process.
8.7.3 To confirm an identification of a compound, the background
corrected mass spectrum of the compound must be obtained from the sample
extract and must be compared with a mass spectrum from a stock or
calibration standard analyzed under the same chromatographic conditions.
At least 25 ng of material should be injected into the GC/MS. The
identification criteria specified in Method 8270 must be met for
qualitative confirmation.
8.7.3.1 Should the MS procedure fail to provide
satisfactory results, additional steps may be taken before
reanalysis. These steps may include the use of alternate packed or
capillary GC columns or additional sample cleanup.
9.0 METHOD PERFORMANCE
9.1 The MDL is defined in Chapter One. The MDLs listed in Table 1 were
obtained by using organic-free reagent water. Details on how to determine MDLs
are given in Chapter One. The MDLs actually achieved in a given analysis will
vary since they depend on instrument sensitivity and matrix effects.
9.2 This method has been tested in a single laboratory by using
organic-free reagent water, sandy loam samples and extracts which were spiked
with the test compounds at one concentration. Single-operator precision and
method accuracy were found to be related to the concentration of compound and the
type of matrix.
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9.3 Single laboratory accuracy data were obtained for chlorinated
hydrocarbons in a clay soil. The spiking concentrations ranged from 500 to 5000
jug/kg, depending on the sensitivity of the analyte to the electron capture
detector. The spiking solution was mixed into the soil during addition and then
immediatly transferred to the extraction device and immersed in the extraction
solvent. The spiked sample was then extracted by Method 3541 (Automated
Soxhlet). The data represents a single determination. Analysis was by capillary
column gas chromatography/electron capture detector following Method 8121 for the
chlorinated hydrocarbons. These data are listed in Table 9 and were taken from
Reference 4.
10.0 REFERENCES
1. Lopez-Avila, V., N.S. Dodhiwala, and J. Milanes, "Single Laboratory
Evaluation of Method 8120, Chlorinated Hydrocarbons", 1988, EPA Contract
Numbers 68-03-3226 and 68-03-3511.
2. Glazer, J.A., G.D. Foerst, G.D. McKee, S.A. Quave, and W.L. Budde, "Trace
Analyses for Wastewaters," Environ. Sci. and Technol. 15:1426-1431, 1981.
3. Lopez-Avila, V.; Baldin, E.; Benedicto, J; Milanes, J.; Beckert, W. F.
"Application of Open-Tubular Columns to SW 846 GC Methods"; final report
to the U.S. Environmental Protection Agency on Contract 68-03-3511;
Mid-Pacific Environmental Laboratory, Mountain View, CA, 1990.
4. Lopez-Avila, V. (Beckert, W., Project Officer), "Development of a Soxtec
Extraction Procedure for Extracting Organic Compounds from Soils and
Sediments", EPA 600/X-91/140, US EPA, Environmental Monitoring Systems
Laboratory-Las Vegas, October 1991.
8121 - 10 Revision 0
September 1994
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TABLE 1
METHOD DETECTION LIMITS FOR CHLORINATED HYDROCARBONS
SINGLE COLUMN METHOD OF ANALYSIS
Compound name
Benzal chloride
Benzotri chloride
Benzyl chloride
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachl orobenzene
Hexachl orobutadi ene
a~Hexachlorocyclohexane (a-BHC)
/3-Hexachlorocyclohexane ((8-BHC)
-y-Hexachlorocyclohexane (7-BHC)
5 -Hexachl orocyclohexane (
-------
TABLE 2
ESTIMATED QUANTITATION LIMIT (EQL) FACTORS FOR VARIOUS MATRICES'
Matrix Factor
Ground water 10
Low-concentration soil by ultrasonic extraction 670
with GPC cleanup
High-concentration soil and sludges by ultrasonic 10,000
extraction
Waste not nriscible with water 100,000
a EQL = [Method detection limit (see Table 1)] x [Factor found in this
table]. For nonaqueous samples, the factor is on a wet-weight basis.
Sample EQLs are highly matrix-dependent. The EQLs listed herein are
provided for guidance and may not always be achievable.
8121 - 12 Revision 0
September 1994
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TABLE 3
GAS CHROMATOGRAPHIC RETENTION TIMES FOR CHLORINATED HYDROCARBONS: SINGLE
COLUMN METHOD OF ANALYSIS
Compound name
Retention time (min)
DB-210"DB-WAXb
Benzal chloride
Benzotrichloride
Benzyl chloride
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachlorobutadiene
a-BHC
K-BHC
6-mt
Hexachl orocycl opentadi ene
Hexachloroethane
Pentachlorobenzene
1,2, 3 ,4-Tetrachl orobenzene
1,2,4,5-Tetrachlorobenzene
1,2,3 , 5-Tetrachl orobenzene
1 , 2 , 4-Tr i chl orobenzene
1,2, 3 -Trichl orobenzene
1, 3, 5-Tri chl orobenzene
6.86
7.85
4.59
13.45
4.44
3.66
3.80
19.23
5.77
25.54
24.07
26.16
8.86
3.35
14.86
11.90
10.18
10.18
6.86
8.14
5.45
15.91
15.44
10.37
23.75
9.58
7.73
8.49
29.16
9.98
33.84
54.30
33.79
c
8.13
23.75
21.17
17.81
17.50
13.74
16.00
10.37
Internal Standards
2,5-Dibromotoluene
1,3,5-Tri bromobenzene
a,a'-Di bromo-meta-xylene
Surrogates
9.55
11.68
18.43
or,2,6-Trichlorotoluene 12.96
1,4-Dichloronaphthalene 17.43
2,3,4,5,6-Pentachlorotoluene 18.96
18.55
22.60
35.94
22.53
26.83
27.91
GC operating conditions: 30 m x 0.53 mm ID DB-210 fused-silica
capillary column; 1 /xm film thickness; carrier gas helium at 10 mL/min;
makeup gas is nitrogen at 40 mL/min; temperature program from 65°C to
175°C (hold 20 minutes) at 4°C/min; injector temperature 220°C; detector
temperature 250°C.
GC operating conditions: 30 m x 0.53 mm ID DB-WAX fused-silica
capillary column; 1 /urn film thickness; carrier gas helium at 10 mL/min;
makeup gas is nitrogen at 40 mL/min; temperature program from 60°C to
170°C (hold 30 minutes) at 4°C/min; injector temperature 200°C; detector
temperature 230°C.
Compound decomposes on-column.
8121 - 13
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September 1994
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TABLE 4
RETENTION TIMES OF THE CHLORINATED HYDROCARBONS'
DUAL COLUMN METHOD OF ANALYSIS
Compound
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Benzyl chloride
1,2-Dichlorobenzene
Hexachloroethane
1,3, 5-Trichl orobenzene
Benzal chloride
1, 2, 4-Trichl orobenzene
1,2, 3 -Tri chl orobenzene
Hexachlorobutadiene
Benzotrichloride
1,2,3 , 5-Tetrachl orobenzene
1,2,4 , 5-Tetrachl orobenzene
Hexachl orocycl opentadi ene
1,2,3 , 4-Tetrachl orobenzene
2-Chl oronaphthal ene
Pentachl orobenzene
a-BHC
Hexachl orobenzene
0-BHC
7-BHC
5-BHC
DB-5
RT(min)
5.82
6.00
6.00
6.64
7.91
10.07
10.27
11.97
13.58
13.88
14.09
19.35
19.35
19.85
21.97
21.77
29.02
34.64
34.98
35.99
36.25
37.39
DB-1701
RT(min)
7.22
7.53
8.47
8.58
8.58
11.55
14.41
14.54
16.93
14.41
17.12
21.85
22.07
21.17
25.71
26.60
31.05
38.79
36.52
43.77
40.59
44.62
Internal Standard
1,3,5-Tribromobenzene 11.83 13.34
Surrogate
1,4-Dichloronaphthalene 15.42 17.71
"The GC operating conditions were as follows: 30 m x 0.53 mm ID DB-5
(0.83-|um film thickness) and 30 m x 0.53 mm ID DB-1701 (1.0 p.m film
thickness) connected to an 8-in injection tee (Supelco Inc.). Temperature
program: 80°C (1.5 min hold) to 125°C (1 min hold) at 2'C/min then to 240eC
(2 min hold) at 5°C/min; injector temperature 250°C; detector temperature
320°C; helium carrier gas 6 mL/min; nitrogen makeup gas 20 mL/min.
8121 - 14 Revision 0
September 1994
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TABLE 5
GC OPERATING CONDITIONS FOR CHLOROHYDROCARBONS
SINGLE COLUMN METHOD OF ANALYSIS
Column 1: DB-210 30 m x 0.53 mm ID fused-silica capillary column
chemically bonded with trifluoropropyl methyl silicone
Carrier gas (He) 10 mL/min
Column temperature:
Initial temperature
Temperature program
Final temperature
Injector temperature 220°C
Detector temperature 250"C
Injection volume 1-2 jtiL
65'C
658C to 175°C at 4°C/min
175°C, hold 20 minutes.
Column 2: DB-WAX 30 m x 0.53 mm ID fused-silica capillary column
chemically bonded with polyethylene glycol
Carrier gas (He) 10 mL/min
Column temperature:
Initial temperature
Temperature program
Final temperature
Injector temperature 200°C
Detector temperature 230°C
Injection volume 1-2 /iL
60°C
60°C to 170°C at 4°C/min
170°C, hold 30 minutes.
8121 - 15
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September 1994
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TABLE 6
GC OPERATING CONDITIONS FOR CHLORINATED HYDROCARBONS
DUAL COLUMN METHOD OF ANALYSIS
Column 1:
Column 2:
Type: DB-1701 (J&W Scientific) or equivalent
Dimensions: 30 m x 0.53 mm ID
Film Thickness: 1.0 (jum)
Type: DB-5 (J&W Scientific) or equivalent
Dimensions: 30 m x 0.53 mm ID
Film Thickness: 0.83
Carrier gas flowrate (mL/min): 6 (Helium)
Makeup gas flowrate (mL/min): 20 (Nitrogen)
Temperature program: 80° C (1.5 mi n hold) to 125°C (1 min hold) at 2°C/min
then to 240°C (2 min hold) at 5°C/min.
Injector temperature: 250°C
Detector temperature: 320°C
Injection volume: 2 juL
Solvent: Hexane
Type of injector: Flash vaporization
Detector type: Dual ECD
Range: 10
Attenuation: 32 (DB-1701J/32 (DB-5)
Type of splitter: Supelco 8-in injection tee
8121 - 16 Revision 0
September 1994
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TABLE 7
SUGGESTED CONCENTRATIONS FOR THE CALIBRATION SOLUTIONS8
Concentration (ng//nL)
Benzal chloride
Benzotrichloride
Benzyl chloride
2-Chloronaphthalene
1,2-Dichlorobenzene
1 , 3 -Di chl orobenzene
1,4-Dichlorobenzene
Hexachl orobenzene
Hexachlorobutadiene
a-BHC
/3-BHC
T-BHC
6-BHC
Hexachl orocycl opentad i ene
Hexachl oroethane
Pentachl orobenzene
1,2,3 , 4-Tetrachl orobenzene
1,2, 4, 5-Tetrachl orobenzene
1 ,2,3,5-Tetrachlorobenzene
1, 2, 4-Tri chl orobenzene
1 , 2 , 3-Tri chl orobenzene
1, 3, 5-Tri chl orobenzene
0.1
0.1
0.1
2.0
1.0
1.0
1.0
0.01
0.01
0.1
0.1
0.1
0.1
0.01
0.01
0.01
0.1
0.1
0.1
0.1
0.1
0.1
0.2
0.2
0.2
4.0
2.0
2.0
2.0
0.02
0.02
0.2
0.2
0.2
0.2
0.02
0.02
0.02
0.2
0.2
0.2
0.2
0.2
0.2
0.5
0.5
0.5
10
5.0
5.0
5.0
0.05
0.05
0.5
0.5
0.5
0.5
0.05
0.05
0.05
0.5
0.5
0.5
0.5
0.5
0.5
0.8
0.8
0.8
16
8.0
8.0
8.0
0.08
0.08
0.8
0.8
0.8
0.8
0.08
0.08
0.08
0.8
0.8
0.8
0.8
0.8
0.8
1.0
1.0
1.0
20
10
10
10
0.1
0.1
1.0
1.0
1.0
1.0
0.1
0.1
0.1
1.0
1.0
1.0
1.0
1.0
1.0
Surrogates
a,2,6-Trichlorotoluene 0.02 0.05 0.1 0.15 0.2
1,4-Dichloronaphthalene 0.2 0.5 1.0 1.5 2.0
2,3,4,5,6-Pentachlorotoluene 0.02 0.05 0.1 0.15 0.2
One or more internal standards should be spiked prior to GC/ECD
analysis into all calibration solutions. The spike concentration of
the internal standards should be kept constant for all calibration
solutions.
8121 - 17
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September 1994
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TABLE 8
ELUTION PATTERNS OF CHLORINATED HYDROCARBONS
FROM THE FLORISIL COLUMN BY ELUTION WITH PETROLEUM ETHER (FRACTION 1)
AND 1:1 PETROLEUM ETHER/DIETHYL ETHER (FRACTION 2)
Compound
Benzal chlorided
Benzotri chloride
Benzyl chloride
2-Chloronaphthalene
1 ,2-Dichlorobenzene
1 , 3-Di chl orobenzene
1,4-Dichlorobenzene
Hexachl orobenzene
Hexachl orobutadi ene
a-BHC
/3-BHC
y-BHC
S-BHC
Hexachl orocycl opentadi ene
Hexachl oroethane
Pentachl orobenzene
1,2,3 , 4-Tetrachl orobenzene
1,2,4 , 5-Tetrachl orobenzene"
1,2,3 , 5-Tetrachl orobenzene6
1 , 2 , 4-Tri chl orobenzene
1,2, 3 -Tri chl orobenzene
1 , 3 , 5-Tri chl orobenzene
Amount
(M9)
10
10
100
200
100
100
100
1.0
1.0
10
10
10
10
1.0
1.0
1.0
10
10
10
10
10
10
Recovery
Fraction 1"
0
0
82
115
102
103
104
116
101
93
100
129
104
102
102
59
96
102
(percent)"
Fraction 2°
0
0
16
95
108
105
71
a Values given represent average values of duplicate experiments.
b Fraction 1 was eluted with 200 mL petroleum ether.
c Fraction 2 was eluted with 200 mL petroleum ether/diethyl ether (1:1).
d This compound coelutes with 1,2,4-trichlorobenzene; separate
experiments were performed with benzal chloride to verify that this
compound is not recovered from the Florisil cleanup in either fraction.
e This pair cannot be resolved on the DB-210 fused-silica capillary
columns.
8121 - 18
Revision 0
September 1994
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TABLE 9
SINGLE LABORATORY ACCURACY DATA FOR THE EXTRACTION OF
CHLORINATED HYDROCARBONS FROM SPIKED CLAY SOIL BY METHOD 3541
(AUTOMATED SOXHLET)"
Compound Name Spike Level % Recovery
jug/kg DB-5 DB-1701
1,3-Dichlorobenzene 5000 b 39
1,2-Dichlorobenzene 5000 94 77
Benzal chloride 500 61 66
Benzotrichloride 500 48 53
Hexachlorocyclopentadiene 500 30 32
Pentachlorobenzene 500 76 73
alpha-BHC 500 89 94
delta-BHC 500 86 b
Hexachlorobenzene 500 84 88
a The operating conditions for the automated Soxhlet were as follows:
immersion time 45 min; extraction time 45 min; the sample size was 10 g
clay soil, extraction solvent, 1:1 acetone/hexane. No equilibration time
following spiking.
b Not able to determine because of interference.
Data taken from Reference 4.
8121 - 19 Revision 0
September 1994
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1
3 ;
•
n
N J
I
21 11
14
Iff
10 ia 1113
ll
10
If
TIMi(mln)
ao
as
30
Figure 1. GC/ECD chromatogram of Method 8121 composite standard analyzed on a
30 m x 0.53 mm ID DB-210 fused-silica capillary column. GC
operating conditions are given in Section 7.4. See Table 3 for
compound Identification.
8121 - 20
Revision 0
September 1994
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4
16
10
ft
13
A
10
15
20
25 30
TIME (min)
35
40
45
50
55
Figure 2. 6C/ECD chromatogram of Method 8121 composite standard analyzed on a
30 ro x 0.53 mm ID DB-WAX fused-silica capillary column. GC
operating conditions are given in Section 7.4. See Table 3 for
compound identification.
8121 - 21
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September 1994
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11 If
OB-1701
i
to
•
T .11 14 It tl IS It tt tt II fl 20
u
JU
Figure 3. GC/ECD chromatogram of chlorinated hydrocarbons analyzed on a DB
5/DB 1701 fused-silica, open-tubular column pair. The GC operating
conditions were as follows: 30 m x 0.53 mm ID DB 5 (0.83 /*m film
thickness) and 30 m x 0.53 mm ID DB 1701 (1.0 urn film thickness)
connected to an 8 In Injection tee (Supelco Inc.). Temperature
program: 80°C (1.5 m1n hold) to 125°C (1 m1n hold) at 2°C/min, then
to 240°C (2 m1n hold) at 5°C/m1n.
8121 - 22
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September 1994
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METHOD 8121
CHLORINATED HYDROCARBONS BY GAS CHROMATOGRAPHY: CAPILLARY COLUMN TECHNIQUE
7.1.1
I
7.12 Add appropriate spiking
coffipoundi to sample prior
to extraction procedure
7.2 Exchange oxtracbon
solvent to hexane during
K-D procedure
7.2.1 Following concentration of
mMhylww chkxid* allow K-0
•ppiratu* to drain and cool
122. InctMW tMnptratun of hot
nwtar btlh; *dd hMww; tttrndi
Snydv cohjtnn; plae* •opwikji on «MMr
§^ rpfiwa from
vMttr b«th; cool
72.3 Remov* column; rint* leak
•nd jointt iMlth hcxin*; adjiMt
extract volume
7 JS Choo*» «ppfopn«t« cleanup
technique, if neceteary;
HUORMH cleanup ie recommended.
723 Will further
proceaiingbe
performed iMtNn
MM days?
72.3 Tranefer extract to
Teflon sealed screw^ap
Refer to Method 3620 or to
Section 7.32
7.3.3 GPC
cleanup
required?
7.3.4 Elemental
sulfur removal
required?
7.3.4 Refer to
Section 7.3 '
7.33 Refer to
Method 3640
8121 - 23
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September 1994
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METHOD 8121
(continued)
O
1
1
7.2.3 Stopper concentrator
and refrigerate
I
7.4.1 Set column 1 conditions
7.4.2 Set column 2 conditions
7.5.1 Refer to Method 8000 for
caiibration techniques; select
lowest point on calibration curve
7.5.2 Choose and perform
internal or external calibration
(refer to Method 8000)
7.6.1 Add internal standard
if necessary
7.6.2 Establish daily retention time
windows, analysis sequence,
dilutions, and identification criteria
O
8121 - 24
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September 1994
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METHOD 8121
(concluded)
0
7.6.3 Record sample volume
injected and resulting peak
sizes
i
7.6.4 Determine identity and
quantity of each component peak
that corresponds to compound
used for calibration
7.6.5
Does peak
exceed working
range of
system?
7.6.5 Dilute extract; reanalyze
7.6.6 Compare standard and
sample retention times;
identify compounds
8121 - 25
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September 1994
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METHOD 8140
ORGANOPHOSPHORUS PESTICIDES
1.0 SCOPE AND APPLICATION
1.1 Method 8140 is a gas chromatographic (GC) method used to determine
the concentration of various organosphosphorus pesticides. Table 1 indicates
compounds that may be determined by this method and lists the method detection
limit for each compound in reagent water. Table 2 lists the practical
quantisation limit (PQL) for other matrices.
1.2 When Method 8140 is used to analyze unfamiliar samples, compound
identifications should be supported by at least two additional qualitative
techniques if mass spectroscopy is not employed. Section 8.4 provides gas
chromatograph/mass spectrometer (GC/MS) criteria appropriate for the
qualitative confirmation of compound identifications.
2.0 SUMMARY OF METHOD
2.1 Method 8140 provides gas chromatographic conditions for the
detection of ppb levels of organophosphorus pesticides. Prior to analysis,
appropriate sample extraction techniques must be used. Both neat and diluted
organic liquids (Method 3580, Waste Dilution) may be analyzed by direct
injection. A 2- to 5-uL aliquot of the extract is injected into a gas
chromatograph, and compounds in the GC effluent are detected with a flame
photometric or thermionic detector.
2.2 If interferences are encountered in the analysis, Method 8140 may
also be performed on extracts that have undergone cleanup using Method 3620
and/or Method 3660.
3.0 INTERFERENCES
3.1 Refer to Methods 3500 (Section 3.5, in particular), 3600, and 8000.
3.2 The use of Florlsil cleanup materials (Method 3620) for some of the
compounds in this method has been demonstrated to yield recoveries less than
85% and is therefore not recommended for all compounds. Refer to Table 2 of
Method 3620 for recoveries of organophosphorous pesticides as a function of
Florisil fractions. Use of phosphorus- or halogen-specific detectors,
however, often obviates the necessity for cleanup for relatively clean sample
matrices. If particular circumstances demand the use of an alternative
cleanup procedure, the analyst must determine the elution profile and
demonstrate that the recovery of each analyte is no less than 85%.
8140 - 1
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Date September 1986
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TABLE 1. GAS CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS FOR
ORGANOPHOSPHOROUS PESTICIDES3
Compound
Azinphos methyl
Bolstar
Chlorpyrifos
Coumaphos
Demeton-0
Demeton-S
Diazinon
Dichlorvos
Disulfoton
Ethoprop
Fensulfothion
Fenthion
Merphos
Mevinphos
Naled
Parathion methyl
Phorate
Ronnel
Stirophos (Tetrachlorvinphos)
Tokuthion (Prothiofos)
Trichloronate
GC
col umn"3
la
la
2
la
la
la
2
Ib, 3
la
2
la
la
2
Ib
3
2
la
2
Ib, 3
la
la
Retention
time
(min)
6.80
4.23
6.16
11.6
2.53
1.16
7.73
0.8, 1.50
2.10
3.02
6.41
3.12
7.45
2.41
3.28
3.37
1.43
5.57
8.52, 5.51
3.40
2.94
Method
detection
limit (ug/L)
1.5
0.15
0.3
1.5
0.25
0.25
0.6
0.1
0.20
0.25
1.5
0.10
0.25
0.3
0.1
0.03
0.15
0.3
5.0
0.5
0.15
Development of Analytical Test Procedures for Organic Pollutants in
Wastewater; Report for EPA Contract 68-03-2711 (in preparation).
bSee Sections 4.2.1 and 7.2 for column descriptions and conditions.
8140 - 2
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Date September 1986
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TABLE 2. DETERMINATION OF PRACTICAL QUANTITATION LIMITS (PQL) FOR VARIOUS
MATRICES3
Matrix Factorb
Ground water 10
Low-level soil by sonication with GPC cleanup 670
High-level soil and sludges by sonication 10,000
Non-water miscible waste 100,000
aSample PQLs are highly matrix-dependent. The PQLs listed herein are
provided for guidance and may not always be achievable.
bPQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For non-
aqueous samples, the factor is on a wet-weight basis.
8140 - 3
Revision
Date September 1986
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3.3 Use of a flame photometric detector 1n the phosphorus mode will
minimize interferences from materials that do not contain phosphorus.
Elemental sulfur, however, may interfere with the determination of certain
organophosphorus pesticides by flame photometric gas chromatography. Sulfur
cleanup using Method 3660 may alleviate this interference.
3.4 A halogen-specific detector (i.e., electrolytic conductivity or
microcoulometric) is very selective for the halogen-containing pesticides and
is recommended for use with dichlorvos, naled, and stirophos.
4.0 APPARATUS AND MATERIALS
4.1 Gas chrpmatograph; Analytical system complete with gas
chromatograph suitable for on-column Injections and all required accessories,
including detectors, column supplies, recorder, gases, and syringes. A data
system for measuring peak areas and/or peak heights is recommended.
4.1.1 Columns:
4.1.1.1 Column la and Ib: 1.8-m x 2-mm I.D. glass, packed
with 5% SP-2401 on Supelcoport, 100/120 mesh (or equivalent).
4.1.1.2 Column 2: 1.8-m x 2-mm I.D. glass, packed with 3% SP-
2401 on Supelcoport, 100/120 mesh (or equivalent).
4.1.1.3 Column 3: 50-cm x l/8-1n O.D. Teflon, packed with 15%
SE-54 on Gas Chrom Q, 100/120 mesh (or equivalent).
4.1.2 Detectors: The following detectors have proven effective in
analysis for the analytes listed 1n Table 1 and were used to develop the
accuracy and precision statements in Section 9.0.
4.1.2.1 Phosphorus-specific: Nitrogen/Phosphorus (N/P),
operated in phosphorus-sensitive mode.
4.1.2.2 Flame Photometric (FPD): FPD is more selective for
phosphorus than the N/P.
4.1.2.3 Halogen-specific: Electrolytic conductivity or
microcoulometric. These are very selective for those pesticides
containing halogen substituents.
4.2 Balance; analytical, capable of accurately weighing to the nearest
0.0001 g.
4.3 Vials; Amber glass, 10- to 15-mL capacity with Teflon-lined screw-
cap.
4.4 Kuderna-Danish (K-D) apparatus;
4.4.1 Concentrator tube: 10-mL, graduated (Kontes K-570050-1025 or
equivalent). Ground-glass stopper is used to prevent evaporation of
extracts
8140 - 4
Revision 0
Date September 1986
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4.4.2 Evaporation flask: 500-mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs.
4.4.3 Snyder column: Three-ball macro (Kontes K-503000-0121 or
equivalent).
4.4.4 Snyder column: Two-ball micro (Kontes K-569001-0219 or
equivalent).
4.5 Boiling chips; Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.6 Water bath; Heated, with concentric ring cover, capable of
temperature control (+5*C). The bath should be used in a hood.
4.7 Microsyringe; 10-uL.
4.8 Syringe; 5-mL.
4.9 Volumetric flasks; 10-, 50-, and 100-mL, ground-glass stopper.
5.0 REAGENTS
5.1 Solvents; Hexane, acetone, isooctane (2,2,4-trimethylpentane)
(pesticide quality or equivalent).
5.2 Stock standard solutions;
5.2.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure material. Dissolve the material in hexane or other
suitable solvent and dilute to volume in a 10-mL volumetric flask.
Larger volumes can be used at the convenience of the analyst. If
compound purity is certified at 96% or greater, the weight can be used
without correction to calculate the concentration of the stock standard.
Commercially prepared stock standards can be used at any concentration if
they are certified by the manufacturer or by an independent source.
5.2.2 Transfer the stock standard solutions into Teflon-sealed
screw-cap bottles. Store at 4*C and protect from light. Stock standard
solutions should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards
from them.
5.2.3 Stock standard solutions must be replaced after one year, or
sooner if comparison with check standards indicates a problem.
5.3 Calibration standards; Calibration standards at a minimum of five
concentration levels for each parameter of interest should be prepared through
dilution of the stock standards with isooctane. One of the concentration
levels should be at a concentration near, but above, the method detection
8140 - 5
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Date September 1986
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limit. The remaining concentration levels should correspond to the expected
range of concentrations found in real samples or should define the working
range of the GC. Calibration standards must be replaced after six months, or
sooner if comparison with check standards indicates a problem.
5.4 Internal standards (If Internal standard calibration is used); To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.4.1 Prepare calibration standards at a minimum of five
concentration levels for each parameter of interest as described 1n
Paragraph 5.3.
5.4.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with hexane or other
suitable solvent.
5.4.3 Analyze each calibration standard according to Section 7.0.
5.5 Surrogate standards; The analyst should monitor the performance of
the extraction,cleanup(when used), and analytical system and the
effectiveness of the method in dealing with each sample matrix by spiking each
sample, standard, and reagent water blank with one or two surrogates (e.g.,
organophosphorous pesticides not expected to be present in the sample)
recommended to encompass the range of the temperature program used in this
method. Deuterated analogs of analytes should not be used as surrogates for
gas chromatographic analysis due to coelutlon problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the Introductory material to this chapter, Organic Analytes,
Section 4.1. Extracts must be stored under refrigeration and analyzed within
40 days of extraction.
7.0 PROCEDURE
7.1 Extraction;
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a
neutral, or as is, pH with methylene chloride, using either Method 3510
or 3520. Solid samples are extracted using either Method 3540 or 3550.
7.1.2 Prior to gas chromatographic analysis, the extraction solvent
may be exchanged to hexane. This is recommended if the detector used is
halogen-specific. The exchange 1s performed during the K-D procedures
listed in all of the extraction methods. The exchange is performed as
follows.
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7.1.2.1 Following K-D of the methylene chloride extract to
1 ml using the macro-Snyder column, allow the apparatus to cool and
drain for at least 10 min.
7.1.2.2 Momentarily remove the Snyder column, add 50 ml of
hexane, a new boiling chip, and reattach the macro-Snyder column.
Concentrate the extract using 1 ml of hexane to prewet the Snyder
column. Place the K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete concentration in 5-10 min. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 1 ml, remove the K-D apparatus and allow it to drain and
cool for at least 10 min.
7.1.2.3 Remove the Snyder column and rinse the flask and its
lower joint into the concentrator tube with 1-2 ml of hexane. A
5-mL syringe is recommended for this operation. Adjust the extract
volume to 10.0 mL. Stopper the concentrator tube and store
refrigerated at 4*C if further processing will not be performed
immediately. If the extract will be stored longer than two days, it
should be transferred to a Teflon-sealed screw-cap vial. Proceed
with gas chromatographic analysis if further cleanup is not
required.
7.2 Gas chromatography conditions (Recommended);
7.2.1 Column la: Set helium carrier gas flow at 30 mL/min flow
rate. Column temperature is set at 150*C for 1 min and then programmed
at 25*C/min to 220'C and held.
7.2.2 Column Ib: Set nitrogen carrier gas flow at 30 mL/min flow
rate. Column temperature is set at 170*C for 2 min and then programmed
at 20«C/nrin to 220*C and held.
7.2.3 Column 2: Set helium carrier gas at 25 mL/min flow rate.
Column temperature is set at 170*C for 7 min and then programmed at
10*C/m1n to 250*C and held.
7.2.4 Column 3: Set nitrogen carrier gas at 30 mL/min flow rate.
Column temperature is set at 100*C and then immediately programmed at
25'C/min to 200*C and held.
7.3 Calibration; Refer to Method 8000 for proper calibration
techniques"!! Use Table 1 and especially Table 2 for guidance on selecting the
lowest point on the calibration curve.
7.3.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these
procedures.
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7.3.2 If cleanup Is performed on the samples, the analyst should
process a series of standards through the cleanup procedure and then
analyze the samples by GC. This will confirm elution patterns and the
absence of interferents from the reagents.
7.4 Gas chromatographic analysis:
7,4.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 uL of internal standard to the sample prior to
injection.
7.4.2 Follow Section 7.6 in Method 8000 for instructions on the
analysis sequence, appropriate dilutions, establishing daily retention
time windows, and identification criteria. Include a mid-level standard
after each group of 10 samples in the analysis sequence.
7.4.3 Examples of chromatograms for various organophosphorous
pesticides are shown in Figures 1 through 4.
7.4.4 Record the sample volume injected and the resulting peak
sizes (in area units or peak heights).
7.4.5 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes. See Section 7.8 of Method 8000 for calculation
equations.
7.4.6 If peak detection and identification are prevented due to
interferences, the hexane extract may undergo cleanup using Method 3620.
The resultant extract(s) may be analyzed by GC directly or may undergo
further cleanup to remove sulfur using Method 3660.
7.5 Cleanup;
7.5.1 Proceed with Method 3620, followed by, if necessary, Method
3660, using the 10-mL hexane extracts obtained from Paragraph 7.1.2.3.
7.5.2 Following cleanup, the extracts should be analyzed by GC, as
described in the previous paragraphs and in Method 8000.
8.0 QUALITY CONTROL.
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the
QC in Method 3600 and in the specific cleanup method.
8.2 Procedures to check the GC system operation are found in Method
8000, Section 8.6.
8140 - 8
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Date September 1986
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Column: 5% SP-2401 on Suptlcoport
Ttmptrature: 170°C 7 Minutes, then
tOOC/Minute to 250°C
D«ttctor: Phosphorus-Specific Flame Photometric
45678
RETENTION TIME (MINUTES)
10
11
12
Figure 1. Gas chromatogram of organophosphorus pesticides (Example 1).
8140 - 9
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Date September 1986
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Column: 3% SP-2401
Program: 170°C 7 Minutes, 10°C/Minute
to250°C
Ofttctor: Phosphorus/Nitrogen
I
7654 32
RETENTION TIME (MINUTES)
Figure 2. Gas chromatogram of organophosphorus pesticides (Example 2).
8140 - 10
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Date September 1986
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Column: 15% SE-54 on Gas Chrom Q
Temperature: 100°C Initial, then
25<>C/Minute to 200°C
Detector: Hall Electrolytic Conductivity-Oxidative Mode
7654321
RETENTION TIME (MINUTES)
Figure 3. Gas chromatogram of organophosphorus pesticides (Example 3).
8140 - 11
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Date September 1986
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Column: 5% SP-2401 on Supelcoport
Temperature: 170°C 2 Minutes, then 20°C/Minute to 22QOC
Detector: Phosphorus-Specific Flame Photometric
3456
RETENTION TIME (MINUTES)
Figure 4. Gas chromatogram of organophosphorus pesticides (Example 4).
8140 - 12
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8.2.1 Select a representative spike concentration for each analyte
to be measured. The quality control check sample concentrate (Method
8000, Section 8.6) should contain each analyte in acetone at a
concentration 1,000 times more concentrated than the selected spike
concentration.
8.2.2 Table 3 indicates Single Operator Accuracy and Precision for
this method. Compare the results obtained with the results given in
Table 3 to determine if the data quality is acceptable.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000, Section 8.10).
8.3.1 If recovery is not within limits, the following procedures
are required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are
a problem or flag the data as "estimated concentration."
8.4 GC/MS confirmation;
8.4.1 GC/MS techniques should be judiciously employed to support
qualitative identifications made with this method. The GC/MS operating
conditions and procedures for analysis are those specified in Method
8270.
8.4.2 When available, chemical ionization mass spectra may be
employed to aid in the qualitative identification process.
8.4.3 Should these MS procedures fail to provide satisfactory
results, additional steps may be taken before reanalysis. These steps
may include the use of alternate packed or capillary GC columns and
additional cleanup.
9.0 METHOD PERFORMANCE
9.1 Single-operator accuracy and precision studies have been conducted
using spiked wastewater samples. The results of these studies are presented
in Table 3.
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Date September 1986
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10.0 REFERENCES
1. Pressley, T.A. and J.E. Longbottoro, "The Determination of Organophosphorus
Pesticides in Industrial and Municipal Wastewater: Method 614," U.S. EPA/EMSL,
Cincinnati, OH, EPA-600/4-82-004, 1982.
2. Burke, J.A., "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists 48, 1037, 1965.
3. U.S. EPA, "Analysis of Volatile Hazardous Substances by GC/MS: Pesticide
Methods Evaluation," Letter Reports 6, 12A, and 14, EPA Contract 68-03-2697,
1982.
4. U.S. EPA, "Method 622, Organophosphorous Pesticides," Environmental
Monitoring and Support Laboratory, Cincinnati, OH 45268.
8140 - 14
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Date September 1986
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TABLE 3. SINGLE-OPERATOR ACCURACY AND PRECISION3
Parameter
Azinphos methyl
Bolstar
Chlorpyrifos
Coumaphos
Demeton
Diazinon
Dichlorvos
Disulfoton
Ethoprop
Fensulfothion
Fenthion
Merphos
Mevinphos
Naled
Parathion methyl
Phorate
Ronnel
Stirophos
Tokuthion
Trichloronate
Average
recovery
72.7
64.6
98.3
109.0
67.4
67.0
72.1
81.9
100.5
94.1
68.7
120.7
56.5
78.0
96.0
62.7
99.2
66.1
64.6
105.0
Standard
deviation
f V \
\ /
18.8
6.3
5.5
12.7
10.5
6.0
7.7
9.0
4.1
17.1
19.9
7.9
7.8
8.1
5.3
8.9
5.6
5.9
6.8
18.6
Spi ke
range
(ug/L)
21-250
4.9-46
1.0-50.5
25-225
11.9-314
5.6
15.6-517
5.2-92
1.0-51.5
23.9-110
5.3-64
1.0-50
15.5-520
25.8-294
0.5-500
4.9-47
1.0-50
30.3-505
5.3-64
20
Number
of
analyses
17
17
18
17
17
7
16
17
18
17
17
18
16
16
21
17
18
16
17
3
Information taken from Reference 4.
8140 - 15
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METHOD B14O
ORGANOPHOSPHORUS PESTICIDES
C
Q
7.1.1
I Choose
appropriate
extraction
procedure
(see Chapter 2)
7.4
Perform GC
analysis (see
Method 8000)
7.1.2
Exchange
i extract-
Ion solvent to
hexane
during K-D
procedures
7.2
Set gas
chrometography
conditions
Is Identlf lca-
tlon C detection
prevented by
interfei—
ences
7.5.1]
Cleanup
using Method
3620 and 3360
If necessary
7
.3
MC
fc
CI
tc
Refer to
thod 8000
ir proper
llbratlon
chnlques
7.3.2
Process
l a series
of standards
through cleanup
procedure:
analyze by GC
8140 - 16
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METHOD 8141A
ORGANOPHOSPHORUS COMPOUNDS BY GAS CHROMATOGRAPHY:
CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8141 is a capillary gas chromatographic (GC) method used to
determine the concentration of organophosphorus (OP) compounds. The fused-
silica, open-tubular columns specified in this method offer improved resolution,
better selectivity, increased sensitivity, and faster analysis than packed
columns. The compounds listed in the table below can be determined by GC using
capillary columns with a flame photometric detector (FPD) or a nitrogen-
phosphorus detector (NPD). Triazine herbicides can also be determined with this
method when the NPD is used. Although performance data are presented for each
of the listed chemicals, it is unlikely that all of them could be determined in
a single analysis. This limitation results because the chemical and
chromatographic behavior of many of these chemicals can result in co-elution.
The analyst must select columns, detectors and calibration procedures for the
specific analytes of interest in a study. Any listed chemical is a potential
method interference when it is not a target analyte.
Compound Name
8141A - 1
CAS Registry No.
OP Pesticides
Aspon,b
Azinphos-methyl
Azinphos-ethyla
Bolstar (Sulprofos)
Carbophenothion8
Chlorfenvinphos8
Chlorpyrifos
Chlorpyrifos methyl8
Coumaphos
Crotoxyphos8
Demeton-0°
Demeton-Sc
Diazinon
Dichlorofenthiona
Dichlorvos (DDVP)
Dicrotophos8
Dimethoate
Dioxathion8'0
Disulfoton
EPN
Ethion"
Ethoprop
Famphur8
Fenitrothion8
Fensulfothion
3244-90-4
86-50-0
2642-71-9
35400-43-2
786-19-6
470-90-6
2921-88-2
5598-13-0
56-72-4
7700-17-6
8065-48-3
8065-48-3
333-41-5
97-17-6
62-73-7
141-66-2
60-51-5
78-34-2
298-04-4
2104-64-5
563-12-2
13194-48-4
52-85-7
122-14-5
115-90-2
Revision 1
September 1994
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Compound Name
CAS Registry No.
Fonophos8
Fenthion
Leptophosa/d
Malathion
Merphos0
Mevinphos6
Monocrotophos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Phosmet8
Phosphamidon8
Ronnel
Stirophos (Tetrachlorovinphos)
Sulfotepp
TEPPd
Terbufos8
Thionazina'b (Zinophos)
Tokuthionb (Protothiofos)
Trichlorfon8
Trichloronateb
Industrial Chemicals
Hexamethylphosphoramide8 (HMPA)
Tri-o-cresylphosphatea'd (TOCP)
Triazine Herbicides (NPD only)
Atrazine8
Simazine3
944-22-9
55-38-9
21609-90-5
121-75-5
150-50-5
7786-34-7
6923-22-4
300-76-5
56-38-2
298-00-0
298-02-2
732-11-6
13171-21-6
299-84-3
22248-79-9
3689-24-5
21646-99-1
13071-79-9
297-97-2
34643-46-4
52-68-6
327-98-0
680-31-9
78-30-8
1912-24-9
122-34-9
a This analyte has been evaluated using a 30-m column only.
b Production discontinued in the U.S., standard not readily available.
c Standards may have multiple components because of oxidation.
d Compound is extremely toxic or neurotoxic.
e Adjacent major/minor peaks can be observed due to cis/trans isomers.
1.2 A dual-column/dual-detector approach may be used for the analysis of
relatively clean extracts. Two 15- or 30-m x 0.53-mm ID fused-silica, open-
tubular columns of different polarities are connected to an injection tee and
each is connected to a detector. Analysts are cautioned regarding the use of a
dual column configuration when their instrument is subject to mechanical stress,
8141A - 2
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September 1994
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when many samples are analyzed over a short time, or when extracts of
contaminated samples are analyzed.
1.3 Two detectors can be used for the listed OP chemicals. The FPD works
by measuring the emission of phosphorus- or sulfur-containing species. Detector
performance is optimized by selecting the proper optical filter and adjusting the
hydrogen and air flows to the flame. The NPD is a flame ionization detector with
a rubidium ceramic flame tip which enhances the response of phosphorus- and
nitrogen-containing analytes. The FPD is more sensitive and more selective, but
is a less common detector in environmental laboratories.
1.4 Table 1 lists method detection limits (MDLs) for the target analytes,
using 15-m columns and FPD, for water and soil matrices. Table 2 lists the
estimated quantitation limits (EQLs) for other matrices. MDLs and EQLs using 30-
m columns will be very similar to those obtained from 15-m columns.
1.5 The use of a 15-m column system has not been fully validated for the
determination of the following compounds. The analyst must demonstrate
chromatographic resolution of all analytes, recoveries of greater than 70
percent, with precision of no more than 15 percent RSD, before data generated on
the 15-m column system can be reported for these, or any additional, analytes:
Azinphos-ethyl Ethion Phosmet
Carbophenothion Famphur Phosphamidon
Chlorfenvinphos HMPA Terbufos
Dioxathion Leptophos TOCP
1.6 When Method 8141 is used to analyze unfamiliar samples, compound
identifications should be supported by confirmatory analysis. Sec. 8.0 provides
gas chromatograph/mass spectrometer (GC/MS) criteria appropriate for the
qualitative confirmation of compound identifications.
1.7 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of capillary gas chromatography and in the
interpretation of chromatograms.
2.0 SUMMARY OF METHOD
2.1 Method 8141 provides gas chromatographic conditions for the detection
of ppb concentrations of organophosphorus compounds. Prior to the use of this
method, appropriate sample preparation techniques must be used. Water samples
are extracted at a neutral pH with methylene chloride by using a separatory
funnel (Method 3510) or a continuous liquid-liquid extractor (Method 3520).
Soxhlet extraction (Method 3540) or automated Soxhlet extraction (Method 3541)
using methylene chloride/acetone (1:1) are used for solid samples. Both neat and
diluted organic liquids (Method 3580, Waste Dilution) may be analyzed by direct
injection. Spiked samples are used to verify the applicability of the chosen
extraction technique to each new sample type. A gas chromatograph with a flame
photometric or nitrogen-phosphorus detector is used for this multiresidue
procedure.
8141A - 3 Revision 1
September 1994
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2.2 Organophosphorus esters and thioesters can hydrolyze under both acid
and base conditions. Samples prepared using acid and base partitioning
procedures are not suitable for analysis by Method 8141.
2.3 Ultrasonic Extraction (Method 3550) is not an appropriate sample
preparation method for Method 8141 and should not be used because of the
potential for destruction of target analytes during the ultrasonic extraction
process.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000, as well as to Sec. 1.1.
3.2 The use of Florisil Cleanup (Method 3620) for some of the compounds
in this method has been demonstrated to yield recoveries less than 85 percent and
is therefore not recommended for all compounds. Refer to Table 2 of Method 3620
for recoveries of organophosphorus compounds. Use of an FPD often eliminates the
need for sample cleanup. If particular circumstances demand the use of an
alternative cleanup procedure, the analyst must determine the elution profile and
demonstrate that the recovery of each analyte is not less than 85 percent.
3.3 The use of Gel Permeation Cleanup (GPC) (Method 3640) for sample
cleanup has been demonstrated to yield recoveries of less than 85 percent for
many method analytes because they elute before bis-(2-ethylhexyl) phthalate.
Method 3640 is therefore not recommended for use with this method, unless
analytes of interest are listed in Method 3640 or are demonstrated to give
greater than 85 percent recovery.
3.4 Use of a flame photometric detector in the phosphorus mode will
minimize interferences from materials that do not contain phosphorus or sulfur.
Elemental sulfur will interfere with the determination of certain
organophosphorus compounds by flame photometric gas chromatography. If Method
3660 is used for sulfur cleanup, only the tetrabutylammonium (TBA)-sulfite option
should be employed, since copper and mercury may destroy OP pesticides. The
stability of each analyte must be tested to ensure that the recovery from the
TBA-sulfite sulfur cleanup step is not less than 85 percent.
3.5 A halogen-specific detector (i.e., electrolytic conductivity or
microcoulometry) is very selective for the halogen-containing compounds and may
be used for the determination of Chlorpyrifos, Ronnel, Coumaphos, Tokuthion,
Trichloronate, Dichlorvos, EPN, Naled, and Stirophos only. Many of the OP
pesticides may also be detected by the electron capture detector (ECD); however,
the ECD is not as specific as the NPD or FPD. The ECD should only be used when
previous analyses have demonstrated that interferences will not adversely effect
quantitation, and that the detector sensitivity is sufficient to meet regulatory
limits.
3.6 Certain analytes will coelute, particularly on 15-m columns (Table
3). If coelution is observed, analysts should (1) select a second column of
different polarity for confirmation, (2) use 30-m x 0.53-mm columns, or (3) use
0.25- or 0.32-mm ID columns. See Figures 1 through 4 for combinations of
compounds that do not coelute on 15-m columns.
8141A - 4 Revision 1
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3.7 The following pairs coeluted on the DB-5/DB-210 30-m column pair:
DB-5 Terbufos/tri-o-cresyl phosphate
Naled/Simazine/Atrazine
Dichlorofenthion/Demeton-0
Trichloronate/Aspon
Bolstar/Stirophos/Carbophenothion
Phosphamidon/Crotoxyphos
Fensulfothion/EPN
DB-210 Terbufos/tri-o-cresyl phosphate
Dichlorofenthion/Phosphamidon
Chlorpyrifos, methyl/Parathion, methyl
Chlorpyrifos/Parathion, ethyl
Aspon/Fenthion
Demeton-0/Dimethoate
Leptophos/Azinphos-methyl
EPN/Phosmet
Famphur/Carbophenothion
See Table 4 for retention times of these compounds on 30-m columns.
3.8 Analytical difficulties encountered for target analytes include:
3.8.1 Tetraethyl pyrophosphate (TEPP) is an unstable diphosphate
which is readily hydrolyzed in water and is thermally labile (TEPP
decomposes at 170°C). Care must be taken to minimize loss during GC
analysis and during sample preparation. Identification of bad standard
lots is difficult since the electron impact (El) mass spectrum of TEPP is
nearly identical to its major breakdown product, triethyl phosphate.
3.8.2 The water solubility of Dichlorvos (DDVP) is 10 g/L at 20°C,
and recovery is poor from aqueous solution.
3.8.3 Naled is converted to Dichlorvos (DDVP) on column by
debromination. This reaction may also occur during sample workup. The
extent of debromination will depend on the nature of the matrix being
analyzed. The analyst must consider the potential for debromination when
Naled is to be determined.
3.8.4 Trichlorfon rearranges and is dehydrochlorinated in acidic,
neutral, or basic media to form Dichlorvos (DDVP) and hydrochloric acid.
If this method is to be used for the determination of organophosphates in
the presence of Trichlorfon, the analyst should be aware of the
possibility of rearrangement to Dichlorvos to prevent misidentification.
3.8.5 Demeton (Systox) is a mixture of two compounds; 0,0-diethyl
0-[2-(ethylthio)ethyl]phosphorothioate (Demeton-0) and 0,0-diethyl S-[2-
(ethylthio)ethyl]phosphorothioate (Demeton-S). Two peaks are observed in
all the chromatograms corresponding to these two isomers. It is
recommended that the early eluting compound (Demeton-S) be used for
quantisation.
8141A - 5 Revision 1
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3.8.6 Dioxathion is a single-component pesticide. However, several
extra peaks are observed in the chromatograms of standards. These peaks
appear to be the result of spontaneous oxygen-sulfur isomerization.
Because of this, Dioxathion is not included in composite standard
mixtures.
3.8.7 Merphos (tributyl phosphorotrithioite) is a single-component
pesticide that is readily oxidized to its phosphorotrithioate (Merphos
oxone). Chromatographic analysis of Merphos almost always results two
peaks (unoxidized Merphos elutes first). As the relative amounts of
oxidation of the sample and the standard are probably different,
quantitation based on the sum of both peaks may be most appropriate.
3.8.8 Retention times of some analytes, particularly Monocrotophos,
may increase with increasing concentrations in the injector. Analysts
should check for retention time shifts in highly contaminated samples.
3.8.9 Many analytes will degrade on reactive sites in the
chromatographic system. Analysts must ensure that injectors and splitters
are free from contamination and are silanized. Columns should be
installed and maintained properly.
3.8.10 Performance of chromatographic systems will degrade with
time. Column resolution, analyte breakdown and baselines may be improved
by column washing (Sec. 7). Oxidation of columns is not reversible.
3.9 Method interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware that lead to discrete
artifacts or elevated baselines in gas chromatograms. All these materials must
be routinely demonstrated to be free from interferences under the conditions of
the analysis by analyzing reagent blanks (Sec. 8.0).
3.10 NP Detector interferences: Triazine herbicides, such as Atrazine
and Simazine, and other nitrogen-containing compounds may interfere.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph: An analytical system complete with a gas
chromatograph suitable for on-column or split/splitless injection, and all
required accessories, including syringes, analytical columns, gases, suitable
detector(s), and a recording device. The analyst should select the detector for
the specific measurement application, either the flame photometric detector or
the nitrogen-phosphorus detector. A data system for measuring peak areas and
dual display of chromatograms is highly recommended.
4.1.1 Capillary Columns (0.53-mm, 0.32-mm, or 0.25-mm ID x 15-m or
30-m length, depending on the resolution required). Columns of 0.53-mm ID
are recommended for most environmental and waste analysis applications.
Dual-column, single-injector analysis requires columns of equal length and
bore. See Sec. 3.0 and Figures 1 through 4 for guidance on selecting the
proper length and diameter for the column(s).
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4.1.1.1 Column 1 - 15- or 30-m x 0.53-mm wide-bore
capillary column, 1.0-fj.m film thickness, chemically bonded with 50%
trifluoropropyl polysiloxane, 50% methyl polysiloxane (DB-210), or
equivalent.
4.1.1.2 Column 2 - 15- or 30-m x 0.53-mm wide-bore
capillary column, 0.83-jum film thickness, chemically bonded with
35% phenyl methyl polysiloxane (DB-608, SPB-608, RTx-35), or
equivalent.
4.1.1.3 Column 3 - 15- or 30-m x 0.53-mm wide-bore
capillary column, 1.0 jum film thickness, chemically bonded with 5%
phenyl polysiloxane, 95% methyl polysiloxane (DB-5, SPB-5, RTx-5),
or equivalent.
4.1.1.4 Column 4 - 15- or 30-m x 0.53-mm ID fused-silica
open-tubular column, chemically bonded with methyl polysiloxane
(DB-1, SPB-1, or equivalent), 1.0-/im or 1.5-jim film thickness.
4.1.1.5 (optional) Column rinsing kit: Bonded-phase column
rinse kit (J&W Scientific, Catalog no. 430-3000 or equivalent).
4.1.2 Splitter: If a dual-column, single-injector configuration is
used, the open tubular columns should be connected to one of the following
splitters, or equivalent:
4.1.2.1 Splitter 1 - J&W Scientific press-fit Y-shaped
glass 3-way union splitter (J&W Scientific, Catalog no. 705-0733).
4.1.2.2 Splitter 2 - Supelco 8-in glass injection tee,
deactivated (Supelco, Catalog no. 2-3665M).
4.1.2.3 Splitter 3 - Restek Y-shaped fused-silica
connector (Restek, Catalog no. 20405).
4.1.3 Injectors:
4.1.3.1 Packed column, 1/4-in injector port with hourglass
liner are recommended for 0.53-mm column. These injector ports can
be fitted with splitters (Sec. 4.0) for dual-column analysis.
4.1.3.2 Split/splitless capillary injectors operated in
the split mode are required for 0.25-mm and 0.32-mm columns.
4.1.4 Detectors:
4.1.4.1 Flame Photometric Detector (FPD) operated in the
phosphorus-specific mode is recommended.
4.1.4.2 Nitrogen-Phosphorus Detector (NPD) operated in the
phosphorus-specific mode is less selective but can detect triazine
herbicides.
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4.1.4.3 Halogen-Specific Detectors (electrolytic
conductivity or microcoulometry) may be used only for a limited
number of halogenated or sulfur-containing analytes (Sec. 3.0).
4.1.4.4 Electron-capture detectors may be used for a
limited number of analytes (Sec. 3.0).
4.1.5 Data system:
4.1.5.1 Data system capable of presenting chromatograms,
retention time, and peak integration data is strongly recommended.
4.1.5.2 Use of a data system that allows storage of raw
chromatographic data is strongly recommended.
5.0 REAGENTS
5.1 Solvents
5.1.1 Isooctane, (CH3)3CCH2CH(CH3)2 - Pesticide qual ity or equivalent.
5.1.2 Hexane, C6H14 - Pesticide quality or equivalent.
5.1.3 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.1.4 Tetrahydrofuran (THF), C4H80 - Pesticide quality or equivalent
(for triazine standards only).
5.1.5 Methyl tert-butyl-ether (MTBE), CH3Ot-C4H9 - Pesticide quality
or equivalent (for triazine standards only).
5.2 Stock standard solutions (1000 mg/L): Can be prepared from pure
standard materials or can be purchased as certified solutions.
5.2.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure compounds. Dissolve the compounds in suitable mixtures
of acetone and hexane and dilute to volume in a 10-mL volumetric flask.
If compound purity is 96 percent or greater, the weight can be used
without correction to calculate the concentration of the stock standard
solution. Commercially prepared stock standard solutions can be used at
any concentration if they are certified by the manufacturer or by an
independent source.
5.2.2 Both Simazine and Atrazine have low solubilities in hexane.
If Simazine and Atrazine standards are required, Atrazine should be
dissolved in MTBE, and Simazine should be dissolved in acetone/MTBE/THF
(1:3:1).
5.2.3 Composite stock standard: This standard can be prepared from
individual stock solutions. The analyst must demonstrate that the
individual analytes and common oxidation products are resolved by the
chromatographic system. For composite stock standards containing less
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than 25 components, take exactly 1 ml of each individual stock solution at
1000 mg/L, add solvent, and mix the solutions in a 25-mL volumetric flask.
For example, for a composite containing 20 individual standards, the
resulting concentration of each component in the mixture, after the volume
is adjusted to 25 ml, will be 40 mg/L. This composite solution can be
further diluted to obtain the desired concentrations. Composite stock
standards containing more than 25 components are not recommended.
5.2.4 Store the standard solutions (stock, composite, calibration,
internal, and surrogate) at 4°C in Teflon-sealed containers in the dark.
All standard solutions should be replaced after two months, or sooner if
routine QC (Sec. 8.0) indicates a problem. Standards for easily
hydrolyzed chemicals including TEPP, Methyl Parathion, and Merphos should
be checked every 30 days.
5.2.5 It is recommended that lots of standards be subdivided and
stored in small vials. Individual vials should be used as working
standards to minimize the potential for contamination or hydrolysis of the
entire lot.
5.3 Calibration standards should be prepared at a minimum of five
concentrations by dilution of the composite stock standard with isooctane or
hexane. The concentrations should correspond to the expected range of
concentrations found in real samples and should bracket the linear range of the
detector. Organophosphorus calibration standards should be replaced after one
or two months, or sooner if comparison with check samples or historical data
indicates that there is a problem. Laboratories may wish to prepare separate
calibration solutions for the easily hydrolyzed standards identified above.
5.4 Internal standard: Internal standards should only be used on well-
characterized samples by analysts experienced in the technique. Use of internal
standards is complicated by co-elution of some OP pesticides and by the
differences in detector response to dissimilar chemicals.
5.4.1 FPD response for organophosphorus compounds is enhanced by the
presence of sulfur atoms bonded to the phosphorus atom. It has not been
established that a thiophosphate can be used as an internal standard for
an OP with a different numbers of sulfur atoms (e.g., phosphorothioates
[P=S] as an internal standard for phosphates [P04]) or phosphorodithioates
[P=S2]).
5.4.2 If internal standards are to be used, the analyst must select
one or more internal standards that are similar in analytical behavior to
the compounds of interest. The analyst must further demonstrate that the
measurement of the internal standard is not affected by method or matrix
interferences.
5.4.3 When 15-m columns are used, it may be difficult to fully
resolve internal standards from target analytes, method interferences and
matrix interferences. The analyst must demonstrate that the measurement
of the internal standard is not affected by method or matrix
interferences.
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5.4.4 The following NPD internal standard has been used for a 30-m
column pair. Make a solution of 1000 mg/L of l-bromo-2-nitrobenzene. For
spiking, dilute this solution to 5 mg/L. Use a spiking volume of 10 /nL/mL
of extract. The spiking concentration of the internal standards should be
kept constant for all samples and calibration standards. Since its FPD
response is small, l-bromo-2-nitrobenzene is not an appropriate internal
standard for that detector. No FPD internal standard is suggested.
5.5 Surrogate standard spiking solutions - The analyst should monitor the
performance of the extraction, cleanup (when used), and analytical system, and
the effectiveness of the method in dealing with each sample matrix, by spiking
each sample, standard, and blank with one or two surrogates (e.g.,
organophosphorus compounds not expected to be present in the sample). If
multiple analytes are to be measured, two surrogates (an early and a late eluter)
are recommended. Deuterated analogs of analytes are not appropriate surrogates
for gas chromatographic/FPD/NPD analysis.
5.5.1 If surrogates are to be used, the analyst must select one or
more compounds that are similar in analytical behavior to the compounds of
interest. The analyst must further demonstrate that the measurement of a
surrogate is not affected by method or matrix interferences. General
guidance on the selection and use of surrogates is provided in Sec. 5.0 of
Method 3500.
5.5.2 Tributyl phosphate and triphenyl phosphate are used as FPD and
NPD surrogates. A volume of 1.0 ml of a 1-M9/L spiking solution (1 ng of
surrogate) is added to each water sample and each soil/sediment sample.
If there is a co-elution problem, 4-chloro-3-nitrobenzo-trifluoride has
also been used as an NPD-only surrogate.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to Chapter Four, "Organic Analytes,"
Sec. 4.0.
6.2 Extracts are to be refrigerated at 4°C and analyzed within 40 days
of extraction. See Sec. 5.2.4 for storage of standards.
6.3 Organophosphorus esters will hydrolyze under acidic or basic
conditions. Adjust samples to a pH of 5 to 8 using sodium hydroxide or sulfuric
acid solution as soon as possible after sample collection. Record the volume
used.
6.4 Even with storage at 4°C and use of mercuric chloride as a
preservative, most OPs in groundwater samples collected for the national
pesticide survey degraded within a 14-day period. Begin sample extraction within
7 days of collection.
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7.0 PROCEDURE
7.1 Extraction and cleanup:
7.1.1 Refer to Chapter Two and Method 8140 for guidance on choosing
the appropriate extraction procedure. In general, water samples are
extracted at a neutral pH with methylene chloride, using either Method
3510 or 3520. Solid samples are extracted using either Method 3540 or
3541 with methylene chloride/acetone (1:1 v/v) or hexane/acetone (1:1 v/v)
as the extraction solvent. Method 3550 is an inappropriate extraction
technique for the target analytes of this method (See Sec. 2.3).
7.1.2 Extraction and cleanup procedures that use solutions below pH
4 or above pH 8 are not appropriate for this method.
7.1.3 If required, the samples may be cleaned up using the Methods
presented in Chapter Four, Sec. 2. Florisil Column Cleanup (Method 3620)
and Sulfur Cleanup (Method 3660, TBA-sulfite option) may have particular
application for OPs. Gel Permeation Cleanup (Method 3640) should not
generally be used for OP pesticides.
7.1.3.1 If sulfur cleanup by Method 3660 is required, do
not use mercury or copper.
7.1.3.2 GPC may only be employed if all target OP
pesticides are listed as analytes of Method 3640, or if the
laboratory has demonstrated a recovery of greater than 85 percent
for target OPs at a concentration not greater than 5 times the
regulatory action level. Laboratories must retain data
demonstrating acceptable recovery.
7.1.4 Prior to gas chromatographic analysis, the extraction solvent
may be exchanged to hexane. The analyst must ensure quantitative transfer
of the extract concentrate. Single-laboratory data indicate that samples
should not be transferred with 100-percent hexane during sample workup, as
the more polar organophosphorus compounds may be lost. Transfer of
organophosphorus esters is best accomplished using methylene chloride or
a hexane/acetone solvent mixture.
7.1.5 Methylene chloride may be used as an injection solvent with
both the FPD and the NPD.
NOTE: Follow manufacturer's instructions as to suitability of using
methylene chloride with any specific detector.
7.2 Gas chromatographic conditions:
7.2.1 Four 0.53-mm ID capillary columns are suggested for the
determination of organophosphates by this method. Column 1 (DB-210 or
equivalent) and Column 2 (SPB-608 or equivalent) of 30-m length are
recommended if a large number of organophosphorus analytes are to be
determined. If superior chromatographic resolution is not required, 15-m
lengths columns may be appropriate. Operating conditions for 15-m columns
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are listed in Table 5. Operating conditions for 30-m columns are listed
in Table 6.
7.2.2 Retention times for analytes on each set of columns are
presented in Tables 3 and 4.
7.3 Calibration: Refer to Method 8000 for proper calibration techniques.
Use Table 5 and Table 6 for establishing the proper operating parameters for the
set of columns being employed in the analyses.
7.4 Gas chromatographic analysis: Method 8000 provides instructions on
the analysis sequence, appropriate dilutions, establishing daily retention time
windows and identification criteria.
7.4.1 Automatic injections of 1 juL are recommended. Hand injections
of no more than 2 /LtL may be used if the analyst demonstrates quantitation
precision of < 10 percent relative standard deviation. The solvent flush
technique may be used if the amount of solvent is kept at a minimum. If
the internal standard calibration technique is used, add 10 juL of internal
standard to each ml of sample prior to injection. Chromatograms of the
target organophosphorus compounds are shown in Figures 1 through 4.
7.4.2 Figures 5 and 6 show chromatograms with and without Simazine,
Atrazine, and Carbophenothion on 30-m columns.
7.5 Record the sample volume injected to the nearest 0.05 pi and the
resulting peak sizes (in area units or peak heights). Using either the internal
or external calibration procedure (Method 8000), determine the identity and
quantity of each component peak in the sample chromatogram which corresponds to
the compounds used for calibration purposes. See Method 8000 for calculation
equations.
7.5.1 If peak detection and identification is prevented by the
presence of interferences, the use of an FPD or further sample cleanup is
required. Before using any cleanup procedure, the analyst must process a
series of calibration standards through the procedure to establish elution
patterns and to determine recovery of target compounds. The absence of
interference from reagents must be demonstrated by routine processing of
reagent blanks through the chosen cleanup procedure. Refer to Sec. 3.0
for interferences.
7.5.2 If the responses exceed the linear range of the system, dilute
the extract and reanalyze. It is recommended that extracts be diluted so
that all peaks are on scale. Overlapping peaks are not always evident
when peaks are off-scale. Computer reproduction of chromatograms,
manipulated to ensure all peaks are on scale over a 100-fold range, are
acceptable if linearity is demonstrated. Peak height measurements are
recommended over peak area integration when overlapping peaks cause errors
in area integration.
7.5.3 If the peak response is less than 2.5 times the baseline noise
level, the validity of the quantitative result may be questionable. The
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analyst should consult with the source of the sample to determine whether
further concentration of the sample extract is warranted.
7.5.4 If partially overlapping or coeluting peaks are found, change
columns or try a GC/MS technique. Refer to Sec. 8.0 and Method 8270.
7.6 Suggested chromatograph maintenance: Corrective measures may require
any one or more of the following remedial actions.
7.6.1 Refer to Method 8000 for general information on the
maintenance of capillary columns and injectors.
7.6.2 Splitter connections: For dual columns which are connected
using a press-fit Y-shaped glass splitter or a Y-shaped fused-silica
connector (J&W Scientific, Restek, or equivalent), clean and deactivate
the splitter. Reattach the columns after cleanly cutting off at least one
foot from the injection port side of the column using a capillary cutting
tool or scribe. The accumulation of high boiling residues can change
split ratios between dual columns and thereby change calibration factors.
7.6.3 Columns will be damaged permanently and irreversibly by
contact with oxygen at elevated temperature. Oxygen can enter the column
during a septum change, when oxygen traps are exhausted, through neoprene
diaphragms of regulators, and through leaks in the gas manifold. Polar
columns including the DB-210 and DB-608 are more prone to oxidation.
Oxidized columns will exhibit baselines that rise rapidly during
temperature programming.
7.6.4 Peak tailing for all components will be exacerbated by dirty
injectors, pre-columns, and glass "Y"s. Additionally, cleaning of this
equipment (or replacement/clipping, as appropriate) will greatly reduce
the peak tailing. Components such as Fensulfothion, Naled, Azinphos-
methyl, and Dimethoate are very good indicators of system performance.
7.7 Detector maintenance:
7.7.1 Older FPDs may be susceptible to stray light in the
photomultiplier tube compartment. This stray light will decrease the
sensitivity and the linearity of the detector. Analysts can check for
leaks by initiating an analysis in a dark room and turning on the lights.
A shift in the baseline indicates that light may be leaking into the
photomultiplier tube compartment. Additional shielding should be applied
to eliminate light leaks and minimize stray light interference.
7.7.2 The bead of the NPD will become exhausted with time, which
will decrease the sensitivity and the selectivity of the detector. The
collector may become contaminated which decreased detector sensitivity.
7.7.3 Both types of detectors use a flame to generate a response.
Flow rates of air and hydrogen should be optimized to give the most
sensitive, linear detector response for target analytes.
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8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Include a mid-level check standard after each group of 10 samples in the analysis
sequence. Quality control to validate sample extraction is covered in Method
3500 and in the extraction method utilized. If extract cleanup was performed,
follow the QC in Method 3600 and in the specific cleanup method.
8.2 Procedures to check the GC system operation are found in Method 8000.
8.3 GC/MS confirmation
8.3.1 GC/MS techniques should be judiciously employed to support
qualitative identifications made with this method. Follow the GC/MS
operating requirements specified in Method 8270.
8.3.2 When available, chemical ionization mass spectra may be
employed to aid in the qualitative identification process.
8.3.3 To confirm an identification of a compound, the background-
corrected mass; spectrum of the compound must be obtained from the sample
extract and must be compared with a mass spectrum from a stock or
calibration standard analyzed under the same chromatographic conditions.
At least 25 ng of material should be injected into the GC/MS. The
following criteria must be met for qualitative confirmation:
8.3.3.1 The qualitative identification of compounds
determined by this method is based on retention time, and on
comparison of the sample mass spectrum, after background correction,
with characteristic ions in a reference mass spectrum. The
reference mass spectrum must be generated by the laboratory using
the conditions of this method. The characteristic ions from the
reference mass spectrum are defined to be the three ions of greatest
relative intensity, or any ions over 30% relative intensity if less
than three such ions occur in the reference spectrum. Compounds
should be identified as present when the criteria below are met.
8.3.3.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographic peak containing ions
specific for the target compound at a compound-specific
retention time will be accepted as meeting this criterion.
8.3.3.1.2 The RRT of the sample component is within
± 0.06 RRT units of the RRT of the standard component.
8.3.3.1.3 The relative intensities of the
characteristic ions agree within 30% of the relative
intensities of these ions in the reference spectrum.
(Example: For an ion with an abundance of 50% in the
reference spectrum, the corresponding abundance in a sample
spectrum can range between 20% and 80%.)
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8.3.3.1.4 Structural isomers that produce very similar
mass spectra should be identified as individual isomers if
they have sufficiently different GC retention times.
Sufficient GC resolution is achieved if the height of the
valley between two isomer peaks is less than 25% of the sum of
the two peak heights. Otherwise, structural isomers are
identified as isomeric pairs.
8.3.3.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce
mass spectra containing ions contributed by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
appropriate selection of analyte spectra and background
spectra is important. Examination of extracted ion current
profiles of appropriate ions can aid in the selection of
spectra, and in qualitative identification of compounds. When
analytes coelute (i.e., only one chromatographic peak is
apparent), the identification criteria can be met, but each
analyte spectrum will contain extraneous ions contributed by
the coeluting compound.
8.3.3.2 For samples containing components not associated
with the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the purpose of the
analyses being conducted. Computer generated library search
routines should not use normalization routines that would
misrepresent the library or unknown spectra when compared to each
other. For example, the RCRA permit or waste delisting requirements
may require the reporting of nontarget analytes. Only after visual
comparison of sample spectra with the nearest library searches will
the mass spectral interpretation specialist assign a tentative
identification. Guidelines for making tentative identification are:
(1) Relative intensities of major ions in the reference
spectrum (ions > 10% of the most abundant ion) should be present in
the sample spectrum.
(2) The relative intensities of the major ions should agree
within ± 20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
between 30 and 70%.)
(3) Molecular ions present in the reference spectrum should
be present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the
reference spectrum should be reviewed for possible background
contamination or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the
sample spectrum should be reviewed for possible subtraction from the
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sample spectrum because of background contamination or coeluting
peaks. Data system library reduction programs can sometimes create
these discrepancies.
8.3.4 Where available, chemical ionization mass spectra may be
employed to aid in the qualitative identification process because of the
extensive fragmentation of organophosphorus pesticides during electron
impact MS processes.
8.3.5 Should the MS procedure fail to provide satisfactory results,
additional steps may be taken before reanalysis. These steps may include
the use of alternate packed or capillary GC columns or additional sample
cleanup.
9.0 METHOD PERFORMANCE
9.1 Estimated MDLs and associated chromatographic conditions for water
and clean soil (uncontaminated with synthetic organics) are listed in Table 1.
As detection limits will vary with the particular matrix to be analyzed, guidance
for determining EQLs is given in Table 2. Recoveries for several method analytes
are provided in Tables 5, 6, and 7.
10.0 REFERENCES
1. Taylor, V.; Mickey, D.M.; Marsden, P.J. "Single Laboratory Validation of
EPA Method 8140"; U.S. Environmental Protection Agency, Environmental
Monitoring Systems Laboratory, Office of Research and Development, Las
Vegas, NV, 1987; EPA-600/4-87-009.
2. Pressley, T.A; Longbottom, J.E. "The Determination of Organophosphorus
Pesticides in Industrial and Municipal Wastewater: Method 614"; U.S.
Environmental Protection Agency, Environmental Monitoring and Support
Laboratory, Cincinnati, OH, 1982; EPA-600/4-82-004.
3. "Analysis of Volatile Hazardous Substances by GC/MS: Pesticide Methods
Evaluation"; Letter Reports 6, 12A, and 14 to the U.S. Environmental
Protection Agency on Contract 68-03-2697, 1982.
4. "Method 622, Organophosphorus Pesticides"; U.S. Environmental Protection
Agency, Environmental Monitoring and Support Laboratory, Cincinnati, OH
45268.
5. Lopez-Avila, V.; Baldin, E.; Benedicto, J; Milanes, J.; Beckert, W. F.
"Application of Open-Tubular Columns to SW-846 GC Methods"; final report
to the U.S. Environmental Protection Agency on Contract 68-03-3511; Mid-
Pacific Environmental Laboratory, Mountain View, CA, 1990.
6. Hatcher, M.D.; Hickey, D.M.; Marsden, P.J.; and Betowski, L.D.;
"Development of a GC/MS Module for RCRA Method 8141"; final report to the
U.S. EPA Environmental Protection Agency on Contract 68-03-1958; S-Cubed,
San Diego, CA, 1988.
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7. Chau, A.S.Y.; Afghan, B.K. Analysis of Pesticides in Water; "Chlorine and
Phosphorus-Containing Pesticides"; CRC: Boca Raton, FL, 1982, Vol. 2, pp
91-113, 238.
8. Hild, J.; Schulte, E; Thier, H.P. "Separation of Organophosphorus
Pesticides and Their Metabolites on Glass-Capillary Columns";
Chromatographia, 1978, 11-17.
9. Luke, M.A.; Froberg, J.E.; Doose, G.M.; Masumoto, H.T. "Improved
Multiresidue Gas Chromatographic Determination of Organophosphorus,
Organonitrogen, and Organohalogen Pesticides in Produce, Using Flame
Photometric and Electrolytic Conductivity Detectors"; J. Assoc. Off, Anal.
Chem. 1981, 1187, 64.
10. Sherma, J.; Berzoa, M. "Analysis of Pesticide Residues in Human and
Environmental Samples"; U.S. Environmental Protection Agency, Research
Triangle Park, NC; EPA-600/8-80-038.
11. Desmarchelier, J.M.; Wustner, D.A.; Fukuto, T.R. "Mass Spectra of
Organophosphorus Esters and Their Alteration Products"; Residue Reviews,
1974, pp 63, 77.
12. Munch, D.J. and Frebis, C.P., "Analyte Stability Studies Conducted during
the National Pesticide Survey", ES & 7", 1992, vol 26, 921-925.
13. T.L. Jones, "Organophosphorus Pesticide Standards: Stability Study", EMSL-
LV Research Report, EPA 600/X-92/040, April, 1992
14. Kotronarou, A., et al., "Decomposition of Parathion in Aqueous Solution by
Ultrasonic Irradiation," £5*7, 1992, Vol. 26, 1460-1462.
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TABLE 1
METHOD DETECTION LIMITS IN A WATER AND A SOIL
MATRIX USING 15-m COLUMNS AND A FLAME PHOTOMETRIC DETECTOR
Compound
Reagent
Water (3510)'
(M9/L)
Soil (3540)
Azinphos-methyl
Bolstar (Sulprofos)
Chlorpyrifos
Coumaphos
Demeton, -0, -S
Diazinon
Dichlorvos (DDVP)
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfothion
Fenthion
Malathion
Merphos
Mevinphos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Ronnel
Sulfotepp
TEPP°
Tetrachlorovinphos
Tokuthion (Protothiofos)c
Trichloronate0
0.10
0.07
0.07
0.20
0.12
0.20
0.80
0.26
0.07
0.04
0.20
0.08
0.08
0.11
0.20
0.50
0.50
0.06
0.12
0.04
0.07
0.07
0.80
0.80
0.07
0.80
5.0
3.5
5.0
10.0
6.0
10.0
40.0
13.0
3.5
2.0
10.0
4.0
5.0
5.5
10.0
25.0
25.0
3.0
6.0
2.0
3.5
3.5
40.0
40.0
5.5
40.0
Sample extracted using Method 3510, Separatory Funnel Liquid-Liquid
Extraction.
Sample extracted using Method 3540, Soxhlet Extraction.
Purity of these standards not established by the EPA Pesticides and
Industrial Chemicals Repository, Research Triangle Park, NC.
8141A - 18
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September 1994
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TABLE 2
DETERMINATION OF ESTIMATED QUANTITATION LIMITS
(EQLs) FOR VARIOUS MATRICES8
Matrix Factor
Ground water (Methods 3510 or 3520) 10b
Low-concentration soil by Soxhlet and no cleanup 10°
Non-water miscible waste (Method 3580) 1000
c
8 EQL = [Method detection limit (see Table 1)] X [Factor found in this table].
For non-aqueous samples, the factor is on a wet-weight basis. Sample EQLs are
highly matrix dependent. The EQLs to be determined herein are for guidance and
may not always be achievable.
b Multiply this factor times the reagent water MDL in Table 1.
c Multiply this factor times the soil MDL in Table 1.
8141A - 19 Revision 1
September 1994
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TABLE 3.
RETENTION TIMES FOR METHOD 8141A ANALYTES
EMPLOYING 15-m COLUMNS
TEPP
Dichlorvos (DDVP)
Mevinphos
Demeton, -0 and -S
Ethoprop
Naled
Phorate
Monochrotophos
Sulfotepp
Dimethoate
Disulfoton
Diazinon
Merphos
Ronnel
Chlorpyrifos
Malathion
Parathion, methyl
Parathion, ethyl
Trichloronate
Tetrachlorovinphos
Tokuthion (Protothiofos)
Fensulfothion
Bolstar^ (Sulprofos)
Famphur*
EPN
Azinphos-methyl
Fenthion
Coumaphos
"Method 8141A has not been fully
Initial temperature
Initial time
Program 1 rate
Program 1 final temp.
Program 1 hold
Program 2 rate
Program 2 final temp.
Program 2 hold
Caoil
Compound
9.63
14.18
18.31
18.62
19.94
20.04
20.11
20.64
23.71
24.27
26.82
29.23
31.17
31.72
31.84
31.85
32.19
34.65
34.67
35.85
36.34
36.40
38.34
38.83
39.83
lary Column
OB-5
6.44
7.91
12.88
15.90
16.48
19.01
17.52
20.11
18.02
20.18
19.96
20.02
21.73
22.98
26.88
28.78
23.71
27.62
28.41
32.99
24.58
35.20
35.08
36.93
37.80
38.04
29.45
38.87
SPB-608
5.12
12.79
18.44
17.24
18.67
17.40
18.19
31.42
19.58
27.96
20.66
19.68
32.44
23.19
25.18
32.58
32.17
33.39
29.95
33.68
39.91
36.80
37.55
37.86
36.71
37.24
28.86
39.47
DB-210
10.66
19.35
36.74
validated for Famphur.
130°C
3 minutes
5°C/min
180°C
10 minutes
2°C/min
250°C
15 minutes
50°C
1 minute
5°C/min
140°C
10 minutes
10°C/min
240°C
10 minutes
50°C
1 minute
5°C/min
140°C
10 minutes
10°C/min
240°C
10 minutes
8141A - 20
Revision 1
September 1994
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TABLE 4.
RETENTION TIMES FOR METHOD 8141A ANALYTES
EMPLOYING 30-m COLUMNS8
Compound
DB-5
RT (min)
DB-210 DB-608
DB-1
Trimethyl phosphate
Dichlorvos (DDVP)
Hexamethyl phosphorami de
Trichlorfon
TEPP
Thionazin
Mevinphos
Ethoprop
Diazinon
Sulfotepp
Terbufos
Tri-o-cresyl phosphate
Naled
Phorate
Fonophos
Disulfoton
Merphos
Oxidized Merphos
Dichlorofenthion
Chlorpyrifos, methyl
Ronnel
Chlorpyrifos
Trichloronate
Aspon
Fenthion
Demeton-S
Demeton-0
Monocrotophos0
Dimethoate
Tokuthion
Malathion
Parathion, methyl
Fenithrothion
Chlorfenvinphos
Parathion, ethyl
Bolstar
Stirophos
Ethion
b
7.45
b
11.22
b
12.32
12.20
12.57
13.23
13.39
13.69
13.69
14.18
12.27
14.44
14.74
14.89
20.25
15.55
15.94
16.30
17.06
17.29
17.29
17.87
11.10
15.57
19.08
18.11
19.29
19.83
20.15
20.63
21.07
21.38
22.09
22.06
22.55
2.36
6.99
7.97
11.63
13.82
24.71
10.82
15.29
18.60
16.32
18.23
18.23
15.85
16.57
18.38
18.84
23.22
24.87
20.09
20.45
21.01
22.22
22.73
21.98
22.11
14.86
17.21
15.98
17.21
24.77
21.75
20.45
21.42
23.66
22.22
27.57
24.63
27.12
6.56
12.69
11.85
18.69
24.03
20.04
22.97
18.92
20.12
23.89
35.16
26.11
26.29
27.33
29.48
30.44
29.14
21.40
17.70
19.62
20.59
33.30
28.87
25.98
32.05
29.29
38.10
33.40
37.61
10.43
14.45
18.52
21.87
19.60
18.78
19.65
21.73
26.23
23.67
24.85
24.63
20.18
19.3
19.87
27.63
24.57
22.97
24.82
29.53
26.90
(continued)
8141A - 21
Revision 1
September 1994
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TABLE 4. (Continued)
Compound
DB-5
RT (min)
DB-210 DB-608
DB-1
Phosphamidon
Crotoxyphos
Leptophos
Fensulfothion
EPN
Phosmet
Azinphos-methyl
Azinphos-ethyl
Famphur
Coumaphos
Atrazine
Simazine
Carbophenothion
Dioxathion
Trithion methyl
Dicrotophos
Internal Standard
1-Bromo-2-nitrobenzene
Surrogates
Tributyl phosphate
Triphenyl phosphate
4-Cl-3-nitrobenzotrifluoride
22.77
22.77
24.62
27.54
27.58
27.89
28.70
29.27
29.41
33.22
13.98
13.85
22.14
d
8.11
5.73
20.09
23.85
31.32
26.76
29.99
29.89
31.25
32.36
27.79
33.64
17.63
17.41
27.92
9.07
5.40
25.88
32.65
44.32
36.58
41.94
41.24
43.33
45.55
38.24
48.02
22.24
36.62
19.33
11.1
33.4
28.58
31.60
32.33
34.82
a The GC operating conditions were as follows:
DB-5 and DB-210 - 30-m x 0.53-mm ID column, DB-5 (1.50- m film thickness) and
DB-210 (1.0- m film thickness). Both connected to a press-fit Y-shaped inlet
splitter. Temperature program: 120°C (3-min hold) to 270°C (10-min hold) at
5°C/min; injector temperature 250°C; detector temperature 300°C; bead temperature
400°C; bias voltage 4.0; hydrogen gas pressure 20 psi; helium carrier gas 6
mL/min; helium makeup gas 20 mL/min.
DB-608 - 30-m x 0.53-mm ID column, DB-608 (1.50- m film thickness) installed in
an 0.25-in packed-column inlet. Temperature program: 110°C (0.5-min hold) to
250°C (4-min hold) at 3°C/min; injector temperature 250°C; helium carrier gas 5
mL/min; flame photometric detector.
DB-1 30-m x 0.32-mm ID column, DB-1 (0.25- m film thickness) split/splitless with
head pressure of 10 psi, split valve closure at 45 sec, injector temp. 250°C,
50°C (1-min hold) to 280°C (2-min hold) at 6°C/min, mass spectrometer full scan
35-550 amu.
b Not detected at 20 ng per injection.
c Retention times may shift to longer times with larger amounts injected (shifts
of over 30 seconds have been observed, Hatcher et. al.)
d Shows multiple peaks; therefore, not included in the composite.
8141A - 22
Revision 1
September 1994
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TABLE 5.
PERCENT RECOVERY OF 27 ORGANOPHOSPHATES BY SEPARATORY FUNNEL EXTRACTION
Compound
Azinphos methyl
Bolstar
Chlorpyrifos
Coumaphos
Demeton
Diazinon
Dichlorvos
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfonthion
Fenthion
Malathion
Merphos
Mevinphos
Monocrotophos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Ronnel
Sulfotep
TEPP
Tetrachlorvinphos
Tokuthion
Trichloroate
Low
126
134
7
103
33
136
80
NR
48
113
82
84
NR
127
NR
NR
NR
NR
101
NR
94
67
87
96
79
NR
NR
Percent Recovery
Medium
143 + 8
141 + 8
89 + 6
90 + 6
67 + 11
121 + 9.5
79 + 11
47 + 3
92 + 7
125 + 9
90 + 6
82 + 12
48 + 10
92 + 6
79
NR
18 + 4
NR
94 + 5
46 + 4
77 + 6
97 + 5
85 + 4
55 + 72
90 + 7
45 + 3
35
High
101
101
86
96
74
82
72
101
84
97
80
96
89
86
81
55
NR
NR
86
44
73
87
83
63
80
90
94
NR = Not recovered.
8141A - 23
Revision 1
September 1994
-------
TABLE 6.
PERCENT RECOVERY OF 27 ORGANOPHOSPHATES BY CONTINUOUS LIQUID-LIQUID EXTRACTION
Percent Recovery
Compound
Azinphos methyl
Bolstar
Chlorpyrifos
Coumaphos
Demeton
Diazinon
Dichlorvos
Dimethoate
Disulfoton
EPN
Ethoprop
Famphur
Fensulfonthion
Fenthion
Malathion
Merphos
Mevinphos
Monocrotophos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Ronnel
Sulfotep
TEPP
Tetrachlorvinphos
Tokuthion
Trichloroate
Low
NR
NR
13
94
38
NR
81
NR
94
NR
39
_.
90
8
105
NR
NR
NR
NR
106
NR
84
82
40
39
56
132
NR
Medium
129
126
82 + 4
79 + 1
23 + 3
128 + 37
32 + 1
10 + 8
69 + 5
104 + 18
76 + 2
63 + 15
67 + 26
32 + 2
87 + 4
80
87
30
NR
81 + 1
50 + 30
63 + 3
83 + 7
77 + 1
18 + 7
70 + 14
32 + 14
NR
High
122
128
88
89
41
118
74
102
81
119
83
--
90
86
86
79
49
1
74
87
43
74
89
85
70
83
90
21
NR = Not recovered.
8141A - 24
Revision 1
September 1994
-------
TABLE 7.
PERCENT RECOVERY OF 27 ORGANOPHOSPHATES BY SOXHLET EXTRACTION
Percent Recovery
Compound
Azinphos methyl
Bolstar
Chlorpyrifos
Coumaphos
Demeton
Diazinon
Dichlorvos
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfonthion
Fenthion
Malathion
Merphos
Mevinphos
Monocrotophos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Ronnel
Sulfotep
TEPP
Tetrachlorvinphos
Tokuthion
Trichloroate
Low
156
102
NR
93
169
87
84
NR
78
114
65
72
NR
100
62
NR
NR
NR
75
NR
75
NR
67
36
50
NR
56
Medium
110 + 6
103 + 15
66 + 17
89 + 11
64 + 6
96 + 3
39 + 21
48 + 7
78 + 6
93 + 8
70 + 7
81 + 18
43 + 7
81+8
53
71
NR
48
80 + 8
41 + 3
77 + 6
83 + 12
72 + 8
34 + 33
81 + 7
40 + 6
53
High
87
79
79
90
75
75
71
98
76
82
75
111
89
81
60
63
NR
NR
80
28
78
79
78
63
83
89
53
NR = Not recovered,
8141A - 25
Revision 1
September 1994
-------
TABLE 8.
SUGGESTED OPERATING CONDITIONS FOR 15-m COLUMNS
Columns 1 and 2 (DB-210 and SPB-608 or their equivalent)
Carrier gas (He) flow rate =
Initial temperature =
Temperature program =
Column 3 (DB-5 or equivalent)
Carrier gas (He) flow rate =
Initial temperature =
Temperature program =
5 mL/min
50°C, hold for 1 minute
50'C to 140°C at 5°C/min, hold for
10 minutes, followed by 140°C to
240°C at 10°C/min, hold for 10
minutes (or a sufficient amount of
time for last compound to elute).
5 mL/min
130°C, hold for 3 minutes
130°C to 180°C at 5°C/min, hold for
10 minutes, followed by 180'C to
250°C at 2°C/min, hold for 15
minutes (or a sufficient amount of
time for last compound to elute).
8141A - 26
Revision 1
September 1994
-------
TABLE 9
SUGGESTED OPERATING CONDITIONS FOR 30-m COLUMNS
Column 1:
Type: DB-210
Dimensions: 30-m x 0.53-mm ID
Film Thickness (urn) : 1.0
Column 2:
Type: DB-5
Dimensions: 30-m x 0.53-mm ID
Film Thickness (nm): 1.5
Carrier gas flowrate (mL/min): 6 (Helium)
Makeup gas flowrate (mL/min): 20 (Helium)
Temperature program: 120°C (3-min hold) to 270°C (10-min hold) at 5°C/min
Injector temperature: 250°C
Detector temperature: 300°C
Injection volume: 2 juL
Solvent: Hexane
Type of injector: Flash vaporization
Detector type: Dual NPD
Range: 1
Attenuation: 64
Type of splitter: Y-shaped or Tee
Data system: Integrator
Hydrogen gas pressure: 20 psi
Bead temperature: 400°C
Bias voltage: 4
8141A - 27 Revision 1
September 1994
-------
TABLE 10
QUANTITATION AND CHARACTERISTIC IONS FOR OP PESTICIDES
Compound Name
Quantitation ions
Characteristic ions
Azinphos-methyl
Bolstar (Sulprofos)
Chlorpyrifos
Coumaphos
Demeton-S
Diazinon
Dichlorvos (DDVP)
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfothion
Fenthion
Malathion
Merphos
Mevinphos
Monocrotophos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Ronnel
Stirophos
Sulfotepp
TEPP
Tokuthion
160
156
197
109
88
137
109
87
88
157
158
293
278
173
209
127
127
109
291
109
75
285
109
322
99
113
77,132
140,143,113,33
97,199,125.314
97,226,362,21
60,114,170
179,152,93,199,304
79,185,145
93,125,58,143
89,60,61,97,142
169,141,63,185
43,97,41,126
97,125,141,109,308
125,109,93,169
125,127,93,158
57,153,41,298
109,67,192
67,97,192,109
145,147,79
97,109,139,155
125,263,79
121,97,47,260
125,287,79,109
329,331,79
97,65,93,121,202
155,127,81,109
43,162,267,309
8141A - 28
Revision 1
September 1994
-------
300.00
250.00
200.00
150.00
100.00
50.00
0.00
..
1
e
2
Tetrachlorovinphos
FensuHothion
V... Ji
. i . . . p . •) i ri | • • i | i . i, i .1 | .1 . , . i . , . ti f i, • , . , ,, , n | , , i ,. , ,, . . . | , . , ,, , ,, , , , , , , , , , , ,, . . . |
3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
Figure 1. Chromatogram of target organophosphorus compounds from a 15-m DB-21C
column with FPD detector. More compounds are shown in Figure 2. See Table 3 for
retention times.
8141A - 29
Revision 1
September 1994
-------
300.00
a.
UJ
250.00
200.00
150.00
100.00
50.00
V
0.00
i>
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
Figure 2. Chromatogram of target organophosphorus compounds from a 15-m DB-210
column with FPD detector. More compounds are shown in Figure 1. See Table 3 for
retention times.
8141A - 30
Revision 1
September 1994
-------
300.00
250.00
200.00-
150.00-
100.00-
50.00-
0.00-J
a.
£
OL
C
Figure 3. Chromatogram of target organophosphorus compounds from a 15-m OB-21
column with NPD detector. More compounds are shown in Figure 4. See Table 3 fo
retention times.
8141A - 31
Revision 1
September 1994
-------
30000-
250.00-
200.00-
150.00-
100.00-
50.00-
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
Figure 4. Chromatogram of target organophosphorus compounds from a 15-m DB-210
column with NPD detector. More compounds are shown in Figure 3. See Table 3 for
retention times.
8141A - 32
Revision 1
September 1994
-------
-r
DB-210
n
DB-5
•4
Figure 5. Chromatogram of target organophosphorus compounds on a 30-m DB-5/DB-21C
column pair with NPD detector, without Simazine, Atrazine and Carbophenothion. See
Table 4 for retention times and for GC operating conditions.
8141A - 33
Revision 1
September 1994
-------
J
is
\J
DB-210
tl
»
II
n
u
«• a
41
•i;
M
U
II
JL
4
•
il
1
1
17
n
u
IM
II
taJJ
1 "
f
•
.11
1
11
M
»
4t
i.
1
DB-5
4
i
4t
44
JU
Figure 6. Chromatogram of target organophosphorus compounds on a 30-m DB-5/DB-210
column pair with NPD detector, with Simazine, Atrazine and Carbophenothion. See
Table 4 for retention times and for GC operating conditions.
8141A - 34
Revision 1
September 1994
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METHOD 8141A
ORGANOPHOSPHORUS COMPOUNDS BY GAS CHROMATOGRAPHY:
CAPILLARY COLUMN TECHNIQUE
( Start )
7.1.1 Refer to Chapter
Two for guidance on
choosing the appropriate
extraction procedure.
7.1.2 Perform
solvent exchange
during K-D
procedures in all
extraction methods.
7.2 Select GC
conditions.
7.3 Refer to Method
8000 for
calibration techniques.
I
7.3.1 Internal or
external
calibration may
be used.
7.4.1 Add internal
standard to sample
if necessary.
7.4.2 Refer to
Method 8000, Sec.
7.6 for instructions
on analysis sequence,
dilutions, retention times,
and identification
criteria.
7.4.3 Inject sample.
7.4.5 Record sample
volume injected and
resulting peak sizes.
I
7.4.6 Determine
identity and
quantity of each
component peak;
refer to Method
8000, Sec. 7.8 for
calculation equations.
7.4.7
Is peak
detection and
identification
prevented by
interfer-
ences?
7.5.1 Perform
appropriate cleanup.
7.5.2 Reanalyze by
GC.
I
( Stop J
8141A - 35
Revision 1
September 1994
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METHOD 8150B
CHLORINATED HERBICIDES BY GAS CHROMATOGRAPHY
1.0 SCOPE AND APPLICATION
1.1 Method 8150 is a gas chromatographic (GC) method for determining
certain chlorinated acid herbicides. The following compounds can be determined
by this method:
Compound Name CAS No.'
2,4-D 94-75-7
2,4-DB 94-82-6
2,4,5-TP (Silvex) 93-72-1
2,4,5-T 93-76-5
Dalapon 75-99-0
Dicamba 1918-00-9
Dichloroprop 120-36-5
Dinoseb 88-85-7
MCPA 94-74-6
MCPP 93-65-2
a Chemical Abstract Services Registry Number.
1.2 Table 1 lists the method detection limit for each compound in
organic-free reagent water. Table 2 lists the estimated quantitation limit (EQL)
for other matrices.
1.3 When Method 8150 is used to analyze unfamiliar samples, compound
identifications should be supported by at least one additional qualitative
technique. This method describes analytical conditions for a second gas
chromatographic column that can be used to confirm measurements made with the
primary column. Sec. 8.4 provides gas chromatograph/mass spectrometer (GC/MS)
criteria appropriate for the qualitative confirmation of compound
identifications.
1.4 Only experienced analysts should be allowed to work with
diazomethane due to the potential hazards associated with its use (the compound
is explosive and carcinogenic).
2.0 SUMMARY OF METHOD
2.1 Method 8150 provides extraction, esterification, and gas
chromatographic conditions for the analysis of chlorinated acid herbicides.
Spiked samples are used to verify the applicability of the chosen extraction
technique to each new sample type. The esters are hydrolyzed with potassium
8150B - 1 Revision 2
September 1994
-------
hydroxide, and extraneous organic material is removed by a solvent wash. After
acidification, the acids are extracted with solvent and converted to their methyl
esters using diazomethane as the derivatizing agent. After excess reagent is
removed, the esters are determined by gas chromatography employing an electron
capture detector, microcoulometric detector, or electrolytic conductivity
detector (Goer!Hz and Lamar, 1967). The results are reported as the acid
equivalents.
2.2 The sensitivity of Method 8150 usually depends on the level of
interferences rather than on instrumental limitations.
3.0 INTERFERENCES
3.1 Refer to Method 8000.
3.2 Method interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware that lead to discrete
artifacts or elevated baselines in gas chromatograms. All these materials must
be routinely demonstrated to be free from interferences under the conditions of
the analysis, by analyzing reagent blanks.
3.2.1 Glassware must be scrupulously cleaned. Clean each piece of
glassware as soon as possible after use by rinsing it with the last
solvent used in it. This should be followed by detergent washing with hot
water and rinses with tap water, then with organic-free reagent water.
Glassware should be solvent-rinsed with acetone and pesticide-quality
hexane. After rinsing and drying, glassware should be sealed and stored
in a clean environment to prevent any accumulation of dust or other
contaminants. Store glassware inverted or capped with aluminum foil.
Immediately prior to use, glassware should be rinsed with the next solvent
to be used.
3.2.2 The use of high purity reagents and solvents helps to minimize
interference problems. Purification of solvents by distillation in all-
glass systems may be required.
3.3 Matrix interferences may be caused by contaminants that are
coextracted from the sample. The extent of matrix interferences will vary
considerably from waste to waste, depending upon the nature and diversity of the
waste being sampled.
3.4 Organic acids, especially chlorinated acids, cause the most direct
interference with the determination. Phenols, including chlorophenols, may also
interfere with this procedure.
3.5 Alkaline hydrolysis and subsequent extraction of the basic solution
remove many chlorinated hydrocarbons and phthalate esters that might otherwise
interfere with the electron capture analysis.
3.6 The herbicides, being strong organic acids, react readily with
alkaline substances and may be lost during analysis. Therefore, glassware and
glass wool must be acid rinsed, and sodium sulfate must be acidified with
sulfuric acid prior to use to avoid this possibility.
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3.7 Sample extracts should be dry prior to methylation or else poor
recoveries will be obtained.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph, analytical system complete with gas
chromatograph suitable for on-column injections and all required
accessories, including detectors, analytical columns, recorder, gases, and
syringes. A data system for measuring peak heights and/or peak areas is
recommended.
4.1.2 Columns
4.1.2.1 Column la and Ib - 1.8 m x 4 mm ID glass, packed
with 1.5% SP-2250/1.95% SP-2401 on Supelcoport (100/120 mesh) or
equivalent.
4.1.2.2 Column 2 - 1.8 m x 4 mm ID glass, packed with 5%
OV-210 on Gas Chrom Q (100/120 mesh) or equivalent.
4.1.2.3 Column 3 - 1.98 m x 2 mm ID glass, packed with
0.1% SP-1000 on 80/100 mesh Carbopack C or equivalent.
4.1.3 Detector - Electron capture (BCD).
4.2 Erlenmeyer flasks - 250 and 500 ml Pyrex, with 24/40 ground glass
joint.
4.3 Beaker - 500 mL.
4.4 Diazomethane generator - Refer to Sec. 7.4 to determine which method
of diazomethane generation should be used for a particular application.
4.4.1 Diazald kit - recommended for the generation of diazomethane
using the procedure given in Sec. 7.4.2 (Aldrich Chemical Co., Cat. No.
210,025-2 or equivalent).
4.4.2 Assemble from two 20 x 150 mm test tubes, two Neoprene rubber
stoppers, and a source of nitrogen. Use Neoprene rubber stoppers with
holes drilled in them to accommodate glass delivery tubes. The exit tube
must be drawn to a point to bubble diazomethane through the sample
extract. The generator assembly is shown in Figure 1. The procedure for
use of this type of generator is given in Sec. 7.4.3.
4.5 Vials - 10 to 15 mL, amber glass, with Teflon lined screw cap or
crimp top.
4.6 Separatory funnel - 2000 mL, 125 mL, and 60 mL.
4.7 Drying column - 400 mm x 20 mm ID Pyrex chromatographic column with
Pyrex glass wool at bottom and a Teflon stopcock.
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NOTE: Fritted glass discs are difficult to decontaminate after highly
contaminated extracts have been passed through. Columns without
frits may be purchased. Use a small pad of Pyrex glass wool to
retain the adsorbent. Prewash the glass wool pad with 50 ml of
acetone followed by 50 ml of elution solvent prior to packing the
column with adsorbent.
4.8 Kuderna-Danish (K-D) apparatus
4.8.1 Concentrator tube - 10 ml, graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts
4.8.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps or
equivalent.
4.8.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.8.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.8.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.9 Boiling chips - Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.10 Water bath - Heated, with concentric ring cover, capable of
temperature control (+ 5°C). The bath should be used in a hood.
4.11 Microsyringe - 10 /xL.
4.12 Wrist shaker - Burrell Model 75 or equivalent.
4.13 Glass wool - Pyrex, acid washed.
4.14 Balance - Analytical, capable of accurately weighing to 0.0001 g.
4.15 Syringe - 5 ml.
4.16 Glass rod.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests.
Unless otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
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5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sulfuric acid solution
5.3.1 ((1:1) (v/v)) - Slowly add 50 ml H2S04 (sp. gr. 1.84) to 50 ml
of organic-free reagent water.
5.3.2 ((1:3) (v/v)) - Slowly add 25 ml H2S04 (sp. gr. 1.84) to 75 ml
of organic-free reagent water.
5.4 Hydrochloric acid ((1:9) (v/v)), HC1. Add one volume of
concentrated HC1 to 9 volumes of organic-free reagent water.
5.5 Potassium hydroxide solution (KOH) - 37% aqueous solution (w/v).
Dissolve 37 g potassium hydroxide pellets in organic-free reagent water, and
dilute to 100 ml.
5.6 Carbitol (Diethylene glycol monoethyl ether), C2H5OCH2CH2OCH2CH2OH.
Available from Aldrich Chemical Co.
5.7 Solvents
5.7.1 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.7.2 Methanol, CH3OH - Pesticide quality or equivalent.
5.7.3 Isooctane, (CH3)3CCH2CH(CH3)2 - Pesticide quality or
equivalent.
5.7.4 Hexane, C6HU - Pesticide quality or equivalent.
5.7.5 Diethyl Ether, C2H5OC2H5. Pesticide quality or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl alcohol preservative must be
added to each liter of ether.
5.8 Sodium sulfate (granular, acidified, anhydrous), Na2S04. Purify by
heating at 400°C for 4 hours in a shallow tray, or by precleaning the sodium
sulfate with methylene chloride. If the sodium sulfate is precleaned with
methylene chloride, a method blank must be analyzed, demonstrating that there is
no interference from the sodium sulfate. Acidify by slurrying 100 g sodium
sulfate with enough diethyl ether to just cover the solid; then add 0.1 ml of
concentrated sulfuric acid and mix thoroughly. Remove the ether under a vacuum.
Mix 1 g of the resulting solid with 5 ml of organic-free reagent water and
measure the pH of the mixture. It must be below a pH of 4. Store at 130°C.
5.9 N-Methyl-N-nitroso-p-toluenesulfonamide (Diazald), CH3C6H4S02N(CH3)NO.
Available from Aldrich Chemical Co.
5.10 Silicic acid. Chromatographic grade, nominal 100 mesh. Store at
130°C.
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5.11 Stock standard solutions - Stock standard solutions can be prepared
from pure standard materials or purchased as certified solutions.
5.11.1 Prepare stock standard solutions by accurately weighing
about 0.0100 g of pure acids. Dissolve the acids in pesticide quality
acetone and dissolve the esters in 10% acetone/isooctane (v/v) and dilute
to volume in a 10 ml volumetric flask. Larger volumes can be used at the
convenience of the analyst. If compound purity is certified at 96% or
greater, the weight can be used without correction to calculate the
concentration of the stock standard. Commercially prepared stock
standards can be used at any concentration if they are certified by the
manufacturer or by an independent source.
5.11.2 Transfer the stock standard solutions into vials with
Teflon lined screw caps or crimp tops. Store at 4°C and protect from
light. Stock standard solutions should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them.
5.11.3 Stock standard solutions of the derivatized acids must
be replaced after 1 year, or sooner, if comparison with check standards
indicates a problem. Stock standard solutions of the free acids degrade
more quickly and should be replaced after two months, or sooner if
comparison with check standards indicates a problem.
5.12 Calibration standards - A minimum of five calibration standards for
each parameter of interest should be prepared through dilution of the stock
standards with diethyl ether. One of the concentrations should be at a
concentration near, but above, the method detection limit. The remaining
concentrations should correspond to the expected range of concentrations found
in real samples or should define the working range of the GC. Calibration
solutions must be replaced after six months, or sooner if comparison with check
standards indicates a problem.
5.13 Internal standards (if internal standard calibration is used) - To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.13.1 Prepare calibration standards at a minimum of five
concentrations for each parameter of interest as described in Sec. 5.12.
5.13.2 To each calibration standard, add a known constant
amount of one or more internal standards, and dilute to volume with
hexane.
5.13.3 Analyze each calibration standard per Sec. 7.0.
5.14 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system, and the effectiveness
of the method in dealing with each sample matrix by spiking each sample,
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standard, and organic-free reagent water blank with one or two herbicide
surrogates (e.g. herbicides that are not expected to be present in the sample).
The surrogates selected should elute over the range of the temperature program
used in this method. 2,4-Dichlorophenylacetic acid (DCAA) is recommended as a
surrogate compound. Deuterated analogs of analytes should not be used as
surrogates for gas chromatographic analysis due to coelution problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Sec. 4.1. Extracts must be stored under refrigeration and analyzed within 40
days of extraction.
7.0 PROCEDURE
7.1 Preparation of waste samples
7.1.1 Extraction
7.1.1.1 Follow Method 3580 except use diethyl ether as the
dilution solvent, acidified anhydrous sodium sulfate, and acidified
glass wool.
7.1.1.2 Transfer 1.0 mL (a lesser volume or a dilution may
be required if herbicide concentrations are high) to a 250 mL ground
glass-stoppered Erlenmeyer flask. Proceed to Sec. 7.2.2 hydrolysis.
7.2 Preparation of soil, sediment, and other solid samples
7.2.1 Extraction
7.2.1.1 To a 500 mL, wide mouth Erlenmeyer flask add 50
g (dry weight as determined in Method 3540, Sec. 7.2.1) of the well
mixed, moist solid sample. Adjust the pH to 2 (See Method 9045)
with concentrated HC1 and monitor the pH for 15 minutes with
occasional stirring. If necessary, add additional HC1 until the pH
remains at 2.
7.2.1.2 Add 20 mL acetone to the flask and mix the
contents with the wrist shaker for 20 minutes. Add 80 mL diethyl
ether to the same flask and shake again for 20 minutes. Decant the
extract and measure the volume of solvent recovered.
7.2.1.3 Extract the sample twice more using 20 mL of
acetone followed by 80 mL of diethyl ether. After addition of each
solvent, the mixture should be shaken with the wrist shaker for
10 minutes and the acetone-ether extract decanted.
7.2.1.4 After the third extraction, the volume of extract
recovered should be at least 75% of the volume of added solvent.
If this is not the case, additional extractions may be necessary.
Combine the extracts in a 2 liter separatory funnel containing
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250 ml of reagent water. If an emulsion forms, slowly add 5 g of
acidified sodium sulfate (anhydrous) until the solvent-water mixture
separates. A quantity of acidified sodium sulfate equal to the
weight of the sample may be added, if necessary.
7.2.1.5 Check the pH of the extract. If it is not at or
below pH 2, add more concentrated HC1 until stabilized at the
desired pH. Gently mix the contents of the separatory funnel for
1 minute and allow the layers to separate. Collect the aqueous
phase in a clean beaker and the extract phase (top layer) in a
500 ml ground glass-stoppered Erlenmeyer flask. Place the aqueous
phase back into the separatory funnel and re-extract using 25 ml of
diethyl ether. Allow the layers to separate and discard the aqueous
layer. Combine the ether extracts in the 500 ml Erlenmeyer flask.
7.2.1.6 An alternative extraction procedure using
ultrasonic extraction can be found in Sec. 7.2 of Method 8151.
7.2.2 Hydrolysis
7..2.2.1 Add 30 ml of organic-free reagent water, 5 ml of
37% KOH, and one or two clean boiling chips to the flask. Place a
three ball Snyder column on the flask, evaporate the diethyl ether
on a water bath, and continue to heat until the hydrolysis step is
completed (usually 1 to 2 hours).
7.2.2.2 Remove the flask from the water bath and allow to
cool. Transfer the water solution to a 125 ml separatory funnel and
extract the basic solutions once with 40 mL and then twice with
20 ml of diethyl ether. Allow sufficient time for the layers to
separate and discard the ether layer each time. The phenoxy-acid
herbicides remain soluble in the aqueous phase as potassium salts.
7.2.3 Solvent cleanup
7.2.3.1 Adjust the pH to 2 by adding 5 ml cold (4°C)
sulfuric acid (1:3) to the separatory funnel. Be sure to check the
pH at this point. Extract the herbicides once with 40 ml and twice
with 20 ml of diethyl ether. Discard the aqueous phase.
7.2.3.2 Combine ether extracts in a 125 ml Erlenmeyer
flask containing 5-7 g of acidified anhydrous sodium sulfate.
Stopper and allow the extract to remain in contact with the
acidified sodium sulfate. If concentration and esterification are
not to be performed immediately, store the sample overnight in the
refrigerator.
NOJE: The drying step is very critical to ensuring complete
esterification. Any moisture remaining in the ether
will result in low herbicide recoveries. The amount of
sodium sulfate is adequate if some free=flowing
crystals are visible when swirling the flask. If all
the sodium sulfate solidifies in a cake, add a few
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additional grams of acidified sodium sulfate and again
test by swirling. The 2 hour drying time is a minimum,
however, the extracts may be held overnight in contact
with the sodium sulfate.
7.2.3.3 Transfer the ether extract, through a funnel
plugged with acid washed glass wool, into a 500 ml K-D flask
equipped with a 10 ml concentrator tube. Use a glass rod to crush
caked sodium sulfate during the transfer. Rinse the Erlenmeyer
flask and column with 20-30 mL of diethyl ether to complete the
quantitative transfer.
7.2.3.4 Add one or two clean boiling chips to the flask
and attach a three ball Snyder column. Prewet the Snyder column by
adding about 1 ml of diethyl ether to the top. Place the apparatus
on a hot water bath (60°-65°C) so that the concentrator tube is
partially immersed in the hot water and the entire lower rounded
surface of the flask is bathed in vapor. Adjust the vertical
position of the apparatus and the water temperature, as required,
to complete the concentration in 15-20 minutes. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 1 mL, remove the K-D apparatus from the water bath and allow
it to drain and cool for at least 10 minutes.
7.2.3.5 Remove the Snyder column and rinse the flask and
its lower joints into the concentrator tube with 1-2 ml of diethyl
ether. A 5 ml syringe is recommended for this operation. Add a
fresh boiling chip, attach a micro Snyder column to the concentrator
tube, and prewet the column by adding 0.5 ml of ethyl ether to the
top. Place the micro K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature as
required to complete concentration in 5-10 minutes. When the
apparent volume of the liquid reaches 0.5 ml, remove the micro K-D
from the bath and allow it to drain and cool. Remove the Snyder
column and add 0.1 ml of methanol. Rinse the walls of the
concentrator tube while adjusting the extract volume to 1.0 ml with
diethyl ether. Proceed to Sec. 7.4 for esterification.
7.3 Preparation of aqueous samples
7.3.1 Extraction
7.3.1.1 Using a 1 liter graduated cylinder, measure 1
liter (nominal) of sample, record the sample volume to the nearest
5 ml, and quantitatively transfer it to the separatory funnel. If
high concentrations are anticipated, a smaller volume may be used
and then diluted with organic-free reagent water to 1 liter. Adjust
the pH to less than 2 with sulfuric acid (1:1).
7.3.1.2 Add 150 ml of diethyl ether to the sample bottle,
seal, and shake for 30 seconds to rinse the walls. Transfer the
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solvent wash to the separatory funnel and extract the sample by
shaking the funnel for 2 minutes with periodic venting to release
excess pressure. Allow the organic layer to separate from the water
layer for a minimum of 10 minutes. If the emulsion interface
between layers is more than one third the size of the solvent layer,
the analyst must employ mechanical techniques to complete the phase
separation. The optimum technique depends upon the sample and may
include stirring, filtration of the emulsion through glass wool,
centrifugation, or other physical methods. Drain the aqueous phase
into a 1 liter Erlenmeyer flask. Collect the solvent extract in a
250 ml ground glass Erlenmeyer flask containing 2 ml of 37% KOH.
Approximately 80 mL of the diethyl ether will remain dissolved in
the aqueous phase.
7.3.1.3 Repeat the extraction two more times using 50 ml
of diethyl ether each time. Combine the extracts in the Erlenmeyer
flask. (Rinse the 1 liter flask with each additional aliquot of
extracting solvent.)
7.3.2 Hydrolysis
7.3.2.1 Add one or two clean boiling chips and 15 ml of
organic-free reagent water to the 250 ml flask and attach a three
ball Snyder column. Prewet the Snyder column by adding about 1 mL
of diethyl ether to the top of the column. Place the apparatus on
a hot water bath (60°-65°C) so that the bottom of the flask is bathed
with hot water vapor. Although the diethyl ether will evaporate in
about 15 minutes, continue heating until the hydrolysis step is
completed (usually 1 to 2 hours). Remove the apparatus and let
stand at room temperature for at least 10 minutes.
7.3.2.2 Transfer the solution to a 60 ml separatory funnel
using 5-10 ml of organic-free reagent water. Wash the basic
solution twice by shaking for 1 minute with 20 ml portions of
diethyl ether. Discard the organic phase. The herbicides remain
in the aqueous phase.
7.3.3 Solvent cleanup
7.3.3.1 Acidify the contents of the separatory funnel to
pH 2 by adding 2 ml of cold (4°C) sulfuric acid (1:3). Test with pH
indicator paper. Add 20 ml diethyl ether and shake vigorously for
2 minutes. Drain the aqueous layer into a 250 mL Erlenmeyer flask,
and pour the organic layer into a 125 mL Erlenmeyer flask containing
about 5-7 g of acidified sodium sulfate. Repeat the extraction
twice more with 10 mL aliquots of diethyl ether, combining all
solvent in the 125 mL flask. Allow the extract to remain in contact
with the sodium sulfate for approximately 2 hours.
NOTE: The drying step is very critical to ensuring complete
esterification. Any moisture remaining in the ether
will result in low herbicide recoveries. The amount of
sodium sulfate is adequate if some free flowing
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crystals are visible when swirling the flask. If all
the sodium sulfate solidifies in a cake, add a few
additional grams of acidified sodium sulfate and again
test by swirling. The 2 hour drying time is a minimum,
however, the extracts may be held overnight in contact
with the sodium sulfate.
7.3.3.2 Transfer the ether extract, through a funnel
plugged with acid washed glass wool, into a 500 ml K-D flask
equipped with a 10 ml concentrator tube. Use a glass rod to crush
caked sodium sulfate during the transfer. Rinse the Erlenmeyer
flask and column with 20-30 ml of diethyl ether to complete the
quantitative transfer.
7.3.3.3 Add one or two clean boiling chips to the flask
and attach a three ball Snyder column. Prewet the Snyder column by
adding about 1 mL of diethyl ether to the top. Place the apparatus
on a hot water bath (60°-65°C) so that the concentrator tube is
partially immersed in the hot water and the entire lower rounded
surface of the flask is bathed in vapor. Adjust the vertical
position of the apparatus and the water temperature, as required,
to complete the concentration in 15-20 minutes. At the proper rate
of distillation the balls of the column will actively chatter, but
the chambers will not flood. When the apparent volume of liquid
reaches 1 ml, remove the K-D apparatus from the water bath and allow
it to drain and cool for at least 10 minutes.
7.3.3.4 Remove the Snyder column and rinse the flask and
its lower joints into the concentrator tube with 1-2 ml of diethyl
ether. A 5 ml syringe is recommended for this operation. Add a
fresh boiling chip, attach a micro Snyder column to the concentrator
tube, and prewet the column by adding 0.5 mL of ethyl ether to the
top. Place the micro K-D apparatus on the water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature as
required to complete concentration in 5-10 minutes. When the
apparent volume of the liquid reaches 0.5 ml, remove the micro K-D
from the bath and allow it to drain and cool. Remove the Snyder
column and add 0.1 ml of methanol. Rinse the walls of the
concentrator tube while adjusting the extract volume to 1.0 mL with
diethyl ether.
7.4 Esterification
7.4.1 Two methods may be used for the generation of diazomethane:
the bubbler method (set up shown in Figure 1) and the Diazald kit method.
The bubbler method is suggested when small batches (10-15) of samples
require esterification. The bubbler method works well with samples that
have low concentrations of herbicides (e.g. aqueous samples) and is safer
to use than the Diazald kit procedure. The Diazald kit method is good for
large quantities of samples needing esterification. The Diazald kit
method is more effective than the bubbler method for soils or samples that
may contain high concentrations of herbicides (e.g., samples such as soils
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that result -in yellow extracts following hydrolysis may be difficult to
handle by the bubbler method). The diazomethane derivatization (U.S. EPA,
1971) procedures, described below, will react efficiently with all of the
chlorinated herbicides described in this method and should be used only by
experienced analysts, due to the potential hazards associated with its
use. The following precautions should be taken:
CAUTION: Diazomethane is a carcinogen and can explode under
certain conditions.
Use a safety screen.
Use mechanical pipetting aides.
Do not heat above 90°C -- EXPLOSION may result.
Avoid grinding surfaces, ground glass joints, sleeve
bearings, glass stirrers -- EXPLOSION may result.
Store away from alkali metals -- EXPLOSION may result.
Solutions of diazomethane decompose rapidly in the
presence of solid materials such as copper powder,
calcium chloride, and boiling chips.
7.4.2 Diazald kit method - Instructions for preparing diazomethane
are provided with the generator kit.
7.4.2.1 Add 2 mL of diazomethane solution and let sample
stand for 10 minutes with occasional swirling.
7.4.2.2 Rinse inside wall of the ampule with several
hundred ^L of diethyl ether. Allow solvent to evaporate
spontaneously at room temperature to about 2 mL.
7.4.2.3 Dissolve the residue in 5 mL of hexane. Analyze
by gas chromatography.
7.4.3 Bubbler method - Assemble the diazomethane bubbler (see
Figure 1).
7.4.3.1 Add 5 mL of diethyl ether to the first test tube.
Add 1 mL of diethyl ether, 1 mL of carbitol, 1.5 mL of 37% KOH, and
0.1-0.2 g Diazald to the second test tube. Immediately place the
exit tube into the concentrator tube containing the sample extract.
Apply nitrogen flow (10 mL/min) to bubble diazomethane through
the extract for 10 minutes or until the yellow color of diazomethane
persists. The amount of Diazald used is sufficient for
esterification of approximately three sample extracts. An
additional 0.1-0.2 g of Diazald may be added (after the initial
Diazald is consumed) to extend the generation of the diazomethane.
There is sufficient KOH present in the original solution to perform
a maximum of approximately 20 minutes of total esterification.
7.4.3.2 Remove the concentrator tube and seal it with a
Neoprene or Teflon stopper. Store at room temperature in a hood for
20 minutes.
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7.4.3.3 Destroy any unreacted diazomethane by adding
0.1-0.2 g silicic acid to the concentrator tube. Allow to stand
until the evolution of nitrogen gas has stopped. Adjust the sample
volume to 10.0 ml with hexane. Stopper the concentrator tube and
store refrigerated if further processing will not be performed
immediately. It is recommended that the methylated extracts be
analyzed immediately to minimize the trans-esterification and other
potential reactions that may occur. Analyze by gas chromatography.
7.5 Gas chromatographic conditions (Recommended)
7.5.1 Column la
Carrier gas (5% methane/95% argon) flow rate: 70 mL/min
Temperature program: 185°C, isothermal.
7.5.2 Column Ib
Carrier gas (5% methane/95% argon) flow rate: 70 mL/min
Initial temperature: 140°C, hold for 6 minutes
Temperature program: 140°C to 200°C at 10°C/min, hold until last
compound has eluted.
7.5.3 Column 2
Carrier gas (5% methane/95% argon) flow rate: 70 mL/min
Temperature program: 185°C, isothermal.
7.5.4 Column 3
Carrier gas (ultra-high purity N2) flow rate: 25 mL/min
Initial temperature: 100°C, no hold
Temperature program: 100°C to 150°C at 10°C/min, hold until last
compound has eluted.
7.6 Calibration - Refer to Method 8000 for proper calibration
techniques. Use Table 1 and especially Table 2 for guidance on selecting the
lowest point on the calibration curve.
7.6.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
7.6.2 The following gas chromatographic columns are recommended for
the compounds indicated:
Analyte Column Analyte Column
Dicamba la,2 Dalapon 3
2,4-D la,2 MCPP Ib
2,4,5-TP la,2 MCPA Ib
2,4,5-T la,2 Dichloroprop Ib
2,4-DB la Dinoseb Ib
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7.7 Gas chromatographic analysis
7.7.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 /xL of internal standard to the sample prior to
injection.
7.7.2 Method 8000 provides instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration check standard after
each group of 10 samples in the analysis sequence.
7.7.3 Examples of chromatograms for various chlorophenoxy acid
herbicides are shown in Figures 2 through 4.
7.7.4 Record the sample volume injected and the resulting peak sizes
(in area units or peak heights).
7.7.5 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes.
7.7.6 If calibration standards have been analyzed in the same manner
as the samples (e.g. have undergone hydrolysis and esterification), then
the calculation of concentration given in Method 8000 should be used.
However, if calibration is done using standards made from methyl ester
compounds (compounds not esterified by application of this method), then
the calculation of concentration must include a correction for the
molecular weight of the methyl ester versus the acid herbicide.
7.7.7 If peak detection and identification are prevented due to
interferences, further cleanup is required. Before using any cleanup
procedure, the analyst must process a series of standards through the
procedure to validate elution patterns and the absence of interferences
from reagents.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the QC
in Method 3600 and in the specific cleanup method.
8.2 Procedures to check the GC system operation are found in Method
8000.
8.2.1 Select a representative spike concentration for each compound
(acid or ester) to be measured. Using stock standards, prepare a quality
control check sample concentrate in acetone 1,000 times more concentrated
than the selected concentrations.
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8.2.2 Table 3 indicates single operator accuracy and precision for
this method. Compare the results obtained with the results given in
Table 3 to determine if the data quality is acceptable.
8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000).
8.3.1 If recovery is not within limits, the following procedures are
required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also,
check instrument performance.
• Recalculate the data and/or reanalyze the extract if
none of the above checks reveal a problem.
• Re-extract and re-analyze the sample if none of the
above are a problem or flag the data as "estimated
concentration".
8.4 GC/MS confirmation
8.4.1 GC/MS techniques should be judiciously employed to support
qualitative identifications made with this method. Refer to Method 8270
for the appropriate GC/MS operating conditions and analysis procedures.
8.4.2 When available, chemical ionization mass spectra may be
employed to aid the qualitative identification process.
8.4.3 Should these MS procedures fail to provide satisfactory
results, additional steps may be taken before reanalysis. These steps may
include the use of alternate packed or capillary GC columns or additional
cleanup.
9.0 METHOD PERFORMANCE
9.1 In a single laboratory, using organic-free reagent water and
effluents from publicly owned treatment works (POTW), the average recoveries
presented in Table 3 were obtained. The standard deviations of the percent
recoveries of these measurements are also included in Table 3.
10.0 REFERENCES
1. U.S. EPA, National Pollutant Discharge Elimination System, Appendix A,
Fed. Reg., 38, No. 75, Pt. II, Method for Chlorinated Phenoxy Acid
Herbicides in Industrial Effluents, Cincinnati, Ohio, 1971.
2. Goerlitz, D.G., and W.L. Lamar, "Determination of Phenoxy Acid Herbicides
in Water by Electron Capture and Microcoulometric Gas Chromatography,"
U.S. Geol. Survey Water Supply Paper, 1817-C, 1967.
8150B - 15 Revision 2
September 1994
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Burke, J.A., "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
U.S. EPA, "Extraction and Cleanup Procedure for the Determination of
Phenoxy Acid Herbicides in Sediment," EPA Toxicant and Analysis Center,
Bay St. Louis, Mississippi, 1972.
"Pesticide Methods Evaluation," Letter Report #33 for EPA Contract No. 68-
03-2697. Available from U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, Ohio 45268.
Eichelberger, J.W., L.E. Harris, and W.L. Budde, "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass
Spectrometry," Analytical Chemistry, 4_7, 995, 1975.
Glaser, J.A. et.al., "Trace Analysis for Wastewaters," Environmental
Science & Technology, 15, 1426, 1981.
Gurka, D.F, Shore, F.L., Pan, S-T, "Single Laboratory Validation of EPA
Method 8150 for Determination of Chlorinated Herbicides in Hazardous
Waste", JAOAC., 69, 970, 1986.
U.S. EPA, "Method 615. The Determination of Chlorinated Herbicides in
Industrial and Municipal Wastewater," Environmental Monitoring and Support
Laboratory, Cincinnati, Ohio, 45268, June 1982.
8150B - 16 Revision 2
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND DETECTION LIMITS
FOR CHLORINATED HERBICIDES
Compound
Retention time (min)"
Col.la Col.Ib Col.2 Col.3
"Column conditions are given in Sees. 4.1 and 7.5.
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION
LIMITS (EQL) FOR VARIOUS MATRICES3
Method
detection
limit (//g/L)
2,4-D
2,4-DB
2,4,5-T
2,4,5-TP (Silvex)
Dalapon
Dicamba
Dichloroprop
Dinoseb
MCPA
MCPP
2.0
4.1
3.4
2.7
-
1.2
-
-
-
-
-
-
-
-
-
4.8
11.2
4.1
3.4
1.6
-
2.4
2.0
5.0
1.0
-
-
-
-
1.2
0.91
0.20
0.17
5.8
0.27
0.65
0.07
249
192
Matrix
Factor
Ground water (based on one liter sample size)
Soil/sediment and other solids
Waste samples
10
200
100,000
aEQL = [Method detection limit (see Table 1)] X [Factor found in this table].
For non-aqueous samples, the factor is on a wet weight basis. Sample EQLs are
highly matrix dependent. The EQLs to be determined herein are provided for
guidance and may not always be achievable.
8150B - 17
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TABLE 3.
SINGLE OPERATOR ACCURACY AND PRECISION8
Compound
2,4-D
Dalapon
2,4-DB
Dicamba
Dichlorprop
Dinoseb
MCPA
MCPP
2,4,5-T
2,4,5-TP
Sample
Type
DW
MW
MW
DW
MW
MW
DW
MW
MW
DW
MW
MW
DW
MW
MW
MW
MW
DW
MW
MW
DW
MW
MW
DW
MW
MW
DW
MW
MW
Spike
10.9
10.1
200
23.4
23.4
468
10.3
10.4
208
1.2
1.1
22.2
10.7
10.7
213
0.5
102
2020
2020
21400
2080
2100
20440
1.1
1.3
25.5
1.0
1.3
25.0
Mean
Recovery
75
77
65
66
96
81
93
93
77
79
86
82
97
72
100
86
81
98
73
97
94
97
95
85
83
78
88
88
72
Standard
deviation
4
4
5
8
13
9
3
3
6
7
9
6
2
3
2
4
3
4
3
2
4
3
2
6
4
5
5
4
5
aAll results based upon seven replicate analyses. Esterification performed using
the bubbler method. Data obtained from reference 8.
DW = ASTM Type II
MW = Municipal water
8150B - 18
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FIGURE 1.
DIAZOMETHANE GENERATOR
glass tubing
nitrogen
rubber stopper
y
tube 1
tube 2
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FIGURE 2.
GAS CHROMATOGRAM OF CHLORINATED HERBICIDES
Column: 1.5% SP-22SO/1.9S% SP-2401 en Supateopon (100/120
Ttmptraturc: tethcrmal it 18S°C
Dtuctor: Electron Capture
0 12345
RETENTION TIME (MINUTES)
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FIGURE 3.
GAS CHROMATOGRAM OF CHLORINATED HERBICIDES
Column: 1J% SP-2250/1.95% SP-2401 on Supateoport (100/120
Program: 140°C for 6 Min, 10°C/Minutt to 200°C
Datactor: Electron Capture
I
468
RETENTION TIME (MINUTES)
10
12
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FIGURE 4.
GAS CHROMATOGRAM OF DALAPON, COLUMN 3
Column: 0.1% 9-1000 on 10/100 Mnh CarboMk C
fcortm: 100°C. 10«C/Min to 180°C
Ovttetor: Eloetron Capture
1 I I
0246
MITENTION TIME (MINUTES)
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METHOD 81BOB
CHLORINATED HERBICIDES BY GAS CHROMATOGRAPHY
7.2.1.1
Adjust sample
pH with HCI.
Solid
Sample
7.2.1.2 Extract
sample with
acetone and
diethyl ether.
7.2.1.3 Extract
twice more.
7.2.1.4
Combine
extracts.
7.2.1.5 Check
pH of extract,
adjust if
necessary.
Separate layara.
7.2.1.5
Re-extract
and discard
aqueous
phase.
7.1.1 Follow
Method 3580 for
extraction, using
diethyl ether,
acidified anhydrous
•odium sulfate and
acidified glass
wool.
7.2.2 Proceed
with
hydrolysis.
7.1.1.2 Use
1.0 ml of
sample for
hydrolysis.
7.2.3 Proceed
with eolvent
cleanup.
7.3.1.1 Adjust
sample pH
with H2S04.
7.3.1.2 Extract
with diethyl
ether.
7.3.1.3
Extract twice more
and combine
extracts.
7.3.2 Proceed
with
hydrolysis.
7.3.3 Proceed
with solvent
cleanup.
V
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METHOD 81SOB
(Continued)
7.4.3 Assemble
diczomethane
bubbler;
generate
diazomethane.
7.4
Choose
method for
esterification
7.4.2 Prepare
diazomethane
according to
kit
instructions.
7.5 Set
chromatographic
conditions.
7.6 Claibrate
according to
Method 8000.
7.6.2 Choose
appropriate
GC column.
7.7 Analyze
by GC (refer
to Method
8000).
7.7.7 Do
interferences
prevent peak
detection?
7.7.7 Process
series of
standards
through system
cleanup.
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METHOD 8151
CHLORINATED HERBICIDES BY GC USING METHYLATION OR PENTAFLUOROBENZYLATION
PERIVATIZATION; CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8151 is a capillary gas chromatographic (GC) method for
determining certain chlorinated acid herbicides and related compounds in aqueous,
soil and waste matrices. Specifically, Method 8151 may be used to determine the
following compounds:
Compound Name CAS No."
2,4-D 94-75-7
2,4-DB 94-82-6
2,4,5-TP (Silvex) 93-72-1
2,4,5-T 93-76-5
Dalapon 75-99-0
Dicamba 1918-00-9
Dichloroprop 120-36-5
Dinoseb 88-85-7
MCPA 94-74-6
MCPP 93-65-2
4-Nitrophenol 100-02-1
Pentachlorophenol 87-86-5
8 Chemical Abstract Services Registry Number.
Because these compounds are produced and used in various forms (i.e., acid,
salt, ester, etc.), Method 8151 describes a hydrolysis step that can be used to
convert herbicide esters into the acid form prior to analysis. Herbicide esters
generally have a half-life of less than one week in soil.
1.2 When Method 8151 is used to analyze unfamiliar samples, compound
identifications should be supported by at least one additional qualitative
technique. Sec. 8.4 provides gas chromatograph/mass spectrometer (GC/MS)
criteria appropriate for the qualitative confirmation of compound
identifications.
1.3 The estimated detection limits for each of the compounds in aqueous
and soil matrices are listed in Table 1. The detection limits for a specific
waste sample may differ from those listed, depending upon the nature of the
interferences and the sample matrix.
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1.4 The following compounds may also be determined using this method:
Compound Name CAS No."
Acifluorfen 50594-66-6
Bentazon 25057-89-0
Chloramben 133-90-4
DCPA diacid" 2136-79-0
3,5-Dichlorobenzoic acid 51-36-5
5-Hydroxydicamba 7600-50-2
Picloram 1918-02-1
8 Chemical Abstract Services Registry Number.
b DCPA monoacid and diacid metabolites included in method scope; DCPA
diacid metabolite used for validation studies. DCPA is a dimethyl
ester.
1.5 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatography and skilled in the
interpretation of gas chromatograms. Each analyst must demonstrate the ability
to generate acceptable results with this method.
1.6 Only experienced analysts should be allowed to work with diazomethane
due to the potential hazards associated with its use (explosive, carcinogenic).
2.0 SUMMARY OF METHOD
2.1 Method 8151 provides extraction, derivatization, and gas
chromatographic conditions for the analysis of chlorinated acid herbicides in
water, soil, and waste samples. An option for the hydrolysis of esters is also
described.
2.1.1 Water samples are extracted with diethyl ether and then
esterified with either diazomethane or pentafluorobenzyl bromide. The
derivatives are determined by gas chromatography with an electron capture
detector (GC/ECD). The results are reported as acid equivalents.
2.1.2 Soil and waste samples are extracted and esterified with
either diazomethane or pentafluorobenzyl bromide. The derivatives are
determined by gas chromatography with an electron capture detector
(GC/ECD). The results are reported as acid equivalents.
2.1.3 If herbicide esters are to be determined using this method,
hydrolysis conditions for the esters in water and soil extracts are
described.
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2.2 The sensitivity of Method 8151 depends on the level of interferences
in addition to instrumental limitations. Table 1 lists the GC/ECD and GC/MS
detection limits that can be obtained in aqueous and soil matrices in the absence
of interferences. Detection limits for a typical waste sample should be higher.
3.0 INTERFERENCES
3.1 Refer to Method 8000.
3.2 Method interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware that lead to discrete
artifacts or elevated baselines in gas chromatograms. All these materials must
be routinely demonstrated to be free from interferences under the conditions of
the analysis, by analyzing reagent blanks.
3.2.1 Glassware must be scrupulously cleaned. Clean each piece of
glassware as soon as possible after use by rinsing it with the last
solvent used in it. This should be followed by detergent washing with hot
water and rinses with tap water, then with organic-free reagent water.
Glassware should be solvent-rinsed with acetone and pesticide-quality
hexane. After rinsing and drying, glassware should be sealed and stored
in a clean environment to prevent any accumulation of dust or other
contaminants. Store glassware inverted or capped with aluminum foil.
Immediately prior to use, glassware should be rinsed with the next solvent
to be used.
3.2.2 The use of high purity reagents and solvents helps to minimize
interference problems. Purification of solvents by distillation in all-
glass systems may be required.
3.3 Matrix interferences may be caused by contaminants that are
coextracted from the sample. The extent of matrix interferences will vary
considerably from waste to waste, depending upon the nature and diversity of the
waste being sampled.
3.4 Organic acids, especially chlorinated acids, cause the most direct
interference with the determination by methylation. Phenols, including
chlorophenols, may also interfere with this procedure. The determination using
pentafluorobenzylation is more sensitive, and more prone to interferences from
the presence of organic acids or phenols than by methylation.
3,5 Alkaline hydrolysis and subsequent extraction of the basic solution
removes many chlorinated hydrocarbons and phthalate esters that might otherwise
interfere with the electron capture analysis. However, hydrolysis may result in
the loss of dinoseb and the formation of aldol condensation products if any
residual acetone remains from the extraction of solids.
3.6 The herbicides, being strong organic acids, react readily with
alkaline substances and may be lost during analysis. Therefore, glassware must
be acid-rinsed and then rinsed to constant pH with organic-free reagent water.
Sodium sulfate must be acidified.
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3.7 Sample extracts should be dry prior to methylation or else poor
recoveries will be obtained.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph
4.1.1 Gas chromatograph - Analytical system complete with gas
chromatograph suitable for Grob-type injection using capillary columns,
and all required accessories including detector, capillary analytical
columns, recorder, gases, and syringes. A data system for measuring peak
heights and/or peak areas is recommended.
4.1.2 Columns
4.1.2.1 Narrow Bore Columns
4.1.2.1.1 Primary Column 1 - 30 m x 0.25 mm, 5%
phenyl/95% methyl silicone (DB-5, J&W Scientific, or
equivalent), 0.25 yum film thickness.
4.1.2.1.2 Primary Column la (GC/MS) - 30 m x 0.32 mm,
5% phenyl/95% methyl silicone, (DB-5, J&W Scientific, or
equivalent), 1 |um film thickness.
4.1.2.1.3 Column 2 - 30 m x 0.25 mm DB-608 (J&W
Scientific or equivalent) with a 25 fj,m film thickness.
4.1.2.1.4 Confirmation Column - 30 m x 0.25 mm, 14%
cyanopropyl phenyl silicone, (DB-1701, J&W Scientific, or
equivalent), 0.25 jttm film thickness.
4.1.2.2 Wide-bore Columns
4.1.2.2.1 Primary Column - 30 m x 0.53 mm DB-608 (J&W
Scientific or equivalent) with 0.83 jum film thickness.
4.1.2.2.2 Confirmation Column - 30 m x 0.53 mm, 14%
cyanopropyl phenyl silicone, (DB-1701, J&W Scientific, or
equivalent), 1.0 urn film thickness.
4.1.3 Detector - Electron Capture Detector (ECD)
4.2 Kuderna-Danish (K-D) apparatus
4.2.1 Concentrator tube - 10 ml graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.2.2 Evaporation flask - 500 mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
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4.2.3 Snyder column - Three ball macro (Kontes K-503000-0121 or
equivalent).
4.2.4 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.2.5 Springs - 1/2 inch (Kontes K-662750 or equivalent).
4.3 Diazomethane Generator: Refer to Sec. 7.5 to determine which method
of diazomethane generation should be used for a particular generation.
4.3.1 Diazald Kit - Recommended for the generation of diazomethane
(Aldrich Chemical Co., Cat No. 210,025-0, or equivalent).
4.3.2 As an alternative, assemble from two 20 mm x 150 mm test
tubes, two Neoprene rubber stoppers, and a source of nitrogen. Use
Neoprene rubber stoppers with holes drilled in them to accommodate glass
delivery tubes. The exit tube must be drawn to a point to bubble
diazomethane through the sample extract. The generator assembly is shown
in Figure 1. The procedure for use of this type of generator is given in
Sec. 7.5.
4.4 Other Glassware
4.4.1 Beaker - 400 mL, thick walled.
4.4.2 Funnel - 75 mm diameter.
4.4.3 Separatory funnel - 500 ml, with Teflon stopcock.
4.4.4 Centrifuge bottle - 500 ml (Pyrex 1260 or equivalent).
4.4.5 Centrifuge bottle - 24/40 500 ml
4.4.6 Continuous Extractor (Hershberg-Wolfe type, Lab Glass No. LG-
6915, or equivalent)
4.4.7 Pipet - Pasteur, glass, disposable (140 mm x 5 mm ID).
4.4.8 Vials - 10 ml, glass, with Teflon lined screw-caps.
4.4.9 Volumetric flasks, Class A - 10 ml to 1000 mL.
4.5 Filter paper - 15 cm diameter (Whatman No. 1 or equivalent).
4.6 Glass Wool - Pyrex, acid washed.
4.7 Boiling chips - Solvent extracted with methylene
chloride,approximately 10/40 mesh (silicon carbide or equivalent).
4.8 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 2°C). The bath should be used in a hood.
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4.9 Balance - Analytical, capable of accurately weighing to 0.0001 g.
4.10 Centrifuge.
4.11 Ultrasonic preparation - A horn-type device equipped with a titanium
tip, or a device that will give equivalent performance, shall be used.
4.11.1 Ultrasonic Disrupter - The disrupter must have a minimum
power wattage of 300 watts, with pulsing capability. A device designed to
reduce the cavitation sound is recommended. Follow the manufacturers
instructions for preparing the disrupter for extraction of samples. Use
a 3/4" horn for most samples.
4.12 Sonabox - Recommended with above disrupters for decreasing cavitation
sound (Heat Systems - Ultrasonics, Inc., Model 432B or equivalent).
4.13 pH paper.
4.14 Silica gel cleanup column (Bond Elut™ - Analytichem, Harbor City, CA
or equivalent).
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free water, as defined in Chapter One.
5.3 Sodium hydroxide solution (0.1 N), NaOH. Dissolve 4 g NaOH in
organic-free reagent water and dilute to 1.0 L.
5.4 Potassium hydroxide solution (37% aqueous solution (w/v)), KOH.
Dissolve 37 g potassium hydroxide pellets in organic-free reagent water and
dilute to 100 ml.
5.5 Phosphate buffer pH = 2.5 (0.1 M). Dissolve 12 g sodium phosphate
(NaH2P04) in organic-free reagent water and dilute to 1.0 L. Add phosphoric acid
to adjust the pH to 2.5.
5.6 N-methyl-N-nitroso-p-toluenesulfonamide (Diazald). High purity,
available from Aldrich Chemical Co. or equivalent.
5.7 Silicic acid, H2Si05. 100 mesh powder, store at 130°C.
5.8 Potassium carbonate, K2C03.
5.9 2,3,4,5,6-Pentafluorobenzyl bromide (PFBBr), C6F5CH2Br. Pesticide
quality or equivalent.
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5.10 Sodium sulfate (granular, acidified, anhydrous), Na2S04. Purify by
heating at 400°C for 4 hours in a shallow tray, or by precleaning the sodium
sulfate with methylene chloride. If the sodium sulfate is precleaned with
methylene chloride, a method blank must be analyzed, demonstrating that there is
no interference from the sodium sulfate. Acidify by slurrying 100 g sodium
sulfate with enough diethyl ether to just cover the solid; then add 0.1 ml of
concentrated sulfuric acid and mix thoroughly. Remove the ether under vacuum.
Mix 1 g of the resulting solid with 5 ml of organic-free reagent water and
measure the pH of the mixture. It must be below a pH of 4. Store the remaining
solid at 130°C.
5.11 Solvents
5.11.1
equivalent.
5.11.2
5.11.3
5.11.4
5.11.5 Diethyl Ether, C2H5OC2H5. Pesticide quality or
equivalent. Must be free of peroxides as indicated by test strips (EM
Quant, or equivalent). Procedures for removal of peroxides are provided
with the test strips. After cleanup, 20 ml of ethyl alcohol preservative
must be added to each liter of ether.
Methylene chloride, CH2C12. Pesticide quality or
Acetone, CH3COCH3. Pesticide quality or equivalent.
Methanol, CH3OH. Pesticide quality or equivalent.
Toluene, C6H5CH3. Pesticide quality or equivalent.
Isooctane, (CH3)3CH2CH(CH3)2. Pesticide quality or
Hexane, C6H14. Pesticide quality or equivalent.
Ethanol, absolute. C2H5OH
Carbitol (diethylene glycol monoethyl ether),
5.11.6
equivalent.
5.11.7
5.11.8
5.11.9
C2H5OCH2CH2OCH2CH20 - optional for producing alcohol-free diazomethane.
5.12 Stock standard solutions (1000 mg/L) - Can be prepared from pure
standard materials or can be purchased as certified solutions.
5.12.1 Prepare stock standard solutions by accurately weighing
about 0.010 g of pure acid. Dissolve the material in pesticide quality
acetone and dilute to volume in a 10 ml volumetric flask. Stocks prepared
from pure methyl esters are dissolved in 10% acetone/isooctane (v/v).
Larger volumes may be used at the convenience of the analyst. If compound
purity is certified at 96% or greater, the weight may be used without
correction to calculate the concentration of the stock standard.
5.12.2 Transfer the stock standard solutions to vials with
Teflon lined screw-caps. Store at 4°C, protected from light. Stock
standard solutions should be checked frequently for signs of degradation
8151 - 7
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or evaporation, especially immediately prior to preparing calibration
standards from them.
5.12.3 Stock standard solutions of the derivatized acids must
be replaced after 1 year, or sooner, if comparison with check standards
indicates a problem. Stock standard solutions of the free acids degrade
more quickly and should be replaced after two months, or sooner if
comparison with check standards indicates a problem.
5.13 Internal Standard Spiking Solution (if internal standard calibration
is used) - To use this approach, the analyst must select one or more internal
standards that are similar in analytical behavior to the compounds of interest.
The analyst must further demonstrate that the measurement of the internal
standard is not affected by method or matrix interferences. The compound 4,4'-
dibromooctafluorobiphenyl (DBOB) has been shown to be an effective internal
standard, but other compounds, such as 1,4-dichlorobenzene, may be used if there
is a DBOB interference.
5.13.1 Prepare an internal standard spiking solution by
accurately weighing approximately 0.0025 g of pure DBOB. Dissolve the
DBOB in acetone and dilute to volume in a 10 ml volumetric flask.
Transfer the internal standard spiking solution to a vial with a Teflon
lined screw-cap, and store at room temperature. Addition of 10 /uL of the
internal standard spiking solution to 10 ml of sample extract results in
a final internal standard concentration of 0.25 /^g/L. The solution should
be replaced if there is a change in internal standard response greater
than 20 percent of the original response recorded.
5.14 Calibration standards - Calibration standards, at a minimum of five
concentrations for each parameter of interest, should be prepared through
dilution of the stock standards with diethyl ether or hexane. One of the
concentrations should be at a concentration near, but above, the method detection
limit. The remaining concentrations should correspond to the expected range of
concentrations found in real samples or should define the working range of the
GC. Calibration solutions must be replaced after six months, or sooner if
comparison with check standards indicates a problem.
5.14.1 Derivatize each calibration standard prepared from free
acids in a 10 ml K-D concentrator tube, according to the procedures
beginning at Sec. 7.5.
5.14.2 Add a known, constant amount of one or more internal
standards to each derivatized calibration standard, and dilute to volume
with the solvent indicated in the derivative option used.
5.15 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and determinative step, and the
effectiveness of the method in dealing with each sample matrix, by spiking each
sample, standard, and blank with one or two herbicide surrogates (e.g.,
herbicides that are not expected to be present in the sample) recommended to
encompass the range of the temperature program used in this method. Deuterated
analogs of analytes should not be used as surrogates in gas chromatographic
8151 - 8 Revision 0
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analysis due to coelution problems. The surrogate standard recommended for use
is 2,4-Dichlorophenylacetic acid (DCAA).
5.15.1 Prepare a surrogate standard spiking solution by
accurately weighing approximately 0.001 g of pure DCAA. Dissolve the DCAA
in acetone, and dilute to volume in a 10 ml volumetric flask. Transfer
the surrogate standard spiking solution to a vial with a Teflon lined
screw-cap, and store at room temperature. Addition of 50 /uL of the
surrogate standard spiking solution to 1 L of sample, prior to extraction,
results in a final concentration in the extract of 0.5 mg/L.
5.16 pH Adjustment Solutions
5.16.1 Sodium hydroxide, NaOH, 6 N.
5.16.2 Sulfuric acid, H2S04, 12 N.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1. 1 L samples should be collected.
6.2 Extracts must be stored under refrigeration (4°C).
7.0 PROCEDURE
7.1 Preparation of High Concentration Waste Samples
7.1.1 Extraction
7.1.1.1 Follow Method 3580, Waste Dilution, with the
following exceptions:
• use diethyl ether as the dilution solvent,
• use acidified anhydrous sodium sulfate, and acidified
glass wool,
• spike the sample with surrogate compound(s) according to
Sec. 5.16.1.
7.1.1.2 If the sample is to be analyzed for both herbicide
esters and acids, then the sample extract must be hydrolyzed. In
this case, transfer 1.0 mL (a smaller volume or a dilution may be
required if herbicide concentrations are large) to a 250 mL ground
glass Erlenmeyer flask. Proceed to Sec. 7.2.1.8. If the analysis
is for acid herbicides only, proceed to Sec. 7.4.5 for
derivatization by diazomethane (if PFB derivatization is selected,
reduce the volume of diethyl ether to 0.1 - 0.5 mL as per Sec. 7.4.2
and then dilute to 4 mL with acetone).
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7.2 Preparation of Soil, Sediment, and Other Solid Samples
7.2.1 Extraction
7.2.1.1 To a 400 ml, thick-wall beaker add 30 g (dry
weight as determined in Method 3540, Sec. 7.2.1) of the well-mixed
solid sample. Adjust the pH to 2 with concentrated hydrochloric
acid or acidify solids in each beaker with 85 ml of 0.1 M phosphate
buffer (pH = 2.5) and thoroughly mix the contents with a glass
stirring rod. Spike the sample with surrogate compound(s) according
to Sec. 5.16.1.
7.2.1.2 The ultrasonic extraction of solids must be
optimized for each type of sample. In order for the ultrasonic
extractor to efficiently extract solid samples, the sample must be
free flowing when the solvent is added. Acidified anhydrous sodium
sulfate should be added to clay type soils (normally 1:1), or any
other solid that is not a free flowing sandy mixture, until a free
flowing mixture is obtained.
7.2.1.3 Add 100 ml of methylene chloride/acetone (1:1
v/v) to the beaker. Perform ultrasonic extraction for 3 minutes,
with output control knob set at 10 (full power) and with mode switch
on Pulse (pulsing energy rather than continuous energy) and percent-
duty cycle knob set at 50% (energy on 50% of time and off 50% of
time). Allow the solids to settle. Transfer the organic layer into
a 500 ml centrifuge bottle.
7.2.1.4 Ultrasonically extract the sample twice more using
100 ml of methylene chloride and the same ultrasonic conditions.
7.2.1.5 Combine the three organic extracts from the sample
in the centrifuge bottle and centrifuge 10 minutes to settle the
fine particles. Filter the combined extract through filter paper
(Whatman #1, or equivalent) containing 7-10 g of acidified sodium
sulfate into a 500 ml 24/40 Erlenmeyer flask. Add 10 g of acidified
anhydrous sodium sulfate. Periodically, vigorously shake the
extract and drying agent and allow the drying agent to remain in
contact with the extract for a minimum of 2 hours. See NOTE in Sec.
7.3.1.6 that emphasizes the need for a dry extract prior to
esterification.
7.2.1.6 Quantitatively transfer the contents of the flask
to a 500-mL Kuderna-Danish flask with a 10-mL concentrator tube
attached. Add boiling chips and attach the macro Snyder column.
Evaporate the extract on the water bath to a volume of approximately
5 ml. Remove the flasks from the water bath and allow them to cool.
7.2.1.7 If hydrolysis or additional cleanup is not
required and the sample is dry, proceed to Sec. 7.4.4 - Nitrogen
Blowdown.
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7.2.1.8 Use this step only if herbicide esters in addition
to herbicide acids are to be determined:
7.2.1.8.1 Add 5 ml of 37% aqueous potassium hydroxide
and 30 ml of water to the extract. Add additional boiling
chips to the flask. Reflux the mixture on a water bath at
60-65°C until the hydrolysis step is completed (usually 1 to
2 hours). Remove the flasks from the water bath and cool to
room temperature. CAUTION - the presence of residual acetone
will result in the formation of aldol condensation products
which will cause GC interference.
7.2.1.8.2 Transfer the hydrolyzed aqueous solution to
a 500 ml separatory funnel and extract the solution three
times with 100 mL portions of methylene chloride. Discard the
methylene chloride phase. At this point the basic (aqueous)
solution contains the herbicide salts.
7.2.1.8.3 Adjust the pH of the solution to <2 with
cold (4°C) sulfuric acid (1:3) and extract once with 40 ml of
diethyl ether and twice with 20 ml portions of ether. Combine
the extracts and pour them through a pre-rinsed drying column
containing 7 to 10 cm of acidified anhydrous sodium sulfate.
Collect the dried extracts in a 500 ml Erlenmeyer flask (with
a 24/40 joint) containing 10 g of acidified anhydrous sodium
sulfate. Periodically, vigorously shake the extract and
drying agent and allow the drying agent to remain in contact
with the extract for a minimum of 2 hours. See NOTE in Sec.
7.3.1.6 that emphasizes the need for a dry extract prior to
esterification. Quantitatively transfer the contents of the
flask to a 500-mL Kuderna-Danish flask with a 10-mL
concentrator tube attached when the extract is known to be
dry.
7.2.1.8.4 Proceed to Sec. 7.4, Extract Concentration.
If additional cleanup is required, proceed to Sec. 7.2.1.9.
7.2.1.9 Use this step if additional cleanup of the non-
hydrolyzed herbicides is required:
7.2.1.9.1 Partition the herbicides by extracting the
methylene chloride from 7.2.1.7 (or diethyl ether from
7.2.1.8.4) with 3 x 15 ml portions of aqueous base prepared
by carefully mixing 30 ml of reagent water into 15 ml of 37%
aqueous potassium hydroxide. Discard the methylene chloride
or ether phase. At this point the basic (aqueous) solution
contains the herbicide salts.
7.2.1.9.2 Adjust the pH of the solution to <2 with
cold (4°C) sulfuric acid (1:3) and extract once with 40 mL of
diethyl ether and twice with 20 mL portions of ether. Combine
the extracts and pour them through a pre-rinsed drying column
containing 7 to 10 cm of acidified anhydrous sodium sulfate.
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Collect the dried extracts in a 500 ml Erlenmeyer flask (with
a 24/40 joint) containing 10 g of acidified anhydrous sodium
sulfate. Periodically, vigorously shake the extract and
drying agent and allow the drying agent to remain in contact
with the extract for a minimum of 2 hours. See NOTE in Sec.
7.3.1.6 that emphasizes the need for a dry extract prior to
esterification. Quantitatively transfer the contents of the
flask to a 500-mL Kuderna-Danish flask with a 10-mL
concentrator tube attached when the extract is known to be
dry.
7.2.1.9.3 Proceed to section 7.4 for extract
concentration.
7.2.1.10 An alternative wrist-shaker extraction procedure
can be found in Sec. 7.2 of Method 8150.
7.3 Preparation of Aqueous Samples
7.3.1 Separatory Funnel
7.3.1.1 Using a graduated cylinder, measure out a 1-L
sample and transfer it into a 2-L separatory funnel. Spike the
sample with surrogate compound(s) according to Sec. 5.15.1.
7.3.1.2 Add 250 g of NaCl to the sample, seal, and shake
to dissolve the salt.
7.3.1.3 Use this step only if herbicide esters in addition
to herbicide acids, are to be determined:
7.3.1.3.1 Add 17 ml of 6 N NaOH to the sample, seal,
and shake. Check the pH of the sample with pH paper; if the
sample does not have a pH greater than or equal to 12, adjust
the pH by adding more 6 N NaOH. Let the sample sit at room
temperature until the hydrolysis step is completed (usually 1
to 2 hours), shaking the separatory funnel and contents
periodically.
7.3.1.3.2 Add 60 ml of methylene chloride to the
sample bottle and rinse both the bottle and the graduated
cylinder. Transfer the methylene chloride to the separatory
funnel and extract the sample by vigorously shaking the funnel
for 2 minutes, with periodic venting to release excess
pressure. Allow the organic layer to separate from the water
phase for a minimum of 10 minutes. If the emulsion interface
between the layers is more than one-third the volume of the
solvent layer, the analyst must employ mechanical techniques
to complete the phase separation. The optimum technique
depends upon the sample, but may include stirring, filtration
through glass wool, centrifugation, or other physical methods.
Discard the methylene chloride phase.
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7.3.1.3.3 Add a second 60 ml volume of methylene
chloride to the separatory funnel and repeat the extraction
procedure a second time, discarding the methylene chloride
layer. Perform a third extraction in the same manner.
7.3.1.4 Add 17 ml of cold (4°C) 12 N sulfuric acid to the
sample (or hydrolyzed sample), seal, and shake to mix. Check the pH
of the sample with pH paper: if the sample does not have a pH less
than or equal to 2, adjust the pH by adding more acid.
7.3.1.5 Add 120 ml diethyl ether to the sample, seal, and
extract the sample by vigorously shaking the funnel for 2 min with
periodic venting to release excess pressure. Allow the organic
layer to separate from the water phase for a minimum of 10 min. If
the emulsion interface between layers is more than one third the
volume of the solvent layer, the analyst must employ mechanical
techniques to complete the phase separation. The optimum techniques
to complete the phase separation depends upon the sample, but may
include stirring, filtration through glass wool, centrifugation, or
other physical methods. Remove the aqueous phase to a 2 L
Erlenmeyer flask and collect the ether phase in a 500 mL Erlenmeyer
flask containing approximately 10 g of acidified anhydrous sodium
sulfate. Periodically, vigorously shake the extract and drying
agent.
7.3.1.6 Return the aqueous phase to the separatory funnel,
add 60 ml of diethyl ether to the sample, and repeat the extraction
procedure a second time, combining the extracts in the 500 ml
Erlenmeyer flask. Perform a third extraction with 60 ml diethyl
ether in the same manner. Allow the extract to remain in contact
with the sodium sulfate for approximately 2 hours.
NOTE: The drying step is very critical to ensuring
complete esterification. Any moisture remaining
in the ether will result in low herbicide
recoveries. The amount of sodium sulfate is
adequate if some free flowing crystals are
visible when swirling the flask. If all of the
sodium sulfate solidifies in a cake, add a few
additional grams of acidified sodium sulfate and
again test by swirling. The 2 hour drying time
is a minimum, however, the extracts may be held
in contact with the sodium sulfate overnight.
7.3.1.7 Pour the dried extract through a funnel plugged
with acid washed glass wool, and collect the extract in the K-D
concentrator. Use a glass rod to crush any caked sodium sulfate
during the transfer. Rinse the Erlenmeyer flask and funnel with 20
to 30 ml of diethyl ether to complete the quantitative transfer.
Proceed to Sec. 7.4 for extract concentration.
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7.4 Extract Concentration
7.4.1 Add one or two clean boiling chips to the flask and attach a
three ball Snyder column. Prewet the Snyder column by adding about 1 ml
of diethyl ether to the top of the column. Place the K-D apparatus on a
hot water bath (15-20°C above the boiling point of the solvent) so that the
concentrator tube is partially immersed in the hot water and the entire
lower rounded surface of the flask is bathed with hot vapor. Adjust the
vertical position of the apparatus and the water temperature, as required,
to complete the concentration in 10-20 minutes. At the proper rate of
distillation the balls of the column will actively chatter, but the
chambers will not flood. When the apparent volume of liquid reaches 1 ml,
remove the K-D apparatus from the water bath and allow it to drain and
cool for at least 10 minutes.
7.4.2 Remove the Snyder column and rinse the flask and its lower
joints into the concentrator tube with 1-2 mL of diethyl ether. The
extract may be further concentrated by using either the micro Snyder
column technique (Sec. 7.4.3) or nitrogen blowdown technique (Sec. 7.4.4).
7.4.3 Micro Snyder Column Technique
7.,4.3.1 Add another one or two clean boiling chips to the
concentrator tube and attach a two ball micro Snyder column. Prewet
the column by adding about 0.5 ml of diethyl ether to the top of the
column. Place the K-D apparatus in a hot water bath so that the
concentrator tube is partially immersed in the hot water. Adjust
the vertical position of the apparatus and the water temperature, as
required, to complete the concentration in 5-10 minutes. At the
proper rate of distillation the balls of the column will actively
chatter, but the chambers will not flood. When the apparent volume
of liquid reaches 0.5 ml, remove the K-D apparatus from the water
bath and allow it to drain and cool for at least 10 minutes. Remove
the Snyder column and rinse the flask and its lower joints with
about 0.2 ml of diethyl ether and add to the concentrator tube.
Proceed to Sec. 7.4.5.
7.4.4 Nitrogen Blowdown Technique
7.4.4.1 Place the concentrator tube in a warm water bath
(approximately 35°C) and evaporate the solvent volume to the
required level using a gentle stream of clean, dry nitrogen
(filtered through a column of activated carbon).
CAUTION: Do not use plasticized tubing between the carbon
trap and the sample.
7.4.4.2 The internal wall of the tube must be rinsed down
several times with diethyl ether during the operation. During
evaporation, the solvent level in the tube must be positioned to
prevent water from condensing into the sample (i.e., the solvent
level should be below the level of the water bath). Under normal
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operating conditions, the extract should not be allowed to become
dry. Proceed to Sec. 7.4.5.
7.4.5 Dilute the extract with 1 mL of isooctane and 0.5 ml of
methanol. Dilute to a final volume of 4 mL with diethyl ether. The
sample is now ready for methylation with diazomethane. If PFB derivation
is being performed, dilute to 4 ml with acetone.
7.5 Esterification - For diazomethane derivatization proceed with Sec.
7.5.1. For PFB derivatization proceed with Sec. 7.5.2.
7.5.1 Diazomethane Derivatization - Two methods may be used for the
generation of diazomethane: the bubbler method (see Figure 1), Sec.
7.5.1.1, and the Diazald kit method, Sec. 7.5.1.2.
CAUTION: Diazomethane is a carcinogen
certain conditions.
and can explode under
The bubbler method is suggested when small batches of samples
(10-15) require esterification. The bubbler method works well with
samples that have low concentrations of herbicides (e.g., aqueous samples)
and is safer to use than the Diazald kit procedure. The Diazald kit
method is good for large quantities of samples needing esterification.
The Diazald kit method is more effective than the bubbler method for soils
or samples that may contain high concentrations of herbicides (e.g.,
samples such as soils that may result in yellow extracts following
hydrolysis may be difficult to handle by the bubbler method). The
diazomethane derivatization (U.S.EPA, 1971) procedures, described below,
will react efficiently with all of the chlorinated herbicides described in
this method and should be used only by experienced analysts, due to the
potential hazards associated with its use. The following precautions
should be taken:
Use a safety screen.
Use mechanical pipetting aides.
Do not heat above 90°C - EXPLOSION may result.
Avoid grinding surfaces, ground-glass joints, sleeve bearings,
and glass stirrers - EXPLOSION may result.
Store away from alkali metals - EXPLOSION may result.
Solutions of diazomethane decompose rapidly in the presence of
solid materials such as copper powder, calcium chloride, and
boiling chips.
7.5.1.1
(see Figure 1).
Bubbler method - Assemble the diazomethane bubbler
7.5.1.1.1 Add 5 mL of diethyl ether to the first test
tube. Add 1 mL of diethyl ether, 1 mL of carbitol, 1.5 mL of
37% KOH, and 0.1-0.2 g of Diazald to the second test tube.
Immediately place the exit tube into the concentrator tube
containing the sample extract. Apply nitrogen flow
(10 mL/min) to bubble diazomethane through the extract for
10 minutes or until the yellow color of diazomethane persists.
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The amount of Diazald used is sufficient for esterification of
approximately three sample extracts. An additional 0.1-0.2 g
of Diazald may be added (after the initial Oiazald is
consumed) to extend the generation of the diazomethane. There
is sufficient KOH present in the original solution to perform
a maximum of approximately 20 minutes of total esterification.
7.5.1.1.2 Remove the concentrator tube and seal it
with a Neoprene or Teflon stopper. Store at room temperature
in a hood for 20 minutes.
7.5.1.1.3 Destroy any unreacted di azomethane by adding
0.1-0.2 g of silicic acid to the concentrator tube. Allow to
stand until the evolution of nitrogen gas has stopped. Adjust
the sample volume to 10.0 ml with hexane. Stopper the
concentrator tube or transfer 1 ml of sample to a GC vial, and
store refrigerated if further processing will not be performed
immediately. Analyze by gas chromatography.
7.5.1.1.4 Extracts should be stored at 4°C away from
light. Preservation study results indicate that most analytes
are stable for 28 days; however, it is recommended that the
methylated extracts be analyzed immediately to minimize the
trans-esterification and other potential reactions that may
occur.
7.5.1.2 Diazald kit method - Instructions for preparing
diazomethane are provided with the generator kit.
7.5.1.2.1 Add 2 ml of diazomethane solution and let
the sample stand for 10 minutes with occasional swirling. The
yellow color of diazomethane should be evident and should
persist for this period.
7.5.1.2.2 Rinse the inside wall of the ampule with 700
/xL of diethyl ether. Reduce the sample volume to
approximately 2 ml to remove excess diazomethane by allowing
the solvent to evaporate spontaneously at room temperature.
Alternatively, 10 mg of silicic acid can be added to destroy
the excess diazomethane.
7.5.1.2.3 Dilute the sample to 10.0 ml with hexane.
Analyze by gas chromatography. It is recommended that the
methylated extracts be analyzed immediately to minimize the
trans-esterification and other potential reactions that may
occur.
7.5.2 PFB Method
7.5.2.1 Add 30 juL of 10% K2C03 and 200 fj.1 of 3% PFBBr in
acetone. Close the tube with a glass stopper and mix on a vortex
mixer. Heat the tube at 60°C for 3 hours.
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7.5.2.2 Evaporate the solution to 0.5 ml with a gentle
stream of nitrogen. Add 2 ml of hexane and repeat evaporation just
to dryness at ambient temperature.
7.5,2.3 Redissolve the residue in 2 ml of toluene:hexane
(1:6) for column cleanup.
7.5.2.4 Top a silica column (Bond Elut™ or equivalent)
with 0.5 cm of anhydrous sodium sulfate. Prewet the column with 5
ml hexane and let the solvent drain to the top of the adsorbent.
Quantitatively transfer the reaction residue to the column with
several rinsings of the toluene:hexane solution (total 2 - 3 ml).
7.5.2.5 Elute the column with sufficient toluene:hexane to
collect 8 ml of eluent. Discard this fraction, which contains
excess reagent.
7.5.2.6 Elute the column with toluene:hexane (9:1) to
collect 8 ml of eluent containing PFB derivatives in a 10 ml
volumetric flask. Dilute to 10 ml with hexane. Analyze by GC/ECD.
7.6 Gas chromatographic conditions (recommended):
7.6.1 Narrow Bore
7.6.1.1 Primary Column 1:
Temperature program: 60°C to 300°C, at 4°C/mir\
Helium carrier flow: 30 cm/sec
Injection volume: 2 ^L, splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
7.6.1.2 Primary Column la:
Temperature program: 60°C to 300°C, at 4°C/min
Helium carrier flow: 30 cm/sec
Injection volume: 2 juL, splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
7.6.1.3 Column 2:
Temperature program: 60°C to 300°C, at 4°C/min
Helium carrier flow: 30 cm/sec
Injection volume: 2 juL, splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
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7.6.1.4 Confirmation Column:
Temperature program: 60°C to 300°C, at 4°C/min
Helium carrier flow: 30 cm/sec
Injection volume: 2 juL, splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
7.6.2 Wide-bore
7.6.2.1 Primary Column:
Temperature program: 0.5 minute at 150°C, 150°C to 270°C at
5°C/min
Helium carrier flow: 7 mL/min
Injection volume: 1 IJ.L
7.6.2.2 Confirmatory Column:
Temperature program: 0.5 minute at 150°C, 150°C to 270°C at
5°C/min
Helium carrier flow: 7 mL/min
Injection volume: 1 /LtL
7.7 Calibration
7.7.1 The procedure for internal or external calibration may be
used. Refer to Method 8000 for a description of each of these procedures.
Use Table 1 for guidance on selecting the lowest point on the calibration
curve.
7.8 Gas chromatographic analysis
7.8.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 /LtL of internal standard to the sample prior to
injection.
7.8.2 Follow Method 8000 for instructions on the analysis sequence,
appropriate dilutions, establishing daily retention time windows, and
identification criteria. Include a mid-concentration standard after each
group of 10 samples in the analysis sequence.
7.8.3 An example of a chromatogram for a methylated chlorophenoxy
herbicide is shown in Figure 2. Tables 2 and 3 present retention times
for the target analytes after esterification, using the diazomethane
derivatization procedure and the PFB derivatization procedure,
respectively.
7.8.4 Record the sample volume injected and the resulting peak sizes
(in area units or peak heights).
7.8.5 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
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in the sample chromatogram which corresponds to the compounds used for
calibration purposes.
7.8.6 If calibration standards have been analyzed in the same manner
as the samples (e.g. have undergone hydrolysis and esterification), then
the calculation of concentration given in Method 8000 should be used.
However, if calibration is performed using standards made from methyl
ester compounds (compounds not esterified by application of this method),
then the calculation of concentration must include a correction for the
molecular weight of the methyl ester versus the acid herbicide.
7.8.7 If peak detection and identification are prevented due to
interferences, further cleanup is required. Before using any cleanup
procedure, the analyst must process a series of standards through the
procedure to validate elution patterns and the absence of interferences
from reagents.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the QC
in Method 3600 and in the specific cleanup method.
8.2 Procedures to check the GC system operation are found in Method 8000.
8.2.1 Select a representative spike concentration for each compound
(acid or ester) to be measured. Using stock standards, prepare a quality
control check sample concentrate, in acetone, that is 1000 times more
concentrated than the selected concentrations. Use this quality control
check sample concentrate to prepare quality control check samples.
8.2.2 Tables 4 and 5 present bias and precision data for water and
clay matrices, using the diazomethane derivatization procedure. Table 6
presents relative recovery data generated using the PFB derivatization
procedure and water samples. Compare the results obtained with the results
given in these Tables to determine if the data quality is acceptable.
8.3 Calculate surrogate standard recovery on all standards, samples,
blanks, and spikes. Determine if the recovery is within limits (limits
established by performing QC procedures outlined in Method 8000).
8.3.1 If recovery is not within limits, the following procedures are
required:
8.3.1.1 Check to be sure there are no errors in
calculations, surrogate solutions and internal standards. Also,
check instrument performance.
8.3.1.2 Recalculate the data and/or reanalyze the extract
if any of the above checks reveal a problem.
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8.3.1.3 Reextract and reanalyze the sample if none of the
above are a problem or flag the data as "estimated concentration."
8.4 GC/MS confirmation
8.4.1 GC/MS techniques should be judiciously employed to support
qualitative identifications made with this method. Refer to Method 8270
for the appropriate GC/MS operating conditions and analysis procedures.
8.4.2 When available, chemical ionization mass spectra may be
employed to aid the qualitative identification process.
8.4.3 Should these MS procedures fail to provide satisfactory
results, additional steps may be taken before reanalysis. These steps may
include the use of alternate packed or capillary GC columns or additional
cleanup.
9.0 METHOD PERFORMANCE
9.1 In single laboratory studies using organic-free reagent water and
clay/still bottom samples, the mean recoveries presented in Tables 4 and 5 were
obtained for diazomethane derivatization. The standard deviations of the percent
recoveries of these measurements are also in Tables 4 and 5.
9.2 Table 6 presents relative recoveries of the target analytes obtained
using the PFB derivatization procedure with spiked water samples.
10.0 REFERENCES
1. Fed. Reg. 1971, 38, No. 75, Pt. II.
2. Goerlitz, D. G.; Lamar, W.L., "Determination of Phenoxy Acid Herbicides in
Water by Electron Capture and Microcoulometric Gas Chromatography,". U.S.
Geol. Survey Water Supply Paper 1967, 1817-C.
3. Burke, J. A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects, J. Assoc. Off Anal. Chem. 1965, 48, 1037.
4. "Extraction and Cleanup Procedures for the Determination of Phenoxy Acid
Herbicides in Sediment"; U.S. Environmental Protection Agency. EPA
Toxicant and Analysis Center: Bay St. Louis, MS, 1972.
5. Shore, F.L.; Amick, E.N.; Pan, S. T. "Single Laboratory Validation of EPA
Method 8151 for the Analysis of Chlorinated Herbicides in Hazardous
Waste"; U.S. Environmental Protection Agency. Environmental Monitoring
Systems Laboratory. Office of Research and Development, Las Vegas, NV,
1985; EPA-60014-85-060.
6. Method 515.1, "Determination of Chlorinated Acids in Water by Gas
Chromatography with an Electron Capture Detector", Revision 4.0, USEPA,
Office of Research and Development, Environmental Monitoring Systems
Laboratory, Cincinnati, Ohio.
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7. Method 1618, "Organo-halide and Organo-phosphorus Pesticides and Phenoxy-
acid Herbicides by Wide Bore Capillary Column Gas Chromatography with
Selective Detectors", Revision A, July 1989, USEPA, Office of Water
Regulations and Standards, Washington, DC.
8. Gurka, D.F, Shore, F.L., Pan, S-T, "Single Laboratory Validation of EPA
Method 8150 for Determination of Chlorinated Herbicides in Hazardous
Waste", JAOAC, 69, 970, 1986.
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Figure 1
DIAZOMETHANE GENERATOR
nitrogen
gloss tubing
rubber stopper
tube I
tube 2
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Figure 2
CHROMATOGRAM OF METHYL ESTERS OF CHLOROPHENOXYACIDS
100.0-
J
1O46
C
813 E
866
RIC-
A
271
Vj
393
200
3:20
L
L 3" LI 479 64
U**^JLJ i , I
• I I " '
693
400
6:40
I
600
10:OO
G
964
H
I
8OO
13:20
10OO
16:40
Scan Tim*
A - Oalapon. methyl ester
B - Oicamba, methyl ester
C : MCPP, methyl estar
D - MCPA. methyl ester
E - Oichlorprop. methyl ester
F = 2.4. D methyl ester
G - Silvex. meihyl ester
H - 2.4.6 T. methyl ester
I = 2.4 DB. methyl ester
J Dinoseb. methyl ether
12OO
2C:OO
8151 - 23
Revision 0
September 1994
-------
TABLE 1
ESTIMATED METHOD DETECTION LIMITS FOR METHOD 8151,
DIAZOMETHANE DERIVATIZATION
Aqueous Samples
Analyte
Acifluorfen
Bentazon
Chloramben
2,4-D
Dalapon
2,4-DB
DCPA diacid"
Dicamba
3,5-Dichlorobenzoic acid
Dichloroprop
Dinoseb
5-Hydroxydicamba
MCPP
MCPA
4-Nitrophenol
Pentachlorophenol
Picloram
2,4,5-T
2,4,5-TP
GC/ECD
Estimated
Detection
Limit8
(M9/L)
0.096
0.2
0.093
0.2
1.3
0.8
0.02
0.081
0.061
0.26
0.19
0.04
0.09d
0.056d
0.13
0.076
0.14
0.08
0.075
Soil Samples
GC/ECD
Estimated
Detection
Limit"
(M9/kg)
4.0
0.11
0.12
0.38
66
43
0.34
0.16
0.28
GC/MS
Estimated
Identification
Limit0
(ng)
1.7
1.25
0.5
0.65
0.43
0.3
0.44
1.3
4.5
a EDL = estimated detection limit; defined as either the MDL (40 CFR Part 136,
Appendix B, Revision 1.11 ), or a concentration of analyte in a sample
yielding a peak in the final extract with signal-to-noise ratio of
approximately 5, whichever value is higher.
b Detection limits determined from standard solutions corrected back to 50 g
samples, extracted and concentrated to 10 mL, with 5 pi injected.
Chromatography using narrow bore capillary column, 0.25 fj.m film,
5% phenyl/95% methyl silicone.
c The minimum amount of analyte to give a Finnigan INCOS FIT value of 800 as
the methyl derivative vs. the spectrum obtained from 50 ng of the respective
free acid herbicide.
d 40 CFR Part 136, Appendix B (49 FR 43234). Chromatography using wide-bore
capillary column.
e DCPA monoacid and diacid metabolites included in method scope; DCPA diacid
metabolite used for validation studies. DCPA is a dimethyl ester.
8151 - 24
Revision 0
September 1994
-------
TABLE 2
RETENTION TIMES (MINUTES) OF METHYL DERIVATIVES OF CHLORINATED HERBICIDES
Megabore Columns
Analyte
Narrow Bore Columns
Primary8 Confirmation8 Primary"
Column Column Column
Wide-bore Columns
Confirmation"
Column
Dalapon
3,5-Dichlorobenzoic acid
4-Nitrophenol
DCAA (surrogate)
Dicamba
Dichloroprop
2,4-D
DBOB (internal std.)
Pentachlorophenol
Chloramben
2,4,5-TP
5-Hydroxydicamba
2,4,5-T
2,4-DB
Dinoseb
Bentazon
Picloram
DCPA diacidc
Acifluorfen
MCPP
MCPA
3.4
18.6
18.6
22.0
22.1
25.0
25.5
27.5
28.3
29.7
29.7
30.0
30.5
32.2
32.4
33.3
34.4
35.8
41.5
4.7
17.7
20.5
14.9
22.6
25.6
27.0
27.6
27.0
32.8
29.5
30.7
30.9
32.2
34.1
34.6
37.5
37.8
42.8
4.39
5.15
5.85
6.97
7.92
8.74
4.24
4.74
4.39
5.46
6.05
7.37
8.20
9.02
4.55
4.94
Primary Column:
Confirmation Column:
Temperature program:
Helium carrier flow:
Injection volume:
Injector temperature:
Detector temperature:
Primary Column:
Confirmatory Column:
Temperature program:
Helium carrier flow:
Injection volume:
5% phenyl/95% methyl silicone
14% cyanopropyl phenyl silicone
60°C to 300°C, at 4°C/min
30 cm/sec
2 nl, splitless, 45 sec delay
250°C
320°C
DB-608
14% cyanopropyl phenyl silicone
0.5 minute at 150°C,
150°C to 270°C, at 5°C/min
7 mL/min
1 ^L
DCPA monoacid and diacid metabolites included in method scope; DCPA diacid
metabolite used for validation studies. DCPA is a dimethyl ester.
8151 - 25
Revision 0
September 1994
-------
TABLE 3
RETENTION TIMES (MINUTES) OF PFB DERIVATIVES OF CHLORINATED HERBICIDES
Herbicide
Gas Chromatographic Column
Thin-film DB-5a
SP-2250*
Thick-film DB-5°
Dalapon
MCPP
Dicamba
MCPA
Dichloroprop
2,4-D
Silvex
2,4,5-T
Dinoseb
2,4-DB
10.41
18.22
18.73
18.88
19.10
19.84
21.00
22.03
22.11
23.85
12.94
22.30
23.57
23.95
24.10
26.33
27.90
31.45
28.93
35.61
13.54
22.98
23.94
24.18
24.70
26.20
29.02
31.36
31.57
35.97
a DB-5 capillary column, 0.25 urn film thickness, 0.25 mm ID x 30 m long.
Column temperature, programmed: 70°C for 1 minute, program 10°C/min. to
240°C, hold for 17 minutes.
b SP-2550 capillary column, 0.25 urn film thickness, 0.25 mm ID x 30 m long.
Column temperature, programmed: 70°C for 1 minute, program 10°C/min. to
240°C, hold for 10 minutes.
c DB-5 capillary column, 1.0 urn film thickness, 0.32 mm ID x 30 m long.
Column temperature, programmed: 70°C for 1 minute, program 10°C/min. to
240°C, hold for 10 minutes.
8151 - 26
Revision 0
September 1994
-------
TABLE 4
ACCURACY AND PRECISION FOR METHOD 8151
DIAZOMETHANE DERIVATIZATION, ORGANIC-FREE REAGENT WATER MATRIX
Analyte
Acifluorfen
Bentazon
Chloramben
2,4-D
Dalapon
2,4-DB
DCPA diacidb
Dicamba
3,5-Dichlorobenzoic acid
Dichloroprop
Dinoseb
5-Hydroxydicamba
4-Nitrophenol
Pentachlorophenol
Picloram
2,4,5-TP
2,4,5-T
Spike
Concentration
(M9/L)
0.2
1
0.4
1
10
4
0.2
0.4
0.6
2
0.4
0.2
1
0.04
0.6
0.4
0.2
Mean8 Standard
Percent Deviation of
Recovery Percent Recovery
121
120
111
131
100
87
74
135
102
107
42
103
131
130
91
117
134
15.7
16.8
14.4
27.5
20.0
13.1
9.7
32.4
16.3
20.3
14.3
16.5
23.6
31.2
15.5
16.4
30.8
Mean percent recovery calculated from 7-8 determinations of spiked
organic-free reagent water.
DCPA monoacid and diacid metabolites included in method scope; DCPA
diacid metabolite used for validation studies. DCPA is a dimethyl ester.
8151 - 27
Revision 0
September 1994
-------
TABLE 5
ACCURACY AND PRECISION FOR METHOD 8151
DIAZOMETHANE DERIVATIZATION, CLAY MATRIX
Analyte
Mean
Percent Recovery8
Linear
Concentration
Rangeb
(ng/g)
Percent
Relative
Standard Deviation0
(n=20)
Dicamba
MCPP
MCPA
Dichloroprop
2,4-D
2,4,5-TP
2,4,5-T
2,4-DB
Dinoseb
95.7
98.3
96.9
97.3
84.3
94.5
83.1
90.7
93.7
0.52
620
620
1.5
1.2
0.42
0.42
4.0
0.82
- 104
- 61,800
- 61,200
- 3,000
- 2,440
- 828
- 828
- 8,060
- 1,620
7.5
3.4
5.3
5.0
5.3
5.7
7.3
7.6
8.7
Mean percent recovery calculated from 10 determinations of spiked clay
and clay/still bottom samples over the linear concentration range.
Linear concentration range was determined using standard solutions and
corrected to 50 g solid samples.
Percent relative standard deviation was calculated using standard
solutions, 10 samples high in the linear concentration range, and 10
samples low in the range.
8151 - 28
Revision 0
September 1994
-------
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METHOD 8151
CHLORINATED HERBICIDES BY GC USING METHYLATION OR PENTAFLUOROBENZYLATION
DERIVATIZATION: CAPILLARY COLUMN TECHNIQUE
Extraction/Hydrolysis of Waste and Soil Samples
NO
1
Concentrate and/or
dilute based on
whether denvatjzatton
is by diazomethane
orPFB
70 Does
sample con
tain a high
cone of
waste''
7.2.1.8.1 AddKOHand
water. Redux for 2 hrs.
Allow to cool.
7.2.1.8.2 Transfer the
hydrolyzed solution to a
sep funnel and extract 3
times with MeCI.
Discard extracts.
72.1.8.3 Acidify and
extract 3 times with
diethyl e»wr. Combine
and dry the extracts 2 hrs.
7.2.1.8.4 Proceed to
Section 7.4 to concentrate
extract
72.1.9.1 Extract 3 limes
withKOH. Discard the
MeCI.
72.192 Acidify and
extract 3 times with
diethyl ether. Combine
and dry the extracts 2 hrs.
72.1 1 Weigh sample
and add to beaker;
add acid and spike;
mix well.
7.2.1.2 Optimize
ultrasonic solid extrac-
tion for each matrix
72.1.3 Add MeCI/
acetone to sample 4
extracts min.;let
settle 4 decant extract
72.1445 Ultra-
sonically extract sample
2 more times with MeCI
72.1.5 Combine organK
extracts, centrifuge, and
filter extract. Dry for
2 hrs
72.1.6 Concentrate
extract to about 5 mL
with Snyder column.
YES
If hydrolysis is not
required, proceed to Section
74.4, Nitrogen Slowdown
72.1.7
Does analysis
ndude herbicide
esters?
8151 - 30
Revision 0
September 1994
-------
METHOD 8151
(continued)
Extraction/Hydrolysis of Aqueous Samples and Extract Concentration
73.1.1 Measure 1 Lot
sample and transfer to
a 2L sep. funnel.
7.3.1.2 AddZSOgNaCI
to sample and shake
to dissolve
7.3.1 3
Does analysis
include heoKXJe esters''
7.3.1.4 Add12Nsulfunc
acid and shake. Add
until pH < 2
7.3.1.5 Adddiethyl
ether to sample and
extract Save both
phases
731.3.1 AddGNNaOHto
sample and shake. Add
until pH> 12. Let stand
1 hr.
73.13.2 AddMeCland
extract by shaking for
2min. Discard MeCI.
73.1.6 Return aqueous phase
to separatory funnel and repeat
extraction 2 more times, combine
extracts, and allow extract to
remain in contact with sodium
sulfate for 2 Mrs.
Does
difficult
emulsion form
>1/3 solvent
volume?
Employ mechanical techniques
to complete phase separation
(e.g. stirring, filtration through
glass wool, centrifugatfon, or
other physical methods).
Discard MeCI
73.1.3.3 Repeat
extraction twice more.
Discard MeCI.
Employ mechanical techniques
to complete phase separation
(e.g. stirring, filtration through
glass wool, centnfugation, or
other physical methods).
Save both phases.
73.1.7 Pour extract
through glass wool and
proceed to Section 7 4 1
7.4.1 Place K-D apparatus
in water bath, concentrate
and cool
742- 7.4.4 Complete
concentration with rmcro-
Snyder column or nitrogen
blow down.
74.5 Dilute extract
with 1 ml isooctane and
0.5 ml methanol
8151 - 31
Revision 0
September 1994
-------
METHOD 8151
(continued)
Extract Derivatization
7.4.5 Dilute extract
to 4 mL with acetone
i
7.5.2.1 Add potassium
carbonate and PFBBr
Close tube, mix & heat
1
752.2 Evaporate with
nitrogen to 0.5 mL. Add
2 mL hexane and repeat
1
752.3 Redissorve the
residue in 2 mL toluene:
hexane (1 6)
1
7.52.4 Load sodium
sulfate / silica cleanup
column with residue.
1 1
7.5.2.5 Bute column
with enough toluene :
hexane to collect 8 mL
eluant
/7.4 5 Witt\
PFB / PFBordlazo- \. Diazo-
* vsmetwnedenva->me*ane
\zattonbe /
x^x
7.5.1.1 Asa
diazomethar
(Figur
i
7.5.1 1.1 Add 5(1
tube. Addl mLd
1 mL carbtot. 1 5
and 0.1 - 0.2 g Oil
2nd tube. Bubble
for 10 min or until
i
BmWethe Bubbler
Mbubbler «— — <
91)
»
nL to 1st test
iethyl ether,
mL ot 37% KOH
izaldtolhe
with nitrogen
yellow persists
7.5.1.1 2 Remove con-
centrator tube and seal
it. Store at room temp.
i
7.5.1.1.3 Addsilicii
concentrator tube a
until nitrogen evolul
stopped. Adjust sai
tolOmLwithhexa
Immediate analysis
(
caodto
nd let stand
km has
mple volume
•w Stopper.
is recommended
i
75^6 Discard 1st fraction
and continue elution with
enough toluene . hexane (1 : i
to collect 8 mL more eluant
Transfer to a 1 0 mL volumetrk
flask and dilute to the mark
with hexane
75.1 1 5 If necessary
store at 4 C in the dark
n for a max of 28 days
-
•
^ 7fi1 A 769 Satftr _^
conditions
7.4.5 Dilute extract
to 4 mL with diethyl
ether
/7.5.1\.
/ WHI the \ DiazaW
S Bubbler or *• Nj^L.
v DiazaW Kit /""""
\me«iodbe/
N-used?/
\ !
7.5.1.2 Read
instructions
\ i
7.5.1.2.1 Add2n
diazomethane sol
Let stand for 10 rr
and swirl
i
kit
nL
utton
tin
75.1.2.2 Rinse ampule with
diethyl ether and evaporate
to 2 mL to remove diazomethane
Alternatively, silicic acid
may be added.
i
75 1.2.3 Dilutes:
to 10mL with hex;
imple
me
8151 - 32
Revision 0
September 1994
-------
METHOD 8151
(continued)
Analysis by Gas Chromatography
7.7 Internal or external
calibration may be uMd
(See method 8000).
7.8.1 Add 10 uL internal
standard to the sample
prior to injection.
7.8.2 See method 8000 for
analysis sequence, appropriate
dilutions, establishing daily
retention time windows, and
identification en tana. Check
stds every 10 samples.
7.8.4 Record volume
infected and the resulting
peak sizes.
7.8.5 Determine the
identity and quantify
it pi
78.6
Have stds
and samples
been prepared and
analyzed (he
way?
Calculate the correction
for molecular weight of
methyl ester vs herbicide
78.6 Calculate con-
centration using procedure
in Method 8000.
7.8.7 Perform further
cleanup if necessary
1
8151 - 33
Revision 0
September 1994
-------
METHOD 8240B
VOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (6C/MS)
1.0 SCOPE AND APPLICATION
1.1 Method 8240 is used to determine volatile organic compounds in a
variety of solid waste matrices. This method is applicable to nearly all types
of samples, regardless of water content, including ground water, aqueous sludges,
caustic liquors, acid liquors, waste solvents, oily wastes, mousses, tars,
fibrous wastes, polymeric emulsions, filter cakes, spent carbons, spent
catalysts, soils, and sediments. The following compounds can be determined by
this method:
Appropriate Technique
Analyte
Acetone
Acetonitrile
Acrolein (Propenal)
Acrylonitrile
Allyl alcohol
Ally! chloride
Benzene
Benzyl chloride
Bromoacetone
Bromochloromethane (I.S.)
Bromodichloromethane
4-Bromofluorobenzene (surr.)
Bromoform
Bromomethane
2-Butanone (MEK)
Carbon disulfide
Carbon tetrachloride
Chloral hydrate
Chlorobenzene
Chlorobenzene-d5 (I.S.)
Chi orodi bromomethane
Chloroethane
2-Chloroethanol
bis-(2-Chloroethyl) sulfide
2-Chloroethyl vinyl ether
Chloroform
Chloromethane
Chloroprene
3-Chloropropionitrile
l,2-Dibromo-3-chloropropane
1,2-Dibromoethane
CAS No.b
67-64-1
75-05-8
107-02-8
107-13-1
107-18-6
107-05-1
71-43-2
100-44-7
598-31-2
74-97-5
75-27-4
460-00-4
75-25-2
74-83-9
78-93-3
75-15-0
56-23-5
302-17-0
108-90-7
3114-55-4
, 124-48-1
75-00-3
107-07-3
505-60-2
110-75-8
67-66-3
74-87-3
126-99-8
542-76-7
96-12-8
106-93-4
Purge-and-Trap
PP
PP
PP
PP
PP
a
a
PP
PP
a
a
a
a
a
PP
PP
a
PP
a
a
a
a
PP
PP
a
a
a
a
ND
PP
a
Direct
Injection
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
pc
pc
a
a
8240B - 1
Revision 2
September 1994
-------
Appropriate Technique
Analyte
Dibromomethane
l,4-Dichloro-2-butene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
l,2-Dichloroethane-d4(surr.)
1,1-Dichloroethene
trans- 1,2-Di chl oroethene
1,2-Dichloropropane
l,3-Dichloro-2-propanol
ci s- 1 , 3-Di chl oropropene
trans- 1,3-Di chl oropropene
1 , 2 , 3 , 4-Di epoxybutarie
1,4-Difluorobenzene (I.S.)
1,4-Dioxane
Epichlorohydrin
Ethanol
Ethyl benzene
Ethylene oxide
Ethyl methacrylate
2-Hexanone
2-Hydroxypropionitrile
lodomethane
Isobutyl alcohol
Malononitrile
Methacrylonitrile
Methyl ene chloride
Methyl iodide
Methyl methacrylate
4-Methyl -2-pentanone
Pentachloroethane
2-Picol ine
Propargyl alcohol
6-Propiolactone
Propionitrile
n-Propylamine
Pyridine
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2,2-Tetrachloroethane
Tetrachl oroethene
Toluene
Toluene-d8 (surr.)
1,1,1-Trichloroethane
1,1,2-Trichloroethane
Trichloroethene
Tri chl orof 1 uoromethane
CAS No.b
74-95-3
764-41-0
75-71-8
75-34-3
107-06-2
107-06-2
75-35-4
156-60-5
78-87-5
96-23-1
10061-01-5
10061-02-6
1464-53-5
540-36-3
123-91-1
106-89-8
64-17-5
100-41-4
75-21-8
97-63-2
591-78-6
78-97-7
74-88-4
78-83-1
109-77-3
126-98-7
75-09-2
74-88-4
80-62-6
108-10-1
76-01-7
109-06-8
107-19-7
57-57-8
107-12-0
107-10-8
110-86-1
100-42-5
630-20-6
79-34-5
127-18-4
108-88-3
2037-26-5
71-55-6
79-00-5
79-01-6
75-69-4
Purge-and-Trap
a
PP
a
a
a
a
a
a
a
PP
a
a
a
a
PP
i
i
a
PP
a
PP
NO
a
PP
PP
PP
a
a
a
PP
i
PP
PP
PP
PP
a
i
a
a
a
a
a
a
a
a
a
a
Direct
Injection
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
pc
a
a
a
a
a
a
a
a
pc
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
8240B - 2
Revision 2
September 1994
-------
Appropriate Technique
Direct
Analyte CAS No.b Purge-and-Trap Injection
1,2,3-Trichloropropane
Vinyl acetate
Vinyl chloride
Xylene (Total)
96-18-4
108-05-4
75-01-4
1330-20-7
a
a
a
a
a
a
a
a
a Adequate response by this technique.
b Chemical Abstract Services Registry Number.
pp Poor purging efficiency resulting in high EQLs.
i Inappropriate technique for this analyte.
pc Poor chromatographic behavior.
surr Surrogate
I.S. Internal Standard
ND Not determined
1.2 Method 8240 can be used to quantitate most volatile organic
compounds that have boiling points below 200°C and that are insoluble or slightly
soluble in water. Volatile water-soluble compounds can be included in this
analytical technique. However, for the more soluble compounds, quantitation
limits are approximately ten times higher because of poor purging efficiency.
The method is also limited to compounds that elute as sharp peaks from a GC
column packed with graphitized carbon lightly coated with a carbowax. Such
compounds include low molecular weight halogenated hydrocarbons, aromatics,
ketones, nitriles, acetates, acrylates, ethers, and sulfides. See Table 1 for
a list of compounds, retention times, and their characteristic ions that have
been evaluated on a purge-and-trap GC/MS system.
1.3 The estimated quantitation limit (EQL) of Method 8240 for an
individual compound is approximately 5 M9/kg (wet weight) for soil/sediment
samples, 0.5 mg/kg (wet weight) for wastes, and 5 /^g/L for ground water (see
Table 2). EQLs will be proportionately higher for sample extracts and samples
that require dilution or reduced sample size to avoid saturation of the detector.
1.4 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of purge-and-trap systems and gas
chromatograph/mass spectrometers, and skilled in the interpretation of mass
spectra and their use as a quantitative tool.
1.5 To increase purging efficiencies of acrylonitrile and acrolein,
refer to Methods 5030 and 8030 for proper purge-and-trap conditions.
2.0 SUMMARY OF METHOD
2.1 The volatile compounds are introduced into the gas chromatograph by
the purge-and-trap method or by direct injection (in limited applications). The
8240B - 3 Revision 2
September 1994
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components are separated via the gas chromatograph and detected using a mass
spectrometer, which is used to provide both qualitative and quantitative
information. The chromatographic conditions, as well as typical mass
spectrometer operating parameters, are given.
2.2 If the above sample introduction techniques are not applicable, a
portion of the sample is dispersed in methanol to dissolve the volatile organic
constituents. A portion of the methanolic solution is combined with organic-free
reagent water in a specially designed purging chamber. It is then analyzed by
purge-and-trap GC/MS following the normal water method.
2.3 The purge-and-trap process - An inert gas is bubbled through the
solution at ambient temperature, and the volatile components are efficiently
transferred from the aqueous phase to the vapor phase. The vapor is swept
through a sorbent column where the volatile components are trapped. After
purging is completed, the sorbent column is heated and backflushed with inert gas
to desorb the components onto a gas chromatographic column. The gas
chromatographic column is heated to elute the components, which are detected with
a mass spectrometer.
3.0 INTERFERENCES
3.1 Interferences purged or coextracted from the samples will vary
considerably from source to source, depending upon the particular sample or
extract being tested. The analytical system, however, should be checked to
ensure freedom from interferences, under the analysis conditions, by analyzing
method blanks.
3.2 Samples can be contaminated by diffusion of volatile organics
(particularly methylene chloride and fluorocarbons) through the septum seal into
the sample during shipment and storage. A trip blank, prepared from organic-free
reagent water and carried through the sampling and handling protocol, can serve
as a check on such contamination.
3.3 Cross contamination can occur whenever high-concentration and low-
concentration samples are analyzed sequentially. Whenever an unusually
concentrated sample is analyzed, it should be followed by the analysis of
organic-free reagent water to check for cross contamination. The purge-and-trap
system may require extensive bake-out and cleaning after a high-concentration
sample.
3.4 The laboratory where volatile analysis is performed should be
completely free of solvents.
3.5 Impurities in the purge gas and from organic compounds out-gassing
from the plumbing ahead of the trap account for the majority of contamination
problems. The analytical system must be demonstrated to be free from
contamination under the conditions of the analysis by running calibration and
reagent blanks. The use of non-TFE plastic coating, non-TFE thread sealants, or
flow controllers with rubber components in the purging device should be avoided.
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4.0 APPARATUS AND MATERIALS
4.1 Microsyringes - 10 /iL, 25 /*L, 100 /A, 250 /iL, 500 /xL, and 1,000 juL.
These syringes should be equipped with a 20 gauge (0.006 in. ID) needle having
a length sufficient to extend from the sample inlet to within 1 cm of the glass
frit in the purging device. The needle length will depend upon the dimensions
of the purging device employed.
4.2 Syringe valve - Two-way, with Luer ends (three each), if applicable
to the purging device.
4.3 Syringe - 5 ml_, gas-tight with shutoff valve.
4.4 Balances - Analytical, 0.0001 g, and top-loading, 0.1 g.
4.5 Glass scintillation vials - 20 ml, with screw caps and Teflon liners
or glass culture tubes with a screw cap and Teflon liner.
4.6 Volumetric flasks, Class A - 10 mL and 100 ml, with ground-glass
stoppers.
4.7 Vials - 2 mL, for GC autosampler.
4.8 Spatula - Stainless steel.
4.9 Disposable pipets - Pasteur.
4.10 Heater or heated oil bath - Should be capable of maintaining the
purging chamber to within 1°C over the temperature range of ambient to 100°C.
4.11 Purge-and-trap device - The purge-and-trap device consists of three
separate pieces of equipment: the sample purger, the trap, and the desorber.
Several complete devices are commercially available.
4.11.1 The recommended purging chamber is designed to accept
5 mL samples with a water column at least 3 cm deep. The gaseous
headspace between the water column and the trap must have a total volume
of less than 15 mL. The purge gas must pass through the water column as
finely divided bubbles with a diameter of less than 3 mm at the origin.
The purge gas must be introduced no more than 5 mm from the base of the
water column. The sample purger, illustrated in Figure 1, meets these
design criteria. Alternate sample purge devices may be utilized, provided
equivalent performance is demonstrated.
4.11.2 The trap must be at least 25 cm long and have an inside
diameter of at least 0.105 in. Starting from the inlet, the trap should
contain the following amounts of adsorbents: 1/3 of 2,6-diphenylene oxide
polymer, 1/3 of silica gel, and 1/3 of coconut charcoal. It is
recommended that 1.0 cm of methyl silicone coated packing be inserted at
the inlet to extend the life of the trap (see Figure 2). If it is not
necessary to analyze for dichlorodifluoromethane or other fluorocarbons
of similar volatility, the charcoal can be eliminated and the polymer
increased to fill 2/3 of the trap. If only compounds boiling above 35°C
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are to be analyzed, both the silica gel and charcoal can be eliminated
and the polymer increased to fill the entire trap. Before initial use,
the trap should be conditioned overnight at 180°C by backflushing with an
inert gas flow of at least 20 mL/min. Vent the trap effluent to the room,
not to the analytical column. Prior to daily use, the trap should be
conditioned for 10 minutes at 180°C with backflushing. The trap may be
vented to the analytical column during daily conditioning. However, the
column must be run through the temperature program prior to analysis of
samples.
4.11.3 The desorber should be capable of rapidly heating the
trap to 180°C for desorption. The polymer section of the trap should not
be heated higher than 180°C, and the remaining sections should not exceed
220°C during bake out mode. The desorber design illustrated in Figure 2
meets these criteria.
4.11.4 The purge-and-trap device may be assembled as a separate
unit or may be coupled to a gas chromatograph, as shown in Figures 3
and 4.
4.11.5 Trap Packing Materials
4.11.5.1 2,6-Diphenylene oxide polymer - 60/80 mesh,
chromatographic grade (Tenax GC or equivalent).
4.11.5.2 Methyl silicone packing - OV-1 (3%) on
Chromosorb-W, 60/80 mesh or equivalent.
4.11.5.3 Silica gel - 35/60 mesh, Davison, grade 15 or
equivalent.
4.11.5.4 Coconut charcoal - Prepare from Barnebey Cheney,
CA-580-26, lot #M-2649, by crushing through 26 mesh screen (or
equivalent).
4.12 Gas chromatograph/mass spectrometer system
4.12.1 Gas chromatograph - An analytical system complete with
a temperature programmable gas chromatograph and all required accessories
including syringes, analytical columns, and gases.
4.12.2 Column - 6 ft x 0.1 in. ID glass, packed with 1% SP-1000
on Carbopack-B (60/80 mesh) or equivalent.
4.12.3 Mass spectrometer - Capable of scanning from 35-260 amu
every 3 seconds or less, using 70 volts (nominal) electron energy in the
electron impact mode and producing a mass spectrum that meets all the
criteria in Table 3 when 50 ng of 4-bromofluorobenzene (BFB) are injected
through the gas chromatograph inlet.
4.12.4 GC/MS interface - Any GC-to-MS interface that gives
acceptable calibration points at 50 ng or less per injection for each of
the analytes and achieves all acceptable performance criteria (see
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Table 3) may be used. GC-to-MS interfaces constructed entirely of glass
or of glass-lined materials are recommended. Glass can be deactivated by
silanizing with dichlorodimethylsilane.
4.12.5 Data system - A computer system that allows the
continuous acquisition and storage on machine readable media of all mass
spectra obtained throughout the duration of the chromatographic program
must be interfaced to the mass spectrometer. The computer must have
software that allows searching any GC/MS data file for ions of a specified
mass and plotting such ion abundances versus time or scan number. This
type of plot is defined as an Extracted Ion Current Profile (EICP).
Software must also be available that allows integrating the abundances in
any EICP between specified time or scan number limits. The most recent
version of the EPA/NIST Mass Spectral Library should also be available.
5.0 REAGENTS
5.1 Reagent grade chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Stock solutions - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standard solutions
in methanol, using assayed liquids or gases, as appropriate.
5.3.1 Place about 9.8 ml of methanol in a 10 ml tared ground-glass-
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 minutes or until all alcohol wetted surfaces have dried. Weigh
the flask to the nearest 0.0001 g.
5.3.2 Add the assayed reference material, as described below.
5.3.2.1 Liquids - Using a 100 /LtL syringe, immediately add
two or more drops of assayed reference material to the flask; then
reweigh. The liquid must fall directly into the alcohol without
contacting the neck of the flask.
5.3.2.2 Gases - To prepare standards for any compounds
that boil below 30°C (e.g. bromomethane, chloroethane,
chloromethane, or vinyl chloride), fill a 5 mL valved gas-tight
syringe with the reference standard to the 5.0 mL mark. Lower the
needle to 5 mm above the methanol meniscus. Slowly introduce the
reference standard above the surface of the liquid. The heavy gas
will rapidly dissolve in the methanol. Standards may also be
prepared by using a lecture bottle equipped with a Hamilton Lecture
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Bottle Septum (186600). Attach Teflon tubing to the side-arm relief
valve and direct a gentle stream of gas into the methanol meniscus.
5.3.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.3.4 Transfer the stock standard solution into a Teflon sealed
screw cap bottle. Store, with minimal headspace, at -10°C to -20°C and
protect from light.
5.3.5 Prepare fresh stock standards for gases weekly or sooner if
comparison with check standards indicates a problem. Reactive compounds
such as 2-chloroethyl vinyl ether and styrene may need to be prepared more
frequently. All other standards must be replaced after six months. Both
gas and liquid standards must be monitored closely by comparison to the
initial calibration curve and by comparison to QC check standards. It may
be necessary to replace the standards more frequently if either check
exceeds a 20% drift.
5.3.6 Optionally, calibration using a certified gaseous mixture can
be accomplished daily utilizing commercially available gaseous analyte
mixture of bromomethane, chloromethane, chloroethane, vinyl chloride,
dichlorodifluoromethane and trichlorofluoromethane in nitrogen. These
mixtures of documented quality are stable for as long as six months
without refrigeration. (VOA-CYL III, RESTEK Corporation, Cat. #20194 or
equivalent).
5.4 Secondary dilution standards - Using stock standard solutions,
prepare in methanol, secondary dilution standards containing the compounds of
interest, either singly or mixed together. Secondary dilution standards must be
stored with minimal headspace and should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them.
5.5 Surrogate standards - The surrogates recommended are toluene-d8,
4-bromofluorobenzene, and l,2-dichloroethane-d4. Other compounds may be used
as surrogates, depending upon the analysis requirements. A stock surrogate
solution in methanol should be prepared as described in Sec. 5.3, and a surrogate
standard spiking solution should be prepared from the stock at a concentration
of 250 M9/10 mL in methanol. Each water sample undergoing GC/MS analysis must
be spiked with 10 juL of the surrogate spiking solution prior to analysis.
5.6 Internal standards - The recommended internal standards are
bromochloromethane, 1,4-difluorobenzene, and chlorobenzene-d5. Other compounds
may be used as internal standards as long as they have retention times similar
to the compounds being detected by GC/MS. Prepare internal standard stock and
secondary dilution standards in methanol using the procedures described in Sees.
5.3 and 5.4. It is recommended that the secondary dilution standard should be
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prepared at a concentration of 25 mg/L of each internal standard compound.
Addition of 10 juL of this standard to 5.0 ml of sample or calibration standard
would be the equivalent of 50 jug/L.
5.7 4-Bromofluorobenzene (BFB) standard - A standard solution containing
25 ng/jLtL of BFB in methanol should be prepared.
5.8 Calibration standards - Calibration standards at a minimum of five
concentrations should be prepared from the secondary dilution of stock standards
(see Sees. 5.3 and 5.4). Prepare these solutions in organic-free reagent water.
One of the concentrations should be at a concentration near, but above, the
method detection limit. The remaining concentrations should correspond to the
expected range of concentrations found in real samples but should not exceed the
working range of the GC/MS system. Each standard should contain each analyte for
detection by this method. It is EPA's intent that all target analytes for a
particular analysis be included in the calibration standard(s). However, these
target analytes may not include the entire List of Analytes (Sec. 1.1) for which
the method has been demonstrated. However, the laboratory shall not report a
quantitative result for a target analyte that was not included in the calibration
standard(s). Calibration standards must be prepared daily.
5.9 Matrix spiking standards - Matrix spiking standards should be
prepared from volatile organic compounds which will be representative of the
compounds being investigated. The suggested compounds are 1,1-dichloroethene,
trichloroethene, chlorobenzene, toluene, and benzene. The standard should be
prepared in methanol, with each compound present at a concentration of
250 jug/10.0 ml.
5.10 Great care must be taken to maintain the integrity of all standard
solutions. It is recommended that all standards in methanol be stored at -10°C
to -20°C in screw cap amber bottles with Teflon liners.
5.11 Methanol, CH3OH. Pesticide quality or equivalent. Store apart from
other solvents.
5.12 Reagent Tetraglyme - Reagent tetraglyme is defined as tetraglyme in
which interference is not observed at the method detection limit of compounds of
interest.
5.12.1 Tetraglyme (tetraethylene glycol dimethyl ether, Aldrich
#17, 240-5 or equivalent), C8H1805. Purify by treatment at reduced pressure
in a rotary evaporator. The tetraglyme should have a peroxide content of
less than 5 ppm as indicated by EM Quant Test Strips (available from
Scientific Products Co., Catalog No. P1126-8 or equivalent).
CAUTION: Glycol ethers are suspected carcinogens. All solvent
handling should be done in a hood while using proper
protective equipment to minimize exposure to liquid and
vapor.
Peroxides may be removed by passing the tetraglyme through a column
of activated alumina. The tetraglyme is placed in a round bottom flask
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equipped with a standard taper joint, and the flask is affixed to a rotary
evaporator. The flask is immersed in a water bath at 90-100°C and a vacuum
is maintained at < 10 mm Hg for at least two hours using a two stage
mechanical pump. The vacuum system is equipped with an all glass trap,
which is maintained in a dry ice/methanol bath. Cool the tetraglyme to
ambient temperature and add 100 mg/L of 2,6-di-tert-butyl-4-methyl-phenol
to prevent peroxide formation. Store the tetraglyme in a tightly sealed
screw cap bottle in an area that is not contaminated by solvent vapors.
5.12.2 In order to demonstrate that all interfering volatiles
have been removed from the tetraglyme, an organic-free reagent
water/tetraglyme blank must be analyzed.
5.13 Polyethylene glycol, H(OCH2CH2)nOH. Free of interferences at the
detection limit of the analytes.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
Samples may be introduced into the GC by either direct injection or purge-
and-trap procedures. Whichever procedure is used, the instrument calibration and
sample introduction must be performed by the same procedure.
Regardless of which sample introduction procedure is employed, establish
GC/MS operating conditions using the following recommendations as guidance.
Recommended GC/MS operating conditions:
Electron energy: 70 volts (nominal).
Mass range: 35-260 amu.
Scan time: To give 5 scans/peak, but not to
exceed 1 sec/scan.
Initial column temperature: 45°C.
Initial column holding time: 3 minutes.
Column temperature program: 8°C/minute.
Final column temperature: 220°C.
Final column holding time: 15 minutes.
Injector temperature: 200-225°C.
Source temperature: According to manufacturer's
specifications.
Transfer line temperature: 250-300°C.
Carrier gas: Hydrogen at 50 cm/sec or helium at 30
cm/sec.
7.1 Direct injection - In very limited applications (e.g. aqueous
cess wastes), direct injection of the sample into the GC/MS system with a 10
syringe may be appropriate. One such application is for verification of the
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alcohol content of an aqueous sample prior to determining if the sample is
ignitable (Methods 1010 or 1020). In this case, it is suggested that direct
injection be used. The detection limit is very high (approximately 10,000 /zg/L);
therefore, it is only permitted when concentrations in excess of 10,000 /xg/L are
expected or for water soluble compounds that do not purge. The system must be
calibrated by direct injection using the procedures described in Sec. 7.2,, but
bypassing the purge-and-trap device.
7.2 Initial calibration for purge-and-trap procedure
7.2.1 Establish the GC/MS operating conditions, using the
recommendations in Sec. 7.0 as guidance.
7.2.2 Each GC/MS system must be hardware tuned to meet the criteria
in Table 3 for a 50 ng injection or purging of 4-bromofluorobenzene (2 juL
injection of the BFB standard). Analyses must not begin until these
criteria are met.
7.2.3 Assemble a purge-and-trap device that meets the specification
in Sec. 4.11. Condition the trap overnight at 180°C in the purge mode
with an inert gas flow of at least 20 mL/min. Prior to use, condition the
trap daily for 10 min while backflushing at 180°C with the column at 220°C.
7.2.4 Connect the purge-and-trap device to a gas chromatograph.
7.2.5 Prepare the final solutions containing the required
concentrations of calibration standards, including surrogate standards,
directly in the purging device (use freshly prepared stock solutions when
preparing the calibration standards for the initial calibration.) Add
5.0 ml of organic-free reagent water to the purging device. The organic-
free reagent water is added to the purging device using a 5 ml glass
syringe fitted with a 15 cm, 20 gauge needle. The needle is inserted
through the sample inlet shown in Figure 1. The internal diameter of the
14 gauge needle that forms the sample inlet will permit insertion of the
20 gauge needle. Next, using a 10 /xL or 25 /nL microsyringe equipped with
a long needle (Sec. 4.1), take a volume of the secondary dilution solution
containing appropriate concentrations of the calibration standards (Sec.
5.6). Add the aliquot of calibration solution directly to the organic-
free reagent water in the purging device by inserting the needle through
the sample inlet. When discharging the contents of the microsyringe, be
sure that the end of the syringe needle is well beneath the surface of the
organic-free reagent water. Similarly, add 10 juL of the internal standard
solution (Sec. 5.4). Close the 2 way syringe valve at the sample inlet.
7.2.6 Carry out the purge-and-trap analysis procedure as described
in Sec. 7.4.1.
7.2.7 Tabulate the area response of the characteristic ions (see
Table 1) against concentration for each compound and each internal
standard. Calculate response factors (RF) for each compound relative to
one of the internal standards. The internal standard selected for the
calculation of the RF for a compound should be the internal standard that
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has a retention time closest to the compound being measured (Sec. 7.5.2).
The RF is calculated as follows:
RF - (AxCis)/(AisCJ
where:
Ax = Area of the characteristic ion for the compound being
measured.
Ais = Area of the characteristic ion for the specific internal
standard.
Cis = Concentration of the specific internal standard.
Cx = Concentration of the compound being measured.
7.2.8 The average RF must be calculated for each compound using the
5 RF values calculated for each compound from the initial (5-point)
calibration curve. A system performance check should be made before this
calibration curve is used. Five compounds (the System Performance Check
Compounds, or SPCCs) are checked for a minimum average relative response
factor. These compounds are chloromethane, 1,1-dichloroethane, bromoform,
1,1,2,2-tetrachloroethane, and chlorobenzene. The minimum acceptable
average RF for these compounds should be 0.300 (>0.10 for bromoform).
These compounds typically have RFs of 0.4-0.6 and are used to check
compound instability and to check for degradation caused by contaminated
lines or active sites in the system. Examples of these occurrences are:
7.2.8.1 Chloromethane - This compound is the most likely
compound to be lost if the purge flow is too fast.
7.2.8.2 Bromoform - This compound is one of the compounds
most likely to be purged very poorly if the purge flow is too slow.
Cold spots and/or active sites in the transfer lines may adversely
affect response. Response of the quantitation ion (m/z 173) is
directly affected by the tuning of BFB at ions m/z 174/176.
Increasing the m/z 174/176 relative to m/z 95 ratio may improve
bromoform response.
7.2.8.3 Tetrachloroethane and 1,1-dichloroethane - These
compounds are degraded by contaminated transfer lines in purge-and-
trap systems and/or active sites in trapping materials.
7.2.9 Using the RFs from the initial calibration, calculate and
record the percent relative standard deviation (%RSD) for all compounds.
The percent RSD is calculated as follows:
SD
%RSD =-_— x 100
RF
where:
RSD = relative standard deviation.
RF = mean of 5 initial RFs for a compound.
SD = standard deviation of average RFs for a compound.
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SD =
where:
N (RFj - RF)
I
1-1 N - 1
RFi = RF for each of the 5 calibration levels
N = Number of RF values (i.e., 5)
The percent relative standard deviation should be less than 15% for
each compound. However, the %RSD for each individual Calibration Check
Compound (CCC) must be less than 30%. Late-eluting compounds usually have
much better agreement. The CCCs are:
1,1-Dichloroethene,
Chloroform,
1,2-Dichloropropane,
Toluene,
Ethyl benzene, and
Vinyl chloride.
7.2.9.1 If a %RSD greater than 30 percent is measured for
any CCC, then corrective action to eliminate a system leak and/or
column reactive sites is required before reattempting calibration.
7.2.10 Linearity - If the %RSD of any compound is 15% or less,
then the relative response factor is assumed to be constant over the
calibration range, and the average relative response factor may be used
for quantitation (Sec. 7.5.2.2).
7.2.10.1 If the %RSD of any compound is greater than 15%,
construct calibration curves of area ratio (A/Ais) versus
concentration using first or higher order regression fit of the five
calibration points. The analyst should select the regression order
which introduces the least calibration error into the quantitation
(Sec. 7.5.2.4). The use of calibration curves is a recommended
alternative to average response factor calibration, and a useful
diagnostic of standard preparation accuracy and absorption activity
in the chromatographic system.
7.2.11 These curves are verified each shift by purging a
performance standard. Recalibration is required only if calibration and
on-going performance criteria cannot be met.
7.3 Daily GC/MS calibration
7.3.1 Prior to the analysis of samples, inject or purge 50 ng of the
4-bromofluorobenzene standard. The resultant mass spectra for the BFB
must meet all of the criteria given in Table 3 before sample analysis
begins. These criteria must be demonstrated each 12 hour shift.
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7.3.2 The initial calibration curve (Sec. 7.2) for each compound of
interest must be checked and verified once every 12 hours of analysis
time. This is accomplished by analyzing a calibration standard that is
at a concentration near the midpoint concentration for the working range
of the GC/MS and checking the SPCC (Sec. 7.3.3) and CCC (Sec. 7.3.4).
7.3.3 System Performance Check Compounds (SPCCs) - A system
performance check must be made each 12 hours. If the SPCC criteria are
met, a comparison of relative response factors is made for all compounds.
This is the same check that is applied during the initial calibration.
If the minimum relative response factors are not met, the system must be
evaluated, and corrective action must be taken before sample analysis
begins. The minimum relative response factor for volatile SPCCs is 0.300
(>0.10 for Bromoform). Some possible problems are standard mixture
degradation, injection port inlet contamination, contamination at the
front end of the analytical column, and active sites in the column or
chromatographic system.
7.3.4 Calibration Check Compounds (CCCs): After the system
performance check is met, CCCs listed in Sec. 7.2.9 are used to check the
validity of the initial calibration.
Calculate the percent drift using the following equation:
C - C
% Drift = —' — x 100
where:
C,
C, = Calibration Check Compound standard concentration.
Cc = Measured concentration using selected quantitation method.
If the percent difference for each CCC is less than 20%, the initial
calibration is assumed to be valid. If the criterion is not met (> 20%
drift), for any one CCC, corrective action must be taken. Problems
similar to those listed under SPCCs could affect this criterion. If no
source of the problem can be determined after corrective action has been
taken, a new five point calibration MUST be generated. This criterion
MUST be met before quantitative sample analysis begins. If the CCCs are
not required analytes by the permit, then all required analytes must meet
the 20% drift criterion.
7.3.5 The internal standard responses and retention times in the
check calibration standard must be evaluated immediately after or during
data acquisition. If the retention time for any internal standard changes
by more than 30 seconds from the last calibration check (12 hours), the
chromatographic system must be inspected for malfunctions and corrections
must be made, as required. If the EICP area for any of the internal
standards changes by a factor of two (- 50% to + 100%) from the last daily
calibration check standard, the mass spectrometer must be inspected for
malfunctions and corrections must be made, as appropriate. When
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corrections are made, reanalysis of samples analyzed while the system was
malfunctioning is necessary.
7.4 GC/MS analysis
7.4.1 Water samples
7.4.1.1 Screening of the sample prior to purge-and-trap
analysis will provide guidance on whether sample dilution is
necessary and will prevent contamination of the purge-and-trap
system. Two screening techniques that can be used are: the
headspace sampler (Method 3810) using a gas chromatograph (GC)
equipped with a photo ionization detector (PID) in series with an
electrolytic conductivity detector (HECD); and extraction of the
sample with hexadecane and analysis of the extract on a GC with a
FID and/or an ECD (Method 3820).
7.4.1.2 All samples and standard solutions must be allowed
to warm to ambient temperature before analysis.
7.4.1.3 Set up the GC/MS system as outlined in Sec. 7.2.1.
7.4.1.4 BFB tuning criteria and daily GC/MS calibration
criteria must be met (Sec. 7.3) before analyzing samples.
7.4.1.5 Adjust the purge gas (helium) flow rate to 25-
40 mL/min on the purge-and-trap device. Optimize the flow rate to
provide the best response for chloromethane and bromoform, if these
compounds are analytes. Excessive flow rate reduces chloromethane
response, whereas insufficient flow reduces bromoform response (see
Sec. 7.2.8).
7.4.1.6 Remove the plunger from a 5 ml syringe and attach
a closed syringe valve. Open the sample or standard bottle, which
has been allowed to come to ambient temperature, and carefully pour
the sample into the syringe barrel to just short of overflowing.
Replace the syringe plunger and compress the sample. Open the
syringe valve and vent any residual air while adjusting the sample
volume to 5.0 ml. This process of taking an aliquot destroys the
validity of the liquid sample for future analysis; therefore, if
there is only one VOA vial, the analyst should fill a second syringe
at this time to protect against possible loss of sample integrity.
This second sample is maintained only until such time when the
analyst has determined that the first sample has been analyzed
properly. Filling one 20 ml syringe would allow the use of only one
syringe. If a second analysis is needed from a syringe, it must be
analyzed within 24 hours. Care must be taken to prevent air from
leaking into the syringe.
7.4.1.7 The following procedure is appropriate for
diluting purgeable samples. All steps must be performed without
delays until the diluted sample is in a gas tight syringe.
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7.4.1.7.1 Dilutions may be made in volumetric flasks
(10 to 100 ml). Select the volumetric flask that will allow
for the necessary dilution. Intermediate dilutions may be
necessary for extremely large dilutions.
7.4.1.7.2 Calculate the approximate volume of organic-
free reagent water to be added to the volumetric flask
selected and add slightly less than this quantity of organic-
free reagent water to the flask.
7.4.1.7.3 Inject the proper aliquot of samples from
the syringe prepared in Sec. 7.4.1.6 into the flask.
Aliquots of less than 1 ml are not recommended. Dilute the
sample to the mark with organic-free reagent water. Cap the
flask, invert, and shake three times. Repeat above procedure
for additional dilutions.
7.4.1.7.4 Fill a 5 ml syringe with the diluted sample
as in Sec. 7.4.1.6.
7.4.1.8 Add 10.0 /zL of surrogate spiking solution (Sec.
5.5) and 10 /iL of internal standard spiking solution (Sec. 5.6)
through the valve bore of the syringe; then close the valve. The
surrogate and internal standards may be mixed and added as a single
spiking solution. The addition of 10 juL of the surrogate spiking
solution to 5 ml of sample is equivalent to a concentration of
50 jitg/L of each surrogate standard.
7.4.1.9 Attach the syringe-syringe valve assembly to the
syringe valve on the purging device. Open the syringe valves and
inject the sample into the purging chamber.
7.4.1.10 Close both valves and purge the sample for
11.0 + 0.1 minutes at ambient temperature.
7.4.1.11 At the conclusion of the purge time, attach the
trap to the chromatograph, adjust the device to the desorb mode, and
begin the gas chromatographic temperature program and GC/MS data
acquisition. Concurrently, introduce the trapped materials to the
gas chromatographic column by rapidly heating the trap to 180°C
while backflushing the trap with inert gas between 20 and 60 mL/min
for 4 minutes. If this rapid heating requirement cannot be met, the
gas chromatographic column must be used as a secondary trap by
cooling it to 30°C (or subambient, if problems persist) instead of
the recommended initial program temperature of 45°C.
7.4.1.12 While the trap is being desorbed into the gas
chromatograph, empty the purging chamber. Wash the chamber with a
minimum of two 5 ml flushes of organic-free reagent water (or
methanol followed by organic-free reagent water) to avoid carryover
of pollutant compounds into subsequent analyses.
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7.4.1.13 After desorbing the sample for 4 minutes,
recondition the trap by returning the purge-and-trap device to the
purge mode. Wait 15 seconds; then close the syringe valve on the
purging device to begin gas flow through the trap. The trap
temperature should be maintained at 180°C. Trap temperatures up to
220°C may be employed; however, the higher temperature will shorten
the useful life of the trap. After approximately 7 minutes, turn
off the trap heater and open the syringe valve to stop the gas flow
through the trap. When cool, the trap is ready for the next sample.
7.4.1.14 If the initial analysis of a sample or a dilution
of the sample has a concentration of analytes that exceeds the
initial calibration range, the sample must be reanalyzed at a higher
dilution. Secondary ion quantitation is allowed only when there are
sample interferences with the primary ion. When a sample is
analyzed that has saturated ions from a compound, this analysis must
be followed by a blank organic-free reagent water analysis. If the
blank analysis is not free of interferences, the system must be
decontaminated. Sample analysis may not resume until a blank can
be analyzed that is free of interferences.
7.4.1.15 For matrix spike analysis, add 10 /j,L of the matrix
spike solution (Sec. 5.9) to the 5 ml of sample to be purged.
Disregarding any dilutions, this is equivalent to a concentration
of 50 /xg/L of each matrix spike standard.
7.4.1.16 All dilutions should keep the response of the
major constituents (previously saturated peaks) in the upper half
of the linear range of the curve. Proceed to Sees. 7.5.1 and 7.5.2
for qualitative and quantitative analysis.
7.4.2 Water miscible liquids
7.4.2.1 Water miscible liquids are analyzed as water
samples after first diluting them at least 50 fold with organic-free
reagent water.
7.4.2.2 Initial and serial dilutions can be prepared by
pipetting 2 ml of the sample to a 100 ml volumetric flask and
diluting to volume with organic-free reagent water. Transfer
immediately to a 5 ml gas tight syringe.
7.4.2.3 Alternatively, prepare dilutions directly in a 5
ml syringe filled with organic-free reagent water by adding at least
20 /iL, but not more than 100 juL of liquid sample. The sample is
ready for addition of internal and surrogate standards.
7.4.3 Sediment/soil and waste samples - It is highly recommended
that all samples of this type be screened prior to the purge-and-trap
GC/MS analysis. The headspace method (Method 3810) or the hexadecane
extraction and screening method (Method 3820) may be used for this
purpose. These samples may contain percent quantities of purgeable
organics that will contaminate the purge-and-trap system, and require
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extensive cleanup and instrument downtime. Use the screening data to
determine whether to use the low-concentration method (0.005-1 nig/kg) or
the high-concentration method (> 1 mg/kg).
7.4.3.1 Low-concentration method - This is designed for
samples containing individual purgeable compounds of < 1 mg/kg. It
is limited to sediment/soil samples and waste that is of a similar
consistency (granular and porous). The low-concentration method is
based on purging a heated sediment/soil sample mixed with organic-
free reagent water containing the surrogate and internal standards.
Analyze all reagent blanks and standards under the same conditions
as the samples. See Figure 5 for an illustration of a low soils
impinger.
7.4.3.1.1 Use a 5 g sample if the expected
concentration is < 0.1 mg/kg or a 1 g sample for expected
concentrations between 0.1 and 1 mg/kg.
7.4.3.1.2 The GC/MS system should be set up as in
Sees. 7.4.1.2-7.4.1.4. This should be done prior to the
preparation of the sample to avoid loss of volatiles from
standards and samples. A heated purge calibration curve must
be prepared and used for the quantitation of all samples
analyzed with the low-concentration method. Follow the
initial and daily calibration instructions, except for the
addition of a 40°C purge temperature.
7.4.3.1.3 Remove the plunger from a 5 ml Luerlock type
syringe equipped with a syringe valve and fill until
overflowing with organic-free reagent water. Replace the
plunger and compress the water to vent trapped air. Adjust
the volume to 5.0 ml. Add 10 juL each of surrogate spiking
solution (Sec. 5.5) and internal standard solution (Sec. 5.6)
to the syringe through the valve. (Surrogate spiking
solution and internal standard solution may be mixed
together.) The addition of 10 /uL of the surrogate spiking
solution to 5 g of sediment/soil is equivalent to 50 ^9/kg of
each surrogate standard.
7.4.3.1.4 The sample (for volatile organics) consists
of the entire contents of the sample container. Do not
discard any supernatant liquids. Mix the contents of the
sample container with a narrow metal spatula. Weigh the
amount determined in Sec. 7.4.3.1.1 into a tared purge
device. Note and record the actual weight to the nearest 0.1
9-
7.4.3.1.5 Determine the percent dry weight of the
soil/sediment sample. This includes waste samples that are
amenable to percent dry weight determination. Other wastes
should be reported on a wet-weight basis.
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7.4.3.1.5.1 Immediately after weighing the sample
for extraction, weigh 5-10 g of the sample into a tared
crucible. Determine the % dry weight of the sample by
drying overnight at 105°C. Allow to cool in a
desiccator before re-weighing. Concentrations of
individual analytes are reported relative to the dry
weight of sample.
WARNING: The drying oven should be contained
in a hood or vented. Significant
laboratory contamination may result
from a heavily contaminated hazardous
waste sample.
% dry weight = g of dry sample x 100
g of sample
7.4.3.1.6 Add the spiked water to the purge device,
which contains the weighed amount of sample, and connect the
device to the purge-and-trap system.
NOTE: Prior to the attachment of the purge device, the
procedures in Sees. 7.4.3.1.4 and 7.4.3.1.6 must
be performed rapidly and without interruption to
avoid loss of volatile organics. These steps
must be performed in a laboratory free of solvent
fumes.
7.4.3.1.7 Heat the sample to 40°C + 1°C and purge the
sample for 11.0 + 0.1 minute.
7.4.3.1.8 Proceed with the analysis as outlined in
Sees. 7.4.1.11-7.4.1.16. Use 5 ml of the same organic-free
reagent water as in the reagent blank. If saturated peaks
occurred or would occur if a 1 g sample were analyzed, the
high-concentration method must be followed.
7.4.3.1.9 For low-concentration sediment/soils add
10 /nL of the matrix spike solution (Sec. 5.9) to the 5 ml of
organic-free reagent water (Sec. 7.4.3.1.3). The
concentration for a 5 g sample would be equivalent to 50
jug/kg of each matrix spike standard.
7.4.3.2 High-concentration method - The method is based on
extracting the sediment/soil with methanol. A waste sample is
either extracted or diluted, depending on its solubility in
methanol. Wastes (i.e. petroleum and coke wastes) that are
insoluble in methanol are diluted with reagent tetraglyme or
possibly polyethylene glycol (PEG). An aliquot of the extract is
added to organic-free reagent water containing internal standards.
This is purged at ambient temperature. All samples with an expected
concentration of > 1.0 mg/kg should be analyzed by this method.
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7.4.3.2.1 The sample (for volatile organics) consists
of the entire contents of the sample container. Do not
discard any supernatant liquids. Mix the contents of the
sample container with a narrow metal spatula. For
sediment/soil and solid wastes that are insoluble in
methanol, weigh 4 g (wet weight) of sample into a tared 20 ml
vial. Use a top loading balance. Note and record the actual
weight to 0.1 gram and determine the percent dry weight of
the sample using the procedure in Sec. 7.4.3.1.5. For waste
that is soluble in methanol, tetraglyme, or PEG, weigh 1 g
(wet weight) into a tared scintillation vial or culture tube
or a 10 ml volumetric flask. (If a vial or tube is used, it
must be calibrated prior to use. Pi pet 10.0 ml of solvent
into the vial and mark the bottom of the meniscus. Discard
this solvent.)
7.4.3.2.2 Quickly add 9.0 ml of appropriate solvent;
then add 1.0 ml of the surrogate spiking solution to the
vial. Cap and shake for 2 minutes.
NOTE: Sees. 7.4.3.2.1 and 7.4.3.2.2 must be performed
rapidly and without interruption to avoid loss of
volatile organics. These steps must be performed
in a laboratory free from solvent fumes.
7.4.3.2.3 Pipet approximately 1 ml of the extract to
a GC vial for storage, using a disposable pipet. The
remainder may be disposed of. Transfer approximately 1 ml of
appropriate solvent to a separate GC vial for use as the
method blank for each set of samples. These extracts may be
stored at 4°C in the dark, prior to analysis. The addition
of a 100 /iL aliquot of each of these extracts in Sec.
7.4.3.2.6 will give a concentration equivalent to 6,200 M9/kg
of each surrogate standard.
7.4.3.2.4 The GC/MS system should be set up as in
Sees. 7.4.1.2-7.4.1.4. This should be done prior to the
addition of the solvent extract to organic-free reagent
water.
7.4.3.2.5 Table 4 can be used to determine the volume
of solvent extract to add to the 5 ml of organic-free reagent
water for analysis. If a screening procedure was followed
(Method 3810 or 3820), use the estimated concentration to
determine the appropriate volume. Otherwise, estimate the
concentration range of the sample from the low-concentration
analysis to determine the appropriate volume. If the sample
was submitted as a high-concentration sample, start with
100 /iL. All dilutions must keep the response of the major
constituents (previously saturated peaks) in the upper half
of the linear range of the curve.
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7.4.3.2.6 Remove the plunger from a 5.0 ml Luerlock
type syringe equipped with a syringe valve and fill until
overflowing with organic-free reagent water. Replace the
plunger and compress the water to vent trapped air. Adjust
the volume to 4.9 ml. Pull the plunger back to 5.0 ml to
allow volume for the addition of the sample extract and of
standards. Add 10 pi of internal standard solution. Also
add the volume of solvent extract determined in Sec.
7.4.3.2.5 and a volume of extraction or dissolution solvent
to total 100 juL (excluding methanol in standards).
7.4.3.2.7 Attach the syringe-syringe valve assembly to
the syringe valve on the purging device. Open the syringe
valve and inject the organic-free reagent water/methanol
sample into the purging chamber.
7.4.3.2.8 Proceed with the analysis as outlined in
Sec. 7.4.1.11-7,4.1.16. Analyze all reagent blanks on the
same instrument as that use for the samples. The standards
and blanks should also contain 100 juL of solvent to simulate
the sample conditions.
7.4.3.2.9 For a matrix spike in the high-concentration
sediment/soil samples, add 8.0 mL of methanol, 1.0 ml of
surrogate spike solution (Sec. 5.5), and 1.0 mL of matrix
spike solution (Sec. 5.9) as in Sec. 7.4.3.2.2. This results
in a 6,200 ^g/kg concentration of each matrix spike standard
when added to a 4 g sample. Add a 100 /^L aliquot of this
extract to 5 ml of organic-free reagent water for purging (as
per Sec. 7.4.3.2.6).
7.5 Data interpretation
7.5.1 Qualitative analysis
7.5.1.1 The qualitative identification of compounds
determined by this method is based on retention time, and on
comparison of the sample mass spectrum, after background correction,
with characteristic ions in a reference mass spectrum. The
reference mass spectrum must be generated by the laboratory using
the conditions of this method. The characteristic ions from the
reference mass spectrum are defined to be the three ions of greatest
relative intensity, or any ions over 30% relative intensity if less
than three such ions occur in the reference spectrum. Compounds
should be identified as present when the criteria below are met.
7.5.1.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographic peak containing ions
specific for the target compound at a compound-specific
retention time will be accepted as meeting this criterion.
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7.5.1.1.2 The RRT of the sample component is within
+ 0.06 RRT units of the RRT of the standard component.
7.5.1.1.3 The relative intensities of the
characteristic ions agree within 30% of the relative
intensities of these ions in the reference spectrum.
(Example: For an ion with an abundance of 50% in the
reference spectrum, the corresponding abundance in a sample
spectrum can range between 20% and 80%.)
7.5.1.1.4 Structural isomers that produce very similar
mass spectra should be identified as individual isomers if
they have sufficiently different GC retention times.
Sufficient GC resolution is achieved if the height of the
valley between two isomer peaks is less than 25% of the sum
of the two peak heights. Otherwise, structural isomers are
identified as isomeric pairs.
7.5.1.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce
mass spectra containing ions contributed by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
appropriate selection of analyte spectra and background
spectra is important. Examination of extracted ion current
profiles of appropriate ions can aid in the selection of
spectra, and in qualitative identification of compounds.
When analytes coelute (i.e., only one chromatographic peak is
apparent), the identification criteria can be met, but each
analyte spectrum will contain extraneous ions contributed by
the coeluting compound.
7.5.1.2 For samples containing components not associated
with the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the type of analyses
being conducted. Guidelines for making tentative identification
are:
(1) Relative intensities of major ions in the reference
spectrum (ions > 10% of the most abundant ion) should be present in
the sample spectrum.
(2) The relative intensities of the major ions should agree
within ± 20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
between 30 and 70%).
(3) Molecular ions present in the reference spectrum should
be present in the sample spectrum.
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(4) Ions present in the sample spectrum but not in the
reference spectrum should be reviewed for possible background
contamination or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the
sample spectrum should be reviewed for possible subtraction from the
sample spectrum because of background contamination or coeluting
peaks. Data system library reduction programs can sometimes create
these discrepancies.
Computer generated library search routines should not use
normalization routines that would misrepresent the library or
unknown spectra when compared to each other. Only after visual
comparison of sample with the nearest library searches will the mass
spectral interpretation specialist assign a tentative
identification.
7.5.2 Quantitative analysis
7.5.2.1 When a compound has been identified, the
quantitation of that compound will be based on the integrated
abundance from the EICP of the primary characteristic ion.
Quantitation will take place using the internal standard technique.
The internal standard used shall be the one nearest the retention
time of that of a given analyte (e.g. see Table 5).
7.5.2.2 When linearity exists, as per Sec. 7.2.10,
calculate the concentration of each identified analyte in the sample
as follows:
Water
(A,)(I.)
concentration (/ig/L) = —
(AJ(RF)(V0)
where:
Ax = Area of characteristic ion for compound being
measured.
Is = Amount of internal standard injected (ng).
A1S = Area of characteristic ion for the internal
standard.
RF = Mean relative response factor for compound being
measured (Sec. 7.2.8).
V0 = Volume of water purged (ml), taking into
consideration any dilutions made.
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Sediment/Soil Sludge (on a dry-weight basis) and Waste
(normally on a wet-weight basis)
(AJd.)(Vt)
concentration (/^g/kg) =
(Ais)(RF)(Vi)(Ws)(D)
where:
Ax> Is> Ais, RF, = Same as for water.
Vt = Volume of total extract (fj.1) (use 10,000 /uL or a
factor of this when dilutions are made).
V- = Volume of extract added (juL) for purging.
Ws = Weight of sample extracted or purged (g).
D = % dry weight of sample/100, or 1 for a wet-weight
basis.
7.5.2.3 Where applicable, an estimate of concentration for
noncalibrated components in the sample should be made. The formulae
given above should be used with the following modifications: The
areas Ax and Ais should be from the total ion chromatograms, and the
RF for the compound should be assumed to be 1. The concentration
obtained should be reported indicating (1) that the value is an
estimate and (2) which internal standard was used to determine
concentration. Use the nearest internal standard free of
interferences.
7.5.2.4 Alternatively, the regression line fitted to the
initial calibration (Sec. 7.2.10.1) may be used for determination
of analyte concentration.
8.0 QUALITY CONTROL
8.1 Each laboratory that uses these methods is required to operate a
formal quality control program. The minimum requirements of this program consist
of an initial demonstration of laboratory capability and an ongoing analysis of
spiked samples to evaluate and document data quality. The laboratory must
maintain records to document the quality of the data generated. Ongoing data
quality checks are compared with established performance criteria to determine
if the results of analyses meet the performance characteristics of the method.
When results of sample spikes indicate atypical method performance, a quality
control reference sample must be analyzed to confirm that the measurements were
performed in an in-control mode of operation.
8.2 Before processing any samples, the analyst should demonstrate,
through the analysis of a method blank, that interferences from the analytical
system, glassware, and reagents are under control. Each time a set of samples
is extracted or there is a change in reagents, a method blank should be processed
as a safeguard against chronic laboratory contamination. The blank samples
should be carried through all stages of sample preparation and measurement.
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8.3 The experience of the analyst performing GC/MS analyses is
invaluable to the success of the methods. Each day that analysis is performed,
the daily calibration standard should be evaluated to determine if the
chromatographic system is operating properly. Questions that should be asked
are: Do the peaks look normal?; Is the response obtained comparable to the
response from previous calibrations? Careful examination of the standard
chromatogram can indicate whether the column is still useable, the injector is
leaking, the injector septum needs replacing, etc. If any changes are made to
the system (e.g. column changed), recalibration of the system must take place.
8.4 Required instrument QC is found in the following section:
8.4.1 The GC/MS system must be tuned to meet the BFB specifications
in Sec. 7.2.2.
8.4.2 There must be an initial calibration of the GC/MS system as
specified in Sec. 7.2.
8.4.3 The GC/MS system must meet the SPCC criteria specified in Step
7.3.3 and the CCC criteria in Sec. 7.3.4, each 12 hours.
8.5 To establish the ability to generate acceptable accuracy and
precision, the analyst must perform the following operations.
8.5.1 A quality control (QC) reference sample concentrate is
required containing each analyte at a concentration of 10 mg/L in
methanol. The QC reference sample concentrate may be prepared from pure
standard materials or purchased as certified solutions. If prepared by
the laboratory, the QC reference sample concentrate must be made using
stock standards prepared independently from those used for calibration.
8.5.2 Prepare a QC reference sample to contain 20 /Ltg/L of each
analyte by adding 200 /xL of QC reference sample concentrate to 100 mL of
water.
8.5.3 Four 5-mL aliquots of the well mixed QC reference sample are
analyzed according to the method beginning in Sec. 7.4.1.
8.5.4 Calculate the average recovery (x) in jug/L, and the standard
deviation of the recovery (s) in jug/L, for each analyte using the four
results.
8.5.5 For each analyte compare s and x with the corresponding
acceptance criteria_for precision and accuracy, respectively, found in
Table 6. If s and x for all analytes meet the acceptance criteria, the
system performance is acceptable and analysis of actual samples can_begin.
If any individual s exceeds the precision limit or any individual x falls
outside the range for accuracy, then the system performance is
unacceptable for that analyte.
NOTE: The large number of analytes in Table 6 present a substantial
probability that one or more will fail at least one of the
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acceptance criteria when all analytes of a given method are
determined.
8.5.6 When one or more of the analytes tested fail at least one of
the acceptance criteria, the analyst must proceed according to Sec.
8.5.6.1 or 8.5.6.2.
8.5.6.1 Locate and correct the source of the problem and
repeat the test for all analytes beginning with Sec. 8.5.2.
8.5.6.2 Beginning with Sec. 8.5.2, repeat the test only
for those analytes that failed to meet criteria. Repeated failure,
however, will confirm a general problem with the measurement system.
If this; occurs, locate and correct the source of the problem and
repeat the test for all compounds of interest beginning with Sec.
8.5.2.
8.6 The laboratory must, on an ongoing basis, analyze a method blank and
a spiked replicate for each analytical batch (up to a maximum of 20
samples/batch) to assess accuracy. For soil and waste samples where detectable
amounts of organics are present, replicate samples may be appropriate in place
of spiked replicates. For laboratories analyzing one to ten samples per month,
at least one spiked sample per month is required.
8.6.1 The concentration of the spike in the sample should be
determined as follows:
8.6.1.1 If, as in compliance monitoring, the concentration
of a specific analyte in the sample is being checked against a
regulatory concentration limit, the spike should be at that limit
or 1 to 5 times higher than the background concentration determined
in Sec. 8.6.2, whichever concentration would be larger.
8.6.1.2 If the concentration of a specific analyte in a
water sample is not being checked against a specific limit, the
spike should be at 20 jug/L or 1 to 5 times higher than the
background concentration determined in Sec. 8.6.2, whichever
concentration would be larger. For other matrices, recommended
spiking concentration is 10 times the EQL.
8.6.2 Analyze one 5-mL sample aliquot to determine the background
concentration (B) of each analyte. If necessary, prepare a new QC
reference sample concentrate (Sec. 8.5.1) appropriate for the background
concentration in the sample. Spike a second 5-mL sample aliquot with
10 fj.L of the QC reference sample concentrate and analyze it to determine
the concentration after spiking (A) of each analyte. Calculate each
percent recovery (p) as 100(A-B)%/T, where T is the known true value of
the spike.
8.6.3 Compare the percent recovery (p) for each analyte in a water
sample with the corresponding QC acceptance criteria found in Table 6.
These acceptance criteria were calculated to include an allowance for
error in measurement of both the background and spike concentrations,
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assuming a spike to background ratio of 5:1. This error will be accounted
for to the extent that the analyst's spike to background ratio approaches
5:1. If spiking was performed at a concentration lower than 20 M9/U the
analyst must use either the QC acceptance criteria presented in Table 6,
or optional QC acceptance criteria calculated for the specific spike
concentration. To calculate optional acceptance criteria for the recovery
of an analyte: (1) Calculate accuracy (x') using the equation found in
Table 7, substituting the spike concentration (T) for C; (2) calculate
overall precision (S') using the equation in Table 7, substituting x' for
x; (3) calculate the range for recovery at the spike concentration as
(100x'/T) + 2.44(100S7T)%.
8.6.4 If any individual p falls outside the designated range for
recovery, that analyte has failed the acceptance criteria. A check
standard containing each analyte that failed the criteria must be analyzed
as described in Sec. 8.7.
8.7 If any analyte in a water sample fails the acceptance criteria for
recovery in Sec. 8.6, a QC reference sample containing each analyte that failed
must be prepared and analyzed.
NOTE: The frequency for the required analysis of a QC reference sample
will depend upon the number of analytes being simultaneously tested,
the complexity of the sample matrix, and the performance of the
laboratory. If the entire list of analytes in Table 6 must be
measured in the sample in Sec. 8.6, the probability that the
analysis of a QC reference sample will be required is high. In this
case, the QC reference sample should be routinely analyzed with the
spiked sample.
8.7.1 Prepare the QC reference sample by adding 10 pi of the QC
reference sample concentrate (Sec. 8.5.1 or 8.6.2) to 5 ml of reagent
water. The QC reference sample needs only to contain the analytes that
failed criteria in the test in Sec. 8.6.
8.7.2 Analyze the QC reference sample to determine the concentration
measured (A) of each analyte. Calculate each percent recovery (ps) as
100(A/T)%, where T is the true value of the standard concentration.
8.7.3 Compare the percent recovery (ps) for each analyte with the
corresponding QC acceptance criteria found in Table 6. Only analytes that
failed the test in Sec. 8.6 need to be compared with these criteria. If
the recovery of any such analyte falls outside the designated range, the
laboratory performance for that analyte is judged to be out of control,
and the problem must be immediately identified and corrected. The result
for that analyte in the unspiked sample is suspect and may not be reported
for regulatory compliance purposes.
8.8 As part of the QC program for the laboratory, method accuracy for
each matrix studied must be assessed and records must be maintained. After the
analysis of five spiked samples _(of the same matrix) as in Sec. 8.6, calculate
the average percent recovery (p) and the standard deviation of the percent
recovery (sp). Express the accuracy assessment as a percent recovery interval
8240B - 27 Revision 2
September 1994
-------
from p - 2sp to p + 2sp. If p = 90% and sp = 10%, for example, the accuracy
interval is expressed as 70-110%. Update the accuracy assessment for each
analyte on a regular basis (e.g., after each five to ten new accuracy
measurements).
8.9 To determine acceptable accuracy and precision limits for surrogate
standards the following procedure should be performed.
8.9.1 For each sample analyzed, calculate the percent recovery of
each surrogate in the sample.
8.9.2 Once a minimum of thirty samples of the same matrix have been
analyzed, calculate the average percent recovery (P) and standard
deviation of the percent recovery (s) for each of the surrogates.
8.9.3 For a given matrix, calculate the upper and lower control
limit for method performance for each surrogate standard. This should be
done as follows:
Upper Control Limit (UCL) = P + 3s
Lower Control Limit (LCL) = P - 3s
8.9.4 For aqueous and soil matrices, these laboratory established
surrogate control limits should, if applicable, be compared with the
control limits listed in Table 8. The limits given in Table 8 are multi-
laboratory performance based limits for soil and aqueous samples, and
therefore, the single-laboratory limits established in Sec. 8.9.3 must
fall within those given in Table 8 for these matrices.
8.9.5 If recovery is not within limits, the following procedures are
required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are
a problem or flag the data as "estimated concentration".
8.9.6 At a minimum, each laboratory should update surrogate recovery
limits on a matrix-by-matrix basis, annually.
8.10 It is recommended that the laboratory adopt additional quality
assurance practices for use with this method. The specific practices that are
most productive depend upon the needs of the laboratory and the nature of the
samples. Field duplicates may be analyzed to assess the precision of the
environmental measurements. When doubt exists over the identification of a peak
on the chromatogram, confirmatory techniques such as gas chromatography with a
dissimilar column or a different ionization mode using a mass spectrometer must
8240B - 28 Revision 2
September 1994
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be used. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 This method was tested by 15 laboratories using organic-free reagent
water, drinking water, surface water, and industrial wastewaters spiked at six
concentrations over the range 5-600 jug/L. Single operator precision, overall
precision, and method accuracy were found to be directly related to the
concentration of the analyte and essentially independent of the sample matrix.
Linear equations to describe these relationships are presented in Table 7.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act, Method 624,"
October 26, 1984.
2. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, July 1985, Revision.
3. Bellar, T.A., and J.J. Lichtenberg, J. Amer. Water Works Assoc., 66(12),
739-744, 1974.
4. Bellar, T.A., and J.J. Lichtenberg, "Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds," in Van Hall, ed., Measurement of Organic Pollutants in Water
and Wastewater, ASTM STP 686, pp. 108-129, 1979.
5. Budde, W.L. and J.W. Eichelberger, "Performance Tests for the Evaluation
of Computerized Gas Chromatography/Mass Spectrometry Equipment and
Laboratories," EPA-600/4-79-020, U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, Ohio 45268,
April 1980.
6. Eichelberger, J.W., L.E. Harris, and W.L. Budde, "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass
Spectrometry Systems," Analytical Chemistry, 47, 995-1000, 1975.
7. "Method Detection Limit for Methods 624 and 625," Olynyk, P., W.L. Budde,
and J.W. Eichelberger, Unpublished report, October 1980.
8. "Interlaboratory Method Study for EPA Method 624-Purgeables," Final Report
for EPA Contract 68-03-3102.
9. "Method Performance Data for Method 624," Memorandum from R. Slater and
T. Pressley, U.S. Environmental Protection Agency, Environmental
Monitoring and Support Laboratory, Cincinnati, Ohio 45268, January 17,
1984.
8240B - 29 Revision 2
September 1994
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10. Gebhart, J.E.; Lucas, S.V.; Naber, S.J.; Berry, A.M.; Danison, T.H.;
Burkholder, H.M. "Validation of SW-846 Methods 8010, 8015, and 8020"; U.S.
Environmental Protection Agency, Environmental Monitoring and Support
Laboratory, Cincinnati, Old 45268, July 1987, Contract No. 68-03-1760.
11. Lucas, S.V.; Kornfeld, R.A. "GC-MS Suitability Testing of RCRA Appendix
VIII and Michigan List Analytes "; U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, OH 45268,
February 20, 1987, Contract No. 68-03-3224.
8240B - 30 Revision 2
September 1994
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TABLE 1.
RETENTION TIMES AND CHARACTERISTIC IONS FOR VOLATILE COMPOUNDS
Compound
Retention
Time (minutes)
Primary Ion Secondary Ion(s)
Ethylene oxide
Chloromethane
Di chl orodi f 1 uoromethane
Bromomethane
Vinyl chloride
Acetonitrile
Chloroethane
Methyl iodide
Methylene chloride
Carbon disulfide
Tri chl orof 1 uoromethane
Propionitrile
Ally! chloride
1,1-Dichloroethene
Bromochloromethane (I.S.)
Ally! alcohol
trans-l,2-Dichloroethene
1,2-Dichloroethane
Propargyl alcohol
Chloroform
l,2-Dichloroethane-d4(surr)
2-Butanone
Methacrylonitrile
Dibromomethane
2-Chloroethanol
b-Propiolactone
Epichlorohydrin
1,1,1 -Tri chloroethane
Carbon tetrachlon'de
1,4-Dioxane
Isobutyl alcohol
Bromod i chl oromethane
Chloroprene
l,2:3,4-Diepoxybutane
1,2-Dichloropropane
Chloral hydrate (b)
cis-l,3-Dichloropropene
Bromoacetone
Trichloroethene
Benzene
trans-l,3-Dichloropropene
1 , 1 , 2-Tri chl oroethane
3-Chloropropionitrile
1,2-Dibromoethane
Pyridine
1.30
2.30
2.47
3.10
3.80
3.97
4.60
5.37
6.40
7.47
8.30
8.53
8.83
9.00
9.30
9.77
10.00
10.10
10.77
11.40
12.10
12.20
12.37
12.53
12.93
13.00
13.10
13.40
13.70
13.70
13.80
14.30
14.77
14.87
15.70
15.77
15.90
16.33
16.50
17.00
17.20
17.20
17.37
18.40
18.57
44
50
85
94
62
41
64
142
84
76
101
54
76
96
128
57
96
62
55
83
65
72
41
93
49
42
57
97
117
88
43
83
53
55
63
82
75
136
130
78
75
97
54
107
79
44, 43, 42
52, 49
85, 87, 101, 103
96, 79
64, 61
41, 40, 39
66, 49
142, 127, 141
49, 51, 86
76, 78, 44
103, 66
54, 52, 55, 40
76, 41, 39, 78
61, 98
49, 130, 51
57, 58, 39
61, 98
64, 98
55, 39, 38, 53
85, 47
102
43, 72
41, 67, 39, 52, 66
93, 174, 95, 172, 176
49, 44, 43, 51, 80
42, 43, 44
57, 49, 62, 51
99, 117
119, 121
88, 58, 43, 57
43, 41, 42, 74
85, 129
53, 88, 90, 51
55, 57, 56
62, 41
44, 84, 86, 111
77, 39
43, 136, 138, 93, 95
95, 97, 132
52, 71
77, 39
83, 85, 99
54, 49, 89, 91
107, 109, 93, 188
79, 52, 51, 50
8240B - 31
Revision 2
September 1994
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TABLE 1.
(Continued)
Compound
Retention
Time (minutes)
Primary Ion Secondary Ion(s)
2-Chloroethyl vinyl ether
2-Hydroxypropionitrile
1,4-Difluorobenzene (I.S.)
Malononitrile
Methyl methacryl ate
Bromoform
1,1,1 , 2-Tetrachl oroethane
l,3-Dichloro-2-propanol
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
1 , 2 , 3-Tr i chl oropropane
l,4-Dichloro-2-butene
n-Propylamine
2-Picoline
Toluene
Ethyl methacryl ate
Chlorobenzene
Pentachl oroethane8
Ethyl benzene
1, 2 -Dibromo-3- chl oropropane
4-Bromofluorobenzene (surr.)
Benzyl chloride
Styrene
bis-(2-Chloroethyl) sulfide(b)
Acetone
Acrolein
Acrylonitrile
Chlorobenzene-d5 (I.S.)
Chlorodibromomethane
1,1-Dichloroethane
Ethanol
2-Hexanone
lodomethane
4-Methyl -2-pentanone
Toluene-d8 (surr.)
Vinyl acetate
Xylene (Total)
18.60
18.97
19.60
19.60
19.77
19.80
20.33
21.83
22.10
22.20
22.20
22.73
23.00
23.20
23.50
23.53
24.60
24.83
26.40
27.23
28.30
29.50
30.83
33.53
--
--
--
--
--
--
--
--
--
--
--
--
_ _
63
44
114
66
69
173
131
79
83
164
75
75
59
93
92
69
112
167
106
157
95
91
104
109
43
56
53
117
129
63
31
43
142
43
98
43
106
65,106
44,43,42,53
63,88
66,39,65,38
69,41,100,39
171,175,252
131,133,117,119,95
79,43,81,49
85,131,133
129,131,166
75,77,110,112,97
75,53,77,124,89
59,41,39
93,66,92,78
91,65
69,41,99,86,114
114,77
167,130,132,165,169
91
157,75,155,77
174,176
91,126,65,128
104,103,78,51,77
111, 158, 160
58
55,58
52,51
82,119
208,206
65,83
45,27,46
58,57, 100
127,141
58,57,100
70,100
86
91
a The base peak at m/e 117 was not used due to an interference at that mass with
a nearly coeluting internal standard, chlorobenzene-d5.
b Response factor judged to be too low (less than 0.02) for practical use.
(I.S.) = Internal Standard
(surr) = Surrogate
8240B - 32
Revision 2
September 1994
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TABLE 2.
ESTIMATED QUANTITATION LIMITS (EQL) FOR VOLATILE ORGANICS
Estimated
Quantitation
Limits"
Ground water
Volatiles /j.g/1
Acetone
Acetonitrile
Ally! chloride
Benzene
Benzyl chloride
Bromodi chl oromethane
Bromoform
Bromomethane
2-Butanone
Carbon disulfide
Carbon tetrachloride
Chlorobenzene
Chlorodibromomethane
Chloroethane
2-Chloroethyl vinyl ether
Chloroform
Chl oromethane
Chloroprene
1 , 2-Di bromo-3-chl oropropane
1,2-Dibromoethane
Dibromomethane
l,4-Dichloro-2-butene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1 Dichloroethene
trans- 1, 2-Di chl oroethene
1, 2-Di chl oropropane
cis-l,3-Dichloropropene
trans-l,3-Dichloropropene
Ethyl benzene
Ethyl methacrylate
2-Hexanone
Isobutyl alcohol
Methacrylonitrile
Methyl ene chloride
Methyl iodide
Methyl methacrylate
4-Methyl -2-pentanone
Pentachloroethane
100
100
5
5
100
5
5
10
100
100
5
5
5
10
10
5
10
5
100
5
5
100
5
5
5
5
5
5
5
5
5
5
50
100
100
5
5
5
50
10
Low Soil/Sediment"
M9/kg
100
100
5
5
100
5
5
10
100
100
5
5
5
10
10
5
10
5
100
5
5
100
5
5
5
5
5
5
5
5
5
5
50
100
100
5
5
50
50
10
8240B - 33 Revision 2
September 1994
-------
TABLE 2.
(Continued)
Estimated
Quantitation
Limits8
Ground water Low Soil/Sedimentb
Volatiles p.g/1
Propionitrile
Styrene
1,1,1, 2 -Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
1,1,1-Trichl oroethane
1 , 1 , 2-Trichl oroethane
Trichloroethene
1,2,3-Trichloropropane
Vinyl acetate
Vinyl chloride
Xylene (Total)
100
5
5
5
5
5
5
5
5
5
50
10
5
100
5
5
5
5
5
5
5
5
5
50
10
5
a Sample EQLs are highly matrix dependent. The EQLs listed herein are provided
for guidance and may not always be achievable.
b EQLs listed for soil/sediment are based on wet weight. Normally data are
reported on a dry weight basis; therefore, EQLs will be higher, based on the
percent dry weight of each sample.
Other Matrices Factor0
Water miscible liquid waste 50
High-concentration soil and sludge 125
Non-water miscible waste 500
CEQL = [EQL for low soil/sediment (see Table 2)] X [Factor found in this
table]. For non-aqueous samples, the factor is on a wet weight basis.
8240B - 34 Revision 2
September 1994
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TABLE 3.
BFB KEY ION ABUNDANCE CRITERIA
Mass Ion Abundance Criteria .
50 15 to 40% of mass 95
75 30 to 60% of mass 95
95 base peak, 100% relative abundance
96 5 to 9% of mass 95
173 less than 2% of mass 174
174 greater than 50% of mass 95
175 5 to 9% of mass 174
176 greater than 95% but less than 101% of mass 174
177 5 to 9% of mass 176
TABLE 4.
QUANTITY OF METHANOL EXTRACT REQUIRED FOR ANALYSIS
OF HIGH-CONCENTRATION SOILS/SEDIMENTS
Approximate Volume of
Concentration Range Methanol Extract
a
500- 10,000 Mg/kg 100 /xL
1,000- 20,000 jugAg 50 ML
5,000-100,000 jug/kg 10 ML
25,000-500,000 Mg/kg 100 ML of 1/50 dilution"
Calculate appropriate dilution factor for concentrations exceeding this
table.
a The volume of methanol added to 5 mL of water being purged should be kept
constant. Therefore, add to the 5 mL syringe whatever volume of methanol
is necessary to maintain a volume of 100 juL added to the syringe.
b Dilute and aliquot of the methanol extract and then take 100 ML for
analysis.
8240B - 35 Revision 2
September 1994
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TABLE 5.
VOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES ASSIGNED
FOR QUANTITATION
Bromochloromethane
Acetone
Acrolein
Acrylonitrile
Bromomethane
Carbon disulfide
Chloroethane
Chloroform
Chioromethane
Di chlorodi f1uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
l,2-Dichloroethane-d4 (surrogate)
1,1-Dichloroethene
trans-l,2-Dichloroethene
lodomethane
Methylene chloride
Tri chlorof1uoromethane
Vinyl chloride
1,4-Di f1uorobenzene
Benzene
Bromodichloromethane
Bromoform
2-Butanone
Carbon tetrachloride
Chlorodi bromomethane
2-Chloroethyl vinyl ether
Dibromomethane
l,4-Dichloro-2-butene
1,2-Dichloropropane
cis-1,3-Dichloropropene
trans-l,3-Dichloropropene
1,1,1-Trichloroethane
1,1,2-Trichloroethane
Trichloroethene
Vinyl acetate
Chlorobenzene-dr
Bromofluorobenzene (surrogate)
Chlorobenzene
Ethyl benzene
Ethyl methacrylate
2-Hexanone
4-Methyl-2-pentanone
Styrene
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
Toluene-d8 (surrogate)
1,2,3-Trichloropropane
Xylene
8240B - 36
Revision 2
September 1994
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TABLE 6.
CALIBRATION AND QC ACCEPTANCE CRITERIA8
Parameter
Benzene
Bromodi chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
2-Chloroethyl vinyl ether
Chloroform
Chl oromethane
Di bromochl oromethane
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans -1, 2-Di chl oroethene
1,2-Dichloropropane
cis-l,3-Dichloropropene
trans-l,3-Dichloropropene
Ethyl benzene
Methyl ene chloride
1,1, 2, 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
1 , 1 , 1 -Tri chl oroethane
1 , 1 , 2-Trichl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
Vinyl chloride
Range
for Q
(M9/L)
12.8-27.2
13.1-26.9
14.2-25.8
2.8-37.2
14.6-25.4
13.2-26.8
D-44.8
13.5-26.5
D-40.8
13.5-26.5
12.6-27.4
14.6-25.4
12.6-27.4
14.5-25.5
13.6-26.4
10.1-29.9
13.9-26.1
6.8-33.2
4.8-35.2
10.0-30.0
11.8-28.2
12.1-27.9
12.1-27.9
14.7-25.3
14.9-25.1
15.0-25.0
14.2-25.8
13.3-26.7
9.6-30.4
0.8-39.2
Q = Concentration measured in QC check
Limit
for s
(M9/L)
6.9
6.4
5.4
17.9
5.2
6.3
25.9
6.1
19.8
6.1
7.1
5.5
7.1
5.1
6.0
9.1
5.7
13.8
15.8
10.4
7.5
7.4
7.4
5.0
4.8
4.6
5.5
6.6
10.0
20.0
sample,
Range
for x
(MgA)
15.2-26.0
10.1-28.0
11.4-31.1
D-41.2
17.2-23.5
16.4-27.4
D-50.4
13.7-24.2
D-45.9
13.8-26.6
11.8-34.7
17.0-28.8
11.8-34.7
14.2-28.4
14.3-27.4
3.7-42.3
13.6-28.4
3.8-36.2
1.0-39.0
7.6-32.4
17.4-26.7
D-41.0
13.5-27.2
17.0-26.6
16.6-26.7
13.7-30.1
14.3-27.1
18.5-27.6
8.9-31.5
D-43.5
in jug/L.
Range
P.Ps
(%)
37-151
35-155
45-169
D-242
70-140
37-160
D-305
51-138
D-273
53-149
18-190
59-156
18-190
59-155
49-155
D-234
54-156
D-210
D-227
17-183
37-162
D-221
46-157
64-148
47-150
52-162
52-150
71-157
17-181
D-251
s_ = Standard deviation of four recovery measurements, in p,g/L.
x = Average recovery
p, ps = Percent recovery
D = Detected; result
for four recovery
measured.
measurements, in /ng/L.
must be greater than zero.
Criteria from 40 CFR Part 136 for Method 624 and were calculated assuming a
QC check sample concentration of 20 ng/l. These criteria are based directly
upon the method performance data in Table 7. Where necessary, the limits for
recovery have been broadened to assure applicability of the limits to
concentrations below those used to develop Table 7.
8240B - 37
Revision 2
September 1994
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TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Parameter
Benzene
Bromodi chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
Chloroethane
2-Chloroethyl vinyl ether8
Chloroform
Chl oromethane
Di bromochl oromethane
1 , 2 -Di chl orobenzeneb
1,3-Dichlorobenzene
l,4-Dichlorobenzeneb
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans - 1 , 2 , -Di chl oroethene
1 , 2-Di chl oropropane8
cis-1 ,3-Dichloropropenea
trans- 1,3-Di chl oropropene"
Ethyl benzene
Methyl ene chloride
1,1,2,2-Tetrachloroethane
Tetrachl oroethene
Toluene
1,1,1-Tri chloroethane
1,1, 2 -Tri chloroethane
Trichloroethene
Trichlorofluoromethane
Vinyl chloride
Accuracy, as
recovery, x'
(M9/L)
0.93C+2.00
1.03C-1.58
1.18C-2.35
l.OOC
1.10C-1.68
0.98C+2.28
1.18C+0.81
l.OOC
0.93C+0.33
1.03C-1.81
1.01C-0.03
0.94C+4.47
1.06C+1.68
0.94C+4.47
1.05C+0.36
1.02C+0.45
1.12C+0.61
1.05C+0.03
l.OOC
l.OOC
l.OOC
0.98C+2.48
0.87C+1.88
0.93C+1.76
1.06C+0.60
0.98C+2.03
1.06C+0.73
0.95C+1.71
1.04C+2.27
0.99C+0.39
l.OOC
Single analyst
precision, sr'
(M9/L)
0.26X-1.74
0.15X+0.59
0.12X+0.34
0.43x
0.12X+0.25
O.lSx-0.09
O.Hx+2.78
0.62x
0.16x+0.22
0.37X+2.14
0.17X-0.18
0.22X-1.45
O.Hx-0.48
0.22X-1.45
0.13X-0.05
0.17x-0.32
0.17X+1.06
0.14X+0.09
0.33x
0.38x
0.25x
0.14X+1.00
0.15X+1.07
0.16X+0.69
0.13X-0.18
0.15X-0.71
0.12X-0.15
0.14x+0.02
O.lSx+0.36
0.33X-1.48
0.48x
Overall
precision,
S' (/*g/L)
0.25x-1.33
0.20x+1.13
0.17X+1.38
0.58x
O.llx+0.37
0.26x-1.92
0.29X+1.75
0.84x
O.lSx+0.16
0.58x+0.43
0.17X+0.49
0.30X-1.20
O.lSx-0.82
0.30x-1.20
0.16X+0.47
0.21X-0.38
0.43X-0.22
0.19X+0.17
0.45x
0.52x
0.34X
0.26X-1.72
0.32X+4.00
0.20X+0.41
0.16X-0.45
0.22x-1.71
0.21X-0.39
O.lSx+0.00
0.12X+0.59
0.34x-0.39
0.65x
x' = Expected recovery for one or more measurements of a sample
containing a concentration of C, in jug/L.
sr' = Expected single analyst standard deviation of measurements at an
average concentration of x, in jug/L-
S' = Expected interlaboratory standard deviation of measurements at an
average concentration found of x, in /xg/L.
C = True value for the concentration, in M9/L-
x = Average recovery found for measurements of samples containing a
concentration of C, in fj.g/1.
a Estimates based upon the performance in a single laboratory.
b Due to chromatographic resolution problems, performance statements for
these isomers are based upon the sums of their concentrations.
8240B - 38
Revision 2
September 1994
-------
TABLE 8.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Low/High Low/High
Surrogate Compound Water Soil/Sediment
4-Bromofluorobenzene 86-115 74-121
l,2-Dichloroethane-d4 76-114 70-121
Toluene-dQ 88-110 81-117
8240B - 39 Revision 2
September 1994
-------
FIGURE 1.
PURGING CHAMBER
FOAMTN*
GOT M W. OJ)
14 MM O.O.
INIET 1M IN. O.O.
EXT 1M M. O.O
10 MM OLA88
MEDIUM HJKJtirt
SAMPLE INIXT
»WAY SYNNOE VALVC
17 CM aO OAUOE SVWNQE NEEOU
• MM O.O flUeSER SEPTUM
INlfT 1M IN O.O.
o o
13X
MOt^CUiAR SICVC
PU*WE GAS FUTCT
PUMQEQAS-
FLOWCONTWX
8240B - 40
Revision 2
September 1994
-------
FIGURE 2.
TRAP PACKINGS AND CONSTRUCTION TO INCLUDE
DESORB CAPABILITY FOR METHOD 8240B
CONSTRUCTION OCTML
8240B - 41
Revision 2
September 1994
-------
FIGURE 3.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - PURGE MODE FOR METHOD 8240B
GAMIER GAS
FLOW CONTROL
PRESSURE
REGULATOR
PURGE QAS
PLOW CONTROL
13X MOLECULAR
SIEVE FILTER
LIQUID INJECTION PORTS
r— COLUMN OVEN
LJUUVr
CONFIRMATORY COLUMN
TO DETECTOR
- ANALYTICAL COLUMN
OPTIONAL 4PORT COLUMN
SELECTION VALVE
TRAP INLET
TRAP
2TC
PURGING
DEVICE
NOTE
ALL LINES BETWEEN TRAP
AMD QC SHOULD BE HEATED
TOID-C.
8240B - 42
Revision 2
September 1994
-------
FIGURE 4.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - DESORB MODE FOR METHOD 8240B
CARRKRGAS
FLOW CONTROL
PRESSURE
REGULATOR
UOWO INJECTION PORTS
-COLUMN OVEN
PURGE GAS
FLOW CONTROL
13X MOLECULAR
SIEVE FILTER
CONFIRMATORY COLUMN
TO DETECTOR
ANALYTICAL COLUMN
OPTIONAL tPORT COLUMN
SELECTION VALVE
TRAP INLET
TRAP
2QO*C
PURQMG
DEVICE
NOTE
ALL LINES BETWEEN TRAP
AND GC SHOULD BE HEATED
TO WC.
8240B - 43
Revision 2
September 1994
-------
FIGURE 5.
LOW SOILS IMPINGER
PURGE INLET FITTING
SAMPLE OUTLET FITTING
9" i 6mm 0 0 CLASS TUBING f fc-
SEPTUM
CAP
40ml VIAL
8240B - 44
Revision 2
September 1994
-------
METHOD 8240B
VOLATILE ORGANICS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS)
Water
Miscible
Liquids
Soil/Sediment
and Waste
Samples
7.1
Select
procedure for
introducing
sample into
GC/MS.
7.4.2.1
Dilute sample
at least 50
fold with
water.
7.4.3 Screen
sample using
Method 3810
or 3820.
Direct
Injection
screening
method for the
waste
matrix.
Water
Samples
rurge-and-trap
7.4.1.1
Screen sample
using Method
3810 or 3820.
7.2.1
Set GC/MS
operating
conditions.
7.4.1.7
Perform
secondary
dilutions.
7.2.4 Connect
purge-and-trap
device to GC.
7.2.6 Perform
purge-and-trap
analysis.
7.4.1.8 Add
internal standard
and surrogate
spiking solutions
7.2.8
Calculate RFs
for 5 SPCCs.
7.4.1.10
Perform
purge-and-trap
procedure.
7.3 Perform
daily
calibration
using SPCCs
and CCCs.
8240B - 45
Revision 2
September 1994
-------
METHOD 8240B
(continued)
7.4.3
Is
concentration
> 1 mg/Kg?
7.4.3.1.1
Choose sample
size based on
estimated
concentration.
7.4.3.1.3 Add
internal standard
and surrogate
spiking solutions.
7.4.3.1.5
Determine
percent dry
weight of
sample.
7.4.3.1.7
Perform
purge-and-trap
procedure.
7.4.3.2 Choose
solvent for
extraction or
dilution. Weigh
sample.
7.4.3.2.2 Add
solvent,
shake.
7.4.3.2.7
Perform
purge-and-trap
procedure.
7.4.1.11
Attach trap
to GC and
perform
analysis.
7.5.1.1 Indentify
analytes by
comparing the
sample retention
time and sample
mass spectra.
7.5.2.2 Calculate
the concentration
of each identified
analyte.
7.5.2.4
Report all
results.
f Stop J
8240B - 46
Revision 2
September 1994
-------
METHOD 8250A
SEMIVOLATILE ORGANIC COMPOUNDS BV GAS CHROMAT06RAPHY/MASS SPECTROMETRY (GC/MS)
1.0 SCOPE AND APPLICATION
1.1 Method 8250 is used to determine the concentration of semivolatile
organic compounds in extracts prepared from all types of solid waste matrices,
soils, and ground water. Direct injection of a sample may be used in limited
applications. The following compounds can be determined by this method:
Appropriate Preparation Techniques
Compounds
CAS Noa 3510
8250A - 1
3520 3540/ 3550 3580
3541
Acenaphthene
Acenaphthene-d10 (I.S.)
Acenaphthylene
Acetophenone
Aldrin
4-Aminobiphenyl
Aniline
Anthracene
Aroclor - 1016 (PCB-1016)
Aroclor - 1221 (PCB-1221)
Aroclor - 1232 (PCB-1232)
Aroclor - 1242 (PCB-1242)
Aroclor - 1248 (PCB-1248)
Aroclor - 1254 (PCB-1254)
Aroclor - 1260 (PCB-1260)
Benzidine
Benzoic acid
Benz(a)anthracene
Benzo(b)f 1 uoranthene
Benzo(k)fluoranthene
Benzo(g,h,i)perylene
Benzo(a)pyrene
Benzyl alcohol
a-BHC
iS-BHC
S-BHC
7-BHC (Lindane)
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
Butyl benzyl phthalate
83-32-9
208-96-8
98-86-2
309-00-2
92-67-1
62-53-3
120-12-7
12674-11-2
11104-28-2
11141-16-5
53469-21-9
12672-29-6
11097-69-1
11096-82-5
92-87-5
65-85-0
56-55-3
205-99-2
207-08-9
191-24-2
50-32-8
100-51-6
319-84-6
319-85-7
319-86-8
58-89-9
111-91-1
111-44-4
108-60-1
117-81-7
101-55-3
85-68-7
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
ND
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
ND
ND
X
X
X
X
X
X
X
X
CP
ND
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
ND
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
Revision 1
September 1994
-------
Compounds
Appropriate Preparation Techniques
CAS No" 3510 3520 3540/ 3550 3580
3541
Chlordane (technical)
4-Chloroaniline
1 -Chi oronaphthal ene
2-Chloronaphthalene
4-Chl oro- 3 -methyl phenol
2-Chlorophenol
4-Chlorophenyl phenyl ether
Chrysene
Chrysene-d12 (I.S.)
4,4'-DDD
4,4'-DDT
4, 4' -DDE
Dibenz(a,j)acridine
Di benz ( a , h ) anthracene
Dibenzofuran
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
l,4-Dichlorobenzene-d4 (I.S)
3,3'-Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorophenol
Dieldrin
Diethyl phthalate
Di methyl ami noazobenzene
7,12-Dimethylbenz(a)-
anthracene
a , o-Dimethyl phenethyl ami ne
2,4-Dimethylphenol
Dimethyl phthalate
4, 6-Dinitro-2-methyl phenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di phenyl ami ne
1, 2 -Di phenyl hydrazine
Di-n-octyl phthalate
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Ethyl methanesulfonate
57-74-9
106-47-8
90-13-1
91-58-7
59-50-7
95-57-8
7005-72-3
218-01-9
72-54-8
50-29-3
72-55-9
224-42-0
53-70-3
132-64-9
84-74-2
95-50-1
541-73-1
106-46-7
3855-82-1
91-94-1
120-83-2
87-65-0
60-57-1
84-66-2
60-11-7
57-97-6
122-09-8
105-67-9
131-11-3
534-52-1
51-28-5
121-14-2
606-20-2
122-39-4
122-66-7
117-84-0
959-98-8
33213-65-9
1031-07-8
72-20-8
7421-93-4
53494-70-5
62-50-0
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP(45)
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
ND
X
X
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
ND
X
X
X
X
X
X
X
X
X
ND
X
ND
X
X
X
X
X
X
X
ND
X
X
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
ND
X
ND
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
ND
X
X
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
8250A - 2
Revision 1
September 1994
-------
Compounds
Appropriate Preparation Techniques
CAS Noa 3510 3520 3540/ 3550 3580
3541
Fluoranthene
Fluorene
2-Fluorobiphenyl (surr.)
2-Fluorophenol (surr.)
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorobutadiene
Hexachlorocyclopentadiene
Hexachloroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Methoxychlor
3-Methyl chol anthrene
Methyl methanesulfonate
2-Methylnaphthalene
2-Methyl phenol
4-Methylphenol
Naphthalene
Naphthalene-d8 (I.S.)
1-Naphthylamine
2-Naphthylamine
2-Nitroanil ine
3-Nitroaniline
4-Nitroaniline
Nitrobenzene
Nitrobenzene-d5 (surr.)
2-Nitrophenol
4-Nitrophenol
N-Nitrosodi butyl ami ne
N-Nitrosodi methyl ami ne
N-Nitrosodiphenylamine
N-Nitrosodi-n-propylamine
N-Nitrosopi peri dine
Pentachl orobenzene
Pentachloronitrobenzene
Pentachlorophenol
Perylene-d12 (I.S.)
Phenacetin
Phenanthrene
Phenanthrene-d10 (I.S.)
Phenol
Phenol -d6 (surr.)
2-Picoline
Pronamide
206-44-0
86-73-7
321-60-8
367-12-4
76-44-8
1024-57-3
118-74-1
87-68-3
77-47-4
67-72-1
193-39-5
78-59-1
72-43-5
56-49-5
66-27-3
91-57-6
95-48-7
106-44-5
91-20-3
1146-65-2
134-32-7
91-59-8
88-74-4
99-09-2
100-01-6
98-95-3
4165-60-0
88-75-5
100-02-7
924-16-3
62-75-9
86-30-6
621-64-7
100-75-4
608-93-5
82-68-8
87-86-5
198-55-0
62-44-2
85-01-8
108-95-2
13127-88-3
109-06-8
23950-58-5
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
OS(44)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
DC(28)
DC(28)
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
X
ND
NO
X
X
ND
ND
X
X
X
X
X
X
X
ND
X
X
X
ND
ND
ND
X
X
ND
X
X
X
X
ND
ND
X
X
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
X
X
X
X
ND
X
X
X
ND
ND
ND
X
X
ND
X
X
X
X
ND
ND
X
X
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
X
ND
ND
X
X
ND
ND
X
X
X
X
X
X
X
ND
X
X
X
ND
ND
ND
X
X
ND
X
X
X
X
ND
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
8250A - 3
Revision 1
September 1994
-------
Appropriate Preparation Techniques
Compounds CAS No" 3510 3520 3540/ 3550 3580
3541
Pyrene
Terphenyl -d14(surr. )
1,2,4,5-Tetrachlorobenzene
2,3,4,6-Tetrachlorophenol
Toxaphene
2,4,6-Tribromopheno'l (surr. )
1,2,4-Trichlorobenzene
2 , 4 , 5-Tri chl orophenol
2,4,6-Trichlorophenol
129-00-0
1718-51-0
95-94-3
58-90-2
8001-35-2
118-79-6
120-82-1
95-95-4
88-06-2
X
X
X
X
X
X
X
X
X
X
X
ND
ND
X
X
X
X
X
X
ND
ND
ND
X
X
X
ND
X
X
X
ND
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
8 Chemical Abstract Service Registry Number.
CP = Nonreproducible chromatographic performance.
DC = Unfavorable distribution coefficient (number in parenthesis is
percent recovery).
ND = Not determined.
OS = Oxidation during storage (number in parenthesis is percent
stability).
X = Greater than 70 percent recovery by this technique.
1.2 Method 8250 can be used to quantitate most neutral, acidic, and
basic organic compounds that are soluble in methylene chloride and capable of
being eluted without derivatization as sharp peaks from a gas chromatographic
packed column. Such compounds include polynuclear aromatic hydrocarbons,
chlorinated hydrocarbons and pesticides, phthalate esters, organophosphate
esters, nitrosamines, haloethers, aldehydes, ethers, ketones, anilines,
pyridines, quinolines, aromatic nitro compounds, and phenols, including
nitrophenols. See Table 1 for a list of compounds and their characteristic ions
that have been evaluated on the specified GC/MS system.
1.3 The following compounds may require special treatment when being
determined by this method. Benzidine can be subject to oxidative losses during
solvent concentration. Also, chromatography is poor. Under the alkaline
conditions of the extraction step, a-BHC, 7-BHC, endosulfan I and II, and endrin
are subject to decomposition. Neutral extraction should be performed if these
compounds are expected and are not being determined by Method 8080.
Hexachlorocyclopentadiene is subject to thermal decomposition in the inlet of the
gas chromatograph, chemical reaction in acetone solution, and photochemical
decomposition. N-nitrosodimethylamine is difficult to separate from the solvent
under the chromatographic conditions described. N-nitrosodiphenylamine
decomposes in the gas chromatographic inlet and cannot be separated from
diphenylamine. Pentachlorophenol, 2,4-dinitrophenol, 4-nitrophenol, 4,6-dinitro-
2-methylphenol, 4-chloro-3-methylphenol, benzoic acid, 2-nitroaniline,
8250A - 4 Revision 1
September 1994
-------
3-nitroaniline, 4-chloroaniline, and benzyl alcohol are subject to erratic
chromatographic behavior, especially if the GC system is contaminated with high
boiling material.
1.4 The estimated quantitation limit (EQL) of Method 8250 for
determining an individual compound is approximately 1 mg/kg (wet weight) for
soil/sediment samples, 1-200 mg/kg for wastes (dependent on matrix and method of
preparation), and 10 /ug/L for ground water samples (see Table 2). EQLs will be
proportionately higher for sample extracts that require dilution to avoid
saturation of the detector.
1.5 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatograph/mass spectrometers and
skilled in the interpretation of mass spectra. Each analyst must demonstrate the
ability to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Prior to using this method, the samples should be prepared for
chromatography using the appropriate sample preparation and cleanup methods.
This method describes chromatographic conditions that will allow for the
separation of the compounds in the extract.
3.0 INTERFERENCES
3.1 Raw GC/MS data from all blanks, samples, and spikes must be
evaluated for interferences. Determine if the source of interference is in the
preparation and/or cleanup of the samples and take corrective action to eliminate
the problem.
3.2 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
sample syringe must be rinsed out between samples with solvent. Whenever an
unusually concentrated sample is encountered, it should be followed by the
analysis of solvent to check for cross contamination.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph/mass spectrometer system
4.1.1 Gas chromatograph - An analytical system complete with a
temperature-programmable gas chromatograph suitable for splitless
injection and all required accessories, including syringes, analytical
columns, and gases.
4.1.2 Columns
4.1.2.1 For base/neutral compound detection - 2 m x 2
mm ID stainless or glass, packed with 3% SP-2250-DB on 100/120 mesh
Supelcoport or equivalent.
8250A - 5 Revision 1
September 1994
-------
4.1.2.2 For acid compound detection - 2 m x 2 mm ID glass,
packed with 1% SP-1240-DA on 100/120 mesh Supelcoport or equivalent.
4.1.3 Mass spectrometer - Capable of scanning from 35 to 500 amu
every 1 second or less, using 70 volts (nominal) electron energy in the
electron impact ionization mode. The mass spectrometer must be capable
of producing a mass spectrum for decafluorotriphenylphosphine (DFTPP)
which meets all of the criteria in Table 3 when 1 /zL of the GC/MS tuning
standard is injected through the GC (50 ng of DFTPP).
4.1.4 GC/MS interface - Any GC-to-MS interface that gives acceptable
calibration points at 50 ng per injection for each compound of interest
and achieves acceptable tuning performance criteria may be used. GC-to-MS
interfaces constructed entirely of glass or glass-lined materials are
recommended. Glass may be deactivated by silanizing with
dichlorodimethylsilane.
4.1.5 Data system - A computer system must be interfaced to the mass
spectrometer. The system must allow the continuous acquisition and
storage on machine-readable media of all mass spectra obtained throughout
the duration of the chromatographic program. The computer must have
software that can search any GC/MS data file for ions of a specific mass
and that can plot such ion abundances versus time or scan number. This
type of plot is defined as an Extracted Ion Current Profile (EICP).
Software must also be available that allows integrating the abundances in
any EICP between specified time or scan-number limits. The most recent
version of the EPA/NIH Mass Spectral Library should also be available.
4.2 Syringe - 10 pi.
5.0 REAGENTS
5.1 Reagent, grade chemicals shall be used in all tests. Unless otherwise
indicated, it is intended that all reagents shall conform to the specifications
of the Committee on Analytical Reagents of the American Chemical Society, where
such specifications are available. Other grades may be used, provided it is first
ascertained that the reagent is of sufficiently high purity to permit its use
without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Stock standard solutions (1000 mg/L) - Standard solutions can be
prepared from pure standard materials or purchased as certified solutions.
5.3.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure material. Dissolve the material in pesticide quality
acetone or other suitable solvent and dilute to volume in a 10 ml
volumetric flask. Larger volumes can be used at the convenience of the
analyst. When compound purity is assayed to be 96% or greater, the weight
may be used without correction to calculate the concentration of the stock
standard. Commercially prepared stock standards may be used at any
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concentration if they are certified by the manufacturer or by an
independent source.
5.3.2 Transfer the stock standard solutions into bottles with Teflon
lined screw-caps or crimp tops. Store at -10°C to -20°C or less and
protect from light. Stock standard solutions should be checked frequently
for signs of degradation or evaporation, especially just prior to
preparing calibration standards from them.
5.3.3 Stock standard solutions must be replaced after 1 year or
sooner if comparison with quality control check samples indicates a
problem.
5.4 Internal standard solutions - The internal standards recommended are
l,4-dichlorobenzene-d4, naphthalene-d8, acenaphthene-d10, phenanthrene-d10,
cnrysene-d12, and perylene-d12. Other compounds may be used as internal standards
as long as the requirements given in Sec. 7.3.2 are met. Dissolve 200 mg of each
compound with a small volume of carbon disulfide. Transfer to a 50 ml volumetric
flask and dilute to volume with methylene chloride so that the final solvent is
approximately 20% carbon disulfide. Most of the compounds are also soluble in
small volumes of methanol, acetone, or toluene, except for perylene-d12. The
resulting solution will contain each standard at a concentration of 4,000 ng//nL.
Each 1 ml sample extract undergoing analysis should be spiked with 10 juL of the
internal standard solution, resulting in a concentration of 40 ng//iL of each
internal standard. Store at -10°C to -20°C or less when not being used.
5.5 GC/MS tuning standard - A methylene chloride solution containing
50 ng//LtL of decafluorotriphenylphosphine (DFTPP) should be prepared. The
standard should also contain 50 ng/juL each of 4,4'-DDT, pentachlorophenol, and
benzidine to verify injection port inertness and GC column performance. Store
at 4°C or less when not being used.
5.6 Calibration standards - Calibration standards at a minimum of five
concentrations should be prepared. One of the calibration standards should be
at a concentration near, but above, the method detection limit; the others should
correspond to the range of concentrations found in real samples but should not
exceed the working range of the GC/MS system. Each standard should contain each
analyte for detection by this method (e.g. some or all of the compounds listed
in Table 1 may be included). Each 1 ml aliquot of calibration standard should
be spiked with 10 juL of the internal standard solution prior to analysis. All
standards should be stored at -10°C to -20°C and should be freshly prepared once
a year, or sooner if check standards indicate a problem. The daily calibration
standard should be prepared weekly and stored at 4°C.
5.7 Surrogate standards - The recommended surrogate standards are
phenol-de, 2-fluorophenol, 2,4,6-tribromophenol, nitrobenzene-d5, 2-
fluorobiphenyl, and p-terphenyl-d14. See Method 3500 for the instructions on
preparing the surrogate standards. Determine what concentration should be in the
blank extracts after all extraction, cleanup, and concentration steps. Inject
this concentration into the GC/MS to determine recovery of surrogate standards
in all blanks, spikes, and sample extracts. Take into account all dilutions of
sample extracts.
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5.8 Matrix spike standards - See Method 3500 for instructions on
preparing the matrix spike standard. Determine what concentration should be in
the blank extracts after all extraction, cleanup, and concentration steps.
Inject this concentration into the GC/MS to determine recovery of standards in
all matrix spikes. Take into account all dilutions of sample extracts.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Sample preparation - Samples must be prepared by one of the
following methods prior to GC/MS analysis.
Matrix Methods
Water 3510, 3520
Soil/sediment 3540, 3541, 3550
Waste 3540, 3541, 3550, 3580
7.1.1 Direct injection - In very limited applications direct
injection of the sample into the GC/MS system with a 10 p.1 syringe may be
appropriate. The detection limit is very high (approximately
10,000 Mg/L); therefore, it is only permitted where concentrations in
excess of 10,000 /Ltg/L are expected. The system must be calibrated by
direct injection.
7.2 Extract cleanup - Extracts may be cleaned up by any of the following
methods prior to GC/MS analysis.
Compounds Methods
Phenols 3630, 3640, 8040a
Phthalate esters 3610, 3620, 3640
Nitrosamines 3610, 3620, 3640
Organochlorine pesticides & PCBs 3620, 3640, 3660
Nitroaromatics and cyclic ketones 3620, 3640
Polynuclear aromatic hydrocarbons 3611, 3630, 3640
Haloethers 3620, 3640
Chlorinated hydrocarbons 3620, 3640
Organophosphorus pesticides 3620
Petroleum waste 3611, 3650
All basic, neutral, and acidic
Priority Pollutants 3640
aMethod 8040 includes a derivatization technique followed by GC/ECD analysis, if
interferences are encountered on GC/FID.
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7.3 Recommended GC/MS operating conditions
Electron energy: 70 volts (nominal)
Mass range: 35-500 amu
Scan time: 1 sec/scan
Injector temperature: 250-300°C
Transfer line temperature: 250-300°C
Source temperature: According to manufacturer's specifications
Injector: Grob-type, splitless
Sample volume: 1-2 Mi-
Carrier gas: Helium at 30 mL/min
Conditions for base/neutral analysis (3% SP-2250-DB):
Initial column temperature and hold time: 50°C for 4 minutes
Column temperature program: 50-300°C at 8°C/min
Final column temperature hold: 300°C for 20 minutes
Conditions for acid analysis (1% SP-1240-DA):
Initial column temperature and hold time: 70°C for 2 minutes
Column temperature program: 70-200°C at 8°C/min
Final column temperature hold: 200°C for 20 minutes
7.4 Initial calibration
7.4.1 Each GC/MS system must be hardware-tuned to meet the criteria
in Table 3 for a 50 ng injection of DFTPP. Analyses should not begin
until all these criteria are met. Background subtraction should be
straightforward and designed only to eliminate column bleed or instrument
background ions. The GC/MS tuning standard should also be used to assess
GC column performance and injection port inertness. Degradation of DDT
to DDE and ODD should not exceed 20% (See Sec. 7.4.5 of Method 8080).
Benzidine and pentachlorophenol should be present at their normal
responses, and no peak tailing should be visible. If degradation is
excessive and/or poor chromatography is noted, the injection port may
require cleaning.
7.4.2 The internal standards selected in Sec. 5.1 should permit most
of the components of interest in a chromatogram to have retention times
of 0.80-1.20 relative to one of the internal standards. Use the base peak
ion from the specific internal standard as the primary ion for
quantitation (see Table 1). If interferences are noted, use the next most
intense ion as the quantitation ion (i.e. for l,4-dichlorobenzene-d4 use
m/z 152 for quantitation).
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7.4.3 Analyze 1 /zL of each calibration standard (containing internal
standards) and tabulate the area of the primary characteristic ion against
concentration for each compound (as indicated in Table 1). Calculate
response factors (RFs) for each compound relative to the internal standard
as follows:
RF - (AxCis)/(AisCJ
where:
Ax = Area of the characteristic ion for the compound being
measured.
Ais = Area of the characteristic ion for the specific internal
standard.
Cx = Concentration of the compound being measured (ng//iL).
Cjs = Concentration of the specific internal standard (ng/juL).
7.4.4 A system performance check must be performed to ensure that
minimum average response factors, calculated as the mean of the 5
individual relative response factors, are met before the calibration curve
is used. For semivolatiles, the System Performance Check Compounds
(SPCCs) are: N-nitroso-di-n-propylamine; hexachlorocyclopentadiene;
2,4-dinitrophenol; and 4-nitrophenol. The minimum acceptable average RF
for these compounds is 0.050. These SPCCs typically have very low RFs
(0.1-0.2) and tend to decrease in response as the chromatographic system
begins to deteriorate or the standard material begins to deteriorate.
They are usually the first to show poor performance. Therefore, they must
meet the minimum requirement when the system is calibrated.
7.4.4.1 The percent relative standard deviation should be
less than 15% for each compound. However, the %RSD for each
individual Calibration Check Compound (CCC) (see Table 4) must be
less than 30%. The relative retention times of each compound in
each calibration run should agree within 0.06 relative retention
time units. Late-eluting compounds usually have much better
agreement.
SD
%RSD = — x 100
RF
where:
RSD = relative standard deviation.
RF = mean of 5 initial RFs for a compound.
SD = standard deviation of average RFs for a compound.
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SD =
N (RFj - RF)
I
1=1 N - 1
2
where:
RFj = RF for each of the 5 calibration levels
N = Number of RF values (i.e., 5)
7.4.4.2 If the %RSD of any CCC is 30% or greater, then the
chromatographic system is too reactive for analysis to begin. Clean
or replace the injector liner and/or capillary column, then repeat
the calibration procedure beginning with Sec. 7.4.
7.4.5 Linearity - If the %RSD of any compound is 15% or less, then
the relative response factor is assumed to be constant over the
calibration range, and the average relative response factor may be used
for quantitation (Sec. 7.7.2).
7.4.5.1 If the %RSD of any compound is greater than 15%,
construct calibration curves of area ratio (A/Ais) versus
concentration using first or higher order regression fit of the five
calibration points. The analyst should select the regression order
which introduces the least calibration error into the quantitation
(Sees. 7.7.2.2 and 7.7.2.3). The use of calibration curves is a
recommended alternative to average response factor calibration, and
a useful diagnostic of standard preparation accuracy and absorption
activity in the chromatographic system.
7.5 Daily GC/MS calibration
7.5.1 Prior to analysis of samples, the GC/MS tuning standard must
be analyzed. A 50 ng injection of DFTPP must result in a mass spectrum
for DFTPP which meets the criteria given in Table 3. These criteria must
be demonstrated during each 12 hour shift.
7.5.2 A calibration standard(s) at mid-concentration containing all
semivolatile analytes, including all required surrogates, must be
analyzed every 12 hours during analysis. Compare the instrument response
factor from the standards every 12 hours with the SPCC (Sec. 7.5.3) and
CCC (Sec. 7.5.4) criteria.
7.5.3 System Performance Check Compounds (SPCCs) - A system
performance check must be made during every 12 hour shift. If the SPCC
criteria are met, a comparison of response factors is made for all
compounds. This is the same check that is applied during the initial
calibration. If the minimum response factors are not met, the system must
be evaluated, and corrective action must be taken before sample analysis
begins. The minimum RF for semivolatile SPCCs is 0.050. Some possible
problems are standard mixture degradation, injection port inlet
contamination, contamination at the front end of the analytical column,
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and active sites in the column or chromatographic system. This check must
be met before analysis begins.
7.5.4 Calibration Check Compounds (CCCs): After the system
performance check is met, CCCs listed in Table 4 are used to check the
validity of the initial calibration.
Calculate the percent drift using:
C - C
% Drift = - - - x 100
where:
C, = Calibration Check Compound standard concentration.
Cc = Measured concentration using selected quantitation method.
If the percent difference for each CCC is less than or equal to 20%,
the initial calibration is assumed to be valid. If the criterion is not
met (> 20% drift) for any one CCC, corrective action must be taken.
Problems similar to those listed under SPCCs could affect this criterion.
If no source of the problem can be determined after corrective action has
been taken, a new five-point calibration must be generated. This
criterion must be met before sample analysis begins. If the CCCs are not
analytes required by the permit, then all required analytes must meet the
20% drift criterion.
7.5.5 The internal standard responses and retention times in the
calibration check standard must be evaluated immediately after or during
data acquisition. If the retention time for any internal standard changes
by more than 30 seconds from the last daily calibration (Sec. 7.4), the
chromatographic system must be inspected for malfunctions and corrections
must be made., as required. If the EICP area for any of the internal
standards changes by a factor of two (-50% to +100%) from the last daily
calibration check standard, the mass spectrometer must be inspected for
malfunctions and corrections must be made, as appropriate.
7.6 GC/MS analysis
7.6.1 It is highly recommended that the extract be screened on a
GC/FID or GC/PID using the same type of column. This will minimize
contamination of the GC/MS system from unexpectedly high concentrations
of organic compounds.
7.6.2 Spike the 1 ml extract obtained from sample preparation with
10 juL of the internal standard solution (Sec. 5.4) just prior to analysis.
7.6.3 Analyze the 1 ml extract by GC/MS using the appropriate column
(as specified in Sec. 4.1.2). The recommended GC/MS operating conditions
to be used are specified in Sec. 7.3.
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7.6.4 If the response for any quantitation ion exceeds the initial
calibration curve range of the GC/MS system, extract dilution must take
place. Additional internal standard must be added to the diluted extract
to maintain the required 40 ng/juL of each internal standard in the
extracted volume. The diluted extract must be reanalyzed.
7.6.5 Perform all qualitative and quantitative measurements as
described in Sec. 7.7. Store the extracts at 4°C, protected from light
in screw-cap vials equipped with unpierced Teflon lined septa.
7.7 Data interpretation
7.7.1 Qualitative analysis
7.7.1.1 The qualitative identification of compounds
determined by this method is based on retention time, and on
comparison of the sample mass spectrum, after background correction,
with characteristic ions in a reference mass spectrum. The
reference mass spectrum must be generated by the laboratory using
the conditions of this method. The characteristic ions from the
reference mass spectrum are defined to be the three ions of greatest
relative intensity, or any ions over 30% relative intensity if less
than three such ions occur in the reference spectrum. Compounds
should be identified as present when the criteria below are met.
7.7.1.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographic peak containing ions
specific for the target compound at a compound-specific
retention time will be accepted as meeting this criterion.
7.7.1.1.2 The RRT of the sample component is within
± 0.06 RRT units of the RRT of the standard component.
7.7.1.1.3 The relative intensities of the
characteristic ions agree within 30% of the relative
intensities of these ions in the reference spectrum.
(Example: For an ion with an abundance of 50% in the
reference spectrum, the corresponding abundance in a sample
spectrum can range between 20% and 80%.)
7.7.1.1.4 Structural isomers that produce very
similar mass spectra should be identified as individual
isomers if they have sufficiently different GC retention
times. Sufficient GC resolution is achieved if the height of
the valley between two isomer peaks is less than 25% of the
sum of the two peak heights. Otherwise, structural isomers
are identified as isomeric pairs.
7.7.1.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce
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mass spectra containing ions contributed by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
appropriate selection of analyte spectra and background
spectra is important. Examination of extracted ion current
profiles of appropriate ions can aid in the selection of
spectra, and in qualitative identification of compounds. When
analytes coelute (i.e., only one chromatographic peak is
apparent), the identification criteria can be met, but each
analyte spectrum will contain extraneous ions contributed by
the coeluting compound.
7.7.1.2 For samples containing components not associated
with the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the purpose of the
analyses being conducted. Computer generated library search
routines should not use normalization routines that would
misrepresent the library or unknown spectra when compared to each
other. For example, the RCRA permit or waste delisting requirements
may require the reporting of nontarget analytes. Only after visual
comparison of sample spectra with the nearest library searches will
the mass spectral interpretation specialist assign a tentative
identification. Guidelines for making tentative identification are:
(1) Relative intensities of major ions in the reference
spectrum (ions > 10% of the most abundant ion) should be present in
the sample spectrum.
(2) The relative intensities of the major ions should agree
within + 20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
between 30 and 70%.)
(3) Molecular ions present in the reference spectrum should
be present in sample the spectrum.
(4) Ions present in the sample spectrum but not in the
reference spectrum should be reviewed for possible background
contamination or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the
sample spectrum should be reviewed for possible subtraction from the
sample spectrum because of background contamination or coeluting
peaks. Data system library reduction programs can sometimes create
these discrepancies.
7.7.2 Quantitative Analysis
7.7.2.1 When a compound has been identified, the
quantitation of that compound will be based on the integrated
abundance from the EICP of the primary characteristic ion.
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7.7.2.2 If the %RSD of a compound's relative response
factor is 15% or less, then the concentration in the extract may be
determined using the average response factor (RF) from initial
calibration data (Sec. 7.4.3) and the following equation:
(Ax x Cis)
Cex (mg/L) =
(Ais x RF)
where Cex is the concentration of the compound in the extract,
and the other terms are as defined in Sec. 7.4.3.
7.7.2.3 Alternatively, the regression line fitted to the
initial calibration (Sec. 7.4.6.1) may be used for determination of
the extract concentration.
7.7.2.4 Compute the concentration of the analyte in the
sample using the equations in Sees. 7.7.2.4.1 and 7.7.2.4.2.
7.7.2.4.1 The concentration of the analyte in the
liquid phase of the sample is calculated using the
concentration of the analyte in the extract and the volume of
liquid extracted, as follows:
Concentration in liquid (^g/L) = |Cex x VeJ
V.
o
where:
Vex = extract volume, in ml
V0 = volume of liquid extracted, in L.
7.7.2.4.2 The concentration of the analyte in the
solid phase of the sample is calculated using the
concentration of the pollutant in the extract and the weight
of the solids, as follows:
Concentration in solid (/ug/kg) = (Cex x VeK)
where:
Vex = extract volume, in mL
Ws = sample weight, in kg.
7.7.2.5 Where applicable, an estimate of concentration for
noncalibrated components in the sample should be made. The formulae
given above should be used with the following modifications: The
areas Ax and Ais should be from the total ion chromatograms and the
RF for the compound should be assumed to be 1. The concentration
obtained should be reported indicating (1) that the value is an
estimate and (2) which internal standard was used to determine
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concentration. Use the nearest internal standard free of
interferences.
7.7.2.6 Quantitation of multicomponent compounds (e.g.
Aroclors) is beyond the scope of Method 8250A. Normally,
quantitation is performed using a GC/ECD by Method 8080.
8.0 QUALITY CONTROL
8.1 Each laboratory that uses these methods is required to operate a
formal quality control program. The minimum requirements of this program consist
of an initial demonstration of laboratory capability and an ongoing analysis of
spiked samples to evaluate and document data quality. The laboratory must
maintain records to document the quality of the data generated. Ongoing data
quality checks are compared with established performance criteria to determine
if the results of analyses meet the performance characteristics of the method.
When results of sample spikes indicate atypical method performance, a quality
control check standard must be analyzed to confirm that the measurements were
performed in an in-control mode of operation.
8.2 Before processing any samples, the analyst should demonstrate,
through the analysis of a reagent water blank, that interferences from the
analytical system, glassware, and reagents are under control. Each time a set
of samples is extracted or there is a change in reagents, a reagent water blank
should be processed as a safeguard against chronic laboratory contamination. The
blank samples should be carried through all stages of the sample preparation and
measurement steps.
8.3 The experience of the analyst performing GC/MS analyses is
invaluable to the success of the methods. Each day that analysis is performed,
the daily calibration standard should be evaluated to determine if the
chromatographic system is operating properly. Questions that should be asked
are: Do the peaks look normal?; Is the response obtained comparable to the
response from previous calibrations? Careful examination of the standard
chromatogram can indicate whether the column is still good, the injector is
leaking, the injector septum needs replacing, etc. If any changes are made to
the system (e.g. column changed), recalibration of the system must take place.
8.4 Required instrument QC is found in the following section:
8.4.1 The GC/MS system must be tuned to meet the DFTPP
specifications in Sec. 7.3.1 and 7.4.1.
8.4.2 There must be an initial calibration of the GC/MS system as
specified in Sec. 7.4.
8.4.3 The GC/MS system must meet the SPCC criteria specified in Sec.
7.5.3 and the CCC criteria in Sec. 7.5.4, each 12 hr.
8.5 To establish the ability to generate acceptable accuracy and
precision, the analyst must perform the following operations.
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8.5.1 A quality control (QC) check sample concentrate is required
containing each analyte at a concentration of 100 mg/L in acetone. The
QC check sample concentrate may be prepared from pure standard materials
or purchased as certified solutions. If prepared by the laboratory, the
QC check sample concentrate must be made using stock standards prepared
independently from those used for calibration.
8.5.2 Using a pipet, prepare QC check samples at a concentration of
100 jug/L by adding 1.00 ml of QC check sample concentrate to each of four
1-L aliquots of organic-free reagent water.
8.5.3 Analyze the well-mixed QC check samples according to the
method beginning in Sec. 7.1 with extraction of the samples.
8.5.4 Calculate the average recovery (x) in /zg/L, and the standard
deviation of the recovery (s) in /xg/L, for each analyte using the four
results.
8.5.5 For each analyte compare s and x with the corresponding
acceptance criteria jfor precision and accuracy, respectively, found in
Table 6. If s and x for all analytes of interest meet the acceptance
criteria, the system performance is acceptable and analysis of actual
samples can begin. If any individual s exceeds the precision limit or any
individual x falls outside the range for accuracy, then the system
performance is unacceptable for that analyte.
NOTE: The large number of analytes in Table 6 present a substantial
probability that one or more will fail at least one of the
acceptance criteria when all analytes of a given method are
analyzed.
8.5.6 When one or more of the analytes tested fail at least one of
the acceptance criteria, the analyst must proceed according to Sees.
8.5.6.1 or 8.5.6.2.
8.5.6.1 Locate and correct the source of the problem and
repeat the test for all analytes of interest beginning with Sec.
8.5.2.
8.5.6.2 Beginning with Sec. 8.5.2, repeat the test only
for those analytes that failed to meet criteria. Repeated failure,
however, will confirm a general problem with the measurement system.
If this occurs, locate and correct the source of the problem and
repeat the test for all compounds of interest beginning with Sec.
8.5.2.
8.6 The laboratory must, on an ongoing basis, analyze a method blank,
a matrix spike, and a matrix spike/duplicate for each analytical batch (up to a
maximum of 20 samples/batch) to assess accuracy. For laboratories analyzing one
to ten samples per month, at least one spiked sample per month is required.
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8.6.1 The concentration of, the spike in the sample should be
determined as follows:
8.6.1.1 If, as in compliance monitoring, the concentration
of a specific analyte in the sample is being checked against a
regulatory concentration limit, the spike should be at that limit
or 1 to 5 times higher than the background concentration determined
in Sec. 8.6.2, whichever concentration would be larger.
8.6.1.2 If the concentration of a specific analyte in the
sample is not being checked against a limit specific to that
analyte, the spike should be at 100 /^g/L or 1 to 5 times higher than
the background concentration determined in Sec. 8.6.2, whichever
concentration would be larger.
8.6.1.3 If it is impractical to determine background
levels before spiking (e.g., maximum holding times will be
exceeded), the spike concentration should be at (1) the regulatory
concentration limit, if any; or, if none (2) the larger of either
5 times higher than the expected background concentration or
100 /xg/L.
8.6.2 Analyze one sample aliquot to determine the background
concentration (B) of each analyte. If necessary, prepare a new QC check
sample concentrate (Sec. 8.5.1) appropriate for the background
concentration in the sample. Spike a second sample aliquot with 1.00 ml
of the QC reference sample concentrate and analyze it to determine the
concentration after spiking (A) of each analyte. Calculate each percent
recovery (p) as 100(A-B)%/T, where T is the known true value of the spike.
8.6.3 Compare the percent recovery (p) for each analyte with the
corresponding QC acceptance criteria found in Table 6. These acceptance
criteria were calculated to include an allowance for error in measurement
of both the background and spike concentrations, assuming a spike to
background ratio of 5:1. This error will be accounted for to the extent
that the analyst's spike to background ratio approaches 5:1. If spiking
was performed at a concentration lower than 100 /ug/U the analyst must use
either the QC acceptance criteria presented in Table 6, or optional QC
acceptance criteria calculated for the specific spike concentration. To
calculate optional acceptance criteria for the recovery of an analyte: (1)
Calculate accuracy (x') using the equation found in Table 7, substituting
the spike concentration (T) for C; (2) calculate overall precision (S')
using the equation in Table 7, substituting x' for x; (3) calculate the
range for recovery at the spike concentration as (100x'/T)
+ 2.44(100S'/T)%.
8.6.4 If any individual p falls outside the designated range for
recovery, that analyte has failed the acceptance criteria. A check
standard containing each analyte that failed the criteria must be analyzed
as described in Sec. 8.7.
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8.7 If any analyte fails the acceptance criteria for recovery in Sec.
8.6, a QC check standard containing each analyte that failed must be prepared and
analyzed.
NOTE: The frequency for the required analysis of a QC check standard will
depend upon the number of analytes being simultaneously tested, the
complexity of the sample matrix, and the performance of the
laboratory. If the entire list of analytes in Table 6 must be
measured in the sample in Sec. 8.6, the probability that the
analysis of a QC check standard will be required is high. In this
case, the QC check standard should be routinely analyzed with the
spiked sample.
8.7.1 Prepare the QC reference sample by adding 1.0 mL of the QC
check sample concentrate (Sec. 8.5.1 or 8.6.2) to 1 L of reagent water.
The QC check standard needs only to contain the analytes that failed
criteria in the test in Sec. 8.6.
8.7.2 Analyze the QC check standard to determine the concentration
measured (A) of each analyte. Calculate each percent recovery (Ps) as
100(A/T)%, where T is the true value of the standard concentration.
8.7.3 Compare the percent recovery (PJ for each analyte with the
corresponding QC acceptance criteria found in Table 6. Only analytes that
failed the test in Sec. 8.6 need to be compared with these criteria. If
the recovery of any such analyte falls outside the designated range, the
laboratory performance for that analyte is judged to be out of control,
and the problem must be immediately identified and corrected. The result
for that analyte in the unspiked sample is suspect and may not be reported
for regulatory compliance purposes.
8.8 As part of the QC program for the laboratory, method accuracy for
each matrix studied must be assessed and records must be maintained. After the
analysis of five spiked samplesJof the same matrix) as in Sec. 8.6, calculate
the average percent recovery (p) and the standard deviation of the percent
recovery (sp). Express the accuracy assessment as a percent recovery interval
from p - 2sp to p + 2sp. If p = 90% and sp = 10%, for example, the accuracy
interval is expressed as 70-110%. Update the accuracy assessment for each
analyte on a regular basis (e.g. after each five to ten new accuracy
measurements).
8.9 To determine acceptable accuracy and precision limits for surrogate
standards the following procedure should be performed.
8.9.1 For each sample analyzed, calculate the percent recovery of
each surrogate in the sample.
8.9.2 Once a minimum of thirty samples of the same matrix have been
analyzed, calculate the average percent recovery (P) and standard
deviation of the percent recovery (s) for each of the surrogates.
8250A - 19 Revision 1
September 1994
-------
8.9.3 For a given matrix, calculate the upper and lower control
limit for method performance for each surrogate standard. This should be
done as follows:
Upper Control Limit (UCL) = P + 3s
Lower Control Limit (LCL) = P - 3s
8.9.4 For aqueous and soil matrices, these laboratory established
surrogate control limits should, if applicable, be compared with the
control limits listed in Table 8. The limits given in Table 8 are multi-
laboratory performance based limits for soil and aqueous samples, and
therefore, the single-laboratory limits established in Step 8.9.3 must
fall within those given in Table 8 for these matrices.
8.9.5 If recovery is not within limits, the following procedures are
required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are
a problem or flag the data as "estimated concentration".
8.9.6 At a minimum, each laboratory should update surrogate recovery
limits on a matrix-by-matrix basis, annually.
8.10 It is recommended that the laboratory adopt additional quality
assurance practices for use with this method. The specific practices that are
most productive depend upon the needs of the laboratory and the nature of the
samples. Field duplicates may be analyzed to assess the precision of the
environmental measurements. When doubt exists over the identification of a peak
on the chromatogram, confirmatory techniques such as gas chromatography with a
dissimilar column or mass spectrometry using other ionization modes must be used.
Whenever possible, the laboratory should analyze standard reference materials and
participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 Method 8250 was tested by 15 laboratories using organic-free reagent
water, drinking water, surface water, and industrial wastewaters spiked at six
concentrations over the range 5-1,300 M9/L. Single operator accuracy and
precision, and method accuracy were found to be directly related to the
concentration of the analyte and essentially independent of the sample matrix.
Linear equations to describe these relationships are presented in Table 7.
8250A - 20 Revision 1
September 1994
-------
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act, Method 625," October 26,
1984.
2. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, July 1985, Revision.
3. Eichelberger, J.W., I.E. Harris, and W.L. Budde, "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass
Spectrometry Systems," Analytical Chemistry, 47, 995-1000, 1975.
4. "Method Detection Limit for Methods 624 and 625," Olynyk, P., W.L. Budde,
and J.W. Eichelberger, Unpublished report, October 1980.
5. "Inter!aboratory Method Study for EPA Method 625-Base/Neutrals, Acids, and
Pesticides," Final Report for EPA Contract 68-03-3102 (in preparation).
6. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
8250A - 21 Revision 1
September 1994
-------
TABLE 1.
CHROMATOGRAPHIC CONDITIONS, METHOD DETECTION LIMITS, AND
CHARACTERISTIC IONS FOR SEMIVOLATILE COMPOUNDS
Compound
Acenaphthene
Acenaphthene-d10 (I.S.)
Acenaphthylene
Acetophenone
Aldrin
4-Aminobiphenyl
Aniline
Anthracene
Aroclor-1016b
Aroclor-1221b
Aroclor-1232b
Aroclor-1242b
Aroclor-1248b
Aroclor-1254b
Aroclor-1260b
Benzidine8
Benzoic acid
Benzo(a)anthracene
Benzo(b)fl uoranthene
Benzo(k)fluoranthene
Benzo (g , h , i ) peryl ene
Benzo(a)pyrene
Benzyl alcohol
a-BHCa
0-BHC
-------
TABLE 1.
(Continued)
Compound
4,4'-DDT
4,4'-DDE
Dibenz(a,j)acridine
Dibenz (a, h) anthracene
Dibenzofuran
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
l,4-Dichlorobenzene-d4 (I.S.)
3 , 3 ' -Di chl orobenz i d i ne
2,4-Dichlorophenol
2,6-Dichlorophenol
Dieldrin
Diethyl phthalate
p-Dimethylaminoazobenzene
7, 12-Dimethylbenz (a) anthracene
a- , a-Di methyl phenethyl ami ne
2,4-Dimethylphenol
Dimethyl phthalate
4,6-Dinitro-2-methylphenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Diphenylamine
1,2-Diphenylhydrazine
Di-n-octyl phthalate
Endosulfan I"
Endosulfan II"
Endosulfan sulfate
Endrin8
Endrin aldehyde
Endrin ketone
Ethyl methanesulfonate
Fluoranthene
Fluorene
2-Fluorobiphenyl (surr.)
2-Fluorophenol (surr.)
Heptachlor
Heptachlor epoxide
Hexachl orobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi enea
Hexachloroethane
Method
Retention Detection Primary Secondary
Time (min) Limit (p.g/1) Ion Ion(s)
29.3
27.2
--
43.2
--
24.7
8.4
7.4
7.8
--
32.2
9.8
--
27.2
20.1
--
--
--
9.4
18.3
16.2
15.9
19.8
18.7
--
--
32.5
26.4
28.6
29.8
27.9
--
--
--
26.5
19.5
--
--
23.4
25.6
21.0
11.4
13.9
8.4
4.7
--
--
2.5
--
2.5
1.9
1.9
4.4
--
16.5
2.7
--
2.5
1.9
--
--
--
2.7
1.6
24
42
5.7
1.9
--
--
2.5
--
--
5.6
--
--
--
2.2
1.9
--
--
1.9
2.2
1.9
0.9
--
1.6
235
246
279
278
168
149
146
146
146
152
252
162
162
79
149
120
256
58
122
163
198
184
165
165
169
77
149
195
337
272
263
67
317
79
202
166
172
112
100
353
284
225
237
117
237,
24,
280,
139,
139
150,
148,
148,
148,
150,
254,
164,
164,
263,
177,
225,
241,
91,
107,
194,
51,
63,
63,
63,
168,
105,
167,
339,
339,
387,
82,
345,
67,
109,
101,
165,
171
64
272,
355,
142,
223,
235,
201,
165
176
277
279
104
111
111
111
115
126
98
98
279
150
77
257
42
121
164
105
154
89
89
167
182
43
341
341
422
81
250
319
97
203
167
274
351
249
227
272
199
8250A - 23
Revision 1
September 1994
-------
TABLE 1.
(Continued)
Compound
Method
Retention Detection
Time (min) Limit (jug/L)
Primary Secondary
Ion Ion(s)
Indeno(l,2,3-cd)pyrene
Isophorone
Methoxychlor
3-Methyl chol anthrene
Methyl methanesulfonate
2-Methyl naphthalene
2-Methyl phenol
4-Methyl phenol
Naphthalene
Naphthalene-d8 (I.S.)
1-Naphthylamine
2-Naphthylamine
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Nitrobenzene
Nitrobenzene-d5 (surr.)
2-Nitrophenol
4-Nitrophenol
N-Nitroso-di-n-butylamine
N-Nitrosodimethylamine"
N-Nitrosodiphenylaminea
N-Nitroso-di-n-propylamine
N-Nitrosopiperidine
Pentachl orobenzene
Pentachloronitrobenzene
Pentachl orophenol
Perylene-d12 (I.S.)
Phenacetin
Phenanthrene
Phenanthrene-d10 (I.S.)
Phenol
Phenol -d6 (surr.)
2-Picoline
Pronamide
Pyrene
Terphenyl-du (surr.)
1,2,4 , 5-Tetrachl orobenzene
2,3,4 , 6-Tetrachl orophenol
42.7
11.9
--
--
--
--
--
--
12.1
--
--
--
--
--
--
11.1
--
6.5
20.3
--
--
20.5
--
._
--
--
17.5
--
--
22.8
--
8.0
--
--
--
27.3
--
--
--
3.7
2.2
--
--
--
--
--
--
1.6
--
--
--
--
--
--
1.9
--
3.6
2.4
--
--
1.9
--
--
--
--
3.6
--
--
5.4
--
1.5
--
--
1.9
--
--
--
276
82
227
268
80
142
108
108
128
136
143
143
65
138
138
77
82
139
139
84
42
169
70
42
250
295
266
264
108
178
188
94
99
93
173
202
244
216
232
138,
95,
228
253,
79,
141
107,
107,
129,
68
115,
115,
92,
108,
108,
123,
128,
109,
109,
57,
74,
168,
130,
114,
252,
237,
264,
260,
109,
179,
94,
65,
42,
66,
175,
200,
122,
214,
230,
227
138
267
65
79
79
127
116
116
138
92
92
65
54
65
65
41
44
167
42
55
248
142
268
265
179
176
80
66
71
92
145
203
212
218
131
8250A - 24
Revision 1
September 1994
-------
TABLE 1.
(Continued)
Compound
Toxapheneb
2,4,6-Tribromophenol (surr.)
1 , 2 , 4-Tri chl orobenzene
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
Retention
Time (min)
25-34
--
11.6
--
11.8
Method
Detection
Limit (jug/L)
_ _.
--
1.9
--
2.7
Primary
Ion
159
330
180
196
196
Secondary
Ion(s)
231, 233
332, 141
182, 145
198, 200
198, 200
aSee Sec. 1.3
bThese compounds are mixtures of various isomers.
(I.S.) = Internal Standard
(surr). = Surrogate
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQL)
FOR VARIOUS MATRICES8
Matrix
Factor
Ground water 10
Low-concentration soil by ultrasonic extraction with GPC cleanup 670
High-concentration soil and sludges by ultrasonic extraction 10,000
Non-water miscible waste 100,000
EQL = [Method detection limit (see Table 1)] X [Factor found in this table].
For non-aqueous samples, the factor is on a wet-weight basis. Sample EQLs
are highly matrix-dependent. The EQLs to be determined herein are provided
for guidance and may not always be achievable.
8250A - 25
Revision 1
September 1994
-------
TABLE 3.
DFTPP KEY IONS AND ION ABUNDANCE CRITERIA8
Mass Ion Abundance Criteria
51 30-60% of mass 198
68 < 2% of mass 69
70 < 2% of mass 69
127 40-60% of mass 198
197 < 1% of mass 198
198 Base peak, 100% relative abundance
199 5-9% of mass 198
275 10-30% of mass 198
365 > 1% of mass 198
441 Present but less than mass 443
442 > 40% of mass 198
443 17-23% of mass 442
"See Reference 3,
8250A - 26 Revision 1
September 1994
-------
TABLE 4.
CALIBRATION CHECK COMPOUNDS
Base/Neutral Fraction Acid Fraction
Acenaphthene 4-Chloro-3-methyl phenol
1,4-Dichlorobenzene 2,4-Dichlorophenol
Hexachlorobutadiene 2-Nitrophenol
N-Nitroso-di-n-phenylamine Phenol
Di-n-octyl phthalate Pentachlorophenol
Benzo(a)pyrene 2,4,6-Tri chlorophenol
Fluoranthene
8250A - 27 Revision 1
September 1994
-------
TABLE 5.
SEMIVOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
l,4-Dichlorobenzene-D4
Naphthalene-dg
Acenaphthene-di
Aniline
Benzyl alcohol
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl)ether
2-Chlorophenol
1,3-Dichlorobenzene
1,4-Dichlorobenzene
1,2-Dichlorobenzene
Ethyl methanesulfonate
2-Fluorophenol (surr.)
Hexachloroethane
Methyl methanesulfonate
2-Methylphenol
4-Methylphenol
N-Nitrosodimethyl amine
N-Nitroso-di-n-propylamine
Phenol
Phenol-de (surr.)
2-Picoline
Acetophenone
Benzoic acid
Bis(2-chloroethoxy)methane
4-Chloroaniline
4-Chloro-3-methyl phenol
2,4-Dichlorophenol
2,6-Dichlorophenol
a,a-Dimethylphenethylamine
2,4-Dimethylphenol
Hexachlorobutadiene
Isophorone
2-Methylnaphthalene
Naphthalene
Nitrobenzene
Nitrobenzene-d8 (surr.)
2-Nitrophenol
N-Nitroso-di-n-butyl amine
N-Nitrosopiperi dine
1,2,4-Trichlorobenzene
Acenaphthene
Acenaphthylene
1-Chloronaphthalene
2-Chloronaphthalene
4-Chlorophenyl
phenyl ether
Dibenzofuran
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Fluorene
2-Fluorobiphenyl
(surr.)
Hexachlorocyclo-
pentadiene
1-Naphthylamine
2-Naphthylamine
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
4-Nitrophenol
Pentachlorobenzene
1,2,4,5-Tetrachloro-
benzene
2,3,4,6-Tetrachloro-
phenol
2,4,6-Tribromophenol
(Surr.)
2,4,6-Trichlorophenol
2,4,5-Trichlorophenol
(surr.) = surrogate
8250A - 28
Revision 1
September 1994
-------
TABLE 5.
SEMIVOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
(Continued)
Phenanthrene-d
10
Chrysene-d12
Perylene-d12
4-Aminobiphenyl
Anthracene
4-Bromophenyl phenyl ether
Di-n-butyl phthalate
4,6-Dinitro-2-methylphenol
Diphenylamine
1,2-Diphenylhydrazine
Fluoranthene
Hexachlorobenzene
N-Nitrosodiphenylamine
Pentachlorophenol
Pentachloronitrobenzene
Phenacetin
Phenanthrene
Pronarm'de
Benzidine
Benzo(a)anthracene
Bis(2-ethylhexyl) phthalate
Butyl benzyl phthalate
Chrysene
3,3'-Dichlorobenzidine
p-Dimethylaminoazobenzene
Pyrene
Terphenyl-d14 (surr.)
Benzo(b)fluoranthene
Benzo(k)f1uoranthene
Benzo(g,h,i)perylene
Benzo(a)pyrene
Dibenz(a,j)acridine
Dibenz(a,h)anthracene
7,12-Dimethylbenz-
(a)anthracene
Di-n-octyl phthalate
Indeno(l,2,3-cd)pyrene
3-Methylcholanthrene
(surr.) = surrogate
8250A - 29
Revision 1
September 1994
-------
TABLE 6.
QC ACCEPTANCE CRITERIA8
Test
cone.
Compound (M9/L)
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benzo(a)anthracene
Benzo(b)fl uoranthene
Benzo(k)fl uoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Butyl benzyl phthalate
6-BHC
5-BHC
Bis(2-chloroethyl) ether
Bi s ( 2 -chl oroethoxy)methane
Bis(Z-chloroisopropyl) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
2-Chl oronaphthal ene
4-Chlorophenyl phenyl ether
Chrysene
4, 4' -ODD
4,4'-DDE
4,4'-DDT
Dibenzo( a, h) anthracene
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1 ,4-Dichlorobenzene
3,3'-Dichlorobenzidine
Dieldrin
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Endosulfan sulfate
Endrin aldehyde
Fl uoranthene
Fluorene
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorobutadiene
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
Limit
for s
(M9/L)
27.6
40.2
39.0
32.0
27.6
38.8
32.3
39.0
58.9
23.4
31.5
21.6
55.0
34.5
46.3
41.1
23.0
13.0
33.4
48.3
31.0
32.0
61.6
70.0
16.7
30.9
41.7
32.1
71.4
30.7
26.5
23.2
21.8
29.6
31.4
16.7
32.5
32.8
20.7
37.2
54.7
24.9
26.3
Range
for x
(M/L)
60.1-132.3
53.5-126.0
7.2-152.2
43.4-118.0
41.8-133.0
42.0-140.4
25.2-145.7
31.7-148.0
D-195.0
D-139.9
41.5-130.6
D-100.0
42.9-126.0
49.2-164.7
62.8-138.6
28.9-136.8
64.9-114.4
64.5-113.5
38.4-144.7
44.1-139.9
D-134.5
19.2-119.7
D-170.6
D-199.7
8.4-111.0
48.6-112.0
16.7-153.9
37.3-105.7
8.2-212.5
44.3-119.3
D-100.0
D-100.0
47.5-126.9
68.1-136.7
18.6-131.8
D-103.5
D-188.8
42.9-121.3
71.6-108.4
D-172.2
70.9-109.4
7.8-141.5
37.8-102.2
Range
P> Ps
(%)
47-145
33-145
D-166
27-133
33-143
24-159
11-162
17-163
D-219
D-152
24-149
D-110
12-158
33-184
36-166
8-158
53-127
60-118
25-158
17-168
D-145
4-136
D-203
D-227
1-118
32-129
D-172
20-124
D-262
29-136
D-114
D-112
39-139
50-158
4-146
D-107
D-209
26-137
59-121
D-192
26-155
D-152
24-116
8250A - 30
Revision 1
September 1994
-------
TABLE 6.
QC ACCEPTANCE CRITERIA8
(Continued)
Compound
Hexachloroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Naphthalene
Nitrobenzene
N-Nitroso-di-n-propylamine
PCB-1260
Phenanthrene
Pyrene
1 , 2 , 4-Tri chl orobenzene
4-Chloro-3-methyl phenol
2-Chlorophenol
2,4-Chlorophenol
2,4-Dimethylphenol
2,4-Dinitrophenol
2-Methyl-4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
s = Standard deviati
x = Average recovery
p, ps = Percent recovery
D = Detected; result
a r\*4 -faln-i •> •Pixnm A f\ TCD D
Test
cone.
(M9/L)
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
Limit
for s
(M9/L)
24.5
44.6
63.3
30.1
39.3
55.4
54.2
20.6
25.2
28.1
37.2
28.7
26.4
26.1
49.8
93.2
35.2
47.2
48.9
22.6
31.7
on of four recovery
for four
measured
must be
I-../.+ IOC 4
recovery
•
Range
for x
(M9/L)
55.2-100.0
D-150.9
46.6-180.2
35.6-119.6
54.3-157.6
13.6-197.9
19.3-121.0
65.2-108.7
69.6-100.0
57.3-129.2
40.8-127.9
36.2-120.4
52.5-121.7
41.8-109.0
D-172.9
53.0-100.0
45.0-166.7
13.0-106.5
38.1-151.8
16.6-100.0
52.4-129.2
measurements,
measurements,
Range
P» Ps
(%)
40-113
D-171
21-196
21-133
35-180
D-230
D-164
54-120
52-115
44-142
22-147
23-134
39-135
32-119
0-191
D-181
29-182
D-132
14-176
5-112
37-144
in M9/L-
in /ng/L.
greater than zero.
?nv. Mn+Un/J
COC T l«» r\ *•. *•» <- <
kA ^ •t' /*\ M •« •% *\ t/»r\ !•*•«** St sJ
directly on the method performance data in Table 7. Where necessary, the
limits for recovery have been broadened to assure applicability of the
limits to concentrations below those used to develop Table 7.
8250A - 31
Revision 1
September 1994
-------
TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Parameter
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benzo(a)anthracene
Chloroethane
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Butyl benzyl phthalate
B-BHC
tJ-BHC
Bis(2-chloroethyl) ether
Bis(2-chloroethoxy)methane
Bis(2-chloroisopropyl) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
2-Chloronaphthalene
4-Chlorophenyl phenyl ether
Chrysene
4,4'-DDD
4,4'-DOE
4,4'-DDT
Dibenzo( a, h) anthracene
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1 , 4-Di chl orobenzene
3,3'-Dichlorobenzidine
Dieldrin
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Endosulfan sulfate
Endrin aldehyde
Fluoranthene
Fluorene
Heptachlor
Heptachlor epoxide
Hexachl orobenzene
Hexachlorobutadiene
Hexachloroethane
Accuracy, as
recovery, x'
(M9/L)
0.96C+0.19
0.89C+0.74
0.78C+1.66
0.80C+0.68
0.88C-0.60
0.99C-1.53
0.93C-1.80
0.87C-1.56
0.90C-0.13
0.98C-0.86
0.66C-1.68
0.87C-0.94
0.29C-1.09
0.86C-1.54
1.12C-5.04
1.03C-2.31
0.84C-1.18
0.91C-1.34
0.89C+0.01
0.91C+0.53
0.93C-1.00
0.56C-0.40
0.70C-0.54
0.79C-3.28
0.88C+4.72
0.59C+0.71
0.80C+0.28
0.86C-0.70
0.73C-1.47
1.23C-12.65
0.82C-0.16
0.43C+1.00
0.20C+1.03
0.92C-4.81
1.06C-3.60
0.76C-0.79
0.39C+0.41
0.76C-3.86
0.81C+1.10
0.90C-0.00
0.87C-2.97
0.92C-1.87
0.74C+0.66
0.71C-1.01
0.73C-0.83
Single analyst
precision, s/
(M9/L)
0.15X-0.12
0.24X-1.06
0.27X-1.28
0.21X-0.32
O.lSx+0.93
0.14X-0.13
0.22X+0.43
0.19X+1.03
0.22X+0.48
0.29x+2.40
O.lSx+0.94
0.20X-0.58
0.34X+0.86
0.35x~0.99
O.lSx+1.34
0.24X+0.28
0.26x+0.73
0.13X+0.66
0.07X+0.52
0.20x-0.94
0.28x+0,13
0.29x-0.32
0.26X-1.17
0.42X+0.19
0.30X+8.51
0.13X+1.16
0.20X+0.47
0.25X+0.68
0.24X+0.23
0.28X+7.33
0.20X-0.16
0.28X+1.44
0.54x+0'.19
0.12x+1.06
0.14X+1.26
O.Zlx+1.19
0.12X+2.47
O.lSx+3.91
0.22X-0.73
0.12X+0.26
0.24X-0.56
0.33x-0.46
0.18X-0.10
0.19X+0.92
0.17X+0.67
Overall
precision,
S' (/ig/L)
0.21X-0.67
0.26X-0.54
0.43X+1.13
0.27X-0.64
0.26X-0.21
0.17X-0.28
0.29X+0.96
0.35X+0.40
0.32X+1.35
0.51X-0.44
0.53X+0.92
0.30X+1.94
0.93X-0.17
0.35X+0.10
0.26X+2.01
0.25X+1.04
0.36X+0.67
0.16X+0.66
0.13X+0.34
0.30X-0.46
0.33X-0.09
0.66X-0.96
0.39X-1.04
0.65X-0.58
0.59X+0.25
0.39X+0.60
0.24X+0.39
0.41X+0.11
0.29X+0.36
0.47X+3.45
0.26X-0.07
0.52X+0.22
1.05X-0.92
O.Zlx+1.50
0.19X+0.35
0.37X+1.19
0.63X-1.03
0.73X-0.62
0.28X-0.60
0.13X+0.61
O.BOx-0.23
0.28X+0.64
0.43X-0.52
0.26X-I-0.49
0.17X+0.80
8250A - 32
Revision 1
September 1994
-------
TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION3
(Continued)
Parameter
Indeno(l,2,3-cd)pyrene
Isophorone
Naphthalene
Nitrobenzene
N-Nitroso-di-n-propylamine
PCB-1260
Phenanthrene
Pyrene
1,2,4-Trichlorobenzene
4-Chl oro-3 -methyl phenol
2-Chlorophenol
2,4-Dichlorophenol
2,4-Dimethylphenol
2,4-Dinitrophenol
2-Methyl-4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
Accuracy, as
recovery, x'
(Mg/L)
0.78C-3.10
1.12C+1.41
0.76C+1.58
1.09C-3.05
1.12C-6.22
0.81C-10.86
0.87C+0.06
0.84C-0.16
0.94C-0.79
0.84C+0.35
0.78C+0.29
0.87C-0.13
0.71C+4.41
0.81C-18.04
1.04C-28.04
0.07C-1.15
0.61C-1.22
0.93C+1.99
0.43C+1.26
0.91C-0.18
Single analyst
precision, s/
(M9/L)
0.29x+1.46
0.27x+0.77
0.21X-0.41
0.19X+0.92
0.27x+0.68
0.35x+3.61
0.12X+0.57
0.16X+0.06
O.lSx+0.85
0.23X+0.75
O.lSx+1.46
O.lSx+1.25
O.lGx+1.21
0.38X+2.36
O.lOx+42.29
0.16X+1.94
0.38X+2.57
0.24X+3.03
0.26X+0.73
0.16X+2.22
Overall
precision,
S' (Mg/L)
O.BOx-0.44
0.33X+0.26
O.SOx-0.68
0.27X+0.21
0.44X+0.47
0.43X+1.82
0.15X+0.25
0.15X+0.31
0.21X+0.39
0.29X+1.31
0.28X+0.97
0.21X+1.28
0.22X+1.31
0.42X+26.29
0.26X+23.10
0.27x+2.60
0.44X+3.24
0.30X+4.33
0.35X+0.58
0.22X+1.81
X'
S'
C
x
Expected recovery for one or more measurements of a sample
containing a concentration of C, in
Expected single analyst standard deviation of measurements at an
average concentration of x, in p.g/1.
Expected interlaboratory standard deviation of measurements at an
average concentration found of x, in
True value for the concentration, in
Average recovery found for measurements of samples containing a
concentration of C, in
8250A - 33
Revision 1
September 1994
-------
TABLE 8.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Low/Medium Low/Medium
Surrogate Compound Water Soil/Sediment
Nitrobenzene-d5 35-114 23-120
2-Fluorobiphenyl 43-116 30-115
Terphenyl-du 33-141 18-137
Phenol-d6 10-94 24-113
2-Fluorophenol 21-100 25-121
2,4,6-Tribromophenol 10-123 19-122
8250A - 34 Revision 1
September 1994
-------
METHOD 8250A
SEMIVOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS)
7.1 Prepare sample
using Method 3540,
3541, or 35SO
7.1 Prepare tampl*
using Method 3610
or 3520
7.1 Prepare (ample
using Method 3540,
3541, 3550, or 3580
7.2 Cleenup
extract
7.3
Recommended
GC/MS
operating
conditions.
7.4
Initial
Calibration.
7.5 Daily
calibration - Tune
GC/MS with TFTPP
and check SPCC &
CCC.
8250A - 35
Revision 1
September 1994
-------
METHOD 8250A
continued
7.6.1 Screen extract
in GC/FID or GC/PID to
eliminate too high
concentration*.
7.6.2 Spike
sample with
internal
standard.
7.6.3 Analyze
extract by GC/MS
using recommended
column and operating
condition*.
7.6.4
Doe*
response exceed
initial calibration
curve
range?
7.6.4 Dilute
extract.
7.7.1 Identify
compound* by
comparing sample
retention time and
sample mass spectra
to standard*.
7.7.2
Quantitate
sample* using
internal ltd.
technique.
7.7.2.4 Report
results.
1
r
C Stop J
8250A - 36
Revision 1
September 1994
-------
METHOD 8260A
VOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS):
CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8260 is used to determine volatile organic compounds in a
variety of solid waste matrices. This method is applicable to nearly all types
of samples, regardless of water content, including ground water, aqueous sludges,
caustic liquors, acid liquors, waste solvents, oily wastes
fibrous wastes, polymeric emulsions, filter cakes, spent
catalysts, soils, and sediments. The following compounds can
this method:
mousses, tars,
carbons, spent
be determined by
Analyte
CAS No.1
Appropriate Technique
Direct
Purge-and-Trap Injection
Acetone
Acetonitrile
Acrolein (Propenal)
Acrylonitrile
Ally! alcohol
Allyl chloride
Benzene
Benzyl chloride
Bromoacetone
Bromochloromethane (I.S.)
Bromodichloromethane
4-Bromofluorobenzene (surr.)
Bromoform
Bromomethane
n-Butanol
2-Butanone (MEK)
Carbon disulfide
Carbon tetrachloride
Chloral hydrate
Chlorobenzene
2 - Chi oro- 1,3 -butadiene
Chi orod i bromomethane
Chloroethane
2-Chloroethanol
bis-(2-Chloroethyl) sulfide
2-Chloroethyl vinyl ether
Chloroform
Chloromethane
Chloroprene
67-64-1
75-05-8
107-02-8
107-13-1
107-18-6
107-05-1
71-43-2
100-44-7
598-31-2
74-97-5
75-27-4
460-00-4
75-25-2
74-83-9
71-36-3
78-93^3
75-15-0
56-23-5
302-17-0
108-90-7
126-99-8
124-48-1
75-00-3
107-07-3
505-60-2
110-75-8
67-66-3
74-87-3
126-99-8
PP
PP
PP
PP
ht
a
a
a
PP
a
a
a
a
a
ht
PP
PP
a
PP
a
a
a
a
PP
PP
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
pc
8260A - 1
Revision 1
September 1994
-------
Appropriate Technique
Analyte
3-Chloropropene
3-Chl oropropi oni tri 1 e
l,2-Dibromo-3-chloropropane
1,2-Dibromoethane
Dibromomethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
cis-l,4-Dichloro-2-butene
trans- l,4-Dichloro-2-butene
Di chl orod i f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans- 1 ,2-Di chl oroethene
1,2-Dichloropropane
l,3-Dichloro-2-propanol
cis-l,3-Dichloropropene
trans-l,3-Dichloropropene
1,2,3,4-Diepoxybutane
Di ethyl ether
1,4-Difluorobenzene (I.S.)
1,4-Dioxane
Epichlorohydrin
Ethanol
Ethyl acetate
Ethyl benzene
Ethylene oxide
Ethyl methacrylate
Hexachlorobutadiene
Hexachloroethane
2-Hexanone
2-Hydroxypropionitrile
lodomethane
Isobutyl alcohol
Isopropyl benzene
Malononitrile
Methacrylonitrile
Methanol
Methylene chloride (DCM)
Methyl methacrylate
4-Methyl-2-pentanone (MIBK)
Naphthalene
Nitrobenzene
2-Nitropropane
CAS No."
107-05-1
542-76-7
96-12-8
106-93-4
74-95-3
95-50-1
541-73-1
106-46-7
1476-11-5
110-57-6
75-71-8
75-34-3
107-06-2
75-35-4
156-60-5
78-87-5
96-23-1
10061-01-5
10061-02-6
1464-53-5
60-29-7
540-36-3
123-91-1
106-89-8
64-17-5
141-78-6
100-41-4
75-21-8
97-63-2
87-68-3
67-72-1
591-78-6
78-97-7
74-88-4
78-83-1
98-82-8
109-77-3
126-98-7
67-56-1
75-09-2
80-62-6
108-10-1
91-20-3
98-95-3
79-46-9
Purge-and-Trap
a
i
PP
a
a
a
a
a
a
PP
a
a
a
a
a
a
PP
a
a
a
a
a
PP
i
i
i
a
PP
a
a
i
PP
i
a
PP
a
PP
PP
i
a
a
PP
a
a
a
Direct
Injection
a
pc
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
pc
a
a
a
a
a
a
a
a
a
a
a
a
8260A - 2
Revision 1
September 1994
-------
Appropriate Technique
Analyte CAS No.b
Pentachloroethane 76-01-7
2-Picoline 109-06-8
Propargyl alcohol 107-19-7
B-Propiolactone 57-57-8
Propionitrile (ethyl cyanide) 107-12-0
n-Propylamine 107-10-8
Pyridine 110-86-1
Styrene 100-42-5
1,1,1 , 2-Tetrachl oroethane 630-20-6
1,1,2,2-Tetrachloroethane 79-34-5
Tetrachloroethene 127-18-4
Toluene 108-88-3
1,2,4-Trichlorobenzene 120-82-1
1,1,1-Trichloroethane 71-55-6
1,1,2-Trichloroethane 79-00-5
Trichloroethene 79-01-6
Trichlorofluoromethane 75-69-4
1,2,3-Trichloropropane 96-18-4
Vinyl acetate 108-05-4
Vinyl chloride 75-01-4
o-Xylene 95-47-6
m-Xylene 108-38-3
p-Xylene 106-42-3
a Adequate response by this technique.
b Chemical Abstract Services Registry Number
ht Method analyte only when purged at 80°C
i Inappropriate technique for this analyte.
pc Poor chromatographic behavior.
pp Poor purging efficiency resulting in high
surr Surrogate
I.S. Internal Standard
Purge-and-Trap
i
PP
PP
PP
ht
a
i
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
.
EQLs.
Direct
Injection
a
a
a
a
pc
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
1.2 Method 8260 can be used to quantitate most volatile organic compounds
that have boiling points below 200°C and that are insoluble or slightly soluble
in water. Volatile water-soluble compounds can be included in this analytical
technique. However, for the more soluble compounds, quantitation limits are
approximately ten times higher because of poor purging efficiency. Such
compounds include low-molecular-weight halogenated hydrocarbons, aromatics,
ketones, nitriles, acetates, acrylates, ethers, and sulfides. See Tables 1 and
2 for lists of analytes and retention times that have been evaluated on a purge-
8260A - 3 Revision 1
September 1994
-------
and-trap GC/MS system. Also, the method detection limits for 25 ml sample
volumes are presented. The following analytes are also amenable to analysis by
Method 8260:
Bromobenzene 1-Chlorohexane
n-Butylbenzene 2-Chlorotoluene
sec-Butyl benzene 4-Chlorotoluene
tert-Butylbenzene Crotonaldehyde
Chioroaceton i tri1e Di bromofluoromethane
1-Chlorobutane cis-l,2-Dichloroethene
1,3-Dichloropropane Methyl-t-butyl ether
2,2-Dichloropropane Pentafluorobenzene
1,1-Dichloropropene n-Propylbenzene
Fluorobenzene 1,2,3-Trichlorobenzene
p-1sopropyltoluene 1,2,4-Trimethylbenzene
Methyl acrylate 1,3,5-Trimethylbenzene
1.3 The estimated quantitation limit (EQL) of Method 8260 for an
individual compound is somewhat instrument dependent. Using standard quadrupole
instrumentation, limits should be approximately 5 jugAg (wet weight) for
soil/sediment samples, 0.5 mg/kg (wet weight) for wastes, and 5 /xg/L for ground
water (see Table 3). Somewhat lower limits may be achieved using an ion trap
mass spectrometer or other instrumentation of improved design. No matter which
instrument is used, EQLs will be proportionately higher for sample extracts and
samples that require dilution or reduced sample size to avoid saturation of the
detector.
1.4 Method 8260 is based upon a purge-and-trap, gas chromatographic/mass
spectrometric (GC/MS) procedure. This method is restricted to use by, or under
the supervision of, analysts experienced in the use of purge-and-trap systems and
gas chromatograph/mass spectrometers, and skilled in the interpretation of mass
spectra and their use as a quantitative tool.
1.5 An additional method for sample introduction is direct injection.
This technique has been tested for the analysis of waste oil diluted with
hexadecane 1:1 (vol/vol) and may have application for the analysis of some
alcohols and aldehydes in aqueous samples.
2.0 SUMMARY OF METHOD
2.1 The volatile compounds are introduced into the gas chromatograph by
the purge-and-trap method or by direct injection (in limited applications).
Purged sample components are trapped in a tube containing suitable sorbent
materials. When purging is complete, the sorbent tube is heated and backflushed
with helium to desorb trapped sample components. The analytes are desorbed
directly to a large bore capillary or cryofocussed on a capillary precolumn
before being flash evaporated to a narrow bore capillary for analysis. The
column is temperature programmed to separate the analytes which are then detected
with a mass spectrometer (MS) interfaced to the gas chromatograph. Wide bore
capillary columns require a jet separator, whereas narrow bore capillary columns
can be directly interfaced to the ion source.
8260A - 4 Revision 1
September 1994
-------
2.2 If the above sample introduction techniques are not applicable, a
portion of the sample is dispersed in solvent to dissolve the volatile organic
constituents. A portion of the solution is combined with organic-free reagent
water in the purge chamber. It is then analyzed by purge-and-trap GC/MS
following the normal water method.
2.3 Analytes eluted from the capillary column are introduced into the
mass spectrometer via a jet separator or a direct connection. Identification of
target analytes is accomplished by comparing their mass spectra with the electron
impact (or electron impact-like) spectra of authentic standards. Quantitation
is accomplished by comparing the response of a major (quantitation) ion relative
to an internal standard with a five-point calibration curve.
2.4 The method includes specific calibration and quality control steps
that replace the general requirements in Method 8000.
3.0 INTERFERENCES
3.1 Major contaminant sources are volatile materials in the laboratory
and impurities in the inert purging gas and in the sorbent trap. The use of non-
polytetrafluoroethylene (PTFE) thread sealants, plastic tubing, or flow
controllers with rubber components should be avoided since such materials out-gas
organic compounds which will be concentrated in the trap during the purge
operation. Analyses of calibration and reagent blanks provide information about
the presence of contaminants. When potential interfering peaks are noted in
blanks, the analyst should change the purge gas source and regenerate the
molecular sieve purge gas filter (Figure 1). Subtracting blank values from
sample results is not permitted. If reporting values not corrected for blanks
result in what the laboratory feels is a false positive for a sample, this should
be fully explained in text accompanying the uncorrected data.
3.2 Interfering contamination may occur when a sample containing low
concentrations of volatile organic compounds is analyzed immediately after a
sample containing high concentrations of volatile organic compounds. The
preventive technique is rinsing of the purging apparatus and sample syringes with
two portions of organic-free reagent water between samples. After analysis of
a sample containing high concentrations of volatile organic compounds, one or
more calibration blanks should be analyzed to check for cross contamination. For
samples containing large amounts of water soluble materials, suspended solids,
high boiling compounds or high concentrations of compounds being determined, it
may be necessary to wash the purging device with a soap solution, rinse it with
organic-free reagent water, and then dry the purging device in an oven at 105°C.
In extreme situations, the whole purge and trap device may require dismantling
and cleaning. Screening of the samples prior to purge and trap GC/MS analysis
is highly recommended to prevent contamination of the system. This is especially
true for soil and waste samples. Screening may be accomplished with an automated
headspace technique or by Method 3820 (Hexadecane Extraction and Screening of
Purgeable Organics).
3.2.1 The low purging efficiency of many analytes from a 25 ml
sample often results in significant concentrations remaining in the sample
purge vessel after analysis. After removal of the analyzed sample aliquot
8260A - 5 Revision 1
September 1994
-------
and three rinses of the purge vessel with analyte free water, it is
required that the empty vessel be subjected to a heated purge cycle prior
to the analysis of another sample in the same purge vessel to reduce
sample to sample carryover.
3.3 Special precautions must be taken to analyze for methylene chloride.
The analytical and sample storage area should be isolated from all atmospheric
sources of methylene chloride. Otherwise random background levels will result.
Since methylene chloride will permeate through PTFE tubing, all gas
chromatography carrier gas lines and purge gas plumbing should be constructed
from stainless steel or copper tubing. Laboratory clothing worn by the analyst
should be clean since clothing previously exposed to methylene chloride fumes
during liquid/liquid extraction procedures can contribute to sample
contamination.
3.4 Samples can be contaminated by diffusion of volatile organics
(particularly methylene chloride and fluorocarbons) through the septum seal into
the sample during shipment and storage. A trip blank prepared from organic-free
reagent water and carried through the sampling and handling protocol can serve
as a check on such contamination.
3.5 Use of sensitive mass spectrometers to achieve lower detection level
will increase the potential to detect laboratory contaminants as interferences.
3.6 Direct injection - Some contamination may be eliminated by baking out
the column between analyses. Changing the injector liner will reduce the
potential for cross-contamination. A portion of the analytical column may need
to be removed in the case of extreme contamination. Use of direct injection will
result in the need for more frequent instrument maintenance.
3.7 If hexadecane is added to samples or petroleum samples are analyzed,
some chromatographic peaks will elute after the target analytes. The oven
temperature program must include a post-analysis bake out period to ensure that
semi -volatile hydrocarbons are volatilized.
4.0 APPARATUS AND MATERIALS
4.1 Purge-and-trap device - aqueous samples, described in Method 5030.
4.2 Purge-and-trap device - solid samples, described in Method 5030.
4.3 Injection port liners (HP catalogue #18740-80200, or equivalent) are
modified for direct injection analysis by placing a 1-cm plug of pyrex wool
approximately 50-60 mm down the length
of the injection port towards the
oven. An 0.53 mm ID column is mounted s«Pt«.m
1 cm into the liner from the oven side
of the injection port, according to
manufacturer's specifications. Modified Injector
8260A - 6 Revision 1
September 1994
-------
4.4 Gas chromatography/mass spectrometer/data system
4.4.1 Gas chromatograph - An analytical system complete with a
temperature-programmable gas chromatograph suitable for splitless
injection or interface to purge-and-trap apparatus. The system includes
all required accessories, including syringes, analytical columns, and
gases. The GC should be equipped with variable constant differential flow
controllers so that the column flow rate will remain constant throughout
desorption and temperature program operation. For some column
configurations, the column oven must be cooled to < 30°C, therefore, a
subambient oven controller may be required. The capillary column should
be directly coupled to the source.
4.4.1.1 Capillary precolumn interface when using cryogenic
cooling - This device interfaces the purge and trap concentrator to
the capillary gas chromatograph. The interface condenses the
desorbed sample components and focuses them into a narrow band on an
uncoated fused silica capillary precolumn. When the interface is
flash heated, the sample is transferred to the analytical capillary
column.
4.4.1.1.1 During the cryofocussing step, the
temperature of the fused silica in the interface is maintained
at -150°C under a stream of liquid nitrogen. After the
desorption period, the interface must be capable of rapid
heating to 250°C in 15 seconds or less to complete the
transfer of analytes.
4.4.2 Gas chromatographic columns
4.4.2.1 Column 1 - 60 m x 0.75 mm ID capillary column
coated with VOCOL (Supelco), 1.5 jttm film thickness, or equivalent.
4.4.2.2 Column 2 - 30 - 75 m x 0.53 mm ID capillary column
coated with DB-624 (J&W Scientific), Rtx-502.2 (RESTEK), or VOCOL
(Supelco), 3 jLtm film thickness, or equivalent.
4.4.2.3 Column 3 - 30 m x 0.25 - 0.32 mm ID capillary
column coated with 95% dimethyl - 5% diphenyl polysiloxane (DB-5,
Rtx-5, SPB-5, or equivalent), 1 jitm film thickness.
4.4.2.4 Column 4 - 60 m x 0.32 mm ID capillary column
coated with DB-624 (J&W Scientific), 1.8 jum film thickness, or
equivalent.
4.4.3 Mass spectrometer - Capable of scanning from 35 to 300 amu
every 2 sec or less, using 70 volts (nominal) electron energy in the
electron impact ionization mode. The mass spectrometer must be capable of
producing a mass spectrum for p-Bromofluorobenzene (BFB) which meets all
of the criteria in Table 4 when 5-50 ng of the GC/MS tuning standard (BFB)
is injected through the GC. To ensure sufficient precision of mass
spectral data, the desirable MS scan rate allows acquisition of at least
five spectra while a sample component elutes from the GC.
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4.4.3.1 The ion trap mass spectrometer may be used if it
is capable of axial modulation to reduce ion-molecule reactions and
can produce electron impact-like spectra that match those in the
EPA/NIST Library. In an ion trap mass spectrometer, because ion-
molecule reactions with water and methanol may produce interferences
that coelute with chloromethane and chloroethane, the base peak for
both of these analytes will be at m/z 49. This ion should be used
as the quantitation ion in this case. The mass spectrometer must be
capable of producing a mass spectrum for BFB which meets all of the
criteria in Table 3 when 5 or 50 ng are introduced.
4.4.4 GC/MS interface - Two alternatives are used to interface the
GC to the mass spectrometer.
4.4.4.1 Direct coupling by inserting the column into the
mass spectrometer is generally used for 0.25-0.32 mm id columns.
4.4.4.2 A separator including an all-glass transfer line
and glass enrichment device or split interface is used with an
0.53 mm column.
4.4.4.3 Any enrichment device or transfer line can be used
if all of the performance specifications described in Sec. 8
(including acceptable calibration at 50 ng or less) can be achieved.
GC-to-MS interfaces constructed entirely of glass or of glass-lined
materials are recommended. Glass can be deactivated by silanizing
with dichlorodimethylsilane.
4.4.5 Data system - A computer system that allows the continuous
acquisition and storage on machine-readable media of all mass spectra
obtained throughout the duration of the chromatographic program must be
interfaced to the mass spectrometer. The computer must have software that
allows searching any GC/MS data file for ions of a specified mass and
plotting such ion abundances versus time or scan number. This type of
plot is defined as an Extracted Ion Current Profile (EICP). Software must
also be available that allows integrating the abundances in any EICP
between specified time or scan-number limits. The most recent version of
the EPA/NIST Mass Spectral Library should also be available.
4.5 Microsyringes - 10, 25, 100, 250, 500, and 1,000 juL.
4.6 Syringe valve - Two-way, with Luer ends (three each), if applicable
to the purging device.
4.7 Syringes - 5, 10, or 25 mL, gas-tight with shutoff valve.
4.8 Balance - Analytical, 0.0001 g, and top-loading, 0.1 g.
4.9 Glass scintillation vials - 20 mL, with Teflon lined screw-caps or
glass culture tubes with Teflon lined screw-caps.
4.10 Vials - 2 mL, for GC autosampler.
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4.11 Disposable pipets - Pasteur.
4.12 Volumetric flasks, Class A - 10 ml and 100 ml, with ground-glass
stoppers.
4.13 Spatula - Stainless steel.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all inorganic reagents shall conform to
the specifications of the Committee on Analytical Reagents of the American
Chemical Society, where such specifications are available. Other grades may be
used, provided it is first ascertained that the reagent is of sufficiently high
purity to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Methanol, CH3OH - Pesticide quality or equivalent, demonstrated to
be free of analytes. Store apart from other solvents.
5.4 Reagent Hexadecane - Reagent hexadecane is defined as hexadecane in
which interference is not observed at the method detection limit of compounds of
interest.
5.4.1 In order to demonstrate that all interfering volatiles have
been removed from the hexadecane, a direct injection blank must be
analyzed.
5.5 Polyethylene glycol, H(OCH2CH2)nOH - Free of interferences at the
detection limit of the target analytes.
5.6 Hydrochloric acid (1:1 v/v), HC1 - Carefully add a measured volume
of concentrated HC1 to an equal volume of organic-free reagent water.
5.7 Stock solutions - Stock solutions may be prepared from pure standard
materials or purchased as certified solutions. Prepare stock standard solutions
in methanol, using assayed liquids or gases, as appropriate.
5.7.1 Place about 9.8 ml of methanol in a 10 ml tared ground-glass-
stoppered volumetric flask. Allow the flask to stand, unstoppered, for
about 10 minutes or until all alcohol-wetted surfaces have dried. Weigh
the flask to the nearest 0.0001 g.
5.7.2 Add the assayed reference material, as described below.
5.7.2.1 Liquids - Using a 100 fj.1 syringe, immediately add
two or more drops of assayed reference material to the flask; then
reweigh. The liquid must fall directly into the alcohol without
contacting the neck of the flask.
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5.7.2.2 Gases - To prepare standards for any compounds
that boil below 30°C (e.g. bromomethane, chloroethane, chloromethane,
or vinyl chloride), fill a 5 ml valved gas-tight syringe with the
reference standard to the 5.0 ml mark. Lower the needle to 5 mm
above the methanol meniscus. Slowly introduce the reference standard
above the surface of the liquid. The heavy gas will rapidly dissolve
in the rnethanol. Standards may also be prepared by using a lecture
bottle equipped with a Hamilton Lecture Bottle Septum (#86600).
Attach Teflon tubing to the side arm relief valve and direct a gentle
stream of gas into the methanol meniscus.
5.7.3 Reweigh, dilute to volume, stopper, and then mix by inverting
the flask several times. Calculate the concentration in milligrams per
liter (mg/L) from the net gain in weight. When compound purity is assayed
to be 96% or greater, the weight may be used without correction to
calculate the concentration of the stock standard. Commercially prepared
stock standards may be used at any concentration if they are certified by
the manufacturer or by an independent source.
5.7.4 Transfer the stock standard solution into a bottle with a
Teflon lined screw-cap. Store, with minimal headspace, at -10°C to -20°C
and protect from light.
5.7.5 Prepare fresh standards for gases weekly or sooner if
comparison with check standards indicates a problem. Reactive compounds
such as 2-chloroethyl vinyl ether and styrene may need to be prepared more
frequently. All other standards must be replaced after six months, or
sooner if comparison with check standards indicates a problem. Both gas
and liquid standards must be monitored closely by comparison to the
initial calibration curve and by comparison to QC check standards. It may
be necessary to replace the standards more frequently if either check
exceeds a 20% drift.
5.7.6 Optionally calibration using a certified gaseous mixture can
be accomplished daily utilizing commercially available gaseous analyte
mixture of bromomethane, chloromethane, chloroethane, vinyl chloride,
dichlorodifluoromethane and trichlorofluoromethane in nitrogen. These
mixtures of documented quality are stable for as long as six months
without refrigeration. (VOA-CYL III, RESTEK Corporation, Cat. #20194 or
equivalent).
5.7.6.1 Preparation of Calibration Standards From a Gas
Mixture
5.7.6.1.1 Before removing the cylinder shipping cap,
be sure the valve is completely closed (turn clockwise). The
contents are under pressure and should be used in a well-
ventilated area.
5.7.6.1.2 Wrap the pipe thread end of the Luer fitting
with Teflon tape. Remove the shipping cap from the cylinder
and replace it with the Luer fitting.
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5.7.6.1.3 Transfer half the working standard containing
other analytes, internal standards, and surrogates to the
purge apparatus.
5.7.6.1.4 Purge the Luer fitting and stem on the gas
cylinder prior to sample removal using the following sequence:
a) Connect either the 100 juL or 500 juL Luer syringe
to the inlet fitting of the cylinder.
b) Make sure the on/off valve on the syringe is in
the open position.
c) Slowly open the valve on the cylinder and
withdraw a full syringe volume.
d) Be sure to close the valve on the cylinder before
you withdraw the syringe from the Luer fitting.
e) Expel the gas from the syringe into a well-
ventilated area.
f) Repeat steps a through e one more time to fully
purge the fitting.
5.7.6.1.5 Once the fitting and stem have been purged,
quickly withdraw the volume of gas you require using steps
5.6.6.1.4(a) through (d). Be sure to close the valve on the
cylinder and syringe before you withdraw the syringe from the
Luer fitting.
5.7.6.1.6 Open the syringe on/off valve for 5 seconds
to reduce the syringe pressure to atmospheric pressure. The
pressure in the cylinder is -30 psi.
5.7.6.1.7 The gas mixture should be quickly transferred
into the reagent water through the female Luer fitting located
above the purging vessel.
NOTE: Make sure the arrow on the 4-way valve is
pointing toward the female Luer fitting when
transferring the sample from the syringe. Be sure
to switch the 4-way valve back to the closed
position before removing the syringe from the
Luer fitting.
5.7.6.1.8 Transfer the remaining half of the working
standard into the purging vessel. This procedure insures that
the total volume of gas mix is flushed into the purging
vessel, with none remaining in the valve or lines.
5.7.6.1.9 Concentration of each compound in the
cylinder is typically 0.0025
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5.7.6.1.10 The following are the recommended gas volumes
spiked into 5 ml of water to produce a typical 5-point
calibration:
Gas Calibration
Volume Concentration
40 ML 20 M9/L
100 ML 50 M9/L
200 juL 100 jug/L
300 ML 150 M9/L
400 ML 200 M9/L
5.7.6.1.11 The following are the recommended gas volumes
spiked into 25 mL of water to produce a typical 5-point
calibration:
Gas Calibration
Volume Concentration
10 ML 1 M9/L
20 ML 2 M9/L
50 ML 5 M9/L
100 ML 10 M9/L
250 ML 25 M9/L
5.8 Secondary dilution standards - Using stock standard solutions,
prepare in methanol, secondary dilution standards containing the compounds of
interest, either singly or mixed together. Secondary dilution standards must be
stored with minimal headspace and should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them. Store in a vial with no headspace for one week only.
5.9 Surrogate standards - The surrogates recommended are toluene-d8,
4-bromofluorobenzene, l,2-dichloroethane-d4, and dibromofluoromethane. Other
compounds may be used as surrogates, depending upon the analysis requirements.
A stock surrogate solution in methanol should be prepared as described above, and
a surrogate standard spiking solution should be prepared from the stock at a
concentration of 50-250 M9/l° ml in methanol. Each water sample undergoing
GC/MS analysis must be spiked with 10 ML of the surrogate spiking solution prior
to analysis.
5.9.1 If a more sensitive mass spectrometer is employed to achieve
lower detection levels, more dilute surrogate solutions may be required.
5.10 Internal standards - The recommended internal standards are
fluorobenzene, chlorobenzene-d5, and l,4-dichlorobenzene-d4. Other compounds may
be used as internal standards as long as they have retention times similar to the
compounds being detected by GC/MS. Prepare internal standard stock and secondary
dilution standards in methanol using the procedures described in Sees. 5.7 and
5.8. It is recommended that the secondary dilution standard should be prepared
at a concentration of 25 mg/L of each internal standard compound. Addition of
10 ML of this standard to 5.0 ml of sample or calibration standard would be the
equivalent of 50 M9/L-
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5.10.1 If a more sensitive mass spectrometer is employed to
achieve lower detection levels, more dilute internal standard solutions
may be required. Area counts of the internal standard peaks should be
between 50-200% of the area of the target analytes in the mid-point
calibration analysis.
5.11 4-Bromofluorobenzene (BFB) standard - A standard solution containing
25 ng//iL of BFB in methanol should be prepared.
5.11.1 If a more sensitive mass spectrometer is employed to
achieve lower detection levels, a more dilute BFB standard solution may be
required.
5.12 Calibration standards - Calibration standards at a minimum of five
concentrations should be prepared from the secondary dilution of stock standards
(see Sees. 5.7 and 5.8). Prepare these solutions in organic-free reagent water.
One of the concentrations should be at a concentration near, but above, the
method detection limit. The remaining concentrations should correspond to the
expected range of concentrations found in real samples but should not exceed the
working range of the GC/MS system. Each standard should contain each analyte for
detection by this method. It is EPA's intent that all target analytes for a
particular analysis be included in the calibration standard(s). However, these
target analytes may not include the entire List of Analytes (Sec. 1.1) for which
the method has been demonstrated. However, the laboratory shall not report a
quantitative result for a target analyte that was not included in the calibration
standard(s). Calibration standards must be prepared daily.
5.13 Matrix spiking standards - Matrix spiking standards should be
prepared from volatile organic compounds which will be representative of the
compounds being investigated. At a minimum, the matrix spike should include 1,1-
dichloroethene, trichloroethene, chlorobenzene, toluene, and benzene. It is
desirable to perform a matrix spike using compounds found in samples. Some
permits may require spiking specific compounds of interest, especially if they
are polar and would not be represented by the above listed compounds. The
standard should be prepared in methanol, with each compound present at a
concentration of 250 jiig/10.0 ml.
5.13.1 If a more sensitive mass spectrometer is employed to
achieve lower detection levels, more dilute matrix spiking solutions may
be required.
5.14 Great care must be taken to maintain the integrity of all standard
solutions. It is recommended all standards in methanol be stored at -10°C to
-20°C in amber bottles with Teflon lined screw-caps.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes, Sec.
4.1.
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7.0 PROCEDURE
7.1 Three alternate methods are provided for sample introduction. All
internal standards, surrogates, and matrix spikes (when applicable) must be added
to samples before introduction.
7.1.1 Direct injection - in very limited application, (e.g.,
volatiles in waste oil or aqueous process wastes) direct injection of
aqueous samples or samples diluted according to Method 3585 may be
appropriate. Direct injection has been used for the analysis of volatiles
in waste oil (diluted 1:1 with hexadecane) and for determining if the
sample is ignitable (aqueous injection, Methods 1010 or 1020). Direct
injection is only permitted for the determination of volatiles at the
toxicity characteristic (TC) regulatory limits, at concentrations in
excess of 10,000 /xg/L, or for water-soluble compounds that do not purge.
7.1.2 Purge-and-trap for aqueous samples, see Method 5030 for
details.
7.1.3 Purge-and-trap for solid samples, see Method 5030 for details.
7.2 Recommended Chromatographic conditions
7.2.1 General:
Injector temperature: 200-225°C
Transfer line temperature: 250-300°C
7.2.2 Column 1 (A sample chromatogram is presented in Figure 5)
Carrier gas (He) flow rate: 15 mL/min
Initial temperature: 10°C, hold for 5 minutes
Temperature program: 6°C/min to 160°C
Final temperature: 160°C, hold until all expected
compounds have eluted.
7.2.3 Column 2, Cryogenic cooling (A sample chromatogram is
presented in Figure 6)
Carrier gas (He) flow rate: 15 mL/min
Initial temperature: 10°C, hold for 5 minutes
Temperature program: 6°C/min to 160°C
Final temperature: 160°C, hold until all expected
compounds have eluted.
7.2.4 Column 2, Non-cryogenic cooling (A sample chromatogram is
presented in Figure 7). It is recommended that carrier gas flow and split
and make-up gases be set using performance of standards as guidance. Set
the carrier gas head pressure to « 10 psi and the split to « 30 mL/min.
Optimize the make-up gas flow for the separator (approximately 30 mL/min)
by injecting BFB, and determining the optimum response when varying the
make-up gas. This will require several injections of BFB. Next, make
several injections of the volatile working standard with all analytes of
8260A - 14 Revision 1
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interest. Adjust the carrier and split to provide optimum chromatography
and response. This is an especially critical adjustment for the volatile
gas analytes. The head pressure should optimize between 8-12 psi and the
split between 20-60 mL/min. The use of the splitter is important to
minimize the effect of water on analyte response, to allow the use of a
larger volume of helium during trap desorption, and to slow column flow.
Initial temperature: 45°C, hold for 2 minutes
Temperature program: 8°C/min to 200°C
Final temperature: 200°C, hold for 6 minutes.
A trap preheated to 150°C prior to trap desorption is required to
provide adequate chromatography of the gas analytes.
7.2.5 Column 3 (A sample chromatogram is presented in Figure 8)
Carrier gas (He) flow rate: 4 mL/min
Initial temperature: 10°C, hold for 5 minutes
Temperature program: 6°C/min to 70°C, then 15°C/min
to 145°C
Final temperature: 145°C, hold until all expected
compounds have eluted.
7.2.6 Direct injection - Column 2
Carrier gas (He) flow rate: 4 mL/min
Column: J&W DB-624, 70m x 0.53 mm
Initial temperature: 40°C, hold for 3 minutes
Temperature program: 8°C/min
Final temperature: 260°C, hold until all expected
compounds have eluted.
Column Bake out (direct inj): 75 minutes
Injector temperature: 200-225°C
Transfer line temperature: 250-300°C
7.2.7 Direct Split Interface - Column 4
Carrier gas (He) flow rate: 1.5 mL/min
Initial temperature: 35°C, hold for 2 minutes
Temperature program: 4°C/min to 50°C
10°C/min to 220°C
Final temperature: 220°C, hold until all expected
compounds have eluted
Split ratio: 100:1
Injector temperature: 125°C
7.3 Initial calibration - the recommended MS operating conditions
Mass range: 35-260 amu
Scan time: 0.6-2 sec/scan
Source temperature: According to manufacturer's specifications
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Ion trap only: Set axial modulation, manifold temperature,
and emission current to manufacturer's
recommendations
7.3.1 Each GC/MS system must be hardware-tuned to meet the criteria
in Table 4 for a 5-50 ng injection or purging of 4-bromofluorobenzene
(2 p.L injection of the BFB standard). Analyses must not begin until these
criteria are met.
7.3.2 Set up the purge-and-trap system as outlined in Method 5030 if
purge-and-trap analysis is to be utilized. A set of at least five
calibration standards containing the method analytes is needed. One
calibration standard should contain each analyte at a concentration
approaching but greater than the method detection limit (Table 1) for that
compound; the other calibration standards should contain analytes at
concentrations that define the range of the method. Calibration should be
done using the sample introduction technique that will be used for
samples. For Method 5030, the purging efficiency for 5 mL of water is
greater than for 25 ml. Therefore, develop the standard curve with
whichever volume of sample that will be analyzed.
7.3.2.1 To prepare a calibration standard for purge-and-
trap or aqueous direct injection, add an appropriate volume of a
secondary dilution standard solution to an aliquot of organic-free
reagent water in a volumetric flask. Use a microsyringe and rapidly
inject the alcoholic standard into the expanded area of the filled
volumetric flask. Remove the needle as quickly as possible after
injection. Mix by inverting the flask three times only. Discard the
contents contained in the neck of the flask. Aqueous standards are
not stable and should be prepared daily. Transfer 5.0 ml (or 25 ml
if lower detection limits are required) of each standard to a gas
tight syringe along with 10 /LtL of internal standard. Then transfer
the contents to a purging device or syringe. Perform purge-and-trap
or direct injection as outlined in Method 5030.
7.3.2.2 To prepare a calibration standard for direct
injection analysis of oil, dilute standards in hexadecane.
7.3.3 Tabulate the area response of the characteristic ions (see
Table 5) against concentration for each compound and each internal
standard. Calculate response factors (RF) for each compound relative to
one of the internal standards. The internal standard selected for the
calculation of the RF for a compound should be the internal standard that
has a retention time closest to the compound being measured (Sec. 7.6.2).
The RF is calculated as follows:
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RF = (AxC,J/(AisCJ
where:
Ax = Area of the characteristic ion for the compound being
measured.
Ais = Area of the characteristic ion for the specific
internal standard.
Cis = Concentration of the specific internal standard.
Cx = Concentration of the compound being measured.
7.3.4 The average RF must be calculated and recorded for each
compound using the five RF values calculated for each compound from the
initial (5-point) calibration curve. A system performance check should be
made before this calibration curve is used. Five compounds (the System
Performance Check Compounds, or SPCCs) are checked for a minimum average
relative response factor. These compounds are chloromethane; 1,1-
dichloroethane; bromoform; 1,1,2,2-tetrachloroethane; and chlorobenzene.
These compounds are used to check compound instability and to check for
degradation caused by contaminated lines or active sites in the system.
Examples of these occurrences are:
7.3.4.1 Chloromethane - This compound is the most likely
compound to be lost if the purge flow is too fast.
7.3.4.2 Bromoform - This compound is one of the compounds
most likely to be purged very poorly if the purge flow is too slow.
Cold spots and/or active sites in the transfer lines may adversely
affect response. Response of the quantitation ion (m/z 173) is
directly affected by the tuning of BFB at ions m/z 174/176.
Increasing the m/z 174/176 ratio relative to m/z 95 may improve
bromoform response.
7.3.4.3 Tetrachloroethane and 1,1-dichloroethane - These
compounds are degraded by contaminated transfer lines in purge-and-
trap systems and/or active sites in trapping materials.
7.3.5 Using the RFs from the initial calibration, calculate and
record the percent relative standard deviation (%RSD) for all compounds.
The percent RSD is calculated as follows:
SD
% RSD = — x 100%
RFX
where:
RSD = Relative standard deviation.
RFX = mean of 5 initial RFs for a compound.
SD = standard deviation of the 5 initial RFs for a compound.
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so =
E
(RFi-RF):
n-1
where:
RFj = RF for each of the 5 calibration levels
N = number of RF values (i.e., 5)
The percent relative standard deviation should be less than 15% for
each compound. However, the %RSD for each individual Calibration Check
Compound (CCC) must be less than 30%. The CCCs are:
1,1-Dichloroethene,
Chloroform,
1,2-Dichloropropane,
Toluene,
Ethyl benzene, and
Vinyl chloride.
7.3.5.1 If a %RSD greater than 30 percent is measured for
any CCC, then corrective action to eliminate a system leak and/or
column reactive sites is required before reattempting calibration.
7.3.6 Linearity - If the %RSD of any compound is 15% or less, then
the relative response factor is assumed to be constant over the
calibration range, and the average relative response factor may be used
for quantitation.
7.3.6.1 If the %RSD of any compound is greater than 15%,
construct calibration curves of area ratio (A/Ais) versus
concentration using first or higher order regression fit of the five
calibration points. The analyst should select the regression order
which introduces the least calibration error into the quantitation.
The use of calibration curves is a recommended alternative to average
response factor calibration (Sec. 7.6.2.4), and a useful diagnostic
of standard preparation accuracy and absorption activity in the
chromatographic system.
7.3.7 These curves are verified each shift by purging a performance
standard. Recalibration is required only if calibration and on-going
performance criteria cannot be met.
7.4 GC/MS calibration verification
7.4.1 Prior to the analysis of samples, inject or purge 5-50 ng of
the 4-bromofluorobenzene standard following Method 5030. The resultant
mass spectra for the BFB must meet all of the criteria given in Table 4
before sample analysis begins. These criteria must be demonstrated each
12-hour shift.
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7.4.2 The initial calibration curve (Sec. 7,3) for each compound of
interest must be checked and verified once every 12 hours during analysis
with the introduction technique used for samples. This is accomplished by
analyzing a calibration standard that is at a concentration near the
midpoint concentration for the working range of the GC/MS by checking the
SPCC and CCC.
7.4.3 System Performance Check Compounds (SPCCs) - A system
performance check must be made each 12 hours. If the SPCC criteria are
met, a comparison of relative response factors is made for all compounds.
This is the same check that is applied during the initial calibration. If
the minimum relative response factors are not met, the system must be
evaluated, and corrective action must be taken before sample analysis
begins. Some possible problems are standard mixture degradation,
injection port inlet contamination, contamination at the front end of the
analytical column, and active sites in the column or chromatographic
system.
7.4.3.1 The minimum relative response factor for volatile
SPCCs are as follows:
Chloromethane 0.10
1,1-Dichloroethane 0.10
Bromoform >0.10
Chlorobenzene 0.30
1,1,2,2-Tetrachloroethane 0.30
7.4.4 Calibration Check Compounds (CCCs) - After the system
performance check is met, CCCs listed in Sec. 7.3.5 are used to check the
validity of the initial calibration.
Calculate the percent drift using the following equation:
% Drift = (C, - CC)/C, x 100
where:
C, = Calibration Check Compound standard concentration.
Cc = Measured concentration using selected quantitation method.
If the percent drift for each CCC is less than 20%, the initial
calibration is assumed to be valid. If the criterion is not met (> 20%
drift), for any one CCC, corrective action must be taken. Problems
similar to those listed under SPCCs could affect this criterion. If no
source of the problem can be determined after corrective action has been
taken, a new five point calibration MUST be generated. This criterion
MUST be met before quantitative sample analysis begins. If the CCCs are
not required analytes by the permit, then all required analytes must meet
the 20% drift criterion.
7.4.5 The internal standard responses and retention times in the
check calibration standard must be evaluated immediately after or during
data acquisition. If the retention time for any internal standard changes
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by more than 30 seconds from the last calibration check (12 hours), the
chromatographic system must be inspected for malfunctions and corrections
must be made, as required. If the EICP area for any of the internal
standards changes by a factor of two (-50% to +100%) from the last daily
calibration check standard, the mass spectrometer must be inspected for
malfunctions and corrections must be made, as appropriate. When
corrections are made, reanalysis of samples analyzed while the system was
malfunctioning is necessary.
7.5 GC/MS analysis
7.5.1 It is highly recommended that the extract be screened on a
headspace-GC/FID (Methods 3810/8015), headspace-GC/PID/ELCD (Methods
3810/8021), or waste dilution-GC/PID/ELCD (Methods 3585/8021) using the
same type of capillary column. This will minimize contamination of the
GC/MS system from unexpectedly high concentrations of organic compounds.
Use of screening is particularly important when this method is used to
achieve low detection levels.
7.5.2 All samples and standard solutions must be allowed to warm to
ambient temperature before analysis. Set up the purge-and-trap system as
outlined in Method 5030 if purge-and-trap introduction will be used.
7.5.3 BFB tuning criteria and GC/MS calibration verification
criteria must be met before analyzing samples.
7.5.3.1 Remove the plunger from a 5 ml syringe and attach
a closed syringe valve. If lower detection limits are required,- use
a 25 ml syringe. Open the sample or standard bottle, which has been
allowed to come to ambient temperature, and carefully pour the sample
into the syringe barrel to just short of overflowing. Replace the
syringe plunger and compress the sample. Open the syringe valve and
vent any residual air while adjusting the sample volume to 5.0 ml.
7.5.4 The process of taking an aliquot destroys the validity of
aqueous and soil samples for future analysis; therefore, if there is only
one VOA vial, the analyst should prepare a second aliquot for analysis at
this time to protect against possible loss of sample integrity. This
second sample is maintained only until such time when the analyst has
determined that the first sample has been analyzed properly. For aqueous
samples, filling one 20 ml syringe would require the use of only one
syringe. If a second analysis is needed from a syringe, it must be
analyzed within 24 hours. Care must be taken to prevent air from leaking
into the syringe.
7.5.4.1 The following procedure is appropriate for
diluting aqueous purgeable samples. All steps must be performed
without delays until the diluted sample is in a gas-tight syringe.
7.5.4.1.1 Dilutions may be made in volumetric flasks
(10 to 100 ml). Select the volumetric flask that will allow
for the necessary dilution. Intermediate dilutions may be
necessary for extremely large dilutions.
8260A - 20 Revision 1
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7.5.4.1.2 Calculate the approximate volume of organic-
free reagent water to be added to the volumetric flask
selected and add slightly less than this quantity of organic-
free reagent water to the flask.
7.5.4.1.3 Inject the proper aliquot of sample from the
syringe into the flask. Aliquots of less than 1 ml are not
recommended. Dilute the sample to the mark with organic-free
reagent water. Cap the flask, invert, and shake three times.
Repeat above procedure for additional dilutions.
7.5.4.1.4 Fill a 5 ml syringe with the diluted sample.
7.5.4.2 Compositing aqueous samples prior to GC/MS
analysis
7.5.4.2.1 Add 5 ml or equal larger amounts of each
sample (up to 5 samples are allowed) to a 25 mL glass syringe.
Special precautions must be made to maintain zero headspace in
the syringe.
7.5.4.2.2 The samples must be cooled at 4°C during this
step to minimize volatilization losses.
7.5.4.2.3 Mix well and draw out a 5 ml aliquot for
analysis.
7.5.4.2.4 Follow sample introduction, purging, and
desorption steps described in Method 5030.
7.5.4.2.5 If less than five samples are used for
compositing, a proportionately smaller syringe may be used
unless a 25 ml sample is to be purged.
7.5.5 Add 10.0 /xL of surrogate spiking solution and 10 /zL of
internal standard spiking solution to each sample. The surrogate and
internal standards may be mixed and added as a single spiking solution.
The addition of 10 /nL of the surrogate spiking solution to 5 ml of sample
is equivalent to a concentration of 50 /ig/L of each surrogate standard.
The addition of 10 pi of the surrogate spiking solution to 5 g of sample
is equivalent to a concentration of 50 M9/kg of each surrogate standard.
7.5.5.1 If a more sensitive mass spectrometer is employed
to achieve lower detection levels, more dilute surrogate and internal
standard solutions may be required.
7.5.6 Perform purge-and-trap or direct injection by Method 5030. If
the initial analysis of sample or a dilution of the sample has a
concentration of analytes that exceeds the initial calibration range, the
sample must be reanalyzed at a higher dilution. Secondary ion
quantitation is allowed only when there are sample interferences with the
primary ion. When a sample is analyzed that has saturated ions from a
compound, this analysis must be followed by a blank organic-free reagent
8260A - 21 Revision 1
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water analysis. If the blank analysis is not free of interferences, the
system must be decontaminated. Sample analysis may not resume until the
blank analysis is demonstrated to be free of interferences.
7.5.6.1. All dilutions should keep the response of the
major constituents (previously saturated peaks) in the upper half of
the linear range of the curve. Proceed to Sees. 7.6.1 and 7.6.2 for
qualitative and quantitative analysis.
7.5.7 For matrix spike analysis, add 10 juL of the matrix spike
solution (Sec. 5.13) to the 5 ml of sample to be purged. Disregarding any
dilutions, this is equivalent to a concentration of 50 p.g/1 of each matrix
spike standard.
7.6 Data interpretation
7.6.1 Qualitative analysis
7.6.1.1 The qualitative identification of compounds
determined by this method is based on retention time, and on
comparison of the sample mass spectrum, after background correction,
with characteristic ions in a reference mass spectrum. The reference
mass spectrum must be generated by the laboratory using the
conditions of this method. The characteristic ions from the
reference mass spectrum are defined to be the three ions of greatest
relative intensity, or any ions over 30% relative intensity if less
than three such ions occur in the reference spectrum. Compounds
should be identified as present when the criteria below are met.
7.6.1.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographic peak containing ions
specific for the target compound at a compound-specific
retention time will be accepted as meeting this criterion.
7.6.1.1.2 The RRT of the sample component is within
± 0.06 RRT units of the RRT of the standard component.
7.6.1.1.3 The relative intensities of the
characteristic ions agree within 30% of the relative
intensities of these ions in the reference spectrum.
(Example: For an ion with an abundance of 50% in the
reference spectrum, the corresponding abundance in a sample
spectrum can range between 20% and 80%.)
7.6.1.1.4 Structural isomers that produce very similar
mass spectra should be identified as individual isomers if
they have sufficiently different GC retention times.
Sufficient GC resolution is achieved if the height of the
valley between two isomer peaks is less than 25% of the sum of
8260A - 22 Revision 1
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the two peak heights. Otherwise, structural isomers are
identified as isomeric pairs.
7.6.1.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce
mass spectra containing ions contributed by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
appropriate selection of analyte spectra and background
spectra is important. Examination of extracted ion current
profiles of appropriate ions can aid in the selection of
spectra, and in qualitative identification of compounds. When
analytes coelute (i.e., only one chromatographic peak is
apparent), the identification criteria can be met, but each
analyte spectrum will contain extraneous ions contributed by
the coeluting compound.
7.6.1.2 For samples containing components not associated
with the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the type of analyses
being conducted. Guidelines for making tentative identification are:
(1) Relative intensities of major ions in the reference
spectrum (ions > 10% of the most abundant ion) should
be present in the sample spectrum.
(2) The relative intensities of the major ions should agree
within ± 20%. (Example: For an ion with an abundance
of 50% in the standard spectrum, the corresponding
sample ion abundance must be between 30 and 70%).
(3) Molecular ions present in the reference spectrum should
be present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the
reference spectrum should be reviewed for possible
background contamination or presence of coeluting
compounds.
(5) Ions present in the reference spectrum but not in the
sample spectrum should be reviewed for possible
subtraction from the sample spectrum because of
background contamination or coeluting peaks. Data
system library reduction programs can sometimes create
these discrepancies.
Computer generated library search routines should not use
normalization routines that would misrepresent the library or unknown
spectra when compared to each other. Only after visual comparison
of sample with the nearest library searches will the mass spectral
interpretation specialist assign a tentative identification.
8260A - 23 Revision 1
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7.6.2 Quantitative analysis
7.6.2.1 When a compound has been identified, the
quantitation of that compound will be based on the integrated
abundance from the EICP of the primary characteristic ion.
Quantitation will take place using the internal standard technique.
The internal standard used shall be the one nearest the retention
time of that of a given analyte.
7.6.2.2 When MS response is linear and passes through the
origin, calculate the concentration of each identified analyte in the
sample as follows:
Water
(A,)(I.)
concentration (jug/L) = zz;
(AJ(RF)(V0)
where:
Ax = Area of characteristic ion for compound being
measured.
Is = Amount of internal standard injected (ng).
Ais = Area of characteristic ion for the internal
standard.
RF = Mean relative response factor for compound being
measured.
V0 = Volume of water purged (ml), taking into
consideration any dilutions made.
Sediment/Soil Sludge (on a dry-weight basis) and Waste
(normally on a wet-weight basis)
(Ax)(U(Vt)
concentration (M9/kg) = =r
(Ais)(RF)(Vi)(WJ(D)
where:
Ax> Is> Ais, RF, = Same as for water.
Vt = Volume of total extract (/A) (use 10,000 juL or a
factor of this when dilutions are made).
Vi = Volume of extract added (juL) for purging.
Ws = Weight of sample extracted or purged (g).
D = % dry weight of sample/100, or 1 for a wet-weight
basis.
7.6.2.3 Where applicable, an estimate of concentration for
noncalibrated components in the sample should be made. The formulae
given above should be used with the following modifications: The
areas Ax and Ais should be from the total ion chromatograms, and the
RF for the compound should be assumed to be 1. The concentration
8260A - 24 Revision 1
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obtained should be reported indicating (1) that the value is an
estimate and (2) which internal standard was used to determine
concentration. Use the nearest internal standard free of
interferences.
7.6.2.4 Alternatively, the regression line fitted to the
initial calibration (Sec. 7.3.6.1) may be used for determination of
analyte concentration.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for general quality control
procedures.
8.2 Additional required instrument QC is found in the Sees. 7.3 and 7.4:
8.2.1 The GC/MS system must be tuned to meet the BFB specifications.
8.2.2 There must be an initial calibration of the GC/MS system
8.2.3 The GC/MS system must meet the SPCC criteria and the CCC
criteria, each 12 hours.
8.3 To establish the ability to generate acceptable accuracy and
precision, the analyst must perform the following operations.
8.3.1 A quality control (QC) reference sample concentrate is
required containing each analyte at a concentration of 10 mg/L or less in
methanol . The QC reference sample concentrate may be prepared from pure
standard materials or purchased as certified solutions. If prepared by
the laboratory, the QC reference sample concentrate must be made using
stock standards prepared independently from those used for calibration.
8.3.2 Prepare a QC reference sample to contain 20 ng/i or less of
each analyte by adding 200 /zL of QC reference sample concentrate to 100 mL
of organic-free reagent water.
8.3.3 Four 5-mL aliquots of the well mixed QC reference sample are
analyzed according to the method beginning in Sec. 7.5.1.
8.3.4 Calculate the average recovery (x) in M9/L, and the standard
deviation of the recovery (s) in /zg/L, for each analyte using the four
results.
8.3.5 Tables 7 and 8 provide single laboratory recovery and
precision data obtained for the method analytes from water. Similar
results from dosed water should be expected by any experienced laboratory.
Compare s and x (Sec. 8.3.4) for each analyte to the single laboratory
recovery and precision data. Results are comparable if the calculated
standard deviation of the recovery does not exceed 2.6 times the single
laboratory RSD or 20%, whichever is greater, and the mean recovery lies
within the interval x ± 3s or x ± 30%, whichever is greater.
8260A - 25 Revision 1
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NOTE: The large number of analytes in Tables 7 and 8 present a
substantial probability that one or more will fail at least
one of the acceptance criteria when all analytes of a given
method are determined.
8.3.6 When one or more of the analytes tested are not comparable to
the data in Table 6 or 7, the analyst must proceed according to Sec.
8.3.6.1 or 8.3.6.2.
8.3.6.1 Locate and correct the source of the problem and
repeat the test for all analytes beginning with Sec. 8.3.2.
8.3.6.2 Beginning with Sec. 8.3.2, repeat the test only
for those analytes that are not comparable. Repeated failure,
however,, will confirm a general problem with the measurement system.
If this occurs, locate and correct the source of the problem and
repeat the test for all compounds of interest beginning with Sec.
8.3.2.
8.4 For aqueous and soil matrices, laboratory established surrogate
control limits should be compared with the control limits listed in Table 8.
8.4.1 If recovery is not within limits, the following procedures are
required.
8.4.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.4.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and re-analyze
the extract.
8.4.1.3 If no problem is found, re-extract and re-analyze
the sample.
8.4.1.4 If, upon re-analysis, the recovery is again not
within limits, flag the data as "estimated concentration".
8.4.2 At a minimum, each laboratory should update surrogate recovery
limits on a matrix-by-matrix basis, annually.
9.0 METHOD PERFORMANCE
9.1 The method detection limit (MDL) is defined as the minimum
concentration of a substance that can be measured and reported with 99%
confidence that the value is above zero. The MDL actually achieved in a given
analysis will vary depending on instrument sensitivity and matrix effects.
9.2 This method has been tested in a single laboratory using spiked
water. Using a wide-bore capillary column, water was spiked at concentrations
between 0.5 and 10 M9/L. Single laboratory accuracy and precision data are
8260A - 26 Revision 1
September 1994
-------
presented for the method analytes in Table 6. Calculated MDLs are presented in
Table 1.
9.3 The method was tested using water spiked at 0.1 to 0.5 jug/L and
analyzed on a cryofocussed narrow-bore column. The accuracy and precision data
for these compounds are presented in Table 7- MDL values were also calculated
from these data and are presented in Table 2.
9.4 Direct injection has been used for the analysis of waste motor oil
samples using a wide-bore column. The accuracy and precision data for these
compounds are presented in Table 10.
10.0 REFERENCES
1. Methods for the Determination of Organic Compounds in Finished Drinking
Water and Raw Source Water Method 524.2; U.S. Environmental Protection
Agency. Office of Research Development, Environmental Monitoring and
Support Laboratory, Cincinnati, OH 1986.
2. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, July 1985, Revision.
3. Bellar, T.A.; J.J. Lichtenberg. vh. Amer. Water Works Assoc. 1974, 66(12),
739-744.
4. Bellar, T.A.; J.J. Lichtenberg. "Semi-Automated Headspace Analysis of
Drinking Waters and Industrial Waters for Purgeable Volatile Organic
Compounds"; in Van Hall, Ed.; Measurement of Organic Pollutants in Water
and Wastewater, ASTM STP 686, pp 108-129, 1979.
5. Budde, W.L.; J.W. Eichelberger. "Performance Tests for the Evaluation of
Computerized Gas Chromatography/Mass Spectrometry Equipment and
Laboratories"; U.S. Environmental Protection Agency. Environmental
Monitoring and Support Laboratory, Cincinnati, OH 45268, April 1980; EPA-
600/4-79-020.
6. Eichelberger, J.W.; L.E. Harris; W.L. Budde. "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass
Spectrometry Systems"; Analytical Chemistry 1975, 47, 995-1000.
7. Olynyk, P.; W.L. Budde; J.W. Eichelberger. "Method Detection Limit for
Methods 624 and 625"; Unpublished report, October 1980.
8. Non Cryogenic Temperatures Program and Chromatogram, Private
Communications; Myron Stephenson and Frank Allen, EPA Region IV
Laboratory, Athens, GA.
9. Marsden, P.; C.L. Helms, B.N. Colby. "Analysis of Volatiles in Waste Oil";
report for B. Lesnik, OSW/EPA under EPA contract 68-W9-001, 6/92.
8260A - 27 Revision 1
September 1994
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10. Methods for the Determination of Organic Compounds in Drinking Water,
Supplement II Method 524.2; U.S. Environmental Protection Agency. Office
of Research and Development, Environmental Monitoring Systems Laboratory,
Cincinnati, OH 1992.
8260A - 28 Revision 1
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TABLE 1.
CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS (MDL)
FOR VOLATILE ORGANIC COMPOUNDS ON WIDE-BORE CAPILLARY COLUMNS
ANALYTE
Dichlorodifluoromethane
Chloromethane
Vinyl Chloride
Bromomethane
Chloroethane
Tri chl orof 1 uoromethane
Acrolein
lodomethane
Acetonitrile
Carbon disulfide
Allyl chloride
Methylene chloride
1,1-Dichloroethene
Acetone
trans-l,2-Dichloroethene
Acrylonitrile
1,1-Dichloroethane
Vinyl acetate
2,2-Dichloropropane
2-Butanone
cis-l,2-Dichloroethene
Propionitrile
Chloroform
Bromochloromethane
Methacrylonitrile
1,1, 1 -Tri chl oroethane
Carbon tetrachloride
1,1-Dichloropropene
Benzene
1,2-Dichloroethane
Trichloroethene
1,2-Dichloropropane
Bromodi chl oromethane
Dibromomethane
Methyl methacryl ate
1,4-Dioxane
2-Chloroethyl vinyl ether
4-Methyl -2-pentanone
trans-l,3-Dichloropropene
Toluene
cis-l,3-Dichloropropene
1,1,2-Tri chl oroethane
RETENTION TIME
(minutes)
Column la
1.35
1.49
1.56
2.19
2.21
2.42
3.19
3.56
4.11
4.11
4.11
4.40
4.57
4.57
4.57
5.00
6.14
6.43
8.10
--
8.25
8.51
9.01
9.19
10.18
11.02
--
11.50
12.09
14.03
14.51
15.39
15.43
15.50
16.17
--
17.32
17.47
18.29
19.38
19.59
Column 2"
0.70
0.73
0.79
0.96
1.02
1.19
2.06
1.57
2.36
2.93
3.80
3.90
4.80
4.38
4.84
5.26
5.29
5.67
5.83
7.27
7.66
8.49
7.93
--
10.00
--
11.05
Column 2'°
3.13
3.40
3.93
4.80
6.20
9.27
7.83
9.90
10.80
11.87
11.93
12.60
12.37
12.83
13.17
13.10
13.50
13.63
14.80
15.20
15.80
15.43
16.70
17.40
17.90
18.30
MDLd
(M9/L)
0.10
0.13
0.17
0.11
0.10
0.08
0.03
0.12
0.06
0.04
0.35
0.12
0.03
0.04
0.08
0.21
0.10
0.04
0.06
0.19
0.04
0.08
0.24
--
0.11
--
0.10
8260A - 29
Revision 1
September 1994
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TABLE 1.
(Continued)
ANALYTE
RETENTION TIME
(minutes)
MDLd
Ethyl methacrylate
2-Hexanone
Tetrachl oroethene
1,3-Dichloropropane
Di bromochl oromethane
1,2-Dibromoethane
1-Chlorohexane
Chl orobenzene
1,1,1 , 2-Tetrachl oroethane
Ethyl benzene
p-Xylene
m-Xylene
o-Xylene
Styrene
Bromoform
Isopropyl benzene (Cumene)
cis-l,4-Dichloro-2-butene
1,1,2 , 2-Tetrachl oroethane
Bromobenzene
1,2,3-Trichloropropane
n-Propyl benzene
2-Chlorotoluene
trans-l,4-Dichloro-2-butene
1 , 3 , 5-Trimethyl benzene
4-Chlorotoluene
Pentachl oroethane
1,2, 4 -Tr i methyl benzene
sec-Butyl benzene
tert-Butyl benzene
p- Isopropyl tol uene
1 , 3-Di chl orobenzene
1,4-Dichlorobenzene
Benzyl chloride
n-Butyl benzene
1,2-Dichlorobenzene
l,2-Dibromo-3-chloropropane
1, 2, 4-Tri chl orobenzene
Hexachlorobutadiene
Naphthalene
1 , 2 , 3-Tri chl orobenzene
Column 1"
20.01
20.30
20.26
20.51
21.19
21.52
--
23.17
23.36
23.38
23.54
23.54
25.16
25.30
26.23
26.37
27.12
27.29
27.46
27.55
27.58
28.19
28.26
28.31
28.33
29.41
29.47
30.25
30.59
30.59
30.56
31.22
32.00
32.23
32.31
35.30
38.19
38.57
39.05
40.01
Column 2b
11.15
11.31
11.85
11.83
13.29
13.01
13.33
13.39
13.69
13.68
14.52
14.60
14.88
15.46
16.35
15.86
16.23
16.41
16.42
16.90
16.72
17.70
18.09
17.57
18.52
18.14
18.39
19.49
19.17
21.08
23.08
23.68
23.52
24.18
Column 2'°
18.60
18.70
19.20
19.40
--
20.67
20.87
21.00
21.30
21.37
22.27
22.40
22.77
23.30
24.07
24.00
24.13
24.33
24.53
24.83
24.77
31.50
26.13
26.60
26.50
26.37
26.60
27.32
27.43
--
31.50
32.07
32.20
32.97
0.14
0.04
0.05
0.06
0.05
0.04
0.05
0.06
0.13
0.05
0.11
0.04
0.12
0.15
0.04
0.03
0.32
0.04
0.04
0.05
0.06
0.13
0.13
0.14
0.12
0.12
0.03
0.11
0.03
0.26
0.04
0.11
0.04
0.03
8260A - 30
Revision 1
September 1994
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TABLE 1.
(Continued)
ANALYTE
RETENTION TIME
(minutes)
Column la Column 2 Column 2'c
MDLd
(M9/L)
INTERNAL STANDARDS/SURROGATES
1,4-Difluorobenzene
Chlorobenzene-d5
l,4-Dichlorobenzene-d4
4-Bromofl uorobenzene
1 , 2-Di chl orobenzene-d4
Dichloroethane-d4
Di bromof 1 uoromethane
Toluene-d8
Pentaf 1 uorobenzene
Fl uorobenzene
13.26
23.10
31.16
27.83 15.71
32.30 19.08
12.08
18.27
13.00 6.27
23.63
27.25
14.06
a Column 1 - 60 meter x 0.75 mm ID VOCOL capillary. Hold at 10°C for 8 minutes,
then program to 180°C at 4°/niin.
b Column 2 - 30 meter x 0.53 mm ID DB-624 wide-bore capillary using cryogenic
oven. Hold at 10°C for 5 minutes, then program to 160°C at 6°/min.
c Column 2' - 30 meter x 0.53 mm ID DB-624 wide-bore capillary, cooling GC oven
to ambient temperatures. Hold at 10°C for 6 minutes, program to 70°C at
10°/min, program to 120°C at 5°/min, then program to 180°C at 8°/min.
d MDL based on a 25 mL sample volume.
8260A - 31
Revision 1
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TABLE 2.
CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS (MDL)
FOR VOLATILE ORGANIC COMPOUNDS ON NARROW-BORE CAPILLARY COLUMNS
ANALYTE
RETENTION TIME
(minutes)
Column 3a
MDLb
Di chl orodi f 1 uoromethane
Chl oromethane
Vinyl chloride
Bromomethane
Chloroethane
Tr i chl orof 1 uoromethane
1,1-Dichloroethene
Methylene chloride
trans- 1,2-Di chl oroethene
1,1-Dichloroethane
cis- 1,2-Di chl oroethene
2,2-Dichloropropane
Chloroform
Bromochl oromethane
1,1,1-Tri chl oroethane
1,2-Dichloroethane
1,1-Dichloropropene
Carbon tetrachloride
Benzene
1,2-Dichloropropane
Trichloroethene
Dibromomethane
Bromodi chl oromethane
Toluene
1,1, 2 -Tri chl oroethane
1,3-Dichloropropane
Di bromochl oromethane
Tetrachl oroethene
1,2-Di bromoethane
Chlorobenzene
1,1,1 , 2-Tetrachl oroethane
Ethyl benzene
p-Xylene
m-Xylene
Bromoform
o-Xylene
Styrene
1,1,2, 2-Tetrachl oroethane
1,2,3-Trichloropropane
Isopropyl benzene
0.88
0.97
1.04
1.29
1.45
1.77
2.33
2.66
3.54
4.03
5.07
5.31
5.55
5.63
6.76
7.00
7.16
7.41
7.41
8.94
9.02
9.09
9.34
11.51
11.99
12.48
12.80
13.20
13.60
14.33
14.73
14.73
15.30
15.30
15.70
15.78
15.78
15.78
16.26
16.42
0.11
0.05
0.04
0.06
0.02
0.07
0.05
0.09
0.03
0.03
0.06
0.08
0.04
0.09
0.04
0.02
0.12
0.02
0.03
0.02
0.02
0.01
0.03
0.08
0.08
0.08
0.07
0.05
0.10
0.03
0.07
0.03
0.06
0.03
0.20
0.06
0.27
0.20
0.09
0.10
8260A - 32
Revision 1
September 1994
-------
TABLE 2.
(Continued)
ANALYTE
RETENTION TIME
(minutes)
Column 3a
MDLb
(M9/L)
Bromobenzene
2-Chlorotoluene
n-Propyl benzene
4-Chlorotoluene
1,3, 5-Trimethyl benzene
tert-Butyl benzene
1 , 2 , 4-Trimethyl benzene
sec-Butyl benzene
1,3-Dichlorobenzene
p-Isopropyl toluene
1 ,4-Dichlorobenzene
1,2-Dichlorobenzene
n-Butyl benzene
l,2-Dibromo-3-chloropropane
1,2,4-Trichlorobenzene
Naphthalene
Hexachlorobutadiene
1,2,3-Trichlorobenzene
16.42
16.74
16.82
16.82
16.99
17.31
17.31
17.47
17.47
17.63
17.63
17.79
17.95
18.03
18.84
19.07
19.24
19.24
0.11
0.08
0.10
0.06
0.06
0.33
0.09
0.12
0.05
0.26
0.04
0.05
0.10
0.50
0.20
0.10
0.10
0.14
a Column 3-30 meter x 0.32 mm ID DB-5 capillary with 1 /im film thickness,
b MDL based on a 25 ml sample volume.
8260A - 33
Revision 1
September 1994
-------
TABLE 3.
ESTIMATED QUANTITATION LIMITS FOR VOLATILE ANALYTES8
Estimated Quantitation Limits
(All Analytes in Table 1}
Ground water Low Soil/Sediment6
/zg/kg
Purging 5 mL of water 5
Purging 25 mL of water 1
Soil/Sediment
Estimated Quantitation Limit (EQL) - The lowest concentration that can be
reliably achieved within specified limits of precision and accuracy during
routine laboratory operating conditions. The EQL is generally 5 to 10
times the MDL. However, it may be nominally chosen within these guidelines
to simplify data reporting. For many analytes the EQL is selected from the
lowest non-zero standard in the calibration curve. Sample EQLs are highly
matrix-dependent. The EQLs listed herein are provided for guidance and may
not always be achievable.
EQLs listed for soil/sediment are based on wet weight. Normally data are
reported on a dry weight basis; therefore, EQLs will be higher, based on
the percent dry weight in each sample.
Other Matrices Factor0
Water miscible liquid waste 50
High-concentration soil and sludge 125
Non-water miscible waste 500
°EQL = [EQL for low soil/sediment (see Table 3)] X [Factor]. For non-aqueous
samples, the factor is on a wet-weight basis.
8260A - 34 Revision 1
September 1994
-------
TABLE 4.
BFB MASS - INTENSITY SPECIFICATIONS (4-BROMOFLUOROBENZENE)'
Mass Intensity Required (relative abundance)
50 15 to 40% of mass 95
75 30 to 60% of mass 95
95 base peak, 100% relative abundance
96 5 to 9% of mass 95
173 less than 2% of mass 174
174 greater than 50% of mass 95
175 5 to 9% of mass 174
176 greater than 95% but less than 101% of mass 174
177 5 to 9% of mass 176
Alternate tuning criteria may be used (e.g. CLP, Method 524.2, or
manufacturers' instructions), provided that method performance is not
adversely affected.
8260A - 35 Revision 1
September 1994
-------
TABLE 5.
CHARACTERISTIC MASSES (M/Z) FOR PURGEABLE ORGANIC COMPOUNDS
Analyte
Primary
Characteristic
Ion
Secondary
Characteristic
Ion(s)
Acetone
Acetonitrile
Acrolein
Acrylonitrile
Allyl alcohol
Ally! chloride
Benzene
Benzyl chloride
Bromoacetone
Bromobenzene
Bromochl oromethane
Bromodichloromethane
Bromoform
Bromomethane
iso-Butanol
n-Butanol
2-Butanone
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon disulfide
Carbon tetrachloride
Chloral hydrate
Chloroacetonitrile
Chlorobenzene
1-Chlorobutane
Chi orodi bromomethane
Chloroethane
2-Chloroethanol
bis-(2-chloroethyl) sulfide
2-Chloroethyl vinyl ether
Chloroform
Chi oromethane
Chloroprene
3-Chloropropionitrile
2-Chlorotoluene
4-Chlorotoluene
1 , 2-Di bromo-3-chl oropropane
Di bromochl oromethane
1,2-Dibromoethane
Di bromomethane
1,2-Dichlorobenzene
l,2-Dichlorobenzene-d4
58
41
56
53
57
76
78
91
136
156
128
83
173
94
74
56
72
91
105
119
76
117
82
48
112
56
129
64(49*)
49
109
63
83
50(49*)
53
54
91
91
75
129
107
93
146
152
43
41, 40, 39
55, 58
52, 51
57, 58, 39
76, 41, 39, 78
-
91, 126, 65, 128
43, 136, 138, 93, 95
77, 158
49, 130
85, 127
175, 254
96
43
41
43, 72
92, 134
134
91, 134
78
119
44, 84, 86, 111
75
77, 114
49
208, 206
66(51*)
49, 44, 43, 51, 80
111, 158, 160
65, 106
85
52(51*)
53, 88, 90, 51
54, 49, 89, 91
126
126
155, 157
127
109, 188
95, 174
111, 148
115, 150
8260A - 36
Revision 1
September 1994
-------
TABLE 5.(continued)
Analyte
Primary
Characteristic
Ion
Secondary
Characteristic
Ion(s)
1,3-Dichlorobenzene
1 , 4-Di chl orobenzene
cis-l,4-Dichloro-2-butene
trans -1, 4-Di chl oro-2-butene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-l,2-Dichloroethene
trans -1,2-Di chl oroethene
1,2-Dichloropropane
1,3-Dichloropropane
2 , 2-Di chl oropropane
l,3-Dichloro-2-propanol
1,1-Dichloropropene
cis-l,3-Dichloropropene
trans- 1,3-Oi chl oropropene
1,2,3,4-Diepoxybutane
Diethyl ether
1,4-Dioxane
Epichlorohydrin
Ethanol
Ethyl acetate
Ethyl benzene
Ethylene oxide
Ethyl methacrylate
Hexachlorobutadiene
Hexachloroethane
2-Hexanone
2-Hydroxypropionitrile
lodomethane
Isobutyl alcohol
Isopropyl benzene
p- I sopropyl toluene
Malononitrile
Methacrylonitrile
Methyl acrylate
Methyl -t-butyl ether
Methylene chloride
Methyl ethyl ketone
Methyl iodide
Methyl methacrylate
4-Methyl -2-pentanone
Naphthalene
Nitrobenzene
146
146
75
53
85
63
62
96
96
96
63
76
77
79
75
75
75
55
74
88
57
31
88
91
44
69
225
201
43
44
142
43
105
119
66
41
55
73
84
72
142
69
100
128
123
HI,
111,
75,
88,
87
65,
98
61,
61,
61,
112
78
97
79,
110,
77,
77,
55,
45,
88,
57,
45,
43,
106
44,
69,
223,
166,
58,
44,
127,
43,
120
134,
66,
41,
85
57
86,
43
142,
69,
43,
-
51,
148
148
53,
75
83
63
98
98
43,
77
39
39
57,
59
58,
49,
27,
45,
43,
41,
227
199,
57,
43,
141
41,
91
39,
67,
49
127,
41,
58,
77
77, 124
81, 49
56
43, 57
62, 51
46
61
42
99, 86,
203
100
42, 53
42, 74
65, 38
39, 52,
141
100, 39
85
, 89
114
66
8260A - 37
Revision 1
September 1994
-------
TABLE 5.(continued)
Primary Secondary
Characteristic Characteristic
Analyte Ion Ion(s)
2-Nitropropane
2-Picoline
Pentachloroethane
Propargyl alcohol
6-Propiolactone
Propionitrile (ethyl cyanide)
n-Propylamine
n-Propyl benzene
Pyridine
Styrene
1,2,3-Trichlorobenzene
1,2,4-Trichlorobenzene
1,1,1 , 2-Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
Toluene
1,1,1-Trichl oroethane
1 , 1 , 2-Trichl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
1 , 2 , 4-Trimethyl benzene
1, 3, 5-Trimethyl benzene
Vinyl acetate
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
INTERNAL STANDARDS/SURROGATES
1,4-Difluorobenzene
Chlorobenzene-d5
l,4-Dichlorobenzene-d4
4-Bromof 1 uorobenzene
Di bromof 1 uoromethane
Dichloroethane-d4
Toluene-d8
Pentafl uorobenzene
Fl uorobenzene
46
93
167
55
42
54
59
91
79
104
180
180
131
83
164
92
97
83
95
151
75
105
105
43
62
106
106
106
114
117
152
95
113
102
98
168
96
93,
167,
55,
42,
54,
59,
120
52
78
182,
182,
133,
131,
129,
91
99,
97,
97,
101,
77
120
120
86
64
91
91
91
115,
174,
77
66, 92, 78
130, 132, 165, 169
39, 38, 53
43, 44
52, 55, 40
41, 39
145
145
119
85
131, 166
61
85
130, 132
153
150
176
* - characteristic ion for an ion trap mass spectrometer (to be used when
ion-molecule reactions are observed)
8260A - 38 Revision 1
September 1994
-------
TABLE 6.
SINGLE LABORATORY ACCURACY AND PRECISION DATA FOR VOLATILE
ORGANIC COMPOUNDS IN WATER DETERMINED WITH A WIDE-
BORE CAPILLARY COLUMN
Analyte
Benzene
Bromobenzene
Bromochl oromethane
Bromodi chl oromethane
Bromoform
Bromomethane
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chl oromethane
2-Chlorotoluene
4-Chlorotoluene
l,2-Dibromo-3-Chloropropane
Di bromochl oromethane
1,2-Dibromoethane
Dibromomethane
1 , 2-Di chl orobenzene
1, 3 -Di chlorobenzene
1,4-Dichlorobenzene
Dichlorodifluoromethane
1 , 1 -Di chl orobenzene
1,2-Dichlorobenzene
1,1-Dichloroethene
cis-1, 2-Di chl oroethene
trans -1, 2-Di chl oroethene
1 , 2-Di chl oropropane
1,3-Dichloropropane
2 , 2-Di chl oropropane
1,1-Dichloropropene
Ethyl benzene
Hexachlorobutadiene
Isopropyl benzene
p-Isopropyl toluene
Methylene chloride
Naphthalene
n-Propyl benzene
Styrene
Cone. Number
Range, of Recovery8
jLtg/L Samples %
0.1
0.1
0.5
0.1
0.5
0.5
0.5
0.5
0.5
0.5
0.1
0.5
0.5
0.5
0.1
0.1
0.5
0.1
0.5
0.5
0.1
0.5
0.2
0.5
0.5
0.1
0.1
0.5
0.1
0.1
0.1
0.5
0.5
0.1
0.5
0.5
0.1
0.1
0.1
0.1
0.1
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 20
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
- 10
-100
- 10
-100
31
30
24
30
18
18
18
16
18
24
31
24
24
23
31
31
24
31
24
24
31
24
31
18
24
31
34
18
30
30
31
12
18
31
18
16
23
30
31
31
39
97
100
90
95
101
95
100
100
102
84
98
89
90
93
90
99
83
92
102
100
93
99
103
90
96
95
94
101
93
97
96
86
98
99
100
101
99
95
104
100
102
Standard
Deviation Percent
of Recovery13 RSD
6.5
5.5
5.7
5.7
6.4
7.8
7.6
7.6
7.4
7.4
5.8
8.0
5.5
8.3
5.6
8.2
16.6
6.5
4.0
5.6
5.8
6.8
6.6
6.9
5.1
5.1
6.3
6.7
5.2
5.9
5.7
14.6
8.7
8.4
6.8
7.7
6.7
5.0
8.6
5.8
7.3
5.7
5.5
6.4
6.1
6.3
8.2
7.6
7.6
7.3
8.8
5.9
9.0
6.1
8.9
6.2
8.3
19.9
7.0
3.9
5.6
6.2
6.9
6.4
7.7
5.3
5.4
6.7
6.7
5.6
6.1
6.0
16.9
8.9
8.6
6.8
7.6
6.7
5.3
8.2
5.8
7.2
8260A - 39
Revision 1
September 1994
-------
TABLE 6.
(Continued)
Analyte
Cone.
Range,
M9/L
Number
of Recovery"
Samples %
Standard
Deviation Percent
of Recovery1" RSD
1,1,1,2-Tetrachloroethane
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
1,2,3-Tri chlorobenzene
1,2,4-Tri chlorobenzene
1,1,1-Trichloroethane
1,1,2-Trichloroethane
Trichloroethene
Tri chlorof1uoromethane
1,2,3-Trichloropropane
1,2,4-Trimethylbenzene
1,3,5-Trimethylbenzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
0.5 -
0.1 -
0.5 -
0.5 -
0.5 -
0.5 -
0.5 -
0.5 -
0.5 -
0.5 -
0.5 -
0.5 -
0.5 -
0.5 -
0.1 -
0.1 -
0.5 -
10
10
10
10
10
10
10
10
10
10
10
10
10
10
31
10
10
24
30
24
18
18
18
18
18
24
24
16
18
23
18
18
31
18
90
91
89
102
109
108
98
104
90
89
108
99
92
98
103
97
104
6.1
5.7
6.0
8.1
9.4
9.0
7.9
7.6
6.5
7.2
15.6
8.0
6.8
6.5
7.4
6.3
8.0
6.8
6.3
6.8
8.0
8.6
8.3
8.1
7.3
7.3
8.1
14.4
8.1
7.4
6.7
7.2
6.5
7.7
a Recoveries were calculated using internal standard method. Internal standard
was fluorobenzene.
b Standard deviation was calculated by pooling data from three concentrations.
8260A - 40
Revision 1
September 1994
-------
TABLE 7.
SINGLE LABORATORY ACCURACY AND PRECISION DATA FOR
VOLATILE ORGANIC COMPOUNDS IN WATER DETERMINED
WITH A NARROW-BORE CAPILLARY COLUMN
Analyte
Benzene
Bromobenzene
Bromochl oromethane
Bromodichloromethane
Bromoform
Bromomethane
n-Butyl benzene
sec-Butyl benzene
tert-Butyl benzene
Carbon tetrachloride
Chlorobenzene
Chloroethane
Chloroform
Chi oromethane
2-Chlorotoluene
4-Chlorotoluene
l,2-Dibromo-3-chloropropane
Di bromochl oromethane
1,2-Dibromoethane
Dibromomethane
1, 2 -Di chlorobenzene
1, 3 -Di chlorobenzene
1, 4 -Di chlorobenzene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-l,2-Dichloroethene
trans-l,2-Dichloroethene
1 ,2-Dichloropropane
1,3-Dichloropropane
2 , 2 -Di chl oropropane
1 , 1-Dichloropropene
Ethyl benzene
Hexachl orobutadi ene
Isopropyl benzene
p- I sopropyl to! uene
Methylene chloride
Naphthalene
n-Propyl benzene
Cone.
M9/L
0.1
0.5
0.5
0.1
0.5
0.5
0.5
0.5
0.5
0.1
0.1
0.1
0.1
0.5
0.5
0.5
0.5
0.1
0.5
0.5
0.1
0.1
0.1
0.1
0.5
0.1
0.1
0.1
0.1
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
0.5
Number
of
Samples
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
Recovery8
%
99
97
97
100
101
99
94
110
110
108
91
100
105
101
99
96
92
99
97
93
97
101
106
99
98
100
95
100
98
96
99
99
102
99
100
102
113
97
98
99
Standard
Deviation
of Recovery
6.2
7.4
5.8
4.6
5.4
7.1
6.0
7.1
2.5
6.8
5.8
5.8
3.2
4.7
4.6
7.0
10.0
5.6
5.6
5.6
3.5
6.0
6.5
8.8
6.2
6.3
9.0
3.7
7.2
6.0
5.8
4.9
7.4
5.2
6.7
6.4
13.0
13.0
7.2
6.6
Percent
RSD
6.3
7.6
6.0
4.6
5.3
7.2
6.4
6.5
2.3
6.3
6.4
5.8
3.0
4.7
4.6
7.3
10.9
5.7
5.8
6.0
3.6
5.9
6.1
8.9
6.3
6.3
9.5
3.7
7.3
6.3
5.9
4.9
7.3
5.3
6.7
6.3
11.5
13.4
7.3
6.7
8260A - 41
Revision 1
September 1994
-------
TABLE 7.
(Continued)
Analyte
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
Toluene
1 , 2 , 3-Tri chl orobenzene
1,2,4-Trichlorobenzene
1,1,1-Trichloroethane
1 , 1 , 2-Tri chl oroethane
Trichloroethene
Tri chl orof 1 uoromethane
1,2, 3-Tri chl oropropane
1,2, 4 -Tri methyl benzene
1 , 3 , 5-Tri methyl benzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
Cone.
M9/L
0.5
0.5
0.5
0.1
0.5
0.5
0.5
0.5
0.5
0.1
0.1
0.5
0.5
0.5
0.1
0.5
0.5
0.5
Number
of
Samples
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
Recovery"
%
96
100
100
96
100
102
91
100
102
104
97
96
96
101
104
106
106
97
Standard
Deviation
of Recovery
19.0
4.7
12.0
5.0
5.9
8.9
16.0
4.0
4.9
2.0
4.6
6.5
6.5
4.2
0.2
7.5
4.6
6.1
Percent
RSD
19.8
4.7
12.0
5.2
5.9
8.7
17.6
4.0
4.8
1.9
4.7
6.8
6.8
4.2
0.2
7.1
4.3
6.3
Recoveries were calculated using internal standard method. Internal standard
was fluorobenzene.
8260A - 42
Revision 1
September 1994
-------
TABLE 8.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Surrogate Compound
4-Bromof 1 uorobenzene"
Di bromof 1 uoromethane"
Toluene-d88
Dichloroethane-d4a
Percent
Low/High
Water
86-115
86-118
88-110
80-120
Recovery
Low/High
Soil/Sediment
74-121
80-120
81-117
80-120
8 Single laboratory data, for guidance only.
TABLE 9.
QUANTITY OF EXTRACT REQUIRED FOR ANALYSIS OF
HIGH-CONCENTRATION SAMPLES
Approximate Volume of
Concentration Range Extract3
500 - 10,000 M9/kg 100 /xL
1,000 - 20,000 jug/kg 50 ML
5,000 - 100,000 M9/kg 10 juL
25,000 - 500,000 /xg/kg 100 /xL of 1/50 dilution6
Calculate appropriate dilution factor for concentrations exceeding this table.
a The volume of solvent added to 5 mL of water being purged should be kept
constant. Therefore, add to the 5 mL syringe whatever volume of solvent
is necessary to maintain a volume of 100 juL added to the syringe.
b Dilute an aliquot of the solvent extract and then take 100 juL for
analysis.
8260A - 43 Revision 1
September 1994
-------
TABLE 10
DIRECT INJECTION ANALYSIS OF NEW OIL AT 5
PPM
Compound
Acetone
Benzene
n-Butanol*,**
iso-Butanol*,**
Carbon tetrachloride
Carbon disulfide**
Chlorobenzene
Chloroform
1,4-Dichlorobenzene
1,2-Dichloroethane
1,1-Dichloroethene
Diethyl ether
Ethyl acetate
Ethyl benzene
Hexachloroethane
Methylene chloride
Methyl ethyl ketone
MIBK
Nitrobenzene
Pyridine
Tetrachloroethene
Recovery (%)
91
86
107
95
86
53
81
84
98
101
97
76
113
83
71
98
79
93
89
31
82
Trichlorofluoromethane 76
l,l,2-Cl3F3ethane
Toluene
Trichloroethene
Vinyl chloride
o-Xylene
m/p-Xylene
* Alternate mass
** TS nuant itatinr
69
73
66
63
83
84
employed
i
%RSD
14.8
21.3
27.8
19.5
44.7
22.3
29.3
29.3
24.9
23.1
45.3
24.3
27.4
30.1
30.3
45.3
24.6
31.4
30.3
35.9
27.1
27.6
29.2
21.9
28.0
35.2
29.5
29.5
Blank
(ppm)
1.9
0.1
0.5
0.9
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0
0.2
0.0
0.0
0.4
0.0
0.0
0.0
0.0
0.0
0.0
0.6
0.0
0.0
0.4
0.6
Spike
(ppm)
5.0
0.5
5.0
5.0
0.5
5.0
5.0
6.0
7.5
0.5
0.7
5.0
5.0
5.0
3.0
5.0
5.0
5.0
2.0
5.0
0.7
5.0
5.0
5.0
0.5
0.2
5.0
10.0
Data are taken from Reference 9.
8260A - 44
Revision 1
September 1994
-------
FIGURE 1.
PURGING DEVICE
FOAMT1U*
BUT 1M IN 0-0.
•— 14MMO.O.
INLET 1*4 IN. 0.0.
EOT m IN. 0.0.
10 MM GLASS fftr
MCOWM POAQ8ITV
SAMPLE INLET
2-WAY SYNNQE VALVC
17 CM » GAUGE SVMNGE NEEDLE
6 MM O.O. BU88EH SEPTUM
INLET 1M IN. O.D.
1/16 IN O.O
/^STAINLESS STEEL
13X
MOLECULAH SIEVE
MJRQE GAS FILTER
PUMOEGAS
FVOWCXJNTIWL
8260A - 45
Revision 1
September 1994
-------
FIGURE 2.
TRAP PACKING AND CONSTRUCTION TO INCLUDE DESORB CAPABILITY
PACKING DETAIL
CONSTRUCTION DETAIL
r 5 MM OLABB WOOL
77 CM SJOCA GEL
1SCMTENAXQC
*• i CM yn, ov-i
9MMQLAMWOOL
14 FT m»OOT
ACSSTANCCtMME
WNMPCDSOUO
OONTIO.ANO
FVK9MCTCM
a 106 m. u>.
ai
STi
8260A - 46
Revision 1
September 1994
-------
FIGURE 3.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - PURGE MODE
CARRIER GAS
FLOW CONTROL
PRESSURE
REGULATOR
PURGE OAS
FLOW CONTROL
13X MOLECULAR
SIEVE FILTER
UOMO INJECTION PORTS
— COLUMN OVEN
UUUV-
CONFIRMATORY COLUMN
TO DETECTOR
• ANALYTICAL COLUMN
OPTIONAL *PORT COLUMN
SELECTION VALVE
= /- TRAP INLET
TRAP
22-C
PURGING
DEVICE
NOTE
ALL LINES BETWEEN TRAP
AND QC SHOULD B€ HCATEO
TO WC.
8260A - 47
Revision 1
September 1994
-------
FIGURE 4.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - DESORB MODE
CARRIER GAS
FLOW CONTROL
PRESSURE
REGULATOR
UOUW INJECTION PORTS
-COLUMN OVEN
JUUV-
CONRRMATORY COLUMN
TO DETECTOR
ANALYTICAL COLUMN
OPTIONAL *PORT COLUMN
SELECTION VALVE
TRAP INLET
PURGE GAS
FLOW CONTROL
13X MOLECULAR
SIEVt FILTER
NOTE:
ALL UNES BETWEEN TRAP
AND GC SHOULD BE HEATED
TOBTC.
8260A - 48
Revision 1
September 1994
-------
CO
o
in o
o: u-
=3 o
o
CO
<
C5
9.
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o.
9
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3N3H130WOTM3WM131
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-------
00
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cc u.
=> o
CJ3
CJ3
O
n:
o
00
•5 8
S
(SI) $ zdfr'T j^
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CD £
rf a)
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Q.
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-------
: t- ac
I UJ UJ
2
0.
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o
SZ HH _
-°?g
en
c en
O i-l
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> J3
tt) E
o: w
••->
a.
Q)
CO
if
cc.
X
CO,
=1
s
aa
••a-
CSJ
in
CM
CO
-------
FIGURE 9.
LOW SOILS IMPINGER
PURGE INLET nrnNC
SAMPLE OUTLET UTTING
E "****„ ••^•^^^^^••^
JSffi
6mm 00 CLASS TUBING
SEPTUM
CAP
40ml VIAL
8260A - 53
Revision 1
September 1994
-------
METHOD 8260A
VOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS)
CAPILLARY COLUMN TECHNIQUE
( Start J
/ 7.1 \
/ Select N. Direct
S procedure N^ Injection ^
V^ for introducing J ^
>v^ sample into /
NGC/MS./
Purge-and-trap [
V
7.2 Set GC/MS
operating
conditions.
J,
7.3.1 Tune
GC/MS system
with BFB.
1
7.3.2 Assemble
purge-and-trap
device and prepare
calibration standards.
1
7.3.2.1 Perform
purge-and-trap
analysis.
k "
7.3.4 Calculate
RFs for
5 SPCCs.
,
,
7.3.5 Calculate
%RSD of RF
for CCCs.
J
1
7.4 Perform
calibration
verification.
^
,
7.5 Perform GC/MS
analysis utilizing
Methods 5030
or 8260.
^
r
7.6.1 Identify
analytes by
comparing the
sample and standard
mass spectra.
7.6.2 Calculate the
concentration of
each identified
analyte.
7.6.2.3 Report
all results.
f Stop J
8260A - 54
Revision 1
September 1994
-------
METHOD 8270B
SEMIVOLATILE ORGANIC COMPOUNDS BY
GAS CHROMATOGRAPHY/MASS SPECTROHETRY (GC/MS): CAPILLARY COLUMN TECHNIQUE
1.0 SCOPE AND APPLICATION
1.1 Method 8270 is used to determine the concentration of semivolatile
organic compounds in extracts prepared from all types of solid waste matrices,
soils, and ground water. Direct injection of a sample may be used in limited
applications. The following compounds can be determined by this method:
Compounds
Appropriate Preparation Techniques
CAS No8 3510
3540/
3520 3541 3550 3580
Acenaphthene
Acenaphthene-d10 (I.S.)
Acenaphthylene
Acetophenone
2-Acetylaminofluorene
l-Acetyl-2-thiourea
Aldrin
2-Aminoanthraquinone
Aminoazobenzene
4-Aminobiphenyl
3-Amino-9-ethylcarbazole
Anilazine
Aniline
o-Anisidine
Anthracene
Aramite
Aroclor - 1016
Aroclor - 1221
Aroclor - 1232
Aroclor - 1242
Aroclor - 1248
Aroclor - 1254
Aroclor - 1260
Azinphos -methyl
Barban
Benzidine
Benzoic acid
Benz(a)anthracene
Benzo(b)fl uoranthene
Benzo ( k) f 1 uoranthene
Benzo (g , h , i ) peryl ene
Benzo(a)pyrene
83-32-9
208-96-8
98-86-2
53-96-3
591-08-2
309-00-2
117-79-3
60-09-3
92-67-1
132-32-1
101-05-3
62-53-3
90-04-0
120-12-7
140-57-8
12674-11-2
11104-28-2
11141-16-5
53469-21-9
12672-29-6
11097-69-1
11096-82-5
86-50-0
101-27-9
92-87-5
65-85-0
56-55-3
205-99-2
207-08-9
191-24-2
50-32-8
X
X
X
X
X
LR
X
X
X
X
X
X
X
X
X
HS(43)
X
X
X
X
X
X
X
HS(62)
LR
CP
X
X
X
X
X
X
X
X
X
NO
ND
ND
X
ND
ND
ND
X
ND
X
ND
X
ND
X
X
X
X
X
X
X
ND
ND
CP
X
X
X
X
X
X
X
X
X
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
ND
X
ND
X
X
X
X
X
X
X
ND
ND
CP
ND
X
X
X
X
X
X
X
X
ND
ND
ND
X
ND
ND
ND
ND
ND
X
ND
X
ND
X
X
X
X
X
X
X
ND
ND
CP
X
X
X
X
X
X
X
X
X
X
X
LR
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
CP
X
X
X
X
X
X
8270B - 1
Revision 2
September 1994
-------
Appropriate Preparation Techniques
Compounds
p-Benzoquinone
Benzyl alcohol
a-BHC
0-BHC
6-BHC
7-BHC (Lindane)
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
Bromoxynil
Butyl benzyl phthalate
2-sec-Butyl-4,6-dinitrophenol
Captafol
Captan
Carbaryl
Carbofuran
Carbophenothion
Chlordane
Chlorfenvinphos
4-Chloroaniline
Chlorobenzilate
5-Chloro-2-methylanil ine
4-Chloro-3-methyl phenol
3-(Chloromethyl )pyridine
hydrochloride
1 -Chi oronaphthal ene
2-Chloronaphthalene
2-Chlorophenol
4-Chloro- 1,2 -phenyl enedi ami ne
4-Chloro-l ,3-phenylenediamine
4-Chlorophenyl phenyl ether
Chrysene
Chrysene-d12 (I.S.)
Coumaphos
p-Cresidine
Crotoxyphos
2-Cyclohexyl-4,6-dinitro-phenol
4, 4' -ODD
4,4'-DDE
4,4'-DDT
Demeton-0
Demeton-S
Diallate (cis or trans)
2,4-Diaminotoluene
CAS Noa
106-51-4
100-51-6
319-84-6
319-85-7
319-86-8
58-89-9
111-91-1
111-44-4
108-60-1
117-81-7
101-55-3
1689-84-5
85-68-7
88-85-7
2425-06-1
133-06-2
63-25-2
1563-66-2
786-19-6
57-74-9
470-90-6
106-47-8
510-15-6
95-79-4
59-50-7
6959-48-4
90-13-1
91-58-7
95-57-8
95-83-0
5131-60-2
7005-72-3
218-01-9
56-72-4
120-71-8
7700-17-6
131-89-5
72-54-8
72-55-9
50-29-3
298-03-3
126-75-0
2303-16-4
95-80-7
3510 3520
OE
X
X
X
X
X
X
X
X
X
X
X
X
X
HS(55)
HS(40)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
HS(68)
X
X
DC,OE(42)
ND
X
X
X
X
X
X
X
X
X
X
ND
X
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
X
ND
X
X
X
X
X
X
X
X
ND
ND
ND
ND
X
X
X
ND
ND
ND
ND
3540/
3541
ND
ND
X
X
X
X
X
X
X
X
X
ND
X
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
X
ND
X
X
X
ND
ND
X
X
X
ND
ND
ND
ND
X
X
X
ND
ND
ND
ND
3550
ND
X
X
X
X
X
X
X
X
X
X
ND
X
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
X
ND
X
X
X
ND
ND
X
X
X
ND
ND
ND
ND
X
X
X
ND
ND
ND
ND
3580
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
ND
X
X
X
X
X
X
LR
X
X
X
X
X
X
X
8270B - 2
Revision 2
September 1994
-------
Compounds
Appropriate Preparation Techniques
CAS No" 3510
3540/
3520 3541 3550 3580
Dibenz(a,j)acridine
Di benz ( a , h) anthracene
Dibenzofuran
Dibenzo(a,e)pyrene
1 , 2-Di bromo-3-chl oropropane
Di-n-butyl phthalate
Dichlone
1 , 2-Di chl orobenzene
1 , 3 -Di chl orobenzene
1,4-Dichlorobenzene
l,4-Dichlorobenzene-d4 (I.S)
3,3'-Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorophenol
Dichlorovos
Dicrotophos
Dieldrin
Diethyl phthalate
Diethylstilbestrol
Diethyl sulfate
Dihydrosaffrole
Dimethoate
3,3' -Dimethoxybenzidine
Dimethyl aminoazobenzene
7,12-Dimethylbenz(a)-
anthracene
3,3' -Dimethyl benzidine
a, a-Dimethyl phenethyl ami ne
2, 4-Dimethyl phenol
Dimethyl phthalate
1,2-Dinitrobenzene
1,3-Dinitrobenzene
1,4-Dinitrobenzene
4, 6-Dinitro- 2 -methyl phenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Dinocap
Dinoseb
Dioxathion
Diphenylamine
5,5-Diphenylhydantoin
1,2-Diphenylhydrazine
Di-n-octyl phthalate
Disulfoton
224-42-0
53-70-3
132-64-9
192-65-4
96-12-8
84-74-2
117-80-6
95-50-1
541-73-1
106-46-7
91-94-1
120-83-2
87-65-0
62-73-7
141-66-2
60-57-1
84-66-2
56-53-1
64-67-5
56312-13-1
60-51-5
119-90-4
60-11-7
57-97-6
119-93-7
122-09-8
105-67-9
131-11-3
528-29-0
99-65-0
100-25-4
534-52-1
51-28-5
121-14-2
606-20-2
39300-45-3
88-85-7
78-34-2
122-39-4
57-41-0
122-66-7
117-84-0
298-04-4
X
X
X
NO
X
X
OE
X
X
X
X
X
X
X
X
X
X
X
AW,OS(67)
LR
NO
HE,HS(31)
X
X
CP(45)
X
ND
X
X
X
X
HE(14)
X
X
X
X
CP,HS(28)
X
ND
X
X
X
X
X
ND
X
X
ND
X
X
ND
X
X
X
X
X
X
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
X
ND
X
X
ND
ND
X
ND
ND
ND
X
ND
X
X
X
X
X
X
ND
ND
ND
X
X
ND
ND
ND
ND
NO
ND
ND
ND
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
X
ND
X
X
ND
ND
X
X
ND
ND
X
ND
X
X
X
X
X
X
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
X
X
X
X
ND
ND
ND
X
ND
X
X
ND
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
ND
X
LR
X
CP
X
X
X
X
X
X
X
X
X
X
X
CP
X
ND
X
X
X
X
X
8270B - 3
Revision 2
September 1994
-------
Compounds
Appropriate Preparation Techniques
CAS No' 3510
3540/
3520 3541 3550 3580
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
EPN
Ethion
Ethyl carbamate
Ethyl methanesulfonate
Ethyl parathion
Famphur
Fensulfothion
Fenthion
Fluchloral in
Fluoranthene
Fluorene
2-Fluorobiphenyl (surr.)
2-Fluorophenol (surr.)
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachl orobutadi ene
Hexachlorocyclopentadiene
Hexachl oroethane
Hexachl orophene
Hexachl oropropene
Hexamethyl phosphorarnide
Hydroquinone
Indeno(l,2,3-cd)pyrene
Isodrin
Isophorone
Isosafrole
Kepone
Leptophos
Malathion
Maleic anhydride
Mestranol
Methapyrilene
Methoxychlor
3-Methyl chol anthrene
4,4'-Methylenebis
(2-chloroaniline)
4,4'-Methylenebis
(N,N-dimethyl ani line)
959-98-8
33213-65-9
1031-07-8
72-20-8
7421-93-4
53494-70-5
2104-64-5
563-12-2
51-79-6
62-50-0
56-38-2
52-85-7
115-90-2
55-38-9
33245-39-5
206-44-0
86-73-7
321-60-8
367-12-4
76-44-8
1024-57-3
118-74-1
87-68-3
77-47-4
67-72-1
70-30-4
1888-71-7
680-31-9
123-31-9
193-39-5
465-73-6
78-59-1
120-58-1
143-50-0
21609-90-5
121-75-5
108-31-6
72-33-3
91-80-5
72-43-5
56-49-5
101-14-4
101-61-1
X
X
X
X
X
X
X
X
DC(28)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
AW,CP(62)
X
X
NO
X
X
X
DC(46)
X
X
HS(5)
HE
X
X
X
X
OE,OS(0)
X
X
X
X
X
X
X
ND
ND
NO
ND
X
ND
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
ND
X
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
ND
X
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
ND
X
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
ND
8270B - 4
Revision 2
September 1994
-------
Compounds
Appropriate Preparation Techniques
CAS Noa 3510
3540/
3520 3541 3550 3580
Methyl methanesulfonate
2-Methylnaphthalene
2-Methyl-5-nitroaniline
Methyl parathion
2-Methyl phenol
3-Methyl phenol
4-Methyl phenol
2 -Methyl pyri dine
Mevinphos
Mexacarbate
Mi rex
Monocrotophos
Naled
Naphthalene
Naphthalene-d8 (I.S.)
1,4-Naphthoquinone
1-Naphthylamine
2-Naphthylamine
Nicotine
5-Nitroacenaphthene
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
5-Nitro-o-anisidine
Nitrobenzene
Nitrobenzene-d5 (surr.)
4-Nitrobiphenyl
Nitrofen
2-Nitrophenol
4-Nitrophenol
5-Nitro-o-toluidine
Nitroquinoline-1 -oxide
N-Nitrosodi butyl ami ne
N-Nitrosodi ethyl ami ne
N-Nitrosodimethyl amine
N-Nitrosomethyl ethyl ami ne
N-Nitrosodi phenyl amine
N-Nitrosodi -n-propyl amine
N-Nitrosomorphol ine
N-Nitrosopi peri dine
N-Nitrosopyrrol idine
Octamethyl pyrophosphoramide
4,4'-Oxydianiline
Parathion
Pentachl orobenzene
66-27-3
91-57-6
99-55-8
298-00-0
95-48-7
108-39-4
106-44-5
109-06-8
7786-34-7
315-18-4
2385-85-5
6923-22-4
300-76-5
91-20-3
130-15-4
134-32-7
91-59-8
54-11-5
602-87-9
88-74-4
99-09-2
100-01-6
99-59-2
98-95-3
92-93-3
1836-75-5
88-75-5
100-02-7
99-55-8
56-57-5
924-16-3
55-18-5
62-75-9
10595-95-6
86-30-6
621-64-7
59-89-2
100-75-4
930-55-2
152-16-9
101-80-4
56-38-2
608-93-5
X
X
X
X
X
X
X
X
X
HE,HS(68)
X
HE
X
X
X
X
OS(44)
X
DE(67)
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
LR
X
X
X
ND
X
X
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
X
X
X
ND
X
X
ND
ND
X
X
ND
ND
ND
ND
X
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
X
X
ND
ND
ND
ND
X
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
ND
ND
ND
ND
ND
X
X
X
ND
X
X
ND
ND
X
X
ND
ND
ND
ND
X
ND
X
X
ND
ND
ND
ND
ND
ND
ND
X
X
ND
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
LR
X
X
X
8270B - 5
Revision 2
September 1994
-------
Compounds
Appropriate Preparation Techniques
CAS No" 3510
3540/
3520 3541 3550 3580
Pentachloronitrobenzene
Pentachlorophenol
Perylene-d12 (I.S.)
Phenacetin
Phenanthrene
Phenanthrene-d10 (I.S.)
Phenobarbital
Phenol
Phenol -d6 (surr.)
1,4-Phenylenediamine
Phorate
Phosalone
Phosmet
Phosphamidon
Phthalic anhydride
2-Picoline
Piperonyl sulfoxide
Pronamide
Propylthiouracil
Pyrene
Pyridine
Resorcinol
Safrole
Strychnine
Sul fall ate
Terbufos
Terphenyl-d14(surr.)
1,2,4,5-Tetrachlorobenzene
2,3,4,6-Tetrachlorophenol
Tetrachlorvinphos
Tetraethyl dithiopyrophosphate
Tetraethyl pyrophosphate
Thionazine
Thiophenol (Benzenethiol )
Toluene diisocyanate
o-Toluidine
Toxaphene
2,4,6-Tribromophenol (surr.)
1 , 2 , 4-Trichl orobenzene
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
Trifluralin
2,4,5-Trimethylaniline
Trimethyl phosphate
82-68-8
87-86-5
62-44-2
85-01-8
50-06-6
108-95-2
106-50-3
298-02-2
2310-17-0
732-11-6
13171-21-6
85-44-9
109-06-8
120-62-7
23950-58-5
51-52-5
129-00-0
110-86-1
108-46-3
94-59-7
60-41-3
95-06-7
13071-79-9
1718-51-0
95-94-3
58-90-2
961-11-5
3689-24-5
107-49-3
297-97-2
108-98-5
584-84-9
95-53-4
8001-35-2
120-82-1
95-95-4
88-06-2
1582-09-8
137-17-7
512-56-1
X
X
X
X
X
X
X
DC(28)
DC(28)
X
X
HS(65)
HS(15)
HE(63)
CP,HE(1)
ND
X
X
LR
X
ND
DC,OE(10)
X
AW,OS(55)
X
X
X
X
X
X
X
X
X
X
HE(6)
X
X
X
X
X
X
X
X
HE(60)
ND
X
X
ND
X
X
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
X
ND
ND
ND
ND
ND
X
X
X
X
X
ND
ND
ND
ND
X
X
ND
X
X
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
ND
X
ND
ND
ND
ND
X
X
ND
X
X
ND
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
X
X
X
X
ND
ND
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
X
CP
ND
X
X
LR
X
ND
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
X
X
8270B - 6
Revision 2
September 1994
-------
Appropriate Preparation Techniques
3540/
Compounds CAS No8 3510 3520 3541 3550 3580
1,3,5-Trinitrobenzene 99-35-4
Tris(2,3-dibromopropyl) phosphate 126-72-7
Tri-p-tolyl phosphate 78-32-0
0,0,0-Triethyl phosphorothioate 126-68-1
X
X
X
X
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
X
LR
X
X
a Chemical Abstract Service Registry Number.
AW = Adsorption to walls of glassware during extraction and storage.
CP = Nonreproducible chromatographic performance.
DC = Unfavorable distribution coefficient (number in parenthesis is percent
recovery).
HE = Hydrolysis during extraction accelerated by acidic or basic conditions
(number in parenthesis is percent recovery).
HS = Hydrolysis during storage (number in parenthesis is percent stability).
LR = Low response.
ND = Not determined.
OE = Oxidation during extraction accelerated by basic conditions (number in
parenthesis is percent recovery).
OS = Oxidation during storage (number in parenthesis is percent stability).
X = Greater than 70 percent recovery by this technique.
1.2 Method 8270 can be used to quantitate most neutral, acidic, and
basic organic compounds that are soluble in methylene chloride and capable of
being eluted without derivatization as sharp peaks from a gas chromatographic
fused-silica capillary column coated with a slightly polar silicone. Such
compounds include polynuclear aromatic hydrocarbons, chlorinated hydrocarbons and
pesticides, phthalate esters, organophosphate esters, nitrosamines, haloethers,
aldehydes, ethers, ketones, anilines, pyridines, quinolines, aromatic nitro
compounds, and phenols, including nitrophenols. See Table 1 for a list of
compounds and their characteristic ions that have been evaluated on the specified
GC/MS system.
1.3 The following compounds may require special treatment when being
determined by this method. Benzidine can be subject to oxidative losses during
solvent concentration. Also, chromatography is poor. Under the alkaline
conditions of the extraction step, a-BHC, 7-BHC, Endosulfan I and II, and Endrin
are subject to decomposition. Neutral extraction should be performed if these
compounds are expected. Hexachlorocyclopentadiene is subject to thermal
decomposition in the inlet of the gas chromatograph, chemical reaction in acetone
solution, and photochemical decomposition. N-nitrosodimethylamine is difficult
to separate from the solvent under the chromatographic conditions described.
N-nitrosodiphenylamine decomposes in the gas chromatographic inlet and cannot be
separated from diphenylamine. Pentachlorophenol, 2,4-dinitrophenol,
8270B - 7 Revision 2
September 1994
-------
4-nitrophenol, 4,6~dinitro-2-methylphenol,4-chloro-3-methylphenol, benzoicacid,
2-nitroaniline, 3-nitroaniline, 4-chloroaniline, and benzyl alcohol are subject
to erratic chromatographic behavior, especially if the GC system is contaminated
with high boiling material.
1.4 The estimated quantitation limit (EQL) of Method 8270 for
determining an individual compound is approximately 1 mg/kg (wet weight) for
soil/sediment samples, 1-200 mg/kg for wastes (dependent on matrix and method of
preparation), and 10 jug/L for ground water samples (see Table 2). EQLs will be
proportionately higher for sample extracts that require dilution to avoid
saturation of the detector.
1.5 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatograph/mass spectrometers and
skilled in the interpretation of mass spectra. Each analyst must demonstrate the
ability to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 Prior to using this method, the samples should be prepared for
chromatography using the appropriate sample preparation and cleanup methods.
This method describes chromatographic conditions that will allow for the
separation of the compounds in the extract and for their qualitative and
quantitative analysis by mass spectrometry.
3.0 INTERFERENCES
3.1 Raw GC/MS data from all blanks, samples, and spikes must be
evaluated for interferences. Determine if the source of interference is in the
preparation and/or cleanup of the samples and take corrective action to eliminate
the problem.
3.2 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
sample syringe must be rinsed out between samples with solvent. Whenever an
unusually concentrated sample is encountered, it should be followed by the
analysis of solvent to check for cross contamination.
4.0 APPARATUS AND MATERIALS
4.1 Gas chromatograph/mass spectrometer system
4.1.1 Gas chromatograph - An analytical system complete with a
temperature-programmable gas chromatograph suitable for splitless
injection and all required accessories, including syringes, analytical
columns, and gases. The capillary column should be directly coupled to
the source.
8270B - 8 Revision 2
September 1994
-------
4.1.2 Column - 30 m x 0.25 mm ID (or 0.32 mm ID) 1 /im film thickness
silicone-coated fused-silica capillary column (J&W Scientific DB-5 or
equivalent).
4.1.3 Mass spectrometer - Capable of scanning from 35 to 500 amu
every 1 sec or less, using 70 volts (nominal) electron energy in the
electron impact ionization mode. The mass spectrometer must be capable
of producing a mass spectrum for decafluorotriphenylphosphine (DFTPP)
which meets all of the criteria in Table 3 when 1 jttL of the GC/MS tuning
standard is injected through the GC (50 ng of DFTPP).
4.1.4 GC/MS interface - Any GC-to-MS interface that gives acceptable
calibration points at 50 ng per injection for each compound of interest
and achieves acceptable tuning performance criteria may be used. For a
narrow-bore capillary column, the interface is usually capillary-direct
into the mass spectrometer source.
4.1.5 Data system - A computer system must be interfaced to the mass
spectrometer. The system must allow the continuous acquisition and
storage on machine-readable media of all mass spectra obtained throughout
the duration of the chromatographic program. The computer must have
software that can search any GC/MS data file for ions of a specific mass
and that can plot such ion abundances versus time or scan number. This
type of plot is defined as an Extracted Ion Current Profile (EICP).
Software must also be available that allows integrating the abundances in
any EICP between specified time or scan-number limits. The most recent
version of the EPA/NIST Mass Spectral Library should also be available.
4.1.6 Guard column (optional) (J&W Deactivated Fused Silica, 0.25
mm ID x 6 m, or equivalent) between the injecti'on port and the analytical
column joined with column joiners (Hewlett Packard No. 5062-3556, or
equivalent).
4.2 Syringe - 10 ml.
4.3 Volumetric flasks, Class A - Appropriate sizes with ground glass
stoppers.
4.4 Balance - Analytical, 0.0001 g.
4.5 Bottles - glass with Teflon-lined screw caps or crimp tops.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests.
Unless otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
8270B - 9 Revision 2
September 1994
-------
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Stock standard solutions (1000 mg/L) - Standard solutions can be
prepared from pure standard materials or purchased as certified solutions.
5.3.1 Prepare stock standard solutions by accurately weighing about
0.0100 g of pure material. Dissolve the material in pesticide quality
acetone or other suitable solvent and dilute to volume in a 10 ml
volumetric flask. Larger volumes can be used at the convenience of the
analyst. When compound purity is assayed to be 96% or greater, the weight
may be used without correction to calculate the concentration of the stock
standard. Commercially prepared stock standards may be used at any
concentration if they are certified by the manufacturer or by an
independent source.
5.3.2 Transfer the stock standard solutions into bottles with Teflon
lined screw-caps. Store at -10°C to -20°C or less and protect from light.
Stock standard solutions should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards from them.
5.3.3 Stock standard solutions must be replaced after 1 year or
sooner if comparison with quality control check samples indicates a
problem.
5.4 Internal standard solutions - The internal standards recommended are
l,4-dichlorobenzene-d4, naphthalene-d8, acenaphthene-d10, phenanthrene-d10,
chrysene-d12, and perylene-d12 (see Table 5). Other compounds may be used as
internal standards as long as the requirements given in Sec. 7.3.2 are met.
Dissolve 0.200 g of each compound with a small volume of carbon disulfide.
Transfer to a 50 ml volumetric flask and dilute to volume with methylene chloride
so that the final solvent is approximately 20% carbon disulfide. Most of the
compounds are also soluble in small volumes of methanol, acetone, or toluene,
except for perylene-d12. The resulting solution will contain each standard at
a concentration of 4,000 ng//xL. Each 1 ml sample extract undergoing analysis
should be spiked with 10 /iL of the internal standard solution, resulting in a
concentration of 40 ng/juL of each internal standard. Store at -10°C to -20°C
or less when not being used.
5.5 GC/MS tuning standard - A methylene chloride solution containing
50 ng//iL of decafluorotriphenylphosphine (DFTPP) should be prepared. The
standard should also contain 50 ng//il_ each of 4,4'-DDT, pentachlorophenol, and
benzidine to verify injection port inertness and GC column performance. Store
at -10°C to -20°C or less when not being used.
5.6 Calibration standards - A minimum of five calibration standards
should be prepared. One of the calibration standards should be at a
concentration near, but above, the method detection limit; the others should
correspond to the range of concentrations found in real samples but should not
exceed the working range of the GC/MS system. Each standard should contain each
analyte for detection by this method (e.g. some or all of the compounds listed
in Table 1 may be included). Each 1 ml aliquot of calibration standard should
8270B - 10 Revision 2
September 1994
-------
be spiked with 10 jxL of the internal standard solution prior to analysis. All
standards should be stored at -10°C to -20°C or less, and should be freshly
prepared once a year, or sooner if check standards indicate a problem. The daily
calibration standard should be prepared weekly and stored at 4°C.
5.7 Surrogate standards - The recommended surrogate standards are
phenol-d6, 2-fluorophenol, 2,4,6-tribromophenol, nitrobenzene-d5,
2-fluorobiphenyl, and p-terphenyl-du. See Method 3500 for the instructions on
preparing the surrogate standards. Determine what concentration should be in the
blank extracts after all extraction, cleanup, and concentration steps. Inject
this concentration into the GC/MS to determine recovery of surrogate standards
in all blanks, spikes, and sample extracts. Take into account all dilutions of
sample extracts.
5.8 Matrix spike standards - See Method 3500 for instructions on
preparing the matrix spike standard. Determine what concentration should be in
the blank extracts after all extraction, cleanup, and concentration steps.
Inject this concentration into the GC/MS to determine recovery of surrogate
standards in all matrix spikes. Take into account all dilutions of sample
extracts.
5.9 Acetone, hexane, methylene chloride, isooctane, carbon disulfide,
toluene, and other appropriate solvents - Pesticide quality or equivalent
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Sample preparation - Samples must be prepared by one of the
following methods prior to GC/MS analysis.
Matrix Methods
Water 3510, 3520
Soil/sediment 3540, 3541, 3550
Waste 3540, 3541, 3550, 3580
7.1.1 Direct injection - In very limited applications direct
injection of the sample into the GC/MS system with a 10 /^L syringe may be
appropriate. The detection limit is very high (approximately
10,000 jug/L); therefore, it is only permitted where concentrations in
excess of 10,000 /ig/L are expected. The system must be calibrated by
direct injection.
8270B - 11 Revision 2
September 1994
-------
7.2 Extract cleanup - Extracts may be cleaned up by any of the following
methods prior to GC/MS analysis.
Compounds Methods
Phenols 3630, 3640, 8040a
Phthalate esters 3610, 3620, 3640
Nitrosamines 3610, 3620, 3640
Organochlorine pesticides & PCBs 3620, 3660
Nitroaromatics and cyclic ketones 3620, 3640
Polynuclear aromatic hydrocarbons 3611, 3630, 3640
Haloethers 3620, 3640
Chlorinated hydrocarbons 3620, 3640
Organophosphorus pesticides 3620
Petroleum waste 3611, 3650
All priority pollutant base,
neutral, and acids 3640
8 Method 8040 includes a derivatization technique followed by GC/ECD
analysis, if interferences are encountered on GC/FID.
7.3 Initial calibration - The recommended GC/MS operating conditions:
Mass range: 35-500 amu
Scan time: 1 sec/scan
Initial temperature: 40°C, hold for 4 minutes
Temperature program: 40-270°C at 10°C/min
Final temperature: 270°C, hold until benzo[g,h,i]perylene has eluted
Injector temperature: 250-300°C
Transfer line temperature: 250-300°C
Source temperature: According to manufacturer's specifications
Injector: Grob-type, splitless
Sample volume: 1-2 /jL
Carrier gas: Hydrogen at 50 cm/sec or helium at 30 cm/sec
(Split injection is allowed if the sensitivity of the mass spectrometer
is sufficient).
7.3.1 Each GC/MS system must be hardware-tuned to meet the criteria
in Table 3 for a 50 ng injection of DFTPP. Analyses should not begin
until all these criteria are met. Background subtraction should be
straightforward and designed only to eliminate column bleed or instrument
background ions. The GC/MS tuning standard should also be used to assess
GC column performance and injection port inertness. Degradation of DDT
to DDE and ODD should not exceed 20%. (See Sec. 8.3.1 of Method 8081 for
the percent breakdown calculation). Benzidine and pentachlorophenol
should be present at their normal responses, and no peak tailing should
be visible. If degradation is excessive and/or poor chromatography is
noted, the injection port may require cleaning. It may also be necessary
to break off the first 6-12 in. of the capillary column. The use of a
guard column (Sec. 4.1.6) between the injection port and the analytical
column may help prolong analytical column performance.
8270B - 12 Revision 2
September 1994
-------
7.3.2 The internal standards selected in Sec. 5.4 should permit most
of the components of interest in a chromatogram to have retention times
of 0.80-1.20 relative to one of the internal standards. Use the base peak
ion from the specific internal standard as the primary ion for
quantitation (see Table 1). If interferences are noted, use the next most
intense ion as the quantitation ion (i.e. for l,4-dichlorobenzene-d4, use
152 m/z for quantitation).
7.3.3 Analyze 1 /j,L of each calibration standard (containing internal
standards) and tabulate the area of the primary characteristic ion against
concentration for each compound (as indicated in Table 1). Figure 1 shows
a chromatogram of a calibration standard containing base/neutral and acid
analytes. Calculate response factors (RFs) for each compound relative to
one of the internal standards as follows:
RF - (AxCis)/(AisCx)
where:
Ax = Area of the characteristic ion for the compound being
measured.
Ais = Area of the characteristic ion for the specific internal
standard.
Cis = Concentration of the specific internal standard (ng//iL).
Cx = Concentration of the compound being measured (ng//ul_).
7.3.4 A system performance check must be performed to ensure that
minimum average RFs are met before the calibration curve is used. For
semivolatiles, the System Performance Check Compounds (SPCCs) are:
N-nitroso-di-n-propylamine;hexachlorocyclopentadiene;2,4-dinitro-phenol;
and 4-nitrophenol. The minimum acceptable average RF for these compounds
is 0.050. These SPCCs typically have very low RFs (0.1-0.2) and tend to
decrease in response as the chromatographic system begins to deteriorate
or the standard material begins to deteriorate. They are usually the
first to show poor performance. Therefore, they must meet the minimum
requirement when the system is calibrated.
7.3.4.1 The percent relative standard deviation (%RSD)
should be less than 15% for each compound. However, the %RSD for
each individual Calibration Check Compound (CCC) (see Table 4) must
be less than 30%. The relative retention times of each compound in
each calibration run should agree within 0.06 relative retention
time units. Late-eluting compounds usually have much better
agreement.
SD
%RSD =—— x 100
RF
where:
RSD = relative standard deviation.
RF = mean of 5 initial RFs for a compound.
SD = standard deviation of average RFs for a compound.
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- RF)'
SD =
1=1 N - 1
where:
RFj = RF for each of the 5 calibration levels
N = Number of RF values (i.e., 5)
7.3.4.2 If the %RSD of any CCC is 30% or greater, then the
chromatographic system is too reactive for analysis to begin. Clean
or replace the injector liner and/or capillary column, then repeat
the calibration procedure beginning with section 7.3.
7.3.5 Linearity - If the %RSD of any compound is 15% or less, then
the relative response factor is assumed to be constant over the
calibration range, and the average relative response factor may be used
for quantitation (Sec. 7.6.2).
7.3.5.1 If the %RSD of any compound is greater than 15%,
construct calibration curves of area ratio (A/Ais) versus
concentration using first or higher order regression fit of the five
calibration points. The analyst should select the regression order
which introduces the least calibration error into the quantitation
(Sec. 7.6.2.2 and 7.6.2.3). The use of calibration curves is a
recommended alternative to average response factor calibration, and
a useful diagnostic of standard preparation accuracy and absorption
activity in the chromatographic system.
7.4 Daily GC/MS calibration
7.4.1 Prior to analysis of samples, the GC/MS tuning standard must
be analyzed. A 50 ng injection of DFTPP must result in a mass spectrum
for DFTPP which meets the criteria given in Table 3. These criteria must
be demonstrated during each 12 hour shift.
7.4.2 A calibration standard(s) at mid-concentration containing all
semivolatile analytes, including all required surrogates, must be
analyzed every 12 hours during analysis. Compare the instrument response
factor from the standards every 12 hours with the SPCC (Sec. 7.4.3) and
CCC (Sec. 7.4.4) criteria.
7.4.3 System Performance Check Compounds (SPCCs): A system
performance check must be made during every 12 hour shift. For each SPCC
compound in the daily calibration a minimum response factor of 0.050 must
be obtained. This is the same check that is applied during the initial
calibration. If the minimum response factors are not met, the system must
be evaluated, and corrective action must be taken before sample analysis
begins. The minimum RF for semivolatile SPCCs is 0.050. Some possible
problems are standard mixture degradation, injection port inlet
contamination, contamination at the front end of the analytical column,
8270B - 14 Revision 2
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and active sites in the column or chromatographic system. This check must
be met before analysis begins.
7.4.4 Calibration Check Compounds (CCCs): After the system
performance check is met, CCCs listed in Table 4 are used to check the
validity of the initial calibration.
Calculate the percent drift using:
C, - Cc
% Drift = - x 100
where:
C| = Calibration Check Compound standard concentration.
Cc = Measured concentration using selected quantisation method.
If the percent difference for each CCC is less than or equal to 20%,
the initial calibration is assumed to be valid. If the criterion is not
met (> 20% drift) for any one CCC, corrective action must be taken.
Problems similar to those listed under SPCCs could affect this criterion.
If no source of the problem can be determined after corrective action has
been taken, a new five-point calibration must be generated. This
criterion must be met before sample analysis begins. If the CCCs are not
analytes required by the permit, then all required analytes must meet the
20% drift criterion.
7.4.5 The internal standard responses and retention times in the
calibration check standard must be evaluated immediately after or during
data acquisition. If the retention time for any internal standard changes
by more than 30 seconds from the last calibration check (12 hours), the
chromatographic system must be inspected for malfunctions and corrections
must be made, as required. If the EICP area for any of the internal
standards changes by a factor of two (-50% to +100%) from the last daily
calibration check standard, the mass spectrometer must be inspected for
malfunctions and corrections must be made, as appropriate. When
corrections are made, reanalysis of samples analyzed while the system was
malfunctioning is required.
7.5 GC/MS analysis
7.5.1 It is highly recommended that the extract be screened on a
GC/FID or GC/PID using the same type of capillary column. This will
minimize contamination of the GC/MS system from unexpectedly high
concentrations of organic compounds.
7.5.2 Spike the 1 ml extract obtained from sample preparation with
10 n\- of the internal standard solution just prior to analysis.
7.5.3 Analyze the 1 ml extract by GC/MS using a 30 m x 0.25 mm (or
0.32 mm) silicone-coated fused-silica capillary column. The volume to be
injected should ideally contain 100 ng of base/neutral and 200 ng of acid
8270B - 15 Revision 2
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surrogates (for a 1 /il_ injection). The recommended GC/MS operating
conditions to be used are specified in Sec. 7.3.
7.5.4 If the response for any quantitation ion exceeds the initial
calibration curve range of the GC/MS system, extract dilution must take
place. Additional internal standard must be added to the diluted extract
to maintain the required 40 ng//uL of each internal standard in the
extracted volume. The diluted extract must be reanalyzed.
7.5.5 Perform all qualitative and quantitative measurements as
described in Sec. 7.6. Store the extracts at 4°C, protected from light
in screw-cap vials equipped with unpierced Teflon lined septa.
7.6 Data interpretation
7.6.1 Qualitative analysis
7.6.1.1 The qualitative identification of compounds
determined by this method is based on retention time, and on
comparison of the sample mass spectrum, after background correction,
with characteristic ions in a reference mass spectrum. The
reference mass spectrum must be generated by the laboratory using
the conditions of this method. The characteristic ions from the
reference mass spectrum are defined to be the three ions of greatest
relative intensity, or any ions over 30% relative intensity if less
than three such ions occur in the reference spectrum. Compounds
should be identified as present when the criteria below are met.
7.6.1.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographic peak containing ions
specific for the target compound at a compound-specific
retention time will be accepted as meeting this criterion.
7.6.1.1.2 The RRT of the sample component is within
± 0.06 RRT units of the RRT of the standard component.
7.6.1.1.3 The relative intensities of the
characteristic ions agree within 30% of the relative
intensities of these ions in the reference spectrum.
(Example: For an ion with an abundance of 50% in the
reference spectrum, the corresponding abundance in a sample
spectrum can range between 20% and 80%.)
7.6.1.1.4 Structural isomers that produce very similar
mass spectra should be identified as individual isomers if
they have sufficiently different GC retention times.
Sufficient GC resolution is achieved if the height of the
valley between two isomer peaks is less than 25% of the sum
of the two peak heights. Otherwise, structural isomers are
identified as isomeric pairs.
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7.6.1.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce
mass spectra containing ions contributed by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
appropriate selection of analyte spectra and background
spectra is important. Examination of extracted ion current
profiles of appropriate ions can aid in the selection of
spectra, and in qualitative identification of compounds.
When analytes coelute (i.e., only one chromatographic peak is
apparent), the identification criteria can be met, but each
analyte spectrum will contain extraneous ions contributed by
the coeluting compound.
7.6.1.2 For samples containing components not associated
with the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the purpose of the
analyses being conducted. Computer generated library search
routines should not use normalization routines that would
misrepresent the library or unknown spectra when compared to each
other. For example, the RCRA permit or waste delisting requirements
may require the reporting of nontarget analytes. Only after visual
comparison of sample spectra with the nearest library searches will
the mass spectral interpretation specialist assign a tentative
identification. Guidelines for making tentative identification are:
(1) Relative intensities of major ions in the reference
spectrum (ions > 10% of the most abundant ion) should be present in
the sample spectrum.
(2) The relative intensities of the major ions should agree
within + 20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
between 30 and 70%.)
(3) Molecular ions present in the reference spectrum should
be present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the
reference spectrum should be reviewed for possible background
contamination or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the
sample spectrum should be reviewed for possible subtraction from the
sample spectrum because of background contamination or coeluting
peaks. Data system library reduction programs can sometimes create
these discrepancies.
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7.6.2 Quantitative analysis
7.6.2.1 When a compound has been identified, the
quantitation of that compound will be based on the integrated
abundance from the EICP of the primary characteristic ion.
7.6.2.2 If the %RSD of a compound's relative response
factor is 15% or less, then the concentration in the extract may be
determined using the average response factor (RF) from initial
calibration data (7.4.5.2) and the following equation:.
(Ax x CJ
Cex (mg/L) =
(Ais x RF)
where Cex is the concentration of the compound in the extract, and
the other terms are as defined in Sec. 7.4.3.
7.6.2.3 Alternatively, the regression line fitted to the
initial calibration (Sec. 7.3.5.1) may be used for determination of
the extract concentration.
7.6.2.4 Compute the concentration of the analyte in the
sample using the equations in Sees. 7.6.2.4.1 and 7.6.2.4.2.
7.6.2.4.1 The concentration of the analyte in the
liquid phase of the sample is calculated using the
concentration of the analyte in the extract and the volume of
liquid extracted, as follows:
Concentration in liquid (Mg/L) = (Cm x Vex)
V
where:
Vex = extract volume, in ml
V0 = volume of liquid extracted, in L.
7.6.2.4.2 The concentration of the analyte in the
solid phase of the sample is calculated using the
concentration of the pollutant in the extract and the weight
of the solids, as follows:
Concentration in solid (M9/kg) = iCex_x_Vexl
where:
Vex = extract volume, in ml
Ws = sample weight, in kg.
7.6.2.5 Where applicable, an estimate of concentration for
noncalibrated components in the sample should be made. The formulae
8270B - 18 Revision 2
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given above should be used with the following modifications: The
areas Ax and A. should be from the total ion chromatograms and the
RF for the compound should be assumed to be 1. The concentration
obtained should be reported indicating (1) that the value is an
estimate and (2) which internal standard was used to determine
concentration. Use the nearest internal standard free of
interferences.
7.6.2.6 Quantitation of multicomponent compounds (e.g.
Aroclors) is beyond the scope of Method 8270. Normally,
quantitation is performed using a GC/ECD by Method 8081.
8.0 QUALITY CONTROL
8.1 Each laboratory that uses these methods is required to operate a
formal quality control program. The minimum requirements of this program consist
of an initial demonstration of laboratory capability and an ongoing analysis of
spiked samples to evaluate and document quality data. The laboratory must
maintain records to document the quality of the data generated. Ongoing data
quality checks are compared with established performance criteria to determine
if the results of analyses meet the performance characteristics of the method.
When results of sample spikes indicate atypical method performance, a quality
control reference sample (Sec. 8.5.1) must be analyzed to confirm that the
measurements were performed in an in-control mode of operation.
8.2 Before processing any samples, the analyst should demonstrate,
through the analysis of a method blank, that interferences from the analytical
system, glassware, and reagents are under control. Each time a set of samples
is extracted or there is a change in reagents, a method blank should be processed
as a safeguard against chronic laboratory contamination. The blanks should be
carried through all stages of sample preparation and measurement.
8.3 The experience of the analyst performing GC/MS analyses is
invaluable to the success of the methods. Each day that analysis is performed,
the daily calibration standard should be evaluated to determine if the
chromatographic system is operating properly. Questions that should be asked
are: Do the peaks look normal?; Is the response obtained comparable to the
response from previous calibrations? Careful examination of the standard
chromatogram can indicate whether the column is still good, the injector is
leaking, the injector septum needs replacing, etc. If any changes are made to
the system (e.g. column changed), recalibration of the system must take place.
8.4 Required instrument QC is found in the following sections
8.4.1 The GC/MS system must be tuned to meet the DFTPP
specifications in Sees. 7.3.1 and 7.4.1.
8.4.2 There must be an initial calibration of the GC/MS system as
specified in Sec. 7.3.
8.4.3 The GC/MS system must meet the SPCC criteria specified in Sec.
7.4.3 and the CCC criteria in Sec. 7.4.4, each 12 hours.
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8.5 To establish the ability to generate acceptable accuracy and
precision, the analyst must perform the following operations.
8.5.1 A quality control (QC) reference sample concentrate is
required containing each base/neutral analyte at a concentration of 100
mg/L and each acid analyte at a concentration of 200 mg/L in acetone or
methanol. (See Sec. 5.5.1 of Method 3500 for minimum requirements.) The
QC reference sample concentrate may be prepared from pure standard
materials or purchased as certified solutions. If prepared by the
laboratory, the QC reference sample concentrate must be made using stock
standards prepared independently from those used for calibration.
8.5.2 Using a pi pet, prepare QC reference samples at a concentration
of 100 jug/L by adding 1.00 ml of QC reference sample concentrate to each
of four 1-L aliquots of water.
8.5.3 Analyze the well-mixed QC reference samples according to the
method beginning in Sec. 7.1 with extraction of the samples.
8.5.4 Calculate the average recovery (x) in jiig/L, and the standard
deviation of the recovery (s) in ng/L, for each analyte of interest using
the four results.
8.5.5 For each analyte compare s and x with the corresponding
acceptance criteria_for precision and accuracy, respectively, found in
Table 6. If s and x for all analytes meet the acceptance criteria, the
system performance is acceptable and analysis of actual samples can_begin.
If any individual s exceeds the precision limit or any individual x falls
outside the range for accuracy, then the system performance is
unacceptable for that analyte.
NOTE: The large number of analytes in Table 6 present a substantial
probability that one or more will fail at least one of the
acceptance criteria when all analytes of a given method are
analyzed.
8.5.6 When one or more of the analytes tested fail at least one of
the acceptance criteria, the analyst must proceed according to Sec.
8.5.6.1 or 8.5.6.2.
8.5.6.1 Locate and correct the source of the problem and
repeat the test for all analytes of interest beginning with Sec.
8.5.2.
8.5.6.2 Beginning with Sec. 8.5.2, repeat the test only
for those analytes that failed to meet criteria. Repeated failure,
however, will confirm a general problem with the measurement system.
If this occurs, locate and correct the source of the problem and
repeat the test for all compounds of interest beginning with Sec.
8.5.2.
8.6 The laboratory must, on an ongoing basis, analyze a method blank,
a matrix spike, and a replicate for each analytical batch (up to a maximum of 20
8270B - 20 Revision 2
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samples/batch) to assess accuracy. For soil and waste samples where detectable
amounts of organics are present, replicate samples may be appropriate in place
of matrix spiked samples. For laboratories analyzing one to ten samples per
month, at least one spiked sample per month is required.
8.6.1 The concentration of the spike in the sample should be
determined as follows:
8.6.1.1 If, as in compliance monitoring, the concentration
of a specific analyte in the sample is being checked against a
regulatory concentration limit, the spike should be at that limit
or 1 to 5 times higher than the background concentration determined
in Sec. 8.6.2, whichever concentration would be larger.
8.6.1.2 If the concentration of a specific analyte in a
water sample is not being checked against a limit specific to that
analyte, the spike should be at 100 jug/L or 1 to 5 times higher than
the background concentration determined in Step 8.6.2, whichever
concentration would be larger. For other matrices, recommended
spiking concentration is 20 times the EQL.
8.6.1.3 If it is impractical to determine background
levels before spiking (e.g. maximum holding times will be exceeded),
the spike concentration should be at (1) the regulatory
concentration limit, if any; or, if none (2) the larger of either
5 times higher than the expected background concentration or 100
jug/L. For other matrices, recommended spiking concentration is 20
times the EQL.
8.6.2 Analyze one sample aliquot to determine the background
concentration (B) of each analyte. If necessary, prepare a new QC
reference sample concentrate (Sec. 8.5.1) appropriate for the background
concentration in the sample. Spike a second sample aliquot with 1.00 ml
of the QC reference sample concentrate and analyze it to determine the
concentration after spiking (A) of each analyte. Calculate each percent
recovery (p) as 100(A-B)%/T, where T is the known true value of the spike.
8.6.3 Compare the percent recovery (p) for each analyte in a water
sample with the corresponding QC acceptance criteria found in Table 6.
These acceptance criteria were calculated to include an allowance for
error in measurement of both the background and spike concentrations,
assuming a spike to background ratio of 5:1. This error will be accounted
for to the extent that the analyst's spike to background ratio approaches
5:1. If spiking was performed at a concentration lower than 100 /^g/L, the
analyst must use either the QC acceptance criteria presented in Table 6,
or optional QC acceptance criteria calculated for the specific spike
concentration. To calculate optional acceptance criteria for the recovery
of an analyte: (1) Calculate accuracy (x') using the equation found in
Table 7, substituting the spike concentration (T) for C; (2) calculate
overall precision (S') using the equation in Table 7, substituting x' for
x; (3) calculate the range for recovery at the spike concentration as
(lOOx'/T) + 2.44(100S'/T)%.
8270B - 21 Revision 2
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8.6.4 If any individual p falls outside the designated range for
recovery, that analyte has failed the acceptance criteria. A check
standard containing each analyte that failed the criteria must be analyzed
as described in Sec. 8.7.
8.7 If any analyte in a sample fails the acceptance criteria for
recovery in Sec. 8.6, a QC reference sample containing each analyte that failed
must be prepared and analyzed.
NOTE: The frequency for the required analysis of a QC reference sample
will depend upon the number of analytes being simultaneously tested,
the complexity of the sample matrix, and the performance of the
laboratory. If the entire list of analytes in Table 6 must be
measured in the sample in Sec. 8.6, the probability that the
analysis of a QC reference sample will be required is high. In this
case, the QC reference sample should be routinely analyzed with the
spiked sample.
8.7.1 Prepare the QC reference sample by adding 1.0 ml of the QC
reference sample concentrate (Sec. 8.5.1 or 8.6.2) to 1 L of water. The
QC reference sample needs only to contain the analytes that failed
criteria in the test in Sec. 8.6.
8.7.2 Analyze the QC reference sample to determine the concentration
measured (A) of each analyte. Calculate each percent recovery (ps) as
100(A/T)%, where T is the true value of the standard concentration.
8.7.3 Compare the percent recovery (pj for each analyte with the
corresponding QC acceptance criteria found in Table 6. Only analytes that
failed the test in Sec. 8.6 need to be compared with these criteria. If
the recovery of any such analyte falls outside the designated range, the
laboratory performance for that analyte is judged to be out of control,
and the problem must be immediately identified and corrected. The
analytical result for that analyte in the unspiked sample is suspect and
may not be reported for regulatory compliance purposes.
8.8 As part of the QC program for the laboratory, method accuracy for
each matrix studied must be assessed and records must be maintained. After the
analysis of five spiked samples _(of the same matrix) as in Sec. 8.6, calculate
the average percent recovery (p) and the standard deviation of the percent
recovery (sp). Express the accuracy assessment as a percent recovery interval
from p - 2sp to p + 2sp. If p = 90% and sp = 10%, for example, the accuracy
interval is expressed as 70-110%. Update the accuracy assessment for each
analyte on a regular basis (e.g. after each five to ten new accuracy
measurements).
8.9 The following procedure should be performed to determine acceptable
accuracy and precision limits for surrogate standards.
8.9.1 For each sample analyzed, calculate the percent recovery of
each surrogate in the sample.
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8.9.2 Once a minimum of thirty samples of the same matrix have been
analyzed, calculate the average percent recovery (P) and standard
deviation of the percent recovery (s) for each of the surrogates.
8.9.3 For a given matrix, calculate the upper and lower control
limit for method performance for each surrogate standard. This should be
done as follows:
Upper Control Limit (UCL) = P + 3s
Lower Control Limit (LCL) = P - 3s
8.9.4 For aqueous and soil matrices, these laboratory-established
surrogate control limits should, if applicable, be compared with the
control limits listed in Table 8. The limits given in Table 8 are multi-
laboratory performance-based limits for soil and aqueous samples, and
therefore, the single-laboratory limits established in Sec. 8.9.3 must
fall within those given in Table 8 for these matrices.
8.9.5 If recovery is not within limits, the following procedures are
required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are
a problem or flag the data as "estimated concentration".
8.9.6 At a minimum, each laboratory should update surrogate recovery
limits on a matrix-by-matrix basis, annually.
8.10 It is recommended that the laboratory adopt additional quality
assurance practices for use with this method. The specific practices that are
most productive depend upon the needs of the laboratory and the nature of the
samples. Field duplicates may be analyzed to assess the precision of the
environmental measurements. When doubt exists over the identification of a peak
on the chromatogram, confirmatory techniques such as gas chromatography with a
dissimilar column, specific element detector, or a mass spectrometer must be
used. Whenever possible, the laboratory should analyze standard reference
materials and participate in relevant performance evaluation studies.
9.0 METHOD PERFORMANCE
9.1 Method 8250 (the packed column version of Method 8270) was tested
by 15 laboratories using organic-free reagent water, drinking water, surface
water, and industrial wastewaters spiked at six concentrations over the range 5-
1,300 jLtg/L. Single operator accuracy and precision, and method accuracy were
found to be directly related to the concentration of the analyte and essentially
8270B - 23 Revision 2
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independent of the sample matrix. Linear equations to describe these
relationships are presented in Table 7.
9.2 Chromatograms from calibration standards analyzed with Day 0 and Day
7 samples were compared to detect possible deterioration of GC performance.
These recoveries (using Method 3510 extraction) are presented in Table 9.
9.3 Method performance data (using Method 3541 Automated Soxhlet
extraction) are presented in Table 10. Single laboratory accuracy and precision
data were obtained for semivolatile organics in a clay soil by spiking at a
concentration of 6 mg/kg for each compound. The spiking solution was mixed into
the soil during addition and then allowed to equilibrate for approximately 1 hr
prior to extraction. The spiked samples were then extracted by Method 3541
(Automated Soxhlet). Three determinations were performed and each extract was
analyzed by gas chromatography/ mass spectrometry following Method 8270. The low
recovery of the more volatile compounds is probably due to volatilization losses
during equilibration. These data are listed in Table 11 and were taken from
Reference 9.
10.0 REFERENCES
1. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act, Method 625," October 26,
1984.
2. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, July 1985, Revision.
3. Eichelberger, J.W., L.E. Harris, and W.L. Budde, "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass
Spectrometry Systems," Analytical Chemistry, 47, 995-1000, 1975.
4. "Method Detection Limit for Methods 624 and 625," Olynyk, P., W.L. Budde,
and J.W. Eichelberger, Unpublished report, October 1980.
5. "Inter!aboratory Method Study for EPA Method 625-Base/Neutrals, Acids, and
Pesticides," Final Report for EPA Contract 68-03-3102 (in preparation).
6. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
7. Lucas, S.V.; Kornfeld, R.A. "GC-MS Suitability Testing of RCRA Appendix
VIII and Michigan List Analytes "; U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, OH 45268,
February 20, 1987, Contract No. 68-03-3224.
8. Engel, T.M.; Kornfeld, R.A.; Warner, J.S.; Andrews, K.D. "Screening of
Semivolatile Organic Compounds for Extractability and Aqueous Stability
by SW-846, Method 3510"; U.S. Environmental Protection Agency,
Environmental Monitoring and Support Laboratory, Cincinnati, OH 45268,
June 5, 1987, Contract 68-03-3224.
8270B - 24 Revision 2
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9. Lopez-Avila, V. (W. Beckert, Project Officer); "Development of a Soxtec
Extraction Procedure for Extraction of Organic Compounds from Soils and
Sediments"; U.S. Environmental Protection Agency. Environmental
Monitoring and Support Laboratory. Las Vegas, NV, October 1991; EPA
600/X-91/140.
8270B - 25 Revision 2
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TABLE 1.
CHARACTERISTIC IONS FOR SEMIVOLATILE COMPOUNDS
Compound
Retention Primary Secondary
Time (min.) Ion Ion(s)
2-Picoline
Aniline
Phenol
Bis(2-chloroethyl) ether
2-Chlorophenol
1,3-Dichlorobenzene
l,4-Dichlorobenzene-d4 (I.S.)
1,4-Dichlorobenzene
Benzyl alcohol
1,2-Dichlorobenzene
N-Ni trosomethylethyl ami ne
Bis(2-chloroisopropyl) ether
Ethyl carbamate
Thiophenol (Benzenethiol)
Methyl methanesulfonate
N-Nitrosodi-n-propylamine
Hexachloroethane
Maleic anhydride
Nitrobenzene
Isophorone
N-Nitrosodiethyl amine
2-Nitrophenol
2,4-Dimethylphenol
p-Benzoquinone
Bis(2-chloroethoxy)methane
Benzoic acid
2,4-Dichlorophenol
Trimethyl phosphate
Ethyl methanesulfonate
1,2,4-Trichlorobenzene
Naphthalene-d8 (I.S.)
Naphthalene
Hexachlorobutadiene
Tetraethyl pyrophosphate
Diethyl sulfate
4-Chloro-3-methylphenol
2-Methylnaphthalene
2-Methylphenol
Hexachloropropene
Hexachlorocyclopentadi ene
N-Nitrosopyrrolidine
Acetophenone
4-Methylphenol
2,4,6-Trichlorophenol
o-Toluidine
3-Methylphenol
3.75'
5.68
5.77
5.82
5.97
6.27
6.35
6.40
6.78
6.
6.
7.
7.
7.
7.
7.
7,
7.
7,
85
97
22
27
42
48
55
65
65
87
8.53
8.70
8.75
.03
,13
9.
9.
9.23
9.
9.
9.
9.
.38
.48
,53
.62
9.67
9.75
9.82
10.43
11.07
11.37
11.68
11.87
12.40
12.45
12.60
12.65
12.67
12.82
12.85
12.87
12.93
93 66,92
93 66,65
94 65,66
93 63,95
128 64,130
146 148,111
152 150,115
146 148,111
108 79,77
146 148,111
88 42,88,43,56
45 77,121
62 62,44,45,74
110 110,66,109,84
80 80,79,65,95
70 42,101,130
117 201,199
54 54,98,53,44
77 123,65
82 95,138
102 102,42,57,44,56
139 109,65
122 107,121
108 54,108,82,80
93 95,123
122 105,77
162 164,98
110 110,79,95,109,140
79 79,109,97,45,65
180 182,145
136 68
128 129,127
225 223,227
99 99,155,127,81,109
139 139,45,59,99,111,125
107 144,142
142 141
107 107,108,77,79,90
213 213,211,215,117,106,141
237 235,272
100 100,41,42,68,69
105 71,105,51,120
107 107,108,77,79,90
196 198,200
106 106,107,77,51,79
107 107,108,77,79,90
8270B - 26
Revision 2
September 1994
-------
TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary
Ion
Secondary
Ion(s)
2-Chloronaphthalene
N-Nitrosopiperi dine
1,4-Phenylenedi ami ne
1-Chloronaphthalene
2-Nitroaniline
5-Chloro-2-methylaniline
Dimethyl phthalate
Acenaphthylene
2,6-Dinitrotoluene
Phthalic anhydride
o-Anisidine
3-Nitroaniline
Acenaphthene-d10 (I.S.)
Acenaphthene
2,4-Dinitrophenol
2,6-Dinitrophenol
4-Chloroaniline
Isosafrole
Dibenzofuran
2,4-Diaminotoluene
2,4-Dinitrotoluene
4-Nitrophenol
2-Naphthylamine
1,4-Naphthoquinone
p-Cresidine
Dichlorovos
Diethyl phthalate
Fluorene
2,4,5-Trimethylaniline
N-Ni trosodi butyl ami ne
4-Chlorophenyl phenyl ether
Hydroquinone
4,6-Dinitro-2-methylphenol
Resorcinol
N-Nitrosodiphenylamine
Safrole
Hexamethyl phosphoramide
3-(Chloromethyl)pyridine hydrochl
Diphenylamine
1,2,4,5-Tetrachlorobenzene
1-Naphthylamine
l-Acetyl-2-thiourea
4-Bromophenyl phenyl ether
Toluene diisocyanate
2,4,5-Trichlorophenol
Hexachlorobenzene
13.30 162 127,164
13.55 114 42,114,55,56,41
13.62 108 108,80,53,54,52
13.65a 162 127,164
13.75 65 92,138
14.28 106 106,141,140,77,89
14.48 163 194,164
14.57 152 151,153
14.62 165 63,89
14.62 104 104,76,50,148
15.00 108 80,108,123,52
15.02 138 108,92
15.05 164 162,160
15.13 154 153,152
15.35 184 63,154
15.47 162 162,164,126,98,63
15.50 127 127,129,65,92
15.60 162 162,131,104,77,51
15.63 168 139
15.78 121 121,122,94,77,104
15.80 165 63,89
15.80 139 109,65
16.00a 143 115,116
16.23 158 158,104,102,76,50,130
16.45 122 122,94,137,77,93
16.48 109 109,185,79,145
16.70 149 177,150
16.70 166 165,167
16.70 120 120,135,134,91,77
16.73 84 84,57,41,116,158
16.78 204 206,141
16.93 110 110,81,53,55
17.05 198 51,105
17.13 110 110,81,82,53,69
17.17 169 168,167
17.23 162 162,162,104,77,103,135
17.33 135 135,44,179,92,42
oride!7.50 92 92,127,129,65,39
17.54a 169 168,167
17.97 216 216,214,179,108,143,218
18.20 143 143,115,89,63
18.22 118 43,118,42,76
18.27 248 250,141
18.42 174 174,145,173,146,132,91
18.47 196 196,198,97,132,99
18.65 284 142,249
8270B - 27
Revision 2
September 1994
-------
TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary Secondary
Ion Ion(s)
Nicotine
Pentachlorophenol
5-Nitro-o-toluidine
Thionazine
4-Nitroaniline
Phenanthrene-d10(i .s.)
Phenanthrene
Anthracene
1,4-Dinitrobenzene
Mevinphos
Naled
1,3-Dinitrobenzene
Diallate (cis or trans)
1,2-Dinitrobenzene
Diallate (trans or cis)
Pentachlorobenzene
5-Nitro-o-anisidine
Pentachloronitrobenzene
4-Nitroquinoline-l-oxide
Di-n-butyl phthalate
2,3,4,6-Tetrachlorophenol
Dihydrosaffrole
Demeton-0
Fluoranthene
1,3,5-Trinitrobenzene
Dicrotophos
Benzidine
Trifluralin
Bromoxynil
Pyrene
Monocrotophos
Phorate
Sulfall ate
Demeton-S
Phenacetin
Dimethoate
Phenobarbital
Carbofuran
Octamethyl pyrophosphoramide
4-Aminobiphenyl
Dioxathion
Terbufos
a,a-Dimethylphenylamine
Pronamide
Aminoazobenzene
Dichlone
18.70 84 84,133,161,162
19.25 266 264,268
19.27 152 77,152,79,106,94
19.35 107 96,107,97,143,79,68
19.37 138 138,65,108,92,80,39
19.55 188 94,80
19.62 178 179,176
19.77 178 176,179
19.83 168 168,75,50,76,92,122
19.90 127 127,192,109,67,164
20.03 109 109,145,147,301,79,189
20.18 168 168,76,50,75,92,122
20.57 86 86,234,43,70
20.58 168 168,50,63,74
20.78 86 86,234,43,70
21.35 250 250,252,108,248,215,254
21.50 168 168,79,52,138,153,77
21.72 237 237,142,214,249,295,265
21.73 174 174,101,128,75,116
21.78 149 150,104
21.88 232 232,131,230,166,234,168
22.42 135 135,64,77
22.72 88 88,89,60,61,115,171
23.33 202 101,203
23.68 75 75,74,213,120,91,63
23.82 127 127,67,72,109,193,237
23.87 184 92,185
23.88 306 306,43,264,41,290
23.90 277 277,279,88,275,168
24.02 202 200,203
24.08 127 127,192,67,97,109
24.10 75 75,121,97,93,260
24.23 188 188,88,72,60,44
24.30 88 88,60,81,89,114,115
24.33 108 180,179,109,137,80
24.70 87 87,93,125,143,229
24.70 204 204,117,232,146,161
24.90 164 164,149,131,122
24.95 135 135,44,199,286,153,243
25.08 169 169,168,170,115
25.25 97 97,125,270,153
25.35 231 231,57,97,153,103
25.43 58 58,91,65,134,42
25.48 173 173,175,145,109,147
25.72 197 92,197,120,65,77
25.77 191 191,163,226,228,135,193
8270B - 28
Revision 2
September 1994
-------
TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary Secondary
Ion Ion(s)
Dinoseb
Disulfoton
Fluchloralin
Mexacarbate
4,4'-Oxydianiline
Butyl benzyl phthalate
4-Nitrobiphenyl
Phosphamidon
2-Cyclohexyl-4,6-Dinitrophenol
Methyl parathion
Carbaryl
Dimethyl aminoazobenzene
Propylthiouracil
Benz(a)anthracene
Chrysene-d12 (I.S.)
3,3'-Dichlorobenzidine
Chrysene
Malathion
Kepone
Fenthion
Parathion
Anilazine
Bis(2-ethylhexyl) phthalate
3,3'-Dimethylbenzidine
Carbophenothion
5-Nitroacenaphthene
Methapyrilene
Isodrin
Captan
Chlorfenvinphos
Crotoxyphos
Phosmet
EPN
Tetrachlorvinphos
Di-n-octyl phthalate
2-Aminoanthraquinone
Barban
Aramite
Benzo(b)fluoranthene
Nitrofen
Benzo(k)fluoranthene
Chlorobenzilate
Fensulfothion
Ethion
Diethylstilbestrol
Famphur
25.83 211 211,163,147,117,240
25.83 88 88,97,89,142,186
25.88 306 306,63,326,328,264,65
26.02 165 165,150,134,164,222
26.08 200 200,108,171,80,65
26.43 149 91,206
26.55 199 199,152,141,169,151
26.85 127 127,264,72,109,138
26.87 231 231,185,41,193,266
27.03 109 109,125,263,79,93
27.17 144 144,115,116,201
27.50 225 225,120,77,105,148,42
27.68 170 170,142,114,83
27.83 228 229,226
27.88 240 120,236
27.88 252 254,126
27.97 228 226,229
28.08 173 173,125,127,93,158
28.18 272 272,274,237,178,143,270
28.37 278 278,125,109,169,153
28.40 109 109,97,291,139,155
28.47 239 239,241,143,178,89
28.47 149 167,279
28.55 212 212,106,196,180
28.58 157 157,97,121,342,159,199
28.73 199 199,152,169,141,115
28.77 97 97,50,191,71
28.95 193 193,66,195,263,265,147
29.47 79 79,149,77,119,117
29.53 267 267,269,323,325,295
29.73 127 127,105,193,166
30.03 160 160,77,93,317,76
30.11 157 157,169,185,141,323
30.27 329 109,329,331,79,333
30.48 149 167,43
30.63 223 223,167,195
30.83' 222 222,51,87,224,257,153
30.92 185 185,191,319,334,197,321
31.45 252 253,125
31.48 283 283,285,202,139,253
31.55 252 253,125
31.77 251 251,139,253,111,141
31.87 293 293,97,308,125,292
32.08 231 231,97,153,125,121
32.15 268 268,145,107,239,121,159
32.67 218 218,125,93,109,217
8270B - 29
Revision 2
September 1994
-------
TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary Secondary
Ion Ion(s)
Tri-p-tolyl phosphate" 32.75 368
Benzo(a)pyrene 32.80 252
Perylene-d12 (I.S.) 33.05 264
7,12-Dimethylbenz(a)anthracene 33.25 256
5,5-Diphenylhydantoin 33.40 180
Captafol 33.47 79
Dinocap 33.47 69
Methoxychlor 33.55 227
2-Acetylaminofluorene 33.58 181
4,4'-Methylenebis(2-chloroaniline) 34.38 231
3,3'-Dimethoxybenzidine 34.47 244
3-Methylcholanthrene 35.07 268
Phosalone 35.23 182
Azinphos-methyl 35.25 160
Leptophos 35.28 171
Mirex 35.43 272
Tris(2,3-dibromopropyl) phosphate 35.68 201
Dibenz(a,j)acridine 36.40 279
Mestranol 36.48 277
Coumaphos 37.08 362
Indeno(l,2,3-cd)pyrene 39.52 276
Dibenz(a,h)anthracene 39.82 278
Benzo(g,h,i)perylene 41.43 276
l,2:4,5-Dibenzopyrene 41.60 302
Strychnine 45.15 334
Piperonyl sulfoxide 46.43 162
Hexachlorophene 47.98 196
Aldrin -- 66
Aroclor-1016 -- 222
Aroclor-1221 -- 190
Aroclor-1232 -- 190
Aroclor-1242 -- 222
Aroclor-1248 -- 292
Aroclor-1254 -- 292
Aroclor-1260 -- 360
a-BHC -- 183
j3-BHC -- 181
£-BHC -- 183
7-BHC (Lindane) -- 183
4,4'-DDD -- 235
4,4'-DDE -- 246
4,4'-DDT -- 235
Dieldrin -- 79
1,2-Diphenylhydrazine -- 77
Endosulfan I -- 195
Endosulfan II -- 337
368,367,107,165,198
253,125
260,265
256,241,239,120
180,104,252,223,209
79,77,80,107
69,41,39
227,228,152,114,274,212
181,180,223,152
231,266,268,140,195
244,201,229
268,252,253,126,134,113
182,184,367,121,379
160,132,93,104,105
171,377,375,77,155,379
272,237,274,270,239,235
137,201,119,217,219,199
279,280,277,250
277,310,174,147,242
362,226,210,364,97,109
138,227
139,279
138,277
302,151,150,300
334,335,333
162,135,105,77
196,198,209,211,406,408
263,220
260,292
224,260
224,260
256,292
362,326
362,326
362,394
181,109
183,109
181,109
181,109
237,165
248,176
237,165
263,279
105,182
339,341
339,341
8270B - 30
Revision 2
September 1994
-------
TABLE 1.
(Continued)
Retention Primary Secondary
Compound Time (min.) Ion Ion(s)
Endosulfan sulfate -- 272 387,422
Endrin -- 263 82,81
Endrin aldehyde , -- 67 345,250
Endrin ketone -- 317 67,319
2-Fluorobiphenyl (surr.) -- 172 171
2-Fluorophenol (surr.) -- 112 64
Heptachlor -- 100 272,274
Heptachlor epoxide -- 353 355,351
Nitrobenzene-d5 (surr.) -- 82 128,54
N-Nitrosodimethylamine -- 42 74,44
Phenol-d6 (surr.) -- 99 42,71
Terphenyl-du (surr.) -- 244 122,212
2,4,6-Tribromophenol (surr.) -- 330 332,141
Toxaphene -- 159 231,233
I.S. = internal standard.
surr. = surrogate.
""Estimated retention times.
bSubstitute for the non-specific mixture, tricresyl phosphate.
8270B - 31 Revision 2
September 1994
-------
TABLE 2.
ESTIMATED QUANTITATION LIMITS (EQLs) FOR SEMIVOLATILE ORGANICS
Estimated
Quantitation
Limits"
Ground water
Semivolatiles M9/L
Acenaphthene
Acenaphthylene
Acetophenone
2-Acetyl ami nof 1 uorene
l-Acetyl-2-thiourea
2-Aminoanthraquinone
Aminoazobenzene
4-Aminobiphenyl
Anilazine
o-Anisidine
Anthracene
Aramite
Azinphos-methyl
Barban
Benz(a)anthracene
Benzo(b)fl uoranthene
Benzo ( k) f 1 uoranthene
Benzole acid
Benzo (g,h,i)perylene
Benzo(a)pyrene
p-Benzoquinone
Benzyl alcohol
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
4-bromophenyl phenyl ether
Bromoxynil
Butyl benzyl phthalate
Captafol
Captan
Carbaryl
Carbofuran
Carbophenothion
Chlorfenvinphos
4-Chloroaniline
Chi orobenzi late
5-Chloro-2 -methyl aniline
4-Chloro-3 -methyl phenol
3-(Chloromethyl)pyridine hydrochloride
2-Chloronaphthalene
2-Chlorophenol
4-Chlorophenyl phenyl ether
Chrysene
Coumaphos
10
10
10
20
1000
20
10
20
100
10
10
20
100
200
10
10
10
50
10
10
10
20
10
10
10
10
10
10
20
50
10
10
10
20
20
10
10
20
100
10
10
10
10
40
Low Soil/Sediment"
M9Ag
660
660
ND
NO
ND
ND
ND
ND
ND
ND
660
ND
ND
ND
660
660
660
3300
660
660
ND
1300
660
660
660
660
ND
660
ND
ND
ND
ND
ND
ND
1300
ND
ND
1300
ND
660
660
660
660
ND
8270B - 32 Revision 2
September 1994
-------
Semivolatiles
p-Cresidine
Crotoxyphos
2-Cyclohexyl-4,6-dinitrophenol
Demeton-0
Demeton-S
Diallate (cis or trans)
Diallate (trans or cis)
2,4-Diaminotoluene
Dibenz(a,j)acridine
Dibenz( a, h) anthracene
Dibenzofuran
Dibenzo(a,e)pyrene
Di-n-butyl phthalate
Dichlone
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1 ,4-Dichlorobenzene
3,3'-Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorophenol
Dichlorovos
Dicrotophos
Diethyl phthalate
Diethylstilbestrol
Diethyl sulfate
Dimethoate
3,3'-Dimethoxybenzidine
Dimethyl ami noazobenzene
7, 12 -Dimethyl benz( a) anthracene
3, 3' -Dimethyl benzi dine
a, a-Dimethyl phenethyl ami ne
2,4-Dimethylphenol
Dimethyl phthalate
1,2-Dinitrobenzene
1,3-Dinitrobenzene
1,4-Dinitrobenzene
4, 6-Dinitro-2-methyl phenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Dinocap
Dinoseb
5,5-Diphenylhydantoin
Di-n-octyl phthalate
TABLE 2.
(Continued)
Ground
M9/1
10
20
100
10
10
10
10
20
10
10
10
10
10
NA
10
10
10
20
10
10
10
10
10
20
100
20
100
10
10
10
ND
10
10
40
20
40
50
50
10
10
100
20
20
10
Estimated
Quantitation
Limits8
water Low Soil/Sediment6
M9/kg
ND
ND
ND
ND
ND
ND
ND
ND
ND
660
660
ND
ND
ND
660
660
660
1300
660
ND
ND
ND
660
ND
ND
ND
ND
ND
ND
ND
ND
660
660
ND
ND
ND
3300
3300
660
660
ND
ND
ND
660
8270B - 33
Revision 2
September 1994
-------
Semi vol at lies
Disulfoton
EPN
Ethion
Ethyl carbamate
Bis(2-ethylhexy1) phthalate
Ethyl methanesulfonate
Famphur
Fensulfothion
Fenthion
Fluchloralin
Fluoranthene
Fluorene
Hexachlorobenzene
Hexachl orobutadi ene
Hexachl orocycl opentadi ene
Hexachl oroethane
Hexachl orophene
Hexachl oropropene
Hexamethyl phosphorami de
Hydroquinone
Indeno(l,2,3-cd)pyrene
Isodrin
Isophorone
Isosafrole
Kepone
Leptophos
Malathion
Maleic anhydride
Mestranol
Methapyrilene
Methoxychlor
3-Methyl chol anthrene
4,4'-Methylenebis(2-ch1oroanil
Methyl methanesulfonate
2-Methyl naphthalene
Methyl parathion
2-Methylphenol
3-Methyl phenol
4-Methylphenol
Mevinphos
Mexacarbate
Mi rex
Monocrotophos
Naled
TABLE 2.
(Continued)
Estimated
Quantitation
Limits"
Ground water Low Soi
M9/L
10
10
10
50
10
20
20
40
10
20
10
10
10
10
10
10
50
10
20
ND
10
20
10
10
20
10
50
NA
20
100
10
10
ine) NA
10
10
10
10
10
10
10
20
10
40
20
l/Sedimentb
MQ/kg
ND
ND
ND
ND
660
ND
ND
ND
ND
ND
660
660
660
660
660
660
ND
ND
ND
ND
660
ND
660
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
660
ND
660
ND
660
ND
ND
ND
ND
ND
8270B - 34
Revision 2
September 1994
-------
Semivolatiles
Naphthalene
1,4-Naphthoquinone
1-Naphthylamine
2-Naphthylamine
Nicotine
5-Nitroacenaphthene
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
5-Nitro-o-anisidine
Nitrobenzene
4-Nitrobiphenyl
Nitrofen
2-Nitrophenol
4-Nitrophenol
5-Nitro-o-toluidine
4-Nitroqui no! ine-1 -oxide
N-Ni trosodi butyl ami ne
N-Ni trosodi ethyl ami ne
N-Ni trosodi phenyl ami ne
N-Ni troso-di-n-propyl ami ne
N-Ni trosopi peri dine
N-Nitrosopyrrolidine
Octamethyl pyrophosphoramide
4,4'-Oxydianiline
Parathion
Pentachlorobenzene
Pentachloronitrobenzene
Pentachlorophenol
Phenacetin
Phenanthrene
Phenobarbital
Phenol
1 , 4- Phenyl ened i ami ne
Phorate
Phosalone
Phosmet
Phosphamidon
Phthalic anhydride
2-Picoline
Piperonyl sulfoxide
Pronamide
Propylthiouracil
Pyrene
TABLE 2.
(Continued)
Estimated
Quantitation
Limits8
Ground water Low Soi
M9/L
10
10
10
10
20
10
50
50
20
10
10
10
20
10
50
10
40
10
20
10
10
20
40
200
20
10
10
20
50
20
10
10
10
10
10
100
40
100
100
ND
100
10
100
10
I/Sediment"
M9/kg
660
ND
ND
ND
ND
ND
3300
3300
ND
ND
660
ND
ND
660
3300
ND
ND
ND
ND
660
660
ND
ND
ND
ND
ND
ND
ND
3300
ND
660
ND
660
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
660
8270B - 35
Revision 2
September 1994
-------
TABLE 2.
(Continued)
Estimated
Quantitation
Limits"
Ground water Low Soil/Sediment
Semivolatiles M9A
Pyridine
Resorcinol
Safrole
Strychnine
Sul fall ate
Terbufos
1 , 2 , 4, 5-Tetrachl orobenzene
2,3,4,6-Tetrachlorophenol
Tetrachlorvinphos
Tetraethyl pyrophosphate
Thionazine
Thiophenol (Benzenethiol )
Toluene diisocyanate
o-Toluidine
1, 2, 4-Trichl orobenzene
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
Trifluralin
2,4,5-Trimethylanil ine
Trimethyl phosphate
1, 3, 5-Tri nitrobenzene
Tris(2,3-dibromopropyl ) phosphate
Tri-p-tolyl phosphate(h)
0,0,0-Triethyl phosphorothioate
ND
100
10
40
10
20
10
10
20
40
20
20
100
10
10
10
10
10
10
10
10
200
10
NT
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
660
660
660
ND
ND
ND
ND
ND
ND
ND
a Sample EQLs are highly matrix-dependent. The EQLs listed herein are provided
for guidance and may not always be achievable.
b EQLs listed for soil/sediment are based on wet weight. Normally data are
reported on a dry weight basis, therefore, EQLs will be higher based on the
% dry weight of each sample. These EQLs are based on a 30 g sample and gel
permeation chromatography cleanup.
ND = Not determined..
NA = Not applicable.
NT = Not tested.
Other Matrices Factor0
High-concentration soil and sludges by sonicator7~1>
Non-water miscible waste 75
°EQL = (EQL for Low Soil/Sediment given above in Table 2) X (Factor).
8270B - 36 Revision 2
September 1994
-------
TABLE 3.
DFTPP KEY IONS AND ION ABUNDANCE CRITERIA8'"
Mass
Ion Abundance Criteria
51
68
70
127
197
198
199
275
365
441
442
443
30-60% of mass 198
< 2% of mass 69
< 2% of mass 69
40-60% of mass 198
< 1% of mass 198
Base peak, 100% relative abundance
5-9% of mass 198
10-30% of mass 198
> 1% of mass 198
Present but less than mass 443
> 40% of mass 198
17-23% of mass 442
a See Reference 3.
b Alternate tuning criteria may be used (e.g., CLP, Method 525, or
manufacturers' instructions), provided that method performance is not
adversely affected.
TABLE 4.
CALIBRATION CHECK COMPOUNDS
Base/Neutral Fraction
Acid Fraction
Acenaphthene
1,4-Dichlorobenzene
Hexachlorobutadi ene
N-Nitrosodiphenylamine
Di-n-octyl phthalate
Fluoranthene
Benzo(a)pyrene
4-Chloro-3-methyl phenol
2,4-Dichlorophenol
2-Nitrophenol
Phenol
Pentachlorophenol
2,4,6-Trichlorophenol
8270B - 37
Revision 2
September 1994
-------
TABLE 5.
SEMIVOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
l,4-Dichlorobenzene-d4 Naphthalene-d8
Acenaphthene-d
10
Aniline
Benzyl alcohol
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl)
ether
2-Chlorophenol
1,3-Dichlorobenzene
1,4-Dichlorobenzene
1,2-Dichlorobenzene
Ethyl methanesulfonate
2-Fluorophenol (surr.)
Hexachloroethane
Methyl methanesulfonate
2-Methylphenol
4-Methylphenol
N-Nitrosodimethylamine
N-Nitroso-di-n-propyl-
amine
Phenol
Phenol-de (surr.)
2-Picoline
Acetophenone
Benzoic acid
Bis(2-chloroethoxy)methane
4-Chloroaniline
4-Chloro-3-methyl phenol
2,4-Dichlorophenol
2,6-Dichlorophenol
a,a-Dimethyl-
phenethylamine
2,4-Dimethyl phenol
Hexachlorobutadi ene
Isophorone
2-Methylnaphthalene
Naphthalene
Nitrobenzene
Nitrobenzene-d8 (surr.)
2-Nitrophenol
N-Nitrosodibutyl amine
N-Nitrosopiperidine
1,2,4-Tri chlorobenzene
Acenaphthene
Acenaphthylene
1-Chloronaphthalene
2-Chloronaphthalene
4-Chlorophenyl
phenyl ether
Dibenzofuran
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Fluorene
2-Fluorobiphenyl
(surr.)
Hexachlorocyclo-
pentadiene
1-Naphthylamine
2-Naphthylamine
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
4-Nitrophenol
Pentachlorobenzene
1,2,4,5-Tetra-
chlorobenzene
2,3,4,6-Tetra-
chlorophenol
2,4,6-Tribromo-
phenol (surr.)
2,4,6-Trichloro-
phenol
2,4,5-Trichloro-
phenol
(surr.) = surrogate
8270B - 38
Revision 2
September 1994
-------
TABLE 5.
(Continued)
Phenanthrene-d
10
Chrysene-d
12
Perylene-d
12
4-Aminobiphenyl
Anthracene
4-Bromophenyl phenyl
ether
Di-n-butyl phthalate
4,6-Dinitro-2-methyl-
phenol
Diphenylamine
Fluoranthene
Hexachlorobenzene
N-Nitrosodiphenylamine
Pentachlorophenol
Pentachloronitrobenzene
Phenacetin
Phenanthrene
Pronamide
Benzidine
Benzo(a)anthracene
Bis(Z-ethylhexyl)
phthalate
Butyl benzyl phthalate
Chrysene
3,3'-Dichlorobenzidine
p-Dimethylaminoazobenzene
Pyrene
Terphenyl-d14 (surr.)
Benzo(b)fluor-
anthene
Benzo(k)fluor-
anthene
Benzo(g,h,i)-
perylene
Benzo(a)pyrene
Dibenz(a,j)acridine
Dibenz(a,h)-
anthracene
7,12-Dimethylbenz-
(a)anthracene
Di-n-octyl phthalate
Indeno(l,2,3-cd)
pyrene
3-Methylchol-
anthrene
(surr.) = surrogate
8270B - 39
Revision 2
September 1994
-------
TABLE 6.
QC ACCEPTANCE CRITERIA8
Compound
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benz(a)anthracene
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Benzyl butyl phthalate
/3-BHC
5-BHC
Bis(2-chloroethyl) ether
Bis(2-chloroethoxy)methane
Bis(2-chloroisopropyl) ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
2 -Chl oronaphthal ene
4-Chlorophenyl phenyl ether
Chrysene
4,4'-DDD
4, 4' -DDE
4,4'-DDT
Dibenzo( a, h) anthracene
Di-n-butyl phthalate
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
3,3'-Dichlorobenzidine
Dieldrin
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Endosulfan sulfate
Endrin aldehyde
Fluoranthene
Fluorene
Heptachlor
Heptachlor epoxide
Hexachl orobenzene
Hexachlorobutadiene
Test
cone.
(M9/L)
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
Limit
for s
27.6
40.2
39.0
32.0
27.6
38.8
32.3
39.0
58.9
23.4
31.5
21.6
55.0
34.5
46.3
41.1
23.0
13.0
33.4
48.3
31.0
32.0
61.6
70.0
16.7
30.9
41.7
32.1
71.4
30.7
26.5
23.2
21.8
29.6
31.4
16.7
32.5
32.8
20.7
37.2
. 54.7
24.9
26.3
Range
for x
(M9/L)
60.1-132.3
53.5-126.0
7.2-152.2
43.4-118.0
41.8-133.0
42.0-140.4
25.2-145.7
31.7-148.0
D-195.0
D-139.9
41.5-130.6
D-100.0
42.9-126.0
49.2-164.7
62.8-138.6
28.9-136.8
64.9-114.4
64.5-113.5
38.4-144.7
44.1-139.9
D-134.5
19.2-119.7
D-170.6
D-199.7
8.4-111.0
48.6-112.0
16.7-153.9
37.3-105.7
8.2-212.5
44.3-119.3
D-100.0
D-100.0
47.5-126.9
68.1-136.7
18.6-131.8
D-103.5
D-188.8
42.9-121.3
71.6-108.4
D-172.2
70.9-109.4
7.8-141.5
37.8-102.2
Range
P, Ps
47-145
33-145
D-166
27-133
33-143
24-159
11-162
17-163
D-219
D-152
24-149
D-110
12-158
33-184
36-166
8-158
53-127
60-118
25-158
17-168
D-145
4-136
D-203
D-227
1-118
32-129
D-172
20-124
D-262
29-136
D-114
D-112
39-139
50-158
4-146
D-107
D-209
26-137
59-121
D-192
26.155
D-152
24-116
8270B - 40
Revision 2
September 1994
-------
Compound
Hexachloroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Naphthalene
Nitrobenzene
N-Nitrosodi-n-propylamine
PCB-1260
Phenanthrene
Pyrene
1 , 2 , 4-Tri chl orobenzene
4-Chl oro-3-methyl phenol
2-Chlorophenol
2,4-Chlorophenol
2, 4-Dimethyl phenol
2,4-Dinitrophenol
2-Methyl -4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
TABLE 6.
(Continued)
Test
cone.
(M9/L)
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
Limit
for s
(M9/L)
24.5
44.6
63.3
30.1
39.3
55.4
54.2
20.6
25.2
28.1
37.2
28.7
26.4
26.1
49.8
93.2
35.2
47.2
48.9
22.6
31.7
Range
for x
(M9/L)
55.2-100.0
D-150.9
46.6-180.2
35.6-119.6
54.3-157.6
13.6-197.9
19.3-121.0
65.2-108.7
69.6-100.0
57.3-129.2
40.8-127.9
36.2-120.4
52.5-121.7
41.8-109.0
D-172.9
53.0-100.0
45.0-166.7
13.0-106.5
38.1-151.8
16.6-100.0
52.4-129.2
Range
P> Ps
(%)
40-113
D-171
21-196
21-133
35-180
D-230
D-164
54-120
52-115
44-142
22-147
23-134
39-135
32-119
D-191
D-181
29-182
D-132
14-176
5-112
37-144
s
X
P, Ps
D
a
Standard deviation of four recovery measurements, in jiig/L.
Average recovery for four recovery measurements, in jug/L.
Percent recovery measured.
Detected; result must be greater than zero.
Criteria from 40 CFR Part 136 for Method 625. These criteria are based
directly on the method performance data in Table 7. Where necessary, the
limits for recovery have been broadened to assure applicability of the
limits to concentrations below those used to develop Table 7.
8270B - 41
Revision 2
September 1994
-------
TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Compound
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benz(a)anthracene
Chloroethane
Benzo(b)fluoranthene
Benzo ( k) f 1 uoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Benzyl butyl phthalate
/3-BHC
5-BHC
Bis(2-chloroethyl) ether
Bis(2-chloroethoxy)methane
Bi s ( 2-chl oroi sopropyl )
ether
Bis(2-ethylhexyl)
phthalate
4-Bromophenyl phenyl
ether
2-Chloronaphthalene
4-Chlorophenyl phenyl
ether
Chrysene
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dibenzo( a, h) anthracene
Di-n-butyl phthalate
1,2-Dichlorobenzene
1 ,3-Dichl orobenzene
1,4-Dichlorobenzene
3,3'-Dichlorobenzidine
Dieldrin
Diethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Endosulfan sulfate
Endrin aldehyde
Fl uoranthene
Accuracy, as
recovery, x'
(M9/L)
0.96C+0.19
0.89C+0.74
0.78C+1.66
0.80C+0.68
0.88C-0.60
0.99C-1.53
0.93C-1.80
0.87C-1.56
0.90C-0.13
0.98C-0.86
0.66C-1.68
0.87C-0.94
0.29C-1.09
0.86C-1.54
1.12C-5.04
1.03C-2.31
0.84C-1.18
0.91C-1.34
0.89C+0.01
0.91C+0.53
0.93C-1.00
0.56C-0.40
0.70C-0.54
0.79C-3.28
0.88C+4.72
0.59C+0.71
0.80C+0.28
0.86C-0.70
0.73C-1.47
1.23C-12.65
0.82C-0.16
0.43C+1.00
0.20C+1.03
0.92C-4.81
1.06C-3.60
0.76C-0.79
0.39C+0.41
0.76C-3.86
0.81C+1.10
Single analyst
precision, s/
(M9/L)
0.15X-0.12
0.24x-1.06
0.27X-1.28
0.21X-0.32
O.lBx+0.93
0.14X-0.13
0.22X+0.43
0.19X+1.03
0.22X+0.48
0.29X+2.40
O.lSx+0.94
0.20X-0.58
0.34X+0.86
0.35X-0.99
0.16X+1.34
0.24X+0.28
0.26X+0.73
0.13X+0.66
0.07X+0.52
0.20X-0.94
0.28X+0.13
0.29X-0.32
0.26X-1.17
0.42X+0.19
0.30X+8.51
0.13X+1.16
0.20X+0.47
0.25X+0.68
0.24x+0.23
0.28X+7.33
0.20X-0.16
0.28X+1.44
0.54x+0.19
0.12X+1.06
0.14X+1.26
0.21X+1.19
0.12X+2.47
O.lSx+3.91
0.22X-0.73
Overall
precision,
S' (M9/L)
0.21x-0.67
0.26x-0.54
0.43X+1.13
0.27x-0.64
0.26x-0.21
O.Ux-0.28
0.29x+0.96
0.35x+0.40
0.32X+1.35
O.Blx-0.44
0.53x+0.92
0.30x+1.94
0.93x-0.17
0.35x+0.10
0.26x+2.01
0.25X+1.04
0.36X+0.67
0.16x+0.66
0.13x+0.34
0.30X-0.46
0.33X-0.09
0.66X-0.96
0.39x-1.04
0.65X-0.58
0.59X+0.25
0.39x+0.60
0.24x+0.39
0.41X+0.11
0.29x+0.36
0.47x+3.45
0.26x-0.07
0.52X+0.22
l.OBx-0.92
0.21x+1.50
0.19x+0.35
0.37x+1.19
0.63x-1.03
0.73x-0.62
0.28x-0.60
8270B - 42
Revision 2
September 1994
-------
Compound
Fluorene
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorobutadiene
Hexachloroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Naphthalene
Nitrobenzene
N-Nitrosodi -n-propylamine
PCB-1260
Phenanthrene
Pyrene
1,2,4-Trichlorobenzene
4-Chloro-3 -methyl phenol
2-Chlorophenol
2,4-Dichlorophenol
2,4-Dimethylphenol
2,4-Dinitrophenol
2-Methyl-4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
TABLE 7.
(Continued)
Accuracy, as
recovery, x'
(M9/L)
0.90C-0.00
0.87C-2.97
0.92C-1.87
0.74C+0.66
0.71C-1.01
0.73C-0.83
0.78C-3.10
1.12C+1.41
0.76C+1.58
1.09C-3.05
1.12C-6.22
0.81C-10.86
0.87C+0.06
0.84C-0.16
0.94C-0.79
0.84C+0.35
0.78C+0.29
0.87C-0.13
0.71C+4.41
0.81C-18.04
1.04C-28.04
0.07C-1.15
0.61C-1.22
0.93C+1.99
0.43C+1.26
0.91C-0.18
Single analyst
precision, s/
(M9/L)
0.12X+0.26
0.24X-0.56
0.33X-0.46
O.lSx-0.10
0.19X+0.92
0.17X+0.67
0.29x+1.46
0.27x+0.77
O.Zlx-0.41
0.19x+0.92
0.27x+0.68
0.35X+3.61
0.12X+0.57
0.16X+0.06
O.lSx+0.85
0.23x+0.75
O.lSx+1.46
0.15X+1.25
0.16X+1.21
0.38X+2.36
O.lOx+42.29
0.16X+1.94
0.38X+2.57
0.24X+3.03
0.26X+0.73
0.16X+2.22
Overall
precision,
S' (M9/L)
0.13X+0.61
O.SOx-0.23
0.28X+0.64
0.43X-0.52
0.26X+0.49
0.17X+0.80
O.SOx-0.44
0.33X+0.26
0.30X-0.68
0.27x+0.21
0.44X+0.47
0.43X+1.82
O.lSx+0.25
O.lBx+0.31
0.21X+0.39
0.29X+1.31
0.28X+0.97
0.21X+1.28
0.22X+1.31
0.42X+26.29
0.26X+23.10
0.27X+2.60
0.44X+3.24
0.30X+4.33
0.35X+0.58
0.22X+1.81
X'
S'
C
x
Expected recovery for one or more measurements of a sample
containing a concentration of C, in M9/L-
Expected single analyst s_tandard deviation of measurements at an
average concentration of x, in M9/L.
Expected interlaboratory standard deviation of measurements at an
average concentration found of x, in M9/L-
True value for the concentration, in M9/L-
Average recovery found for measurements of samples containing a
concentration of C, in M9/L.
8270B - 43
Revision 2
September 1994
-------
TABLE 8.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Surrogate Compound
Nitrobenzene-d5
2-Fluorobiphenyl
Terphenyl-d14
Phenol -d6
2-Fluorophenol
2,4,6-Tribromophenol
Low/High
Water
35-114
43-116
33-141
10-94
21-100
10-123
Low/High
Soil/Sediment
23-120
30-115
18-137
24-113
25-121
19-122
TABLE 9.
EXTRACTION EFFICIENCY AND AQUEOUS STABILITY RESULTS
COMPOUND
PERCENT RECOVERY
ON DAY 0
AVG. RSD
PERCENT RECOVERY
ON DAY 7
AVG. RSD
3-Amino-9-ethylcarbazole
4-Chloro-l,2-phenylenediamine
4-Chloro-l,3-phenylenediamine
1 , 2-Di bromo-3-chl oropropane
2-sec-Butyl-4,6-diriitrophenol
Ethyl parathion
4 ,4' -Methyl enebis(N,N-di methyl aniline)
2-Methyl-5-nitroaniline
2-Methylpyridine
Tetraethyl dithiopyrophosphate
80
91
84
97
99
100
108
99
80
92
8
1
3
2
3
2
4
10
4
7
73
108
70
98
97
103
90
93
83
70
3
4
3
5
6
4
4
4
4
1
Data from Reference 8.
8270B - 44
Revision 2
September 1994
-------
TABLE 10.
AVERAGE PERCENT RECOVERIES AND PERCENT RSDs FOR THE TARGET COMPOUNDS
FROM SPIKED CLAY SOIL AND TOPSOIL BY AUTOMATED SOXHLET EXTRACTION
WITH HEXANE-ACETONE (1:1)'
Clay Soil
Topsoil
Compound name
1,3-Dichlorobenzene
1,2-Dichlorobenzene
Nitrobenzene
Benzal chloride
Benzotrichloride
4-Chloro-2-nitrotoluene
Hexachlorocyclopentadiene
2,4-Dichloronitrobenzene
3 , 4-Di chl oron i trobenzene
Pentachl orobenzene
2, 3, 4, 5-Tetrachl oron i trobenzene
Benefin
alpha-BHC
Hexachl orobenzene
delta-BHC
Heptachlor
Aldrin
Isopropalin
Heptachlor epoxide
trans-Chlordane
Endosulfan I
Dieldrin
2,5-Dichlorophenyl-
4-nitrophenyl ether
Endrin
Endosulfan II
p,p'-DDT
2,3,6-Trichlorophenyl-
4'-nitrophenyl ether
2 , 3 , 4-Tri chl orophenyl -
4'-nitrophenyl ether
Mi rex
Average
percent
recovery
0
0
0
0
0
0
4.1
35.2
34.9
13.7
55.9
62.6
58.2
26.9
95.8
46.9
97.7
102
90.4
90.1
96.3
129
110
102
104
134
110
112
104
Percent
RSD
..
--
--
--
--
--
15
7.6
15
7.3
6.7
4.8
7.3
13
4.6
9.2
12
4.3
4.4
4.5
4.4
4.7
4.1
4.5
4.1
2.1
4.8
4.4
5.3
Average
percent
recovery
0
0
0
0
0
0
7.8
21.2
20.4
14.8
50.4
62.7
54.8
25.1
99.2
49.1
102
105
93.6
95.0
101
104
112
106
105
111
110
112
108
Percent
RSD
..
--
--
--
--
--
23
15
11
13
6.0
2.9
4.8
5.7
1.3
6.3
7.4
2.3
2.4
2.3
2.2
1.9
2.1
3.7
0.4
2.0
2.8
3.3
2.2
The operating conditions for the Soxtec apparatus were as follows:
immersion time 45 min; extraction time 45 min; the sample size was 10 g;
the spiking concentration was 500 ng/g, except for the surrogate compounds
at 1000 ng/g, compounds 23, 27, and 28 at 1500 ng/g, compound 3 at 2000
ng/g> and compounds 1 and 2 at 5000 ng/g.
8270B - 45
Revision 2
September 1994
-------
TABLE 11.
SINGLE LABORATORY ACCURACY AND PRECISION DATA FOR THE EXTRACTION
OF SEMIVOLATILE ORGANICS FROM SPIKED CLAY BY
METHOD 3541 (AUTOMATED SOXHLET)"
Compound name
Phenol
Bis(2-chloroethy1)ether
2-Chlorophenol
Benzyl alcohol
2-Methyl phenol
Bis(2-chloroisopropyl )ether
4-Methyl phenol
N-Nitroso-di-n-propylamine
Nitrobenzene
Isophorone
2-Nitrophenol
2,4-Dimethylphenol
Benzoic acid
Bis(2-chloroethoxy)methane
2,4-Dichlorophenol
1 , 2 , 4-Tr i chl orobenzene
Naphthalene
4-Chloroaniline
4-Chl oro-3-methyl phenol
2-Methyl naphthal ene
Hexachl orocycl opentadi ene
2,4, 6 -Tr i chl orophenol
2,4,5-Trichlorophenol
2-Chloronaphthalene
2-Nitroaniline
Dimethyl phthalate
Acenaphthylene
3-Nitroaniline
Acenaphthene
2,4-Dinitrophenol
4-Nitrophenol
Dibenzofuran
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Diethyl phthalate
4-Chlorophenyl-phenyl ether
Fluorene
4-Nitroaniline
4 , 6 -Di ni tro -2 -methyl phenol
N-Nitrosodiphenylamine
4-Bromophenyl-phenyl ether
Average
percent
recovery
47.8
25.4
42.7
55.9
17.6
15.0
23.4
41.4
28.2
56.1
36.0
50.1
40.6
44.1
55.6
18.1
26.2
55.7
65.1
47.0
19.3
70.2
26.8
61.2
73.8
74.6
71.6
77.6
79.2
91.9
62.9
82.1
84.2
68.3
74.9
67.2
82.1
79.0
63.4
77.0
62.4
Percent
RSD
5.6
13
4.3
7.2
6.6
15
6.7
6.2
7.7
4.2
6.5
5.7
7.7
3.0
4.6
31
15
12
5.1
8.6
19
6.3
2.9
6.0
6.0
5.2
5.7
5.3
4.0
8.9
16
5.9
5.4
5.8
5.4
3.2
3.4
7.9
6.8
3.4
3.0
8270B - 46
Revision 2
September 1994
-------
Table 11. (Continued)
Average
percent Percent
Compound name recovery RSD
Hexachlorobenzene 72.6 3.7
Pentachlorophenol 62.7 6.1
Phenanthrene 83.9 5.4
Anthracene 96.3 3.9
Di-n-butyl phthalate 78.3 40
Fluoranthene 87.7 6.9
Pyrene 102 0.8
Butyl benzyl phthalate 66.3 5.2
3,3'-Dichlorobenzidine 25.2 11
Benzo(a)anthracene 73.4 3.8
Bis(2-ethylhexyl) phthalate 77.2 4.8
Chrysene 76.2 4.4
Di-n-octyl phthalate 83.1 4.8
Benzo(b)fluoranthene 82.7 5.0
Benzo(k)fluoranthene 71.7 4.1
Benzo(a)pyrene 71.7 4.1
Indeno(l,2,3-cd)pyrene 72.2 4.3
Dibenzo(a,h)anthracene 66.7 6.3
Benzo(g,h,i)perylene 63.9 8.0
1,2-Dichlorobenzene 0
1,3-Dichlorobenzene 0
1,4-Dichlorobenzene 0
Hexachloroethane 0
Hexachlorobutadiene 0
a Number of determinations was three. The operating conditions for the
Soxtec apparatus were as follows: immersion time 45 min; extraction time
45 min; the sample size was 10 g clay soil; the spike concentration was
6 mg/kg per compound. The sample was allowed to equilibrate 1 hour after
spiking.
Data taken from Reference 9.
8270B - 47 Revision 2
September 1994
-------
FIGURE 1.
GAS CHROMATOGRAM OF BASE/NEUTRAL AND ACID CALIBRATION STANDARD
: 5lbH*C«w786 kl
CnLl: 51BHS668786 13
SCANS 2t* TO 270e
KIC
06/u7'86 8:26:08
StWPLE: BASE ACID STD, 2UL/;.uHC,. UL
CODS.:
R»«CE: C 1.2780 Lfl££L: N 6. 4.9 CKJHN: M 0- t.d J 6 bkS£: U 2d- 3
JUL
33:20
41:46
8270B - 48
Revision 2
September 1994
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METHOD 8270B
SEMIVOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY
(GC/MS): CAPILLARY COLUMN TECHNIQUE
7.1 Prepare sample
using Method 3540,
3541, or 3550.
7.1 Prepare sample
using Method 3510
or 3520.
7.1 Prepare sample
using Method 3540,
3541, 3550, or 3580.
7.2 Cleanup
extract.
7.3 Set GC/MS
operating conditions;
perform initial
calibration.
7.4 Perform daily
calibration with SPCCs
and CCCs prior to
analysis of samples.
8270B - 49
Revision 2
September 1994
-------
METHOD 8270B
(Continued)
7.5.1 Screen extract
on GC/FID or GC/PID to
eliminate samples that
are too concentrated.
7.5.3 Analyze extract
by GC/MS, using
appropriate fused-silica
capillary column.
7.S.4 Dilute
Extract.
7.5.4
Does response
exceed initial
calibration
curve?
^
No
r
7.6.1 Identify
analyte by comparing
the sample and standard
mass spectra.
^1
r
7.6.2 Calculate
concentration of each
individual analyte;
report results.
>
r
C Stop "\
8270B - 50
Revision 2
September 1994
-------
ERRATA FOR METHOD 8280
In Section 1.1, delete the following text:
"reactor residues" with no replacement.
In Section 1.5, replace the following text:
"the analyst must take necessary precautions to prevent exposure to
himself, or to others, of"
with:
"the analyst must take necessary precautions to prevent human
exposure from" and
delete the following text:
"to be reviewed and approved by EPA's Dioxin Task Force (Contact
Conrad Kleveno, WH 548A, U.S. EPA, 401 M Street S.W., Washington, D.C.
20450)."
In Section 6.3, replace the following text:
"x = measured as in Figure 2"
with:
"x = height of the valley between 2,3,7,8-TCDD and 1,2,3,4-TCDD,
using the column performance check mixture."
In Section 6.9.2, replace "a 2-hr period" with "a 12 hr period".
In Section 7.4, replace "24" with "20".
8280 ERRATA - 1 July 1992
-------
METHOD 8280
THE ANALYSIS OF POLYCHLORINATED DIBENZO-P-DIOXINS
AND POLYCHLORINATED DIBENZOFURANS
1.0 SCOPE AND APPLICATION
1.1 This method is appropriate for the determination of tetra-, penta-,
hexa-, hepta-, and octachlorinated dibenzo-p-dioxins (PCDD's) and dibenzo-
furans (PCDF's) in chemical wastes including still bottoms, fuel oils,
sludges, fly ash, reactor residues, soil and water.
1.2 The sensitivity of this method is dependent upon the level of
interferents within a given matrix. Proposed quantification levels for target
analytes were 2 ppb in soil samples, up to 10 ppb in other solid wastes and
10 ppt in water. Actual values have been shown to vary by homologous series
and, to a lesser degree, by individual isomer. The total detection limit for
each CDD/CDF homologous series is determined by multiplying the detection
limit of a given isomer within that series by the number of peaks which can be
resolved under the gas chromatographic conditions.
1.3 Certain 2,3,7,8-substituted congeners are used to provide
calibration and method recovery information. Proper column selection and
access to reference isomer standards, may in certain cases, provide isomer
specific data. Special instructions are included which measure 2,3,7,8-
substituted congeners.
1.4 This method is recommended for use only by analysts experienced with
residue analysis and skilled in mass spectral analytical techniques.
1.5 Because of the extreme toxicity of these compounds, the analyst must
take necessary precautions to prevent exposure to himself, or to others, of
materials known or believed to contain PCDD's or PCDF's. Typical infectious
waste incinerators are probably not satisfactory devices for disposal of
materials highly contaminated with PCDD's or PCDF's. A laboratory planning to
use these compounds should prepare a disposal plan to be reviewed and approved
by EPA's Dioxin Task Force (Contact Conrad Kleveno, WH-548A, U.S. EPA, 401 M
Street S.W., Washington, D.C. 20450). Additional safety instructions are
outlined in Appendix B.
2.0 SUMMARY OF THE METHOD
2.1 This procedure uses a matrix-specific extraction, analyte-specific
cleanup, and high-resolution capillary column gas chromatography/low
resolution mass spectrometry (HRGC/LRMS) techniques.
2.2 If interferents are encountered, the method provides selected
cleanup procedures to aid the analyst in their elimination. The analysis flow
chart is shown in Figure 1.
8280 - 1
Revision 0
Date September 1986
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Complex
Waste
Sample
(1) Add Internal Standards: 13C12-PCDD's
and 13C12-PCDF's.
(2) Perform matrix-specific extraction.
Sample
Extract
(1) Wash with 20% KOH
(2) Wash with 5% NaCl
(3) Wash with cone. H2S04
(4) Wash with 5% NaCl
(5) Dry extract
(6) Solvent exchange
(7) Alumina column
60% CH2Cl2/hexane
Fraction
(1) Concentrate eluate
(2) Perform carbon column cleanup
(3) Add recovery standard(s)-13C12-l,2,3,4-TCDD
Analyze by GC/MS
Figure 1. Method 8280 flow chart for sample extraction and cleanup as
used for the analysis of PCDD's and PCDF's 1n complex waste samples.
8280 - 2
Revision 0
Date September 1986
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3.0 INTERFERENCES
3.1 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines which may cause
misinterpretation of chromatographic data. All of these materials must be
demonstrated to be free from interferents under the conditions of analysis by
running laboratory method blanks.
3.2 The use of high purity reagents and solvents helps to minimize
interference problems. Purification of solvents by distillation in all glass
systems may be required.
3.3 Interferents co-extracted from the sample will vary considerably
from source to source, depending upon the industrial process being sampled.
PCDD's and PCDF's are often associated with other interfering chlorinated
compounds such as PCB's and polychlorinated diphenyl ethers which may be found
at concentrations several orders of magnitude higher than that of the analytes
of interest. Retention times of target analytes must be verified using
reference standards. These values must correspond to the retention time
windows established in Section 6-3. While certain cleanup techniques are
provided as part of this method, unique samples may require additional cleanup
techniques to achieve the method detection limit (Section 11.6) stated in
Table 8.
3.4 High resolution capillary columns are used to resolve as many PCDD
and PCDF isomers as possible; however, no single column is known to resolve
all of the isomers.
3.5 Aqueous samples cannot be aliquoted from sample containers. The
entire sample must be used and the sample container washed/rinsed out with the
extracting solvent.
4.0 APPARATUS AND MATERIALS
4.1 Sampling equipment for discrete or composite sampling;
4.1.1 Grab sample bottle—amber glass, 1-liter or 1-quart volume.
French or Boston Round design is recommended. The container must be acid
washed and solvent rinsed before use to minimize interferences.
4.1.2 Bottle caps--threaded to screw onto the sample bottles. Caps
must be lined with Teflon. Solvent washed foil, used with the shiny side
toward the sample, may be substituted for Teflon if the sample is not
corrosive. Apply tape around cap to completely seal cap to bottom.
4.1.3 Compositing equipment—automatic or manual compositing
system. No tygon or rubber tubing may be used, and the system must
incorporate glass sample containers for the collection of a minimum of
250 ml. Sample containers must be kept refrigerated after sampling.
4.2 Water bath—heated, with concentric ring cover, capable of
temperature control (+2*C). The bath should be used in a hood.
8280 - 3
Revision 0
Date September 1986
-------
4.3 Gas chromatograph/mass spectrometer data system;
4.3.1 Gas chromatograph: An analytical system with a temperature-
programmable gas chromatograph and all required accessories including
syringes, analytical columns, and gases.
4.3.2 Fused silica capillary columns are required. As shown in
Table 1, three columns were evaluated using a column performance check
mixture containing 1,2,3,4-TCDD, 2,3,7,8-TCDD, 1,2,3,4,7 PeCDD,
1,2,3,4,7,8-HxCDD, 1,2,3,4,6,7,8-HpCDD, OCDD, and 2,3,7,8-TCDF.
The columns include the following: (a) 50-m CP-Sil-88 programmed 60*-
190* at 20*/minute, then 190*-240* at 5*/minute; (b) DB-5 (30-m x 0.25-mm
I.D.; 0.25-um film thickness) programmed 170* for 10 minutes, then 170*-
320* at 8*/minute, hold at 320*C for 20 minutes; (c) 30-m SP-2250
programmed 70*-320* at 10*/minute. Column/conditions (a) provide good
separation of 2,3,7,8-TCDD from the other TCDD's at the expense of longer
retention times for higher homologs. Column/conditions (b) and (c) can
also provide acceptable separation of 2,3,7,8-TCDD. Resolution of
2,3,7,8-TCDD from the other TCDD's is better on column (c), but column
(b) is more rugged, and may provide better separation from certain
classes of interferents. Data presented in Figure 2 and Tables 1 to 8 of
this Method were obtained using a DB-5 column with temperature
programming described in (b) above. However, any capillary column which
provides separation of 2,3,7,8-TCDD from all other TCDD isomers
equivalent to that specified in Section 6.3 may be used; this separation
must be demonstrated and documented using the performance test mixture
described in Paragraph 6.3.
4.3.3 Mass spectrometer: A low resolution instrument is specified,
utilizing 70 volts (nominal) electron energy in the electron impact
ionization mode. The system must be capable of selected ion monitoring
(SIM) for at least 11 ions simultaneously, with a cycle time of 1 sec or
less. Minimum integration time for SIM is 50 ms per m/z. The use of
systems not capable of monitoring 11 ions simultaneously will require the
analyst to make multiple injections.
4.3.4 GC/MS Interface: Any GC-to-MS interface that gives an
acceptable calibration response for each analyte of interest at the
concentration required and achieves the required tuning performance
criteria (see Paragraphs 6.1.-6.3) may be used. GC-to-MS interfaces
constructed of all glass or glass-lined materials are required. Glass
can be deactivated by silanizing with dichlorodimethyl si lane. Inserting
a fused silica column directly into the MS source is recommended; care
must be taken not to expose the end of the column to the electron beam.
4.3.5 Data system: A computer system must be interfaced to the
mass spectrometer. The system must allow for the continuous acquisition
and storage on machine-readable media of all data obtained throughout the
duration of the chromatographic program. The computer must have software
that can search any GC/MS data file for ions of a specific mass and can
plot such ion abundances versus time or scan number. This type of plot
8280 - 4
Revision 0
Date September 1986
-------
Is defined as an Selected Ion Current Profile (SICP). Software must also
be able to integrate the abundance, in any SICP, between specified time
or scan number limits.
4.4 Pipets-Disposable, Pasteur, 150-mm long x 5-mrn I.D. (Fisher
Scientific Company, No. 13-678-6A, or equivalent).
4.4.1 Pipet, disposable, serological 10-mL (American Scientific
Products No. P4644-10, or equivalent) for preparation of the carbon
column specified in Paragraph 4.19.
4.5 Amber glass bottle (500-mL, Teflon-lined screw-cap).
4.6 Reacti-vial 2-mL, amber glass (Pierce Chemical Company). These
should be silanized prior to use.
4.7 500-mL Erlenmeyer flask (American Scientific Products Cat. No. f4295
500fO) fitted with Teflon stoppers (ASP No. S9058-8, or equivalent).
4.8 Wrist Action Shaker (VWR No. 57040-049, or equivalent).
4.9 125-mL and 2-L Separatory Funnels (Fisher Scientific Company,
No. 10-437-5b, or equivalent).
4.10 500-mL Kuderna-Danish fitted with a 10-mL concentrator tube and
3-ball Snyder column (Ace Glass No. 6707-02, 6707-12, 6575-02, or equivalent).
4.11 Teflon boiling chips (Berghof/American Inc., Main St., Raymond, New
Hampshire 03077, No. 15021-450, or equivalent). Wash with hexane prior to
use.
4.12 300-mm x 10.5-mm glass chromatographic column fitted with Teflon
stopcock.
4.13 15-mL conical concentrator tubes (Kontes No. K-288250, or
equivalent).
4.14 Adaptors for concentrator tubes (14/20 to 19/22) (Ace Glass No.
9092-20, or equivalent).
4.15 Nitrogen blowdown apparatus (N-Evap (reg. trademark) Analytical
Evaporator Model 111, Organomation Associates Inc., Northborough,
Massachusetts or equivalent). Teflon tubing connection to trap and gas
regulator is required.
4.16 Microflex conical vials 2.0-mL (Kontes K-749000, or equivalent).
4.17 Filter paper (Whatman No. 54, or equivalent). Glass fiber filters
or glass wool plugs are also recommended.
4.18 Solvent reservoir (125-mL) Kontes; (special order item) 12.5-cm
diameter, compatible with gravity carbon column.
8280 - 5
Revision 0
Date September 1986
-------
4.19 Carbon column (gravity flow); Prepare carbon/silica gel packing
material by mixing5percent (byweight) active carbon AX-21 (Anderson
Development Co., Adrain, Michigan), pre-washed with methanol and dried Iji
vacuo at 110'C and 95 percent (by weight) Silica gel (Type 60, EM reagent 70
to 230 mesh, CMS No. 393-066) followed by activation of the mixture at 130*
for 6 hr. Prepare a 10-mL disposable serologlcal pipet by cutting off each
end to achieve a 4-1n. column. F1re polish both ends; flare If desired.
Insert a glass-wool plug at one end and pack with 1 g of the carbon/siHca gel
mixture. Cap the packing with a glass-wool plug. (Attach reservoir to column
for addition of solvents).
Option: Carbon column (HPLC): A silanized glass HPLC column (10 mm x 7
cm), or equivalent, which contains 1 g of a packing prepared by mixing 5
percent (by weight) active carbon AX-21, (Anderson Development Co., Adrian,
Michigan), washed with methanol and dried iji vacuo at 110'C, and 95 percent
(by weight) 10 urn silica (Spherisorb S10W from Phase Separations, Inc.,
Norwalk, Connecticut). The mixture must then be stirred and sieved through a
38-um screen (U.S. Sieve Designation 400-mesh, American Scientific Products,
No. S1212-400, or equivalent) to remove any clumps.1
4.20 HPLC pump with loop valve (1.0 ml) injector to be used in the
optional carbon column cleanup procedure.
4.21 Dean-Stark trap, 5- or 10-mL with T joints, (Fisher Scientific
Company, No. 09-146-5, or equivalent) condenser and 125-mL flask.
4.22 Continuous liquid-liquid extractor (Hershberg-Wolfe type, Lab Glass
No. LG-6915; or equivalent.).
4.23 Roto-evaporator, R-110. Buchi/Brinkman - American Scientific No.
E5045-10; or equivalent.
5.0 REAGENTS
5.1 Potassium hydroxide (ASC): 20 percent (w/v) in distilled water.
5.2 Sulfuric acid (ACS), concentrated.
5.3 Methylene chloride, hexane, benzene, petroleum ether, methanol,
tridecane, isooctane, toluene, cyclohexane. Distilled in glass or highest
available purity.
5.4 Prepare stock standards in a glovebox from concentrates or neat
materials. The stock solutions (50 ppm) are stored in the dark at 4*C, and
checked frequently for signs of degradation or evaporation, especially just
prior to the preparation of working standards.
1 The carbon column preparation and use is adapted from W. A. Korfmacher,
L. G. Rushing, D. M. Nestorick, H. C. Thompson, Jr., R. K. Mltchum, and J. R.
Kominsky, Journal of High Resolution Chromatography and Chromatography
Communications, 8, 12-19 (1985).
8280 - 6
Revision 0
Date September 1986
-------
5.5 Alumina, neutral, Super 1, Woelm, 80/200 mesh. Store in a sealed
container at room temperature in a desiccator over self-indicating silica gel.
5.6 Prepurified nitrogen gas.
5.7 Anhydrous sodium sulfate (reagent grade): Extracted by manual
shaking with several portions of hexane and dried at 100*C.
5.8 Sodium chloride - (analytical reagent), 5 percent (w/v) in distilled
water.
6.0 CALIBRATION
6.1 Two types of calibration procedures are required. One type, initial
calibration, is required before any samples are analyzed and is required
intermittently throughout sample analyses as dictated by results of routine
calibration procedures described below. The other type, routine calibration,
consists of analyzing the column performance check solution and a
concentration calibration solution of 500 ng/mL (Paragraph 6.2). No samples
are to be analyzed until acceptable calibration as described in Paragraphs 6.3
and 6.6 is demonstrated and documented.
6.2 Initial calibration;
6.2.1 Prepare multi-level calibration standards2 keeping one of
the recovery standards and the internal standard at fixed concentrations (500
ng/mL). Additional internal standards (13Ci2-OCDD 1,000 ng/mL) are
recommended when quantification of the hepta- and octa-isomers is required.
The use of separate internal standards for the PCDF's is also recommended.
Each calibration standard should contain the following compounds:
2,3,7,8-TCDD,
1,2,3,7,8-PeCDD or any available 2,3,7,8,X-PeCDD isomer,
1,2,3,4,7,8-HxCDD or any available 2,3,7,8,X,Y-HxCDD isomer,
1,2,3,4,6,7,8-HpCDD or any available 2,3,7,8,X,Y,Z-HpCDD isomer,
2,3,7,8-TCDF
l,2,3,7,8,PeCDF or any available 2,3,7,8,X-PeCDF isomer,
1,2,3,4,7,8-HxCDF or any available 2,3,7,8,X,YlHxCDF isomer,
1,2,3,4,6,7,8-HpCDF or any available 2,3,7,8,X,Y,Z-HpCDF isomer,
OCDD, OCDF, 13Ci2-2,3,7,8-TCDD, i3Ci2-l,2,3,4-TCDD and i3Ci2-OCDD.
2 13Cj2-labeled analytes are available from Cambridge Isotope Laboratory,
Woburn, Massachusetts. Proper quantification requires the use of a specific
labeled isomer for each congener to be determined. When labeled PCDD's and
PCDF's of each homolog are available, their use will be required consistent
with the technique of isotopic dilution.
8280 - 7
Revision 0
Date September 1986
-------
Recommended concentration levels for standard analytes are 200, 500, 1,000,
2,000, and 5,000 ng/mL. These values may be adjusted in order to insure that
the analyte concentration falls within the calibration range. Two uL
injections of calibration standards should be made. However, some GC/MS
instruments may require the use of a 1-uL injection volume; if this injection
volume 1s used then all injections of standards, sample extracts and blank
extracts must also be made at this injection volume. Calculation of relative
response factors is described in Paragraph 11.1.2. Standards must be analyzed
using the same solvent as used 1n the final sample extract. A wider
calibration range is useful for higher level samples provided it can be
described within the linear range of the method, and the identification
criteria defined in Paragraph 10.4 are met. All standards must be stored in
an isolated refrigerator at 4*C and protected from light. Calibration
standard solutions must be replaced routinely after six months.
6.3 Establish operating parameters for the GC/MS system; the instrument
should be tuned to meet the isotopic ratio criteria listed in Table 3 for
PCDD's and PCDF's. Once tuning and mass calibration procedures have been
completed, a column performance check mixture3 containing the isomers listed
below should be injected into the GC/MS system:
TCDD 1,3,6,8; 1,2,8,9; 2,3,7,8; 1,2,3,4; 1,2,3,7; 1,2,3,9
PeCDD 1,2,4,6,8; 1,2,3,8,9
HxCDD 1,2,3,4,6,9; 1,2,3,4,6,7
HpCDD 1,2,3,4,6,7,8; 1,2,3,4,6,7,9
OCDD 1,2,3,4,6,7,8,9
TCDF 1,3,6,8; 1,2,8,9
PeCDF 1,3,4,6,8; 1,2,3,8,9
HxCDF 1,2,3,4,6,8; 1,2,3,4,8',9
HpCDF 1,2,3,4,6,7,8; 1,2,3,4,7,8,9
OCDF 1,2,3,4,6,7,8,9
Because of the known overlap between the late-eluting tetra-isomers and
the early-eluting penta-isomers under certain column conditions, it may be
necessary to perform two injections to define the TCDD/TCDF and PeCDD/PeCDF
elution windows, respectively. Use of this performance check mixture will
enable the following parameters to be checked: (a) the retention windows for
each of the homologues, (b) the GC resolution of 2,3,7,8-TCDD and 1,2,3,4-
TCDD, and (c) the relative ion abundance criteria listed for PCDD's and PCDF's
in Table 3. GC column performance should be checked dally for resolution and
peak shape using this check mixture.
The chromatographic peak separation between 2,3,7,8-TCDD and 1,2,3,4-TCDD
must be resolved with a valley of <£25 percent, where
Valley Percent = (x/y) (100)
x = measured as in Figure 2
y = the peak height of 2,3,7,8-TCDD
3 Performance check mixtures are available from Brehm Laboratory, Wright
State University, Dayton, Ohio.
8280 - 8
Revision 0
Date September 1986
-------
It 1s the responsibility of the laboratory to verify the conditions
suitable for maximum resolution of 2,3,7,8-TCDD from all other TCDD isomers.
The peak representing 2,3,7,8-TCDD should be labeled and identified as such on
all chromatograms.
6.4 Acceptable SIM sensitivity is verified by achieving a minimum
signal-to-noise ratio of 50:1 for the m/z 320 ion of 2,3,7,8-TCDD obtained
from injection of the 200 ng/mL calibration standard.
6.5 From injections of the 5 calibration standards, calculate the
relative response factors (RRF's) of analytes vs. the appropriate internal
standards, as described in Paragraph 11.1.2. Relative response factors for
the hepta- and octa-chlorinated CDD's and CDF's are to be calculated using the
corresponding 13Cj2-octachlorinated standards.
6.6 For each analyte calculate the mean relative response factor (RRF),
the standard deviation, and the percent relative standard deviation from
triplicate determinations of relative response factors for each calibration
standard solution.
6.7 The percent relative standard deviations (based on triplicate
analysis) of the relative response factors for each calibration standard
solution should not exceed 15 percent. If this condition is not satisfied,
remedial action should be taken.
6.8 The Laboratory must not proceed with analysis of samples before
determining and documenting acceptable calibration with the criteria specified
in Paragraphs 6.3 and 6.7.
6.9 Routine calibration;
6.9.1 Inject a 2-uL aliquot of the column performance check
mixture. Acquire at least five data points for each GC peak and use the
same data acquisition time for each of the ions being monitored.
NOTE: The same data acquisition parameters previously used to
analyze concentration calibration solutions during initial
calibration must be used for the performance check solution.
The column performance check solution must be run at the
beginning and end of a 12 hr period. If the contractor
laboratory operates during consecutive 12-hr periods
(shifts), analysis of the performance check solution at the
beginning of each 12-hr period and at the end of the final
12-hr period is sufficient.
Determine and document acceptable column performance as described in
Paragraph 6.3.
6.9.2 Inject a 2-uL aliquot of the calibration standard solution at
500 ng/mL at the beginning of a 2-hr period. Determine and document
acceptable calibration as specified in Paragraph 6.3, i.e., SIM
sensitivity and relative ion abundance criteria. The measured RRF's of
8280 - 9
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all analytes must be within +30 percent of the mean values established by
Initial analyses of the calibration standard solutions.
7.0 QUALITY CONTROL
7.1 Before processing any samples, the analyst must demonstrate through
the analysis of a method blank that all glassware and reagents are
interferent-free at the method detection limit of the matrix of interest.
Each time a set of samples is extracted, or there is a change in reagents, a
method blank must be processed as a safeguard against laboratory
contamination.
7.2 A laboratory "method blank" must be run along with each analytical
batch (20 or fewer samples). A method blank is performed by executing all of
the specified extraction and cleanup steps, except for the introduction of a
sample. The method blank is also dosed with the Internal standards. For
water samples, one liter of deionized and/or distilled water should be used as
the method blank. Mineral oil may be used as the method blank for other
matrices.
7.3 The laboratory will be expected to analyze performance evaluation
samples as provided by the EPA on a periodic basis throughout the course of a
given project. Additional sample analyses will not be permitted if the
performance criteria are not achieved. Corrective action must be taken and
acceptable performance must be demonstrated before sample analyses can resume.
7.4 Samples may be split with other participating labs on a periodic
basis to ensure interlaboratory consistency. At least one sample per set of
24 must be run 1n duplicate to determine intralaboratory precision.
7.5 Field duplicates (individual samples taken from the same location at
the same time) should be analyzed periodically to determine the total
precision (field and lab).
7.6 Where appropriate, "field blanks" will be provided to monitor for
possible cross-contamination of samples in the field. The typical "field
blank" will consist of uncontaminated soil (background soil taken off-site).
7.7 GC column performance must be demonstrated initially and verified
prior to analyzing any sample in a 12-hr period. The GC column performance
check solution must be analyzed under the same chromatographic and mass
spectrometric conditions used for other samples and standards.
7.8 Before using any cleanup procedure, the analyst must process a
series of calibration standards (Paragraph 6.2) through the procedure to
validate elution patterns and the absence of interferents from reagents. Both
alumina column and carbon column performance must be checked. Routinely check
the 8 percent CH2Cl2/hexane eluate of environmental extracts from the alumina
column for presence of target analytes.
NOTE: This fraction is intended to contain a high level of interferents
and analysis near the method detection limit may not be possible.
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8.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
8.1 Grab and composite samples must be collected in glass containers.
Conventional sampling practices must be followed. The bottle must not be
prewashed with sample before collection. Composite samples should be
collected in glass containers. Sampling equipment must be free of tygon,
rubber tubing, other potential sources of contamination which may absorb the
target analytes.
8.2 All samples must be stored at 4*C, extracted within 30 days and
completely analyzed within 45 days of collection.
9.0 EXTRACTION AND CLEANUP PROCEDURES
9.1 Internal standard addition. Use a sample aliquot of 1 g to 1,000 mL
(typical sample size requirements for each type of matrix are provided in
Paragraph 9.2) of the chemical waste or soil to be analyzed. Transfer the
sample to a tared flask and determine the weight of the sample. Add an
appropriate quantity of 13Ci2-2,3,7,8-TCDD, and any other material which is to
be used as an internal standard, (Paragraph 6.2). All samples should be
spiked with at least one internal standard, for example, 13Ci2-2,3,7,8-TCDD,
to give a concentration of 500 ng/mL in the final concentrated extract. As an
example, a 10 g sample concentrated to a final volume of 100 uL requires the
addition of 50 ng of 13Ci2-2,3,7,8-TCDD, assuming 100% recovery. Adoption of
different calibration solution sets (as needed to achieve different
quantification limits for different congeners) will require a change In the
fortification level. Individual concentration levels for each homologous
series must be specified.
9.2 Extraction
9.2.1 Sludge/fuel oil. Extract aqueous sludge samples by refluxing
a sample (e.g. 2 g) with 50 mL of toluene (benzene) in a 125-mL flask
fitted with a Dean-Stark water separator. Continue refluxing the sample
until all the water has been removed. Cool the sample, filter the
toluene extract through a fiber filter, or equivalent, into a 100-mL
round bottom flask. Rinse the filter with 10 mL of toluene, combine the
extract and rinsate. Concentrate the combined solution to near dryness
using a rotary evaporator at 50*C. Use of an inert gas to concentrate
the extract is also permitted. Proceed with Step 9.2.4.
9.2.2 Still bottom. Extract still bottom samples by mixing a
sample (e.g., 1.0 g) with 10 mL of toluene (benzene) in a small beaker
and filtering the solution through a glass fiber filter (or equivalent)
Into a 50-mL round bottom flask. Rinse the beaker and filter with 10 mL
of toluene. Concentrate the combined toluene solution to near dryness
using a rotary evaporator at 50*C while connected to a water aspirator.
Proceed with Step 9.2.4.
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9.2.3 Fly ash. Extract fly ash samples by placing a sample (e.g.
10 g) and an equivalent amount of anhydrous sodium sulfate in a Soxhlet
extraction apparatus charged with 100 ml of toluene (benzene) and extract
for 16 hr using a three cycle/hour schedule. Cool and filter the toluene
extract through a glass fiber filter paper into a 500-mL round bottom
flask. Rinse the filter with 5 ml of toluene. Concentrate the combined
toluene solution to near dryness using a rotary evaporator at 50*C.
Proceed with Step 9.2.4.
9.2.4 Transfer the residue to a 125-mL separatory funnel using
15 ml of hexane. Rinse the flask with two 5-mL aliquots of hexane and
add the rinses to the funnel. Shake 2 min with 50 ml of 5% NaCl
solution, discard the aqueous layer and proceed with Step 9.3.
9.2.5 Soil. Extract soil samples by placing the sample (e.g. 10 g)
and an equivalent amount of anhydrous sodium sulfate in a 500-mL
Erlenmeyer flask fitted with a Teflon stopper. Add 20 ml of methanol and
80 ml of petroleum ether, in that order, to the flask. Shake on a wrist-
action shaker for two hr. The solid portion of sample should mix freely.
If a smaller soil aliquot is used, scale down the amount of methanol
proportionally.
9.2.5.1 Filter the extract from Paragraph 9.2.5 through a
glass funnel fitted with a glass fiber filter and filled with
anhydrous sodium sulfate into a 500-mL Kuderna-Danish (KD)
concentrator fitted with a 10-mL concentrator tube. Add 50 ml. of
petroleum ether to the Erlenmeyer flask, restopper the flask and
swirl the sample gently, remove the stopper carefully and decant the
solvent through the funnel as above. Repeat this procedure with two
additional 50-mL aliquots of petroleum ether. Wash the sodium
sulfate in the funnel with two additional 5-mL portions of petroleum
ether.
9.2.5.2 Add a Teflon or PFTE boiling chip and a three-ball
Snyder column to the KD flask. Concentrate in a 70*C water bath to
an apparent volume of 10 mL. Remove the apparatus from the water
bath and allow it to cool for 5 min.
9.2.5.3 Add 50 mL of hexane and a new boiling chip to the KD
flask. Concentrate in a water bath to an apparent volume of 10 mL.
Remove the apparatus from the water bath and allow to cool for 5
min.
9.2.5.4 Remove and invert the Snyder column and rinse it down
into the KD with two 1-mL portions of hexane. Decant the contents
of the KD and concentrator tube into a 125-mL separatory funnel.
Rinse the KD with two additional 5-mL portions of hexane, combine.
Proceed with Step 9.3.
9.2.6 Aqueous samples: Mark the water meniscus on the side of the
1-L sample bottle for later determination of the exact sample volume.
8280 - 12
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Pour the entire sample (approximately 1-1} Into a 2-1 separatory funnel.
Proceed with Step 9.2.6.1.
NOTE: A continuous liquid-liquid extractor may be used 1n place of
a separatory funnel when experience with a sample from a
given source Indicates that a serious emulsion problem will
result or an emulsion is encountered using a separatory
funnel. Add 60 ml of methylene chloride to the sample
bottle, seal, and shake for 30 sec to rinse the Inner
surface. Transfer the solvent to the extractor. Repeat the
sample bottle rinse with an additional 50- to 100-mL portion
of methylene chloride and add the rinse to the extractor.
Add 200 to 500 ml of methylene chloride to the distilling
flask; add sufficient reagent water to ensure proper
operation, and extract for 24 hr. Allow to cool, then detach
the distilling flask. Dry and concentrate the extract as
described in Paragraphs 9.2.6.1 and 9.2.6.2. Proceed with
Paragraph 9.2.6.3.
9.2.6.1 Add 60 mL methylene chloride to the sample bottle,
seal and shake 30 sec to rinse the inner surface. Transfer the
solvent to the separatory funnel and extract the sample by shaking
the funnel for 2 min with periodic venting. Allow the organic layer
to separate from the water phase for a minimum of 10 min. If the
emulsion interface between layers is more than one-third the volume
of the solvent layer, the analyst must employ mechanical techniques
to complete the phase separation. Collect the methylene chloride
(3 x 60 ml) directly Into a 500-mL Kuderna-Danish concentrator
(mounted with a 10-mL concentrator tube) by passing the sample
extracts through a filter funnel packed with a glass wool plug and
5 g of anhydrous sodium sulfate. After the third extraction, rinse
the sodium sulfate with an additional 30 ml of methylene chloride to
ensure quantitative transfer.
9.2.6.2 Attach a Snyder column and concentrate the extract on
a water bath until the apparent volume of the liquid reaches 5 ml.
Remove the K-D apparatus and allow it to drain and cool for at least
10 min. Remove the Snyder column, add 50 ml hexane, re-attach the
Snyder column and concentrate to approximately 5 ml. Add a new
boiling chip to the K-D apparatus before proceeding with the second
concentration step.
Rinse the flask and the lower joint with 2 x 5 ml hexane and combine
rinses with extract to give a final volume of about 15 mL.
9.2.6.3 Determine the original sample volume by refilling the
sample bottle to the mark and transferring the liquid to a 1,000-mL
graduated cylinder. Record the sample volume to the nearest 5 ml.
Proceed with Paragraph 9.3.
9.3 In a 250-mL Separatory funnel, partition the solvent (15 ml hexane)
against 40 ml of 20 percent (w/v) potassium hydroxide. Shake for 2 m1n.
8280 - 13
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Remove and discard the aqueous layer (bottom). Repeat the base washing until
no color is visible in the bottom layer (perform base washings a maximum of
four times). Strong base (KOH) is known to degrade certain PCDD/PCDF's,
contact time must be minimized.
9.4 Partition the solvent (15 ml hexane) against 40 ml of 5 percent
(w/v) sodium chloride. Shake for 2 min. Remove and discard aqueous layer
(bottom).
NOTE: Care should be taken due to the heat of neutralization and
hydration.
9.5 Partition the solvent (15 ml hexane) against 40 ml of concentrated
sulfuric acid. Shake for 2 min. Remove and discard the aqueous layer
(bottom). Repeat the acid washings until no color is visible in the acid
layer. (Perform acid washings a maximum of four times.)
9.6 Partition the extract against 40 ml of 5 percent (w/v) sodium
chloride. Shake for 2 min. Remove and discard the aqueous layer (bottom).
Dry the organic layer by pouring through a funnel containing anhydrous sodium
sulfate into a 50-mL round bottom flask, wash the separatory funnel with two
15-mL portions of hexane, pour through the funnel, and combine the hexane
extracts. Concentrate the hexane solution to near dryness with a rotary
evaporator (35*C water bath), making sure all traces of toluene are removed.
(Use of blowdown with an inert gas to concentrate the extract is also
permitted).
9.7 Pack a gravity column (glass 300-mm x 10.5-mm), fitted with a Teflon
stopcock, in the following manner:
Insert a glass-wool plug into the bottom of the column. Add a 4-g layer
of sodium sulfate. Add a 4-g layer of Woelm super 1 neutral alumina. Tap the
top of the column gently. Woelm super 1 neutral alumina need not be activated
or cleaned prior to use but should be stored in a sealed desiccator. Add a 4-
g layer of sodium sulfate to cover the alumina. Elute with 10 ml of hexane
and close the stopcock just prior to the exposure of the sodium sulfate layer
to air. Discard the eluant. Check the column for channeling. If channeling
is present discard the column. Do not tap a wetted column.
9.8 Dissolve the residue from Step 9.6 in 2 ml of hexane and apply the
hexane solution to the top of the column. Elute with enough hexane (3-4 ml)
to complete the transfer of the sample cleanly to the surface of the alumina.
Discard the eluant.
9.8.1 Elute with 10 ml of 8 percent (v/v) methylene chloride in
hexane. Check by GC/MS analysis that no PCDD's or PCDF's are eluted in
this fraction. See Paragraph 9.9.1.
9.8.2 Elute the PCDD's and PCDF's from the column with 15 ml of 60
percent (v/v) methylene chloride in hexane and collect this fraction in a
conical shaped (15-mL) concentrator tube.
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9.9 Carbon column cleanup;
Prepare a carbon column as described in Paragraph 4.18.
9.9.1 Using a carefully regulated stream of nitrogen (Paragraph
4.15), concentrate the 8 percent fraction from the alumina column
(Paragraph 9.8.1) to about 1 ml. Wash the sides of the tube with a small
volume of hexane (1 to 2 ml) and reconcentrate to about 1 ml. Save this
8 percent concentrate for GC/MS analysis to check for breakthrough of
PCDD's and PCDF's. Concentrate the 60 percent fraction (Paragraph 9.8.2)
to about 2 to 3 ml. Rinse the carbon with 5 ml cyclohexane/methylene
chloride (50:50 v/v) 1n the forward direction of flow and then In the
reverse direction of flow. While still in the reverse direction of flow,
transfer the sample concentrate to the column and elute with 10 ml of
cyclohexane/methylene chloride (50:50 v/v) and 5 ml of methylene
chloride/methanol/benzene (75:20:5, v/v). Save all above eluates and
combine (this fraction may be used as a check on column efficiency). Now
turn the column over and in the direction of forward flow elute the
PCDD/PCDF fraction with 20 ml toluene.
NOTE: Be sure no carbon fines are present in the eluant.
9.9.2 Alternate carbon column cleanup. Proceed as in Section 9.9.1
to obtain the 60 percent fraction re-concentrated to 400 uL which is
transferred to an HPLC injector loop (1 ml). The injector loop 1s
connected to the optional column described in Paragraph 4.18. Rinse the
centrifuge tube with 500 uL of hexane and add this rinsate to the
injector loop. Load the combined concentrate and rinsate onto the
column. Elute the column at 2 mL/min, ambient temperature, with 30 ml of
cyclohexane/methylene chloride 1:1 (v/v). Discard the eluant. Backflush
the column with 40 mL toluene to elute and collect PCDD's and PCDF's
(entire fraction). The column is then discarded and 30 ml of
cyclohexane/methylene chloride 1:1 (v/v) is pumped through a new column
to prepare it for the next sample.
9.9.3 Evaporate the toluene fraction to about 1 ml on a rotary
evaporator using a water bath at 50*C. Transfer to a 2.0-mL React1-vial
using a toluene rinse and concentrate to the desired volume using a
stream of N£. The final volume should be 100 uL for soil samples and
500 uL for sludge, still bottom, and fly ash samples; this is provided
for guidance, the correct volume will depend on the relative concentra-
tion of target analytes. Extracts which are determined to be outside the
calibration range for individual analytes must be diluted or a smaller
portion of the sample must be re-extracted. Gently swirl the solvent on
the lower portion of the vessel to ensure complete dissolution of the
PCDD's and PCDF's.
9.10 Approximately 1 hr before HRGC/LRMS analysis, transfer an aliquot
of the extract to a micro-vial (Paragraph 4.16). Add to this sufficient
recovery standard (13Ci2l,2,3,4-TCDD) to give a concentration of 500 ng/mL.
(Example: 36 uL aliquot of extract and 4 uL of recovery standard solution.
Remember to adjust the final result to correct for this dilution. Inject an
appropriate aliquot (1 or 2 uL) of the sample into the GC/MS instrument.
8280 - 15
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10.0 GC/MS ANALYSIS
10.1 When toluene Is employed as the final solvent use of a bonded phase
column from Paragraph 4.3.2 is recommended. Solvent exchange Into trldecane
is required for other liquid phases or nonbonded columns (CP-Sil-88).
NOTE: Chromatographic conditions must be adjusted to account for solvent
boiling points.
10.2 Calculate response factors for standards relative to the internal
standards, 13C12-2,3,7,8-TCDD and 13Ci2-OCDD (see Section 11). Add the
recovery standard (13Ci2-l,2,3,4-TCDD) to the samples prior to injection. The
concentration of the recovery standard in the sample extract must be the same
as that in the calibration standards used to measure the response factors.
10.3 Analyze samples with selected ion monitoring, using all of the ions
listed in Table 2. It is recommended that the GC/MS run be divided into five
selected 1on monitoring sections, namely: (1) 243, 257,, 304, 306, 320, 322,
332, 334, 340, 356,, 376 (TCDD's, TCDF's, 13Ci2-labeled internal and recovery
standards, PeCDD's,, PeCDF's, HxCDE); (2) 277, 293, 306, 332, 338, 340, 342,
354, 356, 358, 410 (peCDD's, PeCDF's, HpCDE); (3) 311, 327, 340, 356, 372,
374, 376, 388, 390,, 392, 446, (HxCDD's, HxCDF's, OCDE); (4) 345, 361, 374,
390, 406, 408, 410r 422, 424, 426, 480 (HpCDD's, HpCDF's, NCDE) and (5) 379,
395, 408, 424, 442,, 444, 458, 460, 470, 472, 514 (OCDD, OCDF, 13Ci2-OCDD,
DCDE). Cycle time not to exceed 1 sec/descriptor. It is recommended that
selected ion monitoring section 1 should be applied during the GC run to
encompass the retention window (determined in Paragraph 6.3) of the first- and
last-eluting tetra-chlorinated isomers. If a response is observed at m/z 340
or 356, then the GC/MS analysis must be repeated; selected ion monitoring
section 2 should then be applied to encompass the retention window of the
first- and last-eluting penta-chlorinated isomers. HxCDE, HpCDE, OCDE, NCDE,
DCDE, are abbreviations for hexa-, hepta-, octa-, nona-, and decachlorinated
diphenyl ether, respectively.
10.4 Identification criteria for PCDD's and PCDF's:
10.4.1 All of the characteristic ions, i.e. quantisation ion,
confirmation ions, listed in Table 2 for each class of PCDD and PCDF,
must be present in the reconstructed ion chromatogram. It is desirable
that the M - COC1 ion be monitored as an additional requirement.
Detection limits will be based on quantitation ions within the molecules
in cluster.
10.4.2 The maximum intensity of each of the specified charac-
teristic ions must coincide within 2 scans or 2 sec.
10.4.3 The relative intensity of the selected, isotopic ions within
the molecular ion cluster of a homologous series of PCDD's of PCDF's must
lie within the range specified in Table 3.
10.4.4 The GC peaks assigned to a given homologous series must have
retention times within the window established for that series by the
column performance solution.
8280 - 16
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10.5 Quantitate the PCDD and PCDF peaks from the response relative to
the appropriate Internal standard. Recovery of each Internal standard) vs.
the recovery standard must be greater than 40 percent. It Is recommended that
samples with recoveries of less than 40 percent or greater than 120 percent be
re-extracted and re-analyzed.
NOTE: These criteria are used to assess method performance; when
properly applied, Isotope dilution techniques are Independent of
Internal standard recovery.
In those circumstances where these procedures do not yield a definitive
conclusion, the use of high resolution mass spectrometry or HRGC/MS/MS is
suggested.
11.0 CALCULATIONS
NOTE: The relative response factors of a given congener within any
homologous series are known to be different. However, for
purposes of these calculations, 1t will be assumed that every
congener within a given series has the same relative response
factor. In order to minimize the effect of this assumption on
risk assessment, a 2,3,7,8-substituted isomer that 1s
commercially available was chosen as representative of each
series. All relative response factor calculations for a given
homologous series are based on that compound.
11.1 Determine the concentration of individual isomers of tetra-, penta,
and hexa-CDD/CDF according to the equation:
Concentration, ng/g = G X$RRF
where :
Qis = "9 of internal standard 13Ci2-2,3,7,8-TCDD, added to the sample
before extraction.
G = g of sample extracted.
As = area of quantisation ion of the compound of interest.
AJS = area of quantitatlon 1on (m/z 334) of the internal standard,
13C12-2,3,7,8-TCDD.
RRF = response factor of the quantitation ion of the compound of
Interest relative to m/z 334 of 13Ci2-2,3,7,8-TCDD.
NOTE: Any dilution factor introduced by following the procedure in
Paragraph 9.10 should be applied to this calculation.
8280 - 17
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11.1.1 Determine the concentration of individual isomers of hepta-
CDD/CDF and the concentration of OCDD and OCDF according to the equation:
Qis x As
Concentration, ng/g = G x x RRF
where :
Qis = n9 °f internal standard c^-OCDD, added to the sample before
extraction.
G = g of sample extracted.
AS = area of quantitation ion of the compound of interest.
Ajs = area of quantitation ion (m/z 472) of the internal standard,
13Ci2-OCDD.
RRF = response factor of the quantitation ion of the compound of
interest relative to m/z 472 of 13Ci2-OCDD.
NOTE: Any dilution factor introduced by following the procedure in
Paragraph 9.10 should be applied to this calculation.
11.1.2 Relative response factors are calculated using data obtained
from the analysis of multi -level calibration standards according to the
equation:
RRF = ^" ^ C
is s
where:
As = area of quantitation ion of the compound of interest.
AIS = area of quantitation ion of the appropriate internal standard
(m/z 334 for 13C12-2,3,7,8-TCDD; m/z 472 for 13C12-OCDD).
C^s = concentration of the appropriate internal standard,
l3Ci2-2,3,7,8-TCDD or 13C12-OCDD)
Cs = concentration of the compound of interest.
11.1.3 The concentrations of unknown isomers of TCDD shall be
calculated using the mean RRF determined for 2,3,7,8-TCDD.
The concentrations of unknown isomers of PeCDD shall be calculated
using the mean RRF determined for 1,2,3,7,8-PeCDD or any available
2,3,7,8,X-PeCDD isomer.
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The concentrations of unknown isomers of HxCDD shall be calculated
using the mean RRF determined for 1,2,3,4,7,8-HxCDD or any available
2,3,7,8,-X,Y-HXCDD isomer.
The concentrations of unknown isomers of HpCDD shall be calculated
using the mean RRF determined for 1,2,3,4,6,7,8-HpCDD or any available
2,3,7,8,X,YrZ-HpCDD isomer.
The concentrations of unknown isomers of TCDF shall be calculated
using the mean RRF determined for 2,3,7,8-TCDF.
The concentrations of unknown isomers of PeCDF shall be calculated
using the mean RRF determined for 1,2,3,7,8-PeCDF or any available
2,3,7,8lX-PeCDF isomer.
The concentrations of unknown isomers of HxCDF shall be calculated
using the mean RRF determined for 1,2,4,7,8-HxCDF or any available
2,3,7,8-X.Y-HxCDF isomer.
The concentrations of unknown isomers of HpCDF shall be calculated
using the mean RRF determined for 1,2,3,4,6,7,8-HpCDF or any available
2I3,7,8lX,Y,Z-HpCDF isomer.
The concentration of the octa-CDD and octa-CDF shall be calculated
using the mean RRF determined for each.
Mean relative response factors for selected PCDD's and PCDF's are
given in Table 4.
11.1.4 Calculate the percent recovery, Rjs, for each internal
standard in the sample extract, using the equation:
Ais x Q
R
1s Ax RFr xQ.s
where:
Ars = Area of quantisation ion (m/z 334) of the recovery standard,
13Ci2-l,2,3,4-TCDD.
Qrs = "9 °f recovery standard, 13Ci2-l,2,3,4-TCDD, added to
extract.
The response factor for determination of recovery is calculated using
data obtained from the analysis of the multi-level calibration standards
according to the equation:
RF _ Ais x Crs
RFr~ ArsxCis
8280 - 19
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where:
Crs = Concentration of the recovery standard, 13Ci2-l,2,3,4-TCDD.
11.1.5 Calculation of total concentration of all Isomers within
each homologous series of PCDD's and PCDF's.
Total concentration = Sum of the concentrations of the Individual
of PCDD's or PCDF's PCDD or PCDF Isomers
11.4 Report results In nanograms per gram; when duplicate and spiked
samples are reanalyzed, all data obtained should be reported.
11.5 Accuracy and Precision. Table 5 gives the precision data for
revised Method 8280 for selected analytes in the matrices shown. Table 6
lists recovery data for the same analyses. Table 2 shows the linear range and
variation of response factors for selected analyte standards. Table 8
provides the method detection limits as measured in specific sample matrices.
11.6 Method Detection Limit. The Method Detection Limit (MDL) is
defined as the minimum concentration of a substance that can be measured and
reported with 99 percent confidence that the value 1s above zero. The
procedure used to determine the MDL values reported in Table 8 was obtained
from Appendix A of EPA Test Methods manual, EPA-600/4-82-057 July 1982,
"Methods for Organic Chemical Analysis of Municipal and Industrial
Wastewater."
11.7 Maximum Holding Time (MHT). Is that time at which a 10 percent
change in the analyte concentration (Ctio) occurs and the precision of the
method of measurement allows the 10 percent change to be statistically
different from the 0 percent change (Cto) at the 90 percent confidence level.
When the precision of the method is not sufficient to statistically
discriminate a 10 percent change in the concentration from 0 percent change,
then the maximum holding time is that time where the percent change in the
analyte concentration (Ctn) is statistically different than the concentration
at 0 percent change (Cto) an(* greater than 10 percent change at the 90 percent
confidence level.
8280 - 20
Revision 0
Date September 1986
-------
TABLE 1. REPRESENTATIVE GAS CHROMATOGRAPH RETENTION TIMES* OF ANALYTES
Analyte
2,3,7,8-TCDF
2,3,7,8-TCDD
1,2,3, 4-TCDD
1,2,3,4,7-PeCDD
1,2,3,4,7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
OCDD
50-m
CP-S11-88
25.2
23.6
24.1
30.0
39.5
57.0
NM
30-m
DB-5
17.8
17.4
17.3
20.1
22.1
24.1
25.6
3— m
SP-2250
26.7
26.7
26.5
28.1
30.6
33.7
NM
*Retention time in min, using temperature programs shown below.
NM = not measured.
Temperature Programs;
CP-Sil-88 60*C-190*C at 20'/nrin; 190*-240* at 5*/min.
DB-5 170*, 10 min; then at 8*/min to 320'C, hold
30 m x 0.25 mm at 320*C 20 min (until OCDD elutes).
Thin film (0.25 urn)
SP-2250 70'-320' at lO'/minute.
Column Manufacturers
CP-Sil-88 Chrompack, Incorporated, Bridgewater, New Jersey
DB-5, J and W Scientific, Incorporated, Rancho Cordova,
California
SP-2250 Supelco, Incorporated, Bellefonte, Pennsylvania
8280 - 21
Revision
Date September 1986
-------
TABLE 2. IONS SPECIFIED3 FOR SELECTED ION MONITORING
FOR PCDD'S AND PCDF'S
Quantitation
ion
Confirmation
ions
M-COC1
PCDD's
13Ci2-Tetra
Tetra
Penta
Hexa
Hepta
Octa
13Ci2-Octa
PCDF's
334
322
356
390
424
460
472
332
320
354;358
388;392
422;426
458
470
257
293
327
361
395
Tetra
Penta
Hexa
Hepta
Octa
306
340
374
408
444
304
338; 342
372; 376
406;410
442
243
277
311
345
379
alons at m/z 376 (HxCDE), 410 (HpCDE), 446 (OCDE), 480 (NCDE) and 514 (DCDE)
are also included in the scan monitoring sections (1) to (5), respectively.
See Paragraph 10.3.
TABLE 3. CRITERIA FOR ISOTOPIC RATIO MEASUREMENTS FOR PCDD'S AND PCDF'S
Selected ions (m/z) Relative intensity
PCDD's
Tetra
Penta
Hexa
Hepta
Octa
PCDF's
320/322
358/356
392/390
426/424
458/460
0.65-0.89
0.55-0.75
0.69-0.93
0.83-1.12
0.75-1.01
Tetra
Penta
Hexa
Hepta
Octa
304/306
342/340
376/374
410/408
442/444
0.65-0.89
0.55-0.75
0.69-0.93
0.83-1.12
0.75-1.01
8280 - 22
Revision 0
Date September 1986
-------
TABLE 4. MEAN RELATIVE RESPONSE FACTORS OF CALIBRATION STANDARDS
Analyte
2,3,7,8-TCDD
1,2,3,7,8-PeCDD
1,2,3,4,7,8-HxCDD
l,2,3,4,6,7,8-HpCDDb
OCDDb
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
l,2,3,4,6,7,8-HpCDFb
OCDFb
13Ci2-2,3,7,8-TCDD
13Ci2-l,2,3,4-TCDD
13Ci2-OCDD
RRFa
1.13
0.70
0.51
1.08
1.30
1.70
1.25
0.84
1.19
1.57
1.00
0.75
1.00
RSD%
(n = 5)
3.9
10.1
6.6
6.6
7.2
8.0
8.7
9.4
3.8
8.6
-
4.6
-
Quantitation ton
(m/z)
322
356
390
424
460
306
340
374
444
408
334
334
472
aThe RRF value is the mean of the five determinations made. Nominal weights
injected were 0.2, 0.5, 1.0, 2.0 and 5.0 ng.
bRRF values for these analytes were determined relative to 13Cj2-OCDD. All
other RRF's were determined relative to 13Ci2-2,3,7,8-TCDD.
Instrument Conditions/Tune - GC/MS system was tuned as specified in
Paragraph 6.3. RRF data was acquired under
SIM control, as specified in Paragraph 10.3.
GC Program - The GC column temperature was programmed as specified in
Paragraph 4.3.2(b).
8280 - 23
Revision
Date September 1986
-------
TABLE 5. PRECISION DATA FOR REVISED METHOD 8280
Compound
2,3,7,8-TCDD
1,2,3,4-TCDD
1,3,6,8-TCDD
1,3,7,9-TCDD
1,3,7,8-TCDD
1,2,7,8-TCDD
1,2,8,9-TCDD
Analyte
Matrix3
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
level (ng/g)
Native
NDb
378
ND
ND
487
ND
ND
ND
38.5
ND
ND
ND
ND
19.1
227
ND
ND
ND
58.4
ND
ND
ND
ND
16.0
422
ND
ND
ND
2.6
ND
ND
ND
ND
ND
ND
Native
+ spike
5.0
378
125
46
487
5.0
25.0
125
38.5
2500
2.5
25.0
125
19.1
2727
2.5
25.0
125.0
58.4
2500
5.0
25.0
125
16.0
2920
5.0
25.0
125
2.6
2500
5.0
25.0
125
46
2500
N
4
4
4
2
4
3
4
4
4
4
4
4
4
2
2
4
4
4
2
2
4
4
4
4
2
4
4
4
3
2
4
4
4
2
2
Percent
RSD
4.4
2.8
4.8
-
24
1.7
1.1
9.0
7.9
-
7.0
5.1
3.1
-
-
19
2.3
6.5
-
-
7.3
1.3
5.8
3.5
-
7.7
9.0
7.7
23
-
10
0.6
1.9
-
-
8280 - 24
Revision 0
Date September 1986
-------
TABLE 5 (Continued)
Compound
1,2,3,4,7-PeCDD
1,2,3,7,8-PeCDD
1,2,3,4,7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
1,2,7,8-TCDF
1,2,3,7, 8-PeCDF
1,2,3,4,7,8-HxCDF
Analyte
Matrix3
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge0
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom3
clay
soil
sludge
fly ash
still bottom
level (ng/g)
Native
ND
ND
ND
25.8
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
8760
ND
ND
ND
ND
ND
7.4
ND
ND
ND
ND
ND
25600
ND
ND
13.6
24.2
ND
Native
+ spike
5.0
25.0
125
25.8
2500
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
8780
-
-
5.0
25.0
125
7.4
2500
5.0
25.0
125
46
28100
5.0
25.0
139
24.2
2500
N
4
4
4
2
2
4
4
4
2
2
4
4
4
2
2
4
4
4
_
-
4
4
4
3
2
4
4
4
2
2
4
4
4
4
2
Percent
RSD
10
2.8
4.6
6.9
-
25
20
4.7
_
-
38
8.8
3.4
-
-
^
_
-
_
-
3.9
1.0
7.2
7.6
-
6.1
5.0
4.8
—
-
26
6.8
5.6
13.5
-
8280 - 25
Revision 0
Date September 1986
-------
TABLE 5. (Continued)
Compound
OCDF
Analyte
Matr1xa
clay
soil
sludge
fly ash
still bottom
level (ng/g)
Native Percent
Native + spike N RSD
ND -
ND -
192 317 4 3.3
ND -
ND -
amatrix types:
clay: pottery clay.
soil: Times Beach, Missouri, soil blended to forma homogeneous sample.
This sample was analyzed as a performance evaluation sample for the Contract
Laboratory Program (CLP) in April 1983. The results from EMSL-LV and 8
contract laboratories using the CLP protocol were 305.8 ng/g 2,3,7,8-TCDD
with a standard deviation of 81.0.
fly ash: ash from a municipal incinerator; resource recovery ash No. 1.
still bottom: distillation bottoms (tar) from 2,4-dichlorophenol production.
sludge: sludge from cooling tower which received both creosote and
pentachl orophenol 1 c wastewaters .
Cleanup of clay, soil and fly ash samples was through alumina column only.
(Carbon column not used.)
- not detected at concentration Injected (final volume 0.1 mL or greater).
GEstimated concentration out of calibration range of standards.
8280 - 26
Revision 0
Date September 1986
-------
TABLE 6. RECOVERY DATA FOR REVISED METHOD 8280
Compound
2,3,7,8-TCDD
1,2,3,4-TCDD
1,3,6,8-TCDD
1,3,7,9-TCDD
1,3,7,8-TCDD
1,2,7,8-TCDD
1,2,8,9-TCDD
Matr1xa
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
Nat1veb
(ng/g)
ND
378
ND
ND
487
ND
ND
ND
38.5
ND
ND
ND
ND
19.1
227
ND
ND
ND
58.4
ND
ND
ND
ND
16.0
615
ND
ND
ND
2.6
ND
ND
ND
ND
ND
ND
Sp1kedc
level
(ng/g)
5.0
-
125
46
-
5.0
25.0
125
46
2500
2.5
25.0
125
46
2500
2.5
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
Mean
percent
recovery
61.7
-
90.0
90.0
-
67.0
60.3
73.1
105.6
93.8
39.4
64.0
64.5
127.5
80.2
68.5
61.3
78.4
85.0
91.7
68.0
79.3
78.9
80.2
90.5
68.0
75.3
80.4
90.4
88.4
59.7
60.3
72.8
114.3
81.2
8280 - 27
Revision 0
Date September 1986
-------
TABLE 6. (Continued)
Compound
1,2,3,4,7-PeCDD
1,2,3,7,8-PeCDD
1,2,3,4, 7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
2,3,7,8-TCDD
(C-13)
1,2,7, 8-TCDF
1,2,3,7,8-PeCDF
Matrix3
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soi 1
sludge"
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
Nativeb
(ng/g)
ND
ND
ND
25.8
NO
NO
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
8780
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
7.4
ND
ND
ND
ND
ND
25600
Spikedc
level
(ng/g)
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
125
-
-
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
5.0
25.0
125
46
2500
Mean
percent
recovery
58.4
62.2
79.2
102.4
81.8
61.7
68.4
81.5
104.9
84.0
46.8
65.0
81.9
125.4
89.1
ND
ND
-
-
64.9
78.8
78.6
88.6
69.7
65.4
71.1
80.4
90.4
104.5
57.4
64.4
84.8
105.8
-
8280 - 28
Revision 0
Date September 1986
-------
TABLE 6. (Continued)
Compound
1,2,3,4,7,8-HxCDF
OCDF
Matrixa
clay
soil
si udge
fly ash
still bottom
clay
soil
sludge
fly ash
still bottom
Nativeb
(ng/g)
ND
ND
13.6
24.2
ND
ND
ND
192
ND
ND
Spiked0
level
(ng/g)
5.0
25.0
125
46
2500
_
-
125
-
—
Mean
percent
recovery
54.2
68.5
82.2
91.0
92.9
_
-
86.8
-
-
amatrix types:
clay: pottery clay.
soil: Times Beach, Missouri soil blended to form a homogeneous sample. This
sample was analyzed as a performance evaluation sample for the Contract
Laboratory Program (CLP) in April 1983. The results from EMSL-LV and 8
contract laboratories using the CLP protocol were 305.8 ng/g 2,3,7,8-TCDD
with a standard deviation of 81.0.
fly ash: ash from a municipal incinerator: resource recovery ash No. 1.
still bottom: distillation bottoms (tar) from 2,4-dichlorophenol production.
sludge: sludge from cooling tower which received both creosote and
pentachlorophenol wastewaters.
The clay, soil and fly ash samples were subjected to alumina column cleanup,
no carbon column was used.
volume of concentrate 0.1 mL or greater, ND means below quantification
limit, 2 or more samples analyzed.
cAmount of analyte added to sample, 2 or more samples analyzed.
^Esti mated concentration out of calibration range of standards.
8280 - 29
Revision 0
Date September 1986
-------
TABLE 7. LINEAR RANGE AND VARIATIOIN OF RESPONSE FACTORS
Analyte Linear range tested (pg) n&
l,2,7,8-TCDFa
2,3,7,8-TCDDa
2,3,7,8-TCDF
50-6000
50-7000
300-4000
8
7
5
Mean RF
1.634
0.721
2.208
%RSD
12.0
11.9
7.9
Response factors for these analytes were calculated using 2,3,7,8-TCDF as the
internal standard. The response factors for 2,3,7,8-TCDF were calculated vs.
13C12-1,2,3,4-TCDD,
^Each value of n represents a different concentration level.
8280 - 30
Revision
Date September 1986
-------
TABLE 8. METHOD DETECTION LIMITS OF C12 - LABELED PCDD'S and PCDF'S
IN REAGENT WATER (PPT) AND ENVIRONMENTAL SAMPLES (PPB)
13C -Labeled
Analyte
2,3,7,8-TCDD
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
OCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
1,2,3,4,7,8-HxCDF
Reagent
Water3
0.44
1.27
2.21
2.77
3.93
0.63
1.64
2.53
Missouri
Soil5
0.17
0.70
1.25
1.87
2.35
0.11
0.33
0.83
Flyg
Ash
0.07
0.25
0.55
1.41
2.11
0.06
0.16
0.30
Industrial
Sludge0
0.82
1.34
2.30
4.65
6.44
0.46
0.92
2.17
Still-
Bottom
1.81
2.46
6.21
4.59
10.1
0.26
1.61
2.27
Fuel
Oil4
0.75
2.09
5.02
8.14
23.2
0.48
0.80
2.09
Fuel C
Sawdus
0.13
0.18
0.36
0.51
1.48
0.40
0.43
2.22
a
.Sample size 1 ,000 mL.
Sample size 10 g.
.Sample size 2 g.
Sample size 1 g.
Note: The final sample-extract volume was 100 uL for all samples.
Matrix types used in MDL Study:
- Reagent water: distilled, deionized laboratory water.
- Missouri soil: soil blended to form a homogeneous sample.
- Fly-ash: alkaline ash recovered from the electrostatic precipitator of
a coal-burning power plant.
- Industrial sludge: sludge from cooling tower which received creosotic
and pentachlorophenolic wastewaters. Sample was ca. 70 percent water,
mixed with oil and sludge.
- Still-bottom: distillation bottoms (tar) from 2,4-dichlorophenol
production.
- Fuel oil: wood-preservative solution from the modified Thermal Process
tanks. Sample was an oily liquid (>90 percent oil) containing no
water.
- Fuel oil/Sawdust: sawdust was obtained as a very fine powder from the
local lumber yard. Fuel oil (described above) was mixed at the 4
percent (w/w) level.
Procedure used for the Determination of Method Detection Limits was obtained
from "Methods for Organic Chemical Analysis of Municipal and Industrial
Wastewater" Appendix A, EPA-600/4-82-057, July 1982. Using this procedure,
the method detection limit is defined as the minimum concentration of a
substance that can be measured and reported with 99 percent confidence that
the value is above zero.
8280 - 31
Revision 0
Date September 1986
-------
aaoi-8're'z
dd01-87.'C'3
0>
en
I
10
§
O
U
o
I
s
s
2
o
at
CM
3
CD
U.
I
O
8280 - 32
Revision o
Date September 1986
-------
METHOD B3BO
POLYCHLOHINATEO OIBENZO-P-DIOXIINS AND POLYCHLORINATEO OI8ENZONFUHANS
f Start J
e. i
Perform Initial
calibration on
GC/MS system
6.9
1O.2
Calculate
response
factors for
standards
Oo routine
calibration
10.3
Analyze
samples with
selected Ion
monitoring
9.2
Extract
•ample using
appropriate
method for the
waste matrix
9.9
Prepare
carbon column;
do carbon
column cleanup
10.5
Quantltate PCOD
and PCDF peaks
Yes
o
11.01
Determine
concentrations
•nd report
results
f Stop J
8280 - 33
Revision Q
Date September 1986
-------
APPENDIX A
SIGNAL-TO-NOISE DETERMINATION METHODS
MANUAL DETERMINATION
This method corresponds to a manual determination of the S/N from a GC/MS
signal, based on the measurement of its peak height relative to the baseline
noise. The procedure is composed of four steps as outlined below. (Refer to
Figure 1 for the following discussion).
1. Estimate the peak-to-peak noise (N) by tracing the two lines (Ej and
£2) defining the noise envelope. The lines should pass through the
estimated statistical mean of the positive and the negative peak
excursions as shown in Figure 1. In addition, the signal offset (0)
should be set high enough such that negative-going noise (except for
spurious negative spikes) is recorded.
2. Draw the line (C) corresponding to the mean noise between the
segments defining the noise envelope.
3. Measure the height of the GC/MS signal (S) at the apex of the peak
relative to the mean noise C. For noisy GC/MS signals, the average
peak height should be measured from the estimated mean apex signal D
between £3 and £4.
4. Compute the S/N.
This method of S/N measurement is a conventional, accepted method of
noise measurement in analytical chemistry.
INTERACTIVE COMPUTER GRAPHICAL METHOD
This method calls for the measurement of the GC/MS peak area using the
computer data system and Eq. 1:
A/t
S/N = Aj/2t + Af/2t
where t is the elution time window (time interval, t2~t2, at the base of the
peak used to measure the peak area A). (Refer to Figure 2, for the following
discussion).
AI and Ar correspond to the areas of the noise level in a region to the
left (AI) and to the right (Ar) of the GC peak of interest.
8280 - A - 1
Revision
Date September 1986
-------
The procedure to determine the S/N 1s as follows:
1. Estimate the average negative peak excursions of the noise (I.e.,
the low segment-E2-of the noise envelope). Line £2 should pass
through the estimated statistical mean of the negative-going noise
excursions. As stated earlier, 1t 1s Important to have the signal
offset (0) set high enough such that negative-going noise 1s
j recorded„
2. Using the cross-hairs of the video display terminal, measure the
peak area (A) above a baseline corresponding to the mean negative
noise value (£2) and between the time tj and t2 where the GC/MS peak
intersects the baseline, £2. Make note of the time width t=t2-ti.
3. Following a similar procedure as described above, measure the area
of the noise 1n a region to the left (Aj) and to the right (Ar) of
the GC/MS signal using a time window twice the size of t, that Is,
2 x t.
The analyst must sound judgement in regard to the proper selection of
Interference-free regions 1n the measurement of AI and Ar. It is not
recommended to perform these noise measurements (Aj and Ar) in remote regions
exceeding ten time widths (lot).
4. Compute the S/N using Eq. 1.
NOTE: If the noise does not occupy at least 10 percent of the vertical
axis (i.e., the noise envelope cannot be defined accurately), then
1t is necessary to amplify the vertical axis so that the noise
occupies 20 percent of the terminal display (see Figure 3).
8280 - A - 2
Revision
Date September 1986
-------
FIGURE CAPTIONS
Figure 1. Manual determination of S/N.
The peak height (S) is measured between the mean noise (lines C and
D). These mean signal values are obtained by tracing the line
between the baseline average noise extremes, E\ and £21 and between
the apex average noise extremes, £3 and £4, at the apex of the
signal. Note, it is imperative that the instrument's interface
amplifier electronic's zero offset be set high enough such that
negative-going baseline noise is recorded.
Figure 2. Interactive determination of S/N.
The peak area (A) is measured above the baseline average negative
noise £2 and between times tj and t2- The noise is obtained from
the areas AI and Ar measured to the left and to the right of the
peak of interest using time windows Tj and Tr (Ti=Tr=2t).
Figure 3. Interactive determination of S/N.
A) Area measurements without amplification of the vertical axis.
Note that the noise cannot be determined accurately by visual
means. B) Area measurements after amplification (10X) of the
vertical axis so that the noise level occupies approximately 20
percent of the display, thus enabling a better visual estimation of
the baseline noise, Ej, £2, and C.
8280 - A - 3
Revision 0
Date September 1986
-------
O
(O
c.
0>
•M
O
c
(O
2!
3
D)
8280 - A - 4
Revision 0
Date September 1986
-------
t =
100 n
90
80
70-
60-
50-
40-
30*
20-
10-
0
= 558.10
= 88.55
i i r
25:30 26:00 26:30
27:00 27:30 28:00
17 sec.
Figure 2. Interactive Determination of S/N.
8280 - A - 5
Revision p
Date September 1986
-------
100-
90-
80-
70-
60-
50-
40-
30-
20-
10-
0-
A = 686.41
= 17.18
Ar= 13.32
25:30 26:00 36:30 27:00 27:30 28:00
A = 706.59
Ar = 41.88
26:30 26:00 26:30 27:00 27:30 28:00
Figure 3. Interactive Determination of S/N.
8280 - A - 6
Revision 0
Date September 1986
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APPENDIX B
RECOMMENDED SAFETY AND HANDLING PROCEDURES FOR PCDD'S/PCDF'S
1. The human toxicology of PCDD/PCDF is not well defined at present,
although the 2,3,7,8-TCDD isomer has been found to be acnegenic, carcinogenic,
and teratogenic in the course of laboratory animal studies. The 2,3,7,8-TCDD
is a solid at room temperature, and has a relatively low vapor pressure. The
solubility of this compound in water is only about 200 parts-per-trillion, but
the solubility in various organic solvents ranges from about 0.001 perent to
0.14 percent. The physical properties of the 135 other tetra- through octa-
chlorinated PCDD/PCDF have not been well established, although it is presumed
that the physical properties of these congeners are generally similar to those
of the 2,3,7,8-TCDD isomer. On the basis of the available toxicological and
physical property data for TCDD, this compound, as well as the other PCDD and
PCDF, should be handled only by highly trained personnel who are thoroughly
versed in the appropriate procedures, and who understand the associated risks.
2. PCDD/PCDF and samples containing these are handled using essentially
the same techniques as those employed 1n handling radioactive or infectious
materials. Well-ventilated, controlled-access laboratories are required, and
laboratory personel entering these laboratories should wear appropriate safety
clothing, including disposable coveralls, shoe covers, gloves, and face and
head masks. During analytical operations which may give rise to aerosols or
dusts, personnel should wear respirators equipped with activated carbon
filters. Eye protection equipment (preferably full face shields) must be worn
at all times while working in the analytical laboratory with PCDD/PCDF.
Various types of gloves can be used by personnel, depending upon the
analytical operation being accomplished. Latex gloves are generally utilized,
and when handling samples thought to be particularly hazardous, an additional
set of gloves are also worn beneath the latex gloves (for example, Playtex
gloves supplied by American Scientific Products, Cat. No. 67216). Bench-tops
and other work surfaces in the laboratory should be covered with plastic-
backed absorbent paper during all analytical processing. When finely divided
samples (dusts, soils, dry chemicals) are processed, removal of these from
sample contaners, as well as other operations, including weighing,
transferring, and mixing with solvents, should all be accomplished within a
glove box. Glove boxes, hoods and the effluents from mechanical vacuum pumps
and gas chromatographs on the mass spectrometers should be vented to the
atmosphere preferably only after passing through HEPA particulate filters and
vapor-sorbing charcoal.
3. All laboratory ware, safety clothing, and other items potentially
contaminated with PCDD/PCDF in the course of analyses must be carefully
secured and subjected to proper disposal. When feasible, liquid wastes are
concentrated, and the residues are placed in approved steel hazardous waste
drums fitted with heavy gauge polyethylene liners. Glass and combustible
items are compacted using a dedicated trash compactor used only for hazardous
waste materials and then placed 1n the same type of disposal drum. Disposal
of accumulated wastes 1s periodically accomplished by high temperature
incineration at EPA-aproved facilities.
8280 - B - 1
Revision 0
Date September 1986
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4. Surfaces of laboratory benches, apparatus and other appropriate areas
should be periodically subjected to surface wipe tests using solvent-wetted
filter paper which Is then analyzed to check for PCDD/PCDF contamination 1n
the laboratory. Typically, if the detectable level of TCDD or TCDF from such
a test is greater than 50 ng/m2, this indicates the need for decontamination
of the laboratory. A typical action limit 1n terms of surface contamination
of the other PCDD/PCDF (summed) is 500 ng/m2. In the event of a spill within
the laboratory, absorbent paper is used to wipe up the spilled material and
this is then placed into a hazardous waste drum. The contaminated surface is
subsequently cleaned thoroughly by washing with appropriate solvents
(methylene chloride followed by methanol) and laboratory detergents. This is
repeated until wipe tests Indicate that the levels of surface contamination
are below the limits cited.
5. In the unlikely event that analytical personnel experience skin
contact with PCDD/PCDF or samples containing these, the contaminated skin
area should Immediately be thoroughly scurbbed using mild soap and water.
Personnel involved in any such accident should subsequently be taken to the
nearest medical facility, preferably a facility whose staff is knowledgeable
in the toxicology of chlorinated hydrocarbons. Again, disposal of
contaminated clothing is accomplished by placing it in hazardous waste drums.
6. It is desirable that personnel working in laboratories where
PCDD/PCDF are handled be given periodic physical examinations (at least
yearly). Such examinations should Include specialized tests, such as those
for urinary porphyrlns and for certain blood parameters which, based upon
published clinical observations, are appropriate for persons who may be
exposed to PCDD/PCDF. Periodic facial photographs to document the onset of
dermatologic problems are also advisable.
8280 - B - 2
Revision 0
Date September 1986
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Page 1 of 2
DIOXIN SAMPLE DATA SUMMARY FORM 8280-1
LAB NAME CONTRACT No.
CASE No.
QUANTITY FOUND (ng/g)
SAMPLE NO. FILE NAME TCDD PeCDD HxCDD HpCDD OCDD
DATA RELEASE AUTHORIZED BY
8280 - B - 3
Revision 0
Date September 1986
-------
Page 2 of 2
DIOXIN SAMPLE DATA SUMMARY FORM 8280-1
LAB NAME CONTRACT No.
CASE No.
QUANTITY FOUND (ng/g)
SAMPLE NO. FILE NAME TCDF PeCDF HxCDF HpCDF OCDF
8280 - B - 4
Revision
Date September 1986
-------
Page 1 of 2
DIOXIN SAMPLE DATA SUMMARY FORM 8280-1-W
LAB NAME CONTRACT No.
CASE No.
QUANTITY FOUND (ug/L)
SAMPLE NO. FILE NAME TCDD PeCDD HxCDD HpCDD OCDD
DATA RELEASE AUTHORIZED BY
8280 - B - 5
Revision
Date September 1986
-------
Page 2 of 2
DIOXIN SAMPLE DATA SUMMARY FORM 8280-1-W
LAB NAME CONTRACT No.
CASE No.
QUANTITY FOUND (ug/L)
SAMPLE NO. FILE NAME TCDF PeCDF HxCDF HpCDF OCDF
8280 - B - 6
Revision
Date September 1986
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DIOXIN RAW SAMPLE DATA FORM 8280-2
LAB NAME ANALYST(s) CASE No.
SAMPLE No. TYPE OF SAMPLE CONTRACT No.
SAMPLE SIZE % MOISTURE FINAL EXTRACT VOLUME
EXTRACTION METHOD ALIQUOT USED FOR ANALYSIS
CLEAN UP OPTION
CONCENTRATION FACTOR DILUTION FACTOR
DATE EXTRACTED DATA ANALYZED
VOLUME 13Ci2-l,2,3,4-TCDD ADDED TO SAMPLE VOLUME
VOLUME INJECTED Wt 13Ci2-l,2,3,4-TCDD ADDED
Wt 13Ci2-2,3,7,8-TCDD ADDED 13Ci2-2,3,7,8-TCDD % RECOVERY
Wt 13Ci2-2,3,7,8-OCDD ADDED 13C12-OCDD % RECOVERY
13Ci2-2,3,7,8-TCDD RRF 13Ci2-OCDD RRF
13Ci2-2,3,7,8-TCDD
AREA 332 AREA 334 RATIO 332/334 _
13Ci2-OCDD AREA 470 AREA 472 RATIO 470/472
RT 2,3,7,8-TCDD (Standard) RT 2,3,7,8-TCDD (Sample)
13C12-2,3,7,8-TCDD - 13Ci2-l,2,3,4-TCDD Percent Valley
8280 - B - 7
Revision 0
Date September 1986
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DIOXIN INITIAL CALIBRATION STANDARD DATA SUMMARY
FORM 8280-3
CASE No.
Lab Name
Date of Initial Calibration
Relative to
Contract No.
Analyst(s)
CALIBRATION
STANDARD
RRF
1
RRF RRF RRF
2 34
or 13Ci2-l,2r3,4-TCDD_
RRF
5
MEAN %RSD
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
8280 - B - 8
Revision 0
Date September 1986
-------
FORM 8280-3 (Continued)
CONCENTRATIONS IN PG/UL
1 2345
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
8280 - B - 9
Revision
Date September 1986
-------
DIOXIN CONTINUING CALIBRATION SUMMARY
FORM 8280-4
CASE No.
Lab Name
Date of Initial Calibration
Relative to 13Ci2-2,3,7,8-TCDD_
Contract No.
Analyst(s)
or 13C12-1,2,3,4-TCDD
COMPOUND
RRF
RRF
%D
TCDD
PeCDD
HxCDD
HpCDD
OCDD
TCDF
PeCDF
HxCDF
HpCDF
OCDF
8280 - B - 10
Revision 0
Date September 1986
-------
DIOXIN RAW SAMPLE DATA FORM 8280-5-A
LAB NAME ANALYST(s) CASE No.
CONTRACT No. SAMPLE No.
TCDD REQUIRED 320/322 RATIO WINDOW IS 0.65 - 0.89
QUANTITATED FROM 2,3,7,8-TCDD 1,2,3,4-TCDD RRF
SCAN I RRT AREA AREA AREA 320/ CONFIRM
322 320 257 322 AS TCDD
Y/N CONC,
TOTAL TCDD
TCDF REQUIRED 304/306 RATIO WINDOW IS 0.65 - 0.89
QUANTITATED FROM 2,3,7,8-TCDD 1,2,3,4-TCDD RRF
SCAN # RRT AREA AREA AREA 304/ CONFIRM
306 304 243 306 AS TCDD
_^_^_ Y/N CONC,
TOTAL TCDD
8280 - B - 11
Revision
Date September 1986
-------
DIOXIN RAW SAMPLE DATA FORM 8280-5-B
LAB NAME
ANALYST(s)
CASE No.
CONTRACT No.
SAMPLE No.
PeCDD REQUIRED 320/322 RATIO WINDOW IS 0.55 - 0.75
QUANTITATED FROM 2,3,7,8-TCDD
SCAN # RRT AREA AREA
356 358
1,2,3,4-TCDD
AREA
354
AREA
293
3587
356
RRF
CONFIRM
AS PeCDD
Y/N
CONC.
TOTAL PeCDD
PeCDF REQUIRED 342/340 RATIO WINDOW IS 0.55 - 0.75
QUANTITATED FROM 2,3,7,8-TCDD 1,2,3,4-TCDD
SCAN
RRT
AREA
340
AREA
342
AREA
338
AREA
277
342/
340
RRF
CONFIRM
AS PeCDF
Y/N
CONC.
TOTAL PeCDF
8280 - B - 12
Revision 0
Date September 1986
-------
DIOXIN RAW SAMPLE DATA FORM 8280-5-C
LAB NAME
ANALYST(s)
CASE No.
CONTRACT No.
SAMPLE No.
HxCDD REQUIRED 392/390 RATIO WINDOW IS 0.69 - 0.93
QUANTITATED FROM 2,3,7,8-TCDD
SCAN # RRT AREA AREA
390 392
1,2,3,4-TCDD
AREA
388
AREA
327
392/
390
RRF
CONFIRM
AS HxCDD
Y/N
CONC.
TOTAL HxCDD
HxCDF REQUIRED 376/374
QUANTITATED FROM 2,3,7
SCAN # RRT AREA
376
RATIO WINDOW IS
,8-TCDD
AREA
374
AREA
372
0.69 - 0.93
1,2,3, 4-TCDD
AREA
311
376/
374
RRF
CONFIRM
AS HxCDF
Y/N CONC.
TOTAL HxCDF
8280 - B - 13
Revision 0
Date September 1986
-------
DIOXIN RAW SAMPLE DATA FORM 8280-5-D
LAB NAME
ANALYST(s)
CASE No.
CONTRACT No.
SAMPLE No.
HpCDD REQUIRED 426/444 RATIO WINDOW IS 0.83 - 1.12
QUANTITATED FROM 2,3,7,8-TCDD
SCAN I RRT AREA AREA
424 426
1,2,3,4-TCDD
AREA
422
AREA
361
4267
424
RRF
CONFIRM
AS HpCDD
Y/N
CONC.
TOTAL HpCDD
HpCDF REQUIRED 410/408
QUANTITATED FROM 2,3,7
SCAN # RRT AREA
408
RATIO WINDOW IS
,8-TCDD
AREA AREA
410 406
0.83 - 1.12
1,2,3,4-TCDD
AREA
345
410/
408
RRF
CONFIRM
AS HpCDF
Y/N CONC.
TOTAL HpCDF
8280 - B - 14
Revision 0
Date September 1986
-------
DIOXIN RAW SAMPLE DATA FORM 8280-5-E
LAB NAME ANALYST(s) CASE No..
CONTRACT No. SAMPLE No.
OCDD REQUIRED 458/460 RATIO WINDOW IS 0.75 - 1.01
QUANTITATED FROM 2,3,7,8-TCDD 1,2,3,4-TCDD RRF
SCAN I RRT AREA AREA AREA 458/ CONFIRM
460 458 395 460 AS OCDD
Y/N CONC.
TOTAL OCDD
OCDF REQUIRED 442/444 RATIO WINDOW IS 0.75 - 1.01
QUANTITATED FROM 2,3,7,8-TCDD 1,2,3,4-TCDD RRF
SCAN # RRT AREA AREA AREA 442/ CONFIRM
444 442 379 444 AS OCDF
Y/N CONC.
TOTAL OCDF
8280 - B - 15
Revision
Date September 1986
-------
DIOXIN SYSTEM PERFORMANCE CHECK ANALYSIS FORM 8280-6
LAB NAME
CASE No.
BEGINNING DATE
ENDING DATE
TIME
TIME
CONTRACT No..
ANALYST(s)
PC SOLUTION IDENTIFIER
PCDD's
ISOTOPIC RATIO CRITERIA MEASUREMENT
IONS
RATIOED
RATIO AT
BEGINNING OF
12 HOUR PERIOD
RATIO AT
END OF 12 ACCEPTABLE
HOUR PERIOD WINDOW
Tetra
320/322
0.65-0.89
Penta
358/356
0.55-0.75
Hexa
392/390
0.69-0.93
Hepta
426/424
0.83-1.12
Octa
458/460
0.75-1.01
PCDF's
Tetra
304/306
0.65-0.89
Penta
342-340
0.55-0.75
Hexa
376-374
0.69-0.93
Hepta
410/408
0.83-1.12
Octa
442/444
0.75-1.01
Ratios out of criteria
PCDD
PCDF
Beginning
_ out of
out of
End
out of
out of
NOTE: One form is required for each 12 hour period samples are analyzed.
8280 - B - 16
Revision 0
Date September 1986
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METHOD 8290
POLYCHLORINATED DIBENZODIOXINS (PCDDs) AND POLYCHLORINATED DIBENZOFURANS
(PCDFs)BY HIGH-RESOLUTION GAS CHROMATOGRAPHY/HIGH-RESOLUTION
MASS SPECTROMETRY (HRGC/HRMS)
1.0 SCOPE AND APPLICATION
1.1 This method provides procedures for the detection and quantitative
measurement of polychlorinated dibenzo-p-dioxins (tetra- through octachlorinated
homologues; PCDDs), and polychlorinated dibenzofurans (tetra- through
octachlorinated homologues; PCDFs) in a variety of environmental matrices and at
part-per-trillion (ppt) to part-per-quadrillion (ppq) concentrations. The
following compounds can be determined by this method:
Compound Name
CAS No"
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)
1,2,3,7 , 8-Pentachl orodi benzo-p-di oxi n (PeCDD)
1,2,3,6,7,8-Hexachlorodibenzo-p-dioxin (HxCDD)
1,2, 3, 4, 7, 8-Hexachl orodi benzo-p-di oxi n (HxCDD)
1,2, 3, 7, 8, 9-Hexachl orodi benzo-p-di oxi n (HxCDD)
1,2, 3, 4, 6, 7, 8-Heptachl orodi benzo-p-di oxi n (HpCDD)
1,2,3,4,6,7,8,9-Octachlorodibenzo-p-dioxin (OCDD)
2,3,7,8-Tetrachlorodibenzofuran (TCDF)
1,2,3,7,8-Pentachlorodibenzofuran (PeCDF)
2,3,4,7,8-Pentachlorodibenzofuran (PeCDF)
1,2,3,6,7, 8-Hexachl orodi benzof uran (HxCDF )
1,2, 3, 7, 8, 9-Hexachl orodi benzofuran (HxCDF)
1,2,3,4,7 , 8-Hexachl orodi benzof uran (HxCDF)
2, 3, 4, 6, 7, 8-Hexachl orodi benzof uran (HxCDF)
1,2,3,4,6,7 , 8-Heptachl orod i benzofuran ( HpCDF)
1,2,3,4,7,8 , 9-Heptachl orod i benzofuran (HpCDF)
1 , 2 , 3 , 4 , 6 , 7 , 8 , 9 -Octachl orod i benzofuran (OCDF)
1746-01-6
40321-76-4
57653-85-7
39227-28-6
19408-74-3
35822-39-4
3268-87-9
51207-31-9
57117-41-6
57117-31-4
57117-44-9
72918-21-9
70648-26-9
60851-34-5
67562-39-4
55673-89-7
39001-02-0
a Chemical Abstract Service Registry Number
1.2 The analytical method calls for the use of high-resolution gas
chromatography and high-resolution mass spectrometry (HRGC/HRMS) on purified
sample extracts. Table 1 lists the various sample types covered by this
analytical protocol, the 2,3,7,8-TCDD-based method calibration limits (MCLs), and
other pertinent information. Samples containing concentrations of specific
congeneric analytes (PCDDs and PCDFs) considered within the scope of this method
that are greater than ten times the upper MCLs must be analyzed by a protocol
designed for such concentration levels, e.g., Method 8280. An optional method
for reporting the analytical results using a 2,3,7,8-TCDD toxicity equivalency
factor (TEF) is described.
8290 - 1
Revision 0
September 1994
-------
1.3 The sensitivity of this method is dependent upon the level of inter-
ferences within a given matrix. The calibration range of the method for a I L
water sample is 10 to 2000 ppq for TCDD/TCDF and PeCDD/PeCDF, and 1.0 to 200 ppt
for a 10 g soil, sediment, fly ash, or tissue sample for the same analytes
(Table 1). Analysis of a one-tenth aliquot of the sample permits measurement of
concentrations up to 10 times the upper MCL. The actual limits of detection and
quantitation will differ from the lower MCL, depending on the complexity of the
matrix.
1.4 This method is designed for use by analysts who are experienced with
residue analysis and skilled in HRGC/HRMS.
1.5 Because of the extreme toxicity of many of these compounds, the
analyst must take the necessary precautions to prevent exposure to materials
known or believed to contain PCDDs or PCDFs. It is the responsibility of the
laboratory personnel to ensure that safe handling procedures are employed. Sec.
11 of this method discusses safety procedures.
2.0 SUMMARY OF METHOD
e specific
2.1 This procedure uses matrix specific extraction, analyt
cleanup, and HRGC/HRMS analysis techniques.
2.2 If interferences are encountered, the method provides selected
cleanup procedures to aid the analyst in their elimination. A simplified
analysis flow chart is presented at the end of this method.
2.3 A specified amount (see Table 1) of soil, sediment, fly ash, water,
sludge (including paper pulp), still bottom, fuel oil, chemical reactor residue,
fish tissue, or human adipose tissue is spiked with a solution containing
specified amounts of each of the nine isotopically (13C12) labeled PCDDs/PCDFs
listed in Column 1 of Table 2. The sample is then extracted according to a
matrix specific extraction procedure. Aqueous samples that are judged to contain
1 percent or more solids, and solid samples that show an aqueous phase, are
filtered, the solid phase (including the filter) and the aqueous phase extracted
separately, and the extracts combined before extract cleanup. The extraction
procedures are:
a) Toluene: Soxhlet extraction for soil, sediment, fly ash, and paper
pulp samples;
b) Methylene chloride: liquid-liquid extraction for water samples;
c) Toluene: Dean-Stark extraction for fuel oil, and aqueous sludge
samples;
d) Toluene extraction for still bottom samples;
e) Hexane/methylene chloride: Soxhlet extraction or methylene
chloride: Soxhlet extraction for fish tissue samples; and
f) Methylene chloride extraction for human adipose tissue samples.
8290 - 2
Revision 0
September 1994
-------
g) As an option, all solid samples (wet or dry) may be extracted with
toluene using a Soxhlet/Dean Stark extraction system.
The decision for the selection of an extraction procedure for chemical
reactor residue samples is based on the appearance (consistency, viscosity) of
the samples. Generally, they can be handled according to the procedure used for
still bottom (or chemical sludge) samples.
2.4 The extracts are submitted to an acid-base washing treatment and
dried. Following a solvent exchange step, the extracts are cleaned up by column
chromatography on alumina, silica gel, and activated carbon.
2.4.1 The extracts from adipose tissue samples are treated with
silica gel impregnated with sulfuric acid before chromatography on acidic
silica gel, neutral alumina, and activated carbon.
2.4.2 Fish tissue and paper pulp extracts are subjected to an acid
wash treatment only, prior to chromatography on alumina and activated
carbon.
2.5 The preparation of the final extract for HRGC/HRMS analysis is
accomplished by adding 10 to 50 /xL (depending on the matrix) of a nonane
solution containing 50 pg/juL of the recovery standards 13C12-1,2,3,4-TCDD and
13C12-l,2,3,7,8,9-HxCDD (Table 2). The former is used to determine the percent
recoveries of tetra- and pentachlorinated PCDD/PCDF congeners, while the latter
is used to determine the percent recoveries of the hexa-, hepta- and
octachlorinated PCDD/PCDF congeners.
2.6 Two juL of the concentrated extract are injected into an HRGC/HRMS
system capable of performing selected ion monitoring at resolving powers of at
least 10,000 (10 percent valley definition).
2.7 The identification of OCDD and nine of the fifteen 2,3,7,8-
substituted congeners (Table 3), for which a 13C-labeled standard is available
in the sample fortification and recovery standard solutions (Table 2), is based
on their elution at their exact retention time (within 0.005 retention time units
measured in the routine calibration) and the simultaneous detection of the two
most abundant ions in the molecular ion region. The remaining six 2,3,7,8-
substituted congeners (i.e., 2,3,4,7,8-PeCDF; 1,2,3,4,7,8-HxCDD; 1,2,3,6,7,8-
HxCDF; 1,2,3,7,8,9-HxCDF; 2,3,4,6,7,8-HxCDF, and 1,2,3,4,7,8,9-HpCDF), for which
no carbon-labeled internal standards are available in the sample fortification
solution, and all other PCDD/PCDF congeners are identified when their relative
retention times fall within their respective PCDD/PCDF retention time windows,
as established from the routine calibration data, and the simultaneous detection
of the two most abundant ions in the molecular ion region. The identification
of OCDF is based on its retention time relative to 13C12-OCDD and the simultaneous
detection of the two most abundant ions in the molecular ion region.
Identification also is based on a comparison of the ratios of the integrated ion
abundance of the molecular ion species to their theoretical abundance ratios.
2.8 Quantitation of the individual congeners, total PCDDs and total PCDFs
is achieved in conjunction with the establishment of a multipoint (five points)
8290 - 3 Revision 0
September 1994
-------
calibration curve for each homologue, during which each calibration solution is
analyzed once.
3.0 INTERFERENCES
3.1 Solvents, reagents, glassware and other sample processing hardware
may yield discrete artifacts or elevated baselines that may cause misinter-
pretation of the chromatographic data (see references 1 and 2.) All of these
materials must be demonstrated to be free from interferants under the conditions
of analysis by performing laboratory method blanks. Analysts should avoid using
PVC gloves.
3.2 The use of high purity reagents and solvents helps minimize
interference problems. Purification of solvents by distillation in all-glass
systems may be necessary.
3.3 Interferants coextracted from the sample will vary considerably from
matrix to matrix. PCDDs and PCDFs are often associated with other interfering
chlorinated substances such as polychlorinated biphenyls (PCBs), polychlorinated
diphenyl ethers (PCDPEs), polychlorinated naphthalenes, and polychlorinated
alkyldibenzofurans, that may be found at concentrations several orders of
magnitude higher than the analytes of interest. Retention times of target
analytes must be verified using reference standards. These values must
correspond to the retention time windows established in Sec. 8.1.1.3. While
cleanup techniques are provided as part of this method, unique samples may
require additional cleanup steps to achieve lower detection limits.
3.4 A high-resolution capillary column (60 m DB-5, J&W Scientific, or
equivalent) is used in this method. However, no single column is known to
resolve all isomers. The 60 m DB-5 GC column is capable of 2,3,7,8-TCDD isomer
specificity (Sec. 8.1.1). In order to determine the concentration of the
2,3,7,8-TCDF (if detected on the DB-5 column), the sample extract must be
reanalyzed on a column capable of 2,3,7,8-TCDF isomer specificity (e.g., DB-225,
SP-2330, SP-2331, or equivalent).
4.0 APPARATUS AND MATERIALS
4.1 High-Resolution Gas Chromatograph/High-Resolution Mass
Spectrometer/Data System (HRGC/HRMS/DS) - The GC must be equipped for temperature
programming, and all required accessories must be available, such as syringes,
gases, and capillary columns.
4.1.1 GC Injection Port - The GC injection port must be designed for
capillary columns. The use of splitless injection techniques is
recommended. On column 1 juL injections can be used on the 60 m DB-5
column. The use of a moving needle injection port is also acceptable.
When using the method described in this protocol, a 2 juL injection volume
is used consistently (i.e., the injection volumes for all extracts,
blanks, calibration solutions and the performance check samples are 2 /LtL).
One ^L injections are allowed; however, laboratories must remain
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consistent throughout the analyses by using the same injection volume at
all times.
4.1.2 Gas Chromatograph/Mass Spectrometer (GC/MS) Interface - The
GC/MS interface components should withstand 350°C. The interface must be
designed so that the separation of 2,3,7,8-TCDD from the other TCDD
isomers achieved in the gas chromatographic column is not appreciably
degraded. Cold spots or active surfaces (adsorption sites) in the GC/MS
interface can cause peak tailing and peak broadening. It is recommended
that the GC column be fitted directly into the mass spectrometer ion
source without being exposed to the ionizing electron beam. Graphite
ferrules should be avoided in the injection port because they may adsorb
the PCDDs and PCDFs. Vespel™, or equivalent, ferrules are recommended.
4.1.3 Mass Spectrometer - The static resolving power of the
instrument must be maintained at a minimum of 10,000 (10 percent valley).
4.1.4 Data System - A dedicated data system is employed to control
the rapid multiple-ion monitoring process and to acquire the data.
Quantitation data (peak areas or peak heights) and SIM traces (displays of
intensities of each ion signal being monitored including the lock-mass ion
as a function of time) must be acquired during the analyses and stored.
Quantitations may be reported based upon computer generated peak areas or
upon measured peak heights (chart recording). The data system must be
capable of acquiring data at a minimum of 10 ions in a single scan. It is
also recommended to have a data system capable of switching to different
sets of ions (descriptors) at specified times during an HRGC/HRMS
acquisition. The data system should be able to provide hard copies of
individual ion chromatograms for selected gas chromatographic time
intervals. It should also be able to acquire mass spectral peak profiles
(Sec. 8.1.2.3) and provide hard copies of peak profiles to demonstrate the
required resolving power. The data system should permit the measurement
of noise on the base line.
NOTE: The detector ADC zero setting must allow peak-to-peak measure-
ment of the noise on the base line of every monitored channel
and allow for good estimation of the instrument resolving
power. In Figure 2, the effect of different zero settings on
the measured resolving power is shown.
4.2 GC Columns
4.2.1 In order to have an isomer specific determination for 2,3,7,8-
TCDD and to allow the detection of OCDD/OCDF within a reasonable time
interval in one HRGC/HRMS analysis, use of the 60 m DB-5 fused silica
capillary column is recommended. Minimum acceptance criteria must be
demonstrated and documented (Sec. 8.2.2). At the beginning of each 12
hour period (after mass resolution and GC resolution are demonstrated)
during which sample extracts or concentration calibration solutions will
be analyzed, column operating conditions must be attained for the required
separation on the column to be used for samples. Operating conditions
known to produce acceptable results with the recommended column are shown
in Sec. 7.6.
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4.2.2 Isomer specificity for all 2,3,7,8-substituted PCDDs/PCDFs
cannot be achieved on the 60 m DB-5 GC column alone. In order to
determine the proper concentrations of the individual 2,3,7,8-substituted
congeners, the sample extract must be reanalyzed on another GC column that
resolves the isomers.
4.2.3 30 m DB-225 fused silica capillary column, (J&W Scientific) or
equivalent.
4.3 Miscellaneous Equipment and Materials - The following list of items
does not necessarily constitute an exhaustive compendium of the equipment needed
for this analytical method.
4.3.1 Nitrogen evaporation apparatus with variable flow rate.
4.3.2 Balances capable of accurately weighing to 0.01 g and
0.0001 g.
4.3.3 Centrifuge.
4.3.4 Water bath, equipped with concentric ring covers and capable
of being temperature controlled within + 2°C.
4.3.5 Stainless steel or glass container large enough to hold
contents of one pint sample containers.
4.3.6 Glove box.
4.3.7 Drying oven.
4.3.8 Stainless steel spoons and spatulas.
4.3.9 Laboratory hoods,
4.3.10 Pipets, disposable, Pasteur, 150 mm long x 5 mm ID.
4.3.11 Pipets, disposable, serological, 10 ml, for the
preparation of the carbon columns specified in Sec. 7.5.3.
4.3.12 Reaction vial, 2 ml, silanized amber glass (Reacti-vial,
or equivalent).
4.3.13 Stainless steel meat grinder with a 3 to 5 mm hole size
inner plate.
4.3.14 Separatory funnels, 125 ml and 2000 ml.
4.3.15 Kuderna-Danish concentrator, 500 ml, fitted with 10 ml
concentrator tube and three ball Snyder column.
4.3.16 Teflon™ or carborundum (silicon carbide) boiling chips
(or equivalent), washed with hexane before use.
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NOTE: Teflon™ boiling chips may float in methylene chloride, may
not work in the presence of any water phase, and may be
penetrated by nonpolar organic compounds.
4.3.17 Chromatographic columns, glass, 300 mm x 10.5 mm, fitted
with Teflon™ stopcock.
4.3.18 Adapters for concentrator tubes.
4.3.19 Glass fiber filters, 0.70 urn, Whatman GFF, or
equivalent.
4.3.20 Dean-Stark trap, 5 or 10 ml, with T-joints, condenser
and 125 ml flask.
4.3.21 Continuous liquid-liquid extractor.
4.3.22 All glass Soxhlet apparatus, 500 ml flask.
4.3.23 Soxhlet/Dean Stark extractor (optional), all glass, 500
ml flask.
4.3.24 Glass funnels, sized to hold 170 ml of liquid.
4.3.25 Desiccator.
4.3.26 Solvent reservoir (125 ml), Kontes; 12.35 cm diameter
(special order item), compatible with gravity carbon column.
4.3.27 Rotary evaporator with a temperature controlled water
bath.
4.3.28 High speed tissue homogenizer, equipped with an EN-8
probe, or equivalent.
4.3.29 Glass wool, extracted with methylene chloride, dried and
stored in a clean glass jar.
4.3.30 Extraction jars, glass, 250 ml, with teflon lined screw
cap.
4.3.31 Volumetric flasks, Class A - 10 ml to 1000 ml.
4.3.32 Glass vials, 1 dram (or metric equivalent).
NOTE: Reuse of glassware should be minimized to avoid the risk of
contamination. All glassware that is reused must be
scrupulously cleaned as soon as possible after use, according
to the following procedure: Rinse glassware with the last
solvent used in it. Wash with hot detergent water, then rinse
with copious amounts of tap water and several portions of
organic-free reagent water. Rinse with high purity acetone
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and hexane and store it inverted or capped with solvent rinsed
aluminum foil in a clean environment.
5.0 REAGENTS AND STANDARD SOLUTIONS
5.1 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.2 Column Chromatography Reagents
5.2.1 Alumina, neutral, 80/200 mesh (Super 1, Woelm®, or
equivalent). Store in a sealed container at room temperature, in a
desiccator, over self-indicating silica gel.
5.2.2 Alumina, acidic AG4, (Bio Rad Laboratories catalog #132-1240,
or equivalent). Soxhlet extract with methylene chloride for 24 hours if
blanks show contamination, and activate by heating in a foil covered glass
container for 24 hours at 190°C. Store in a glass bottle sealed with a
Teflon™ lined screw cap.
5.2.3 Silica gel, high purity grade, type 60, 70-230 mesh; Soxhlet
extract with methylene chloride for 24 hours if blanks show contamination,
and activate by heating in a foil covered glass container for 24 hours at
190°C. Store in a glass bottle sealed with a Teflon™ lined screw cap.
5.2.4 Silica gel impregnated with sodium hydroxide. Add one part
(by weight) of 1 M NaOH solution to two parts (by weight) silica gel
(extracted and activated) in a screw cap bottle and mix with a gl
until free of lumps. Store in a glass bottle sealed with a Teflon1
.ass rod
™ lined
screw cap.
5.2.5 Silica gel impregnated with 40 percent (by weight) sulfuric
acid. Add two parts (by weight) concentrated sulfuric acid to three parts
(by weight) silica gel (extracted and activated), mix with a glass rod
until free of lumps, and store in a screw capped glass bottle. Store in
a glass bottle sealed with a Teflon™ lined screw cap.
5.2.6 Celite 545® (Supelco), or equivalent.
5.2.7 Active carbon AX-21 (Anderson Development Co., Adrian, MI), or
equivalent, prewashed with methanol and dried in vacuo at 110°C. Store in
a glass bottle sealed with a Teflon™ lined screw cap.
5.3 Reagents
5.3.1 Sulfuric acid, H2S04, concentrated, ACS grade, specific gravity
1.84.
5.3.2 Potassium hydroxide, KOH, ACS grade, 20 percent (w/v) in
organic-free reagent water.
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5.3.3 Sodium chloride, NaCl, analytical reagent, 5 percent (w/v) in
organic-free reagent water.
5.3.4 Potassium carbonate, K2C03, anhydrous, analytical reagent.
5.4 Desiccating agent
5.4.1 Sodium sulfate (powder, anhydrous), Na2S04. Purify by heating
at 400°C for 4 hours in a shallow tray, or by precleaning the sodium
sulfate with methylene chloride. If the sodium sulfate is precleaned with
methylene chloride, a method blank must be analyzed, demonstrating that
there is no interference from the sodium sulfate.
5.5 Solvents
5.5.1 Methylene chloride, CH2C12. High purity, distilled in glass
or highest available purity.
5.5.2 Hexane, CeH14. High purity, distilled in glass or highest
available purity.
5.5.3 Methanol, CH3OH. High purity, distilled in glass or highest
available purity.
5.5.4 Nonane, C9H20. High purity, distilled in glass or highest
available purity.
5.5.5 Toluene, C6H5CH3. High purity, distilled in glass or highest
available purity.
5.5.6 Cyclohexane, C6H12,
available purity.
5.5.7 Acetone,
available purity.
CH3COCH3.
High purity, distilled in glass or highest
High purity, distilled in glass or highest
5.6 High-Resolution Concentration Calibration Solutions (Table 5) - Five
nonane solutions containing unlabeled (totaling 17) and carbon-labeled (totaling
11) PCDDs and PCDFs at known concentrations are used to calibrate the instrument.
The concentration ranges are homologue dependent, with the lowest values for the
tetrachlorinated dioxin and furan (1.0 pg/^L) and the highest values for the
octachlorinated congeners (1000 pg//iL).
5.6.1 Depending on the availability of materials, these high-
resolution concentration calibration solutions may be obtained from the
Environmental Monitoring Systems Laboratory, U.S. EPA, Cincinnati, Ohio.
However, additional secondary standards must be obtained from commercial
sources, and solutions should be prepared in the analyst's laboratory. It
is the responsibility of the laboratory to ascertain that the calibration
solutions received (or prepared) are indeed at the appropriate
concentrations before they are used to analyze samples.
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5.6.2 Store the concentration calibration solutions in 1 ml
mini vials at room temperature in the dark.
5.7 GC Column Performance Check Solution - This solution contains the
first and last eluting isomers for each homologous series from tetra- through
heptachlorinated congeners. The solution also contains a series of other TCDD
isomers for the purpose of documenting the chromatographic resolution. The
13Cl2-2,3,7,8-TCDD is also present. The laboratory is required to use nonane as
the solvent and adjust the volume so that the final concentration does not exceed
100 pg/juL per congener. Table 7 summarizes the qualitative composition (minimum
requirement) of this performance evaluation solution.
5.8 Sample Fortification Solution - This nonane solution contains the
nine internal standards at the nominal concentrations that are listed in Table 2.
The solution contains at least one carbon-labeled standard for each homologous
series, and it is used to measure the concentrations of the native substances.
(Note that 13C12-OCDf: is not present in the solution.)
5.9 Recovery Standard Solution - This nonane solution contains two
recovery standards, 13C12-1,2,3,4-TCDD and 13C12-l,2,3,7,8,9-HxCDD, at a nominal
concentration of 50 pg//iL per compound. 10 to 50 juL of this solution will be
spiked into each sample extract before the final concentration step and HRGC/HRMS
analysis.
5.10 Matrix Spike Fortification Solution - Solution used to prepare the
MS and MSD samples. It contains all unlabeled analytes listed in Table 5 at con-
centrations corresponding to the HRCC 3.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
6.2 Sample Collection
6.2.1 Sample collection personnel should, to the extent possible,
homogenize samples in the field before filling the sample containers.
This should minimize or eliminate the necessity for sample homogenization
in the laboratory. The analyst should make a judgment, based on the
appearance of the sample, regarding the necessity for additional mixing.
If the sample is clearly not homogeneous, the entire contents should be
transferred to a glass or stainless steel pan for mixing with a stainless
steel spoon or spatula before removal of a sample portion for analysis.
6.2.2 Grab and composite samples must be collected in glass
containers. Conventional sampling practices must be followed. The bottle
must not be prewashed with sample before collection. Sampling equipment
must be free of potential sources of contamination.
6.3 Grinding or Blending of Fish Samples - If not otherwise specified by
the U.S. EPA, the whole fish (frozen) should be blended or ground to provide a
homogeneous sample. The use of a stainless steel meat grinder with a 3 to 5 mm
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hole size inner plate is recommended. In some circumstances, analysis of fillet
or specific organs of fish may be requested by the U.S. EPA. If so requested,
the above whole fish requirement is superseded.
6.4 Storage and Holding Times - All samples, except fish and adipose
tissue samples, must be stored at 4°C in the dark, extracted within 30 days and
completely analyzed within 45 days of extraction. Fish and adipose tissue
samples must be stored at -20°C in the dark, extracted within 30 days and
completely analyzed within 45 days of collection. Whenever samples are analyzed
after the holding time expiration date, the results should be considered to be
minimum concentrations and should be identified as such.
NOTE: The holding times listed in Sec. 6.4 are recommendations. PCDDs and
PCDFs are very stable in a variety of matrices, and holding times
under the conditions listed in Sec. 6.4 may be as high as a year for
certain matrices. Sample extracts, however, should always be
analyzed within 45 days of extraction.
6.5 Phase Separation - This is a guideline for phase separation for very
wet (>Z5 percent water) soil, sediment and paper pulp samples. Place a 50 g
portion in a suitable centrifuge bottle and centrifuge for 30 minutes at
2,000 rpm. Remove the bottle and mark the interface level on the bottle.
Estimate the relative volume of each phase. With a disposable pipet, transfer
the liquid layer into a clean bottle. Mix the solid with a stainless steel
spatula and remove a portion to be weighed and analyzed (percent dry weight
determination, extraction). Return the remaining solid portion to the original
sample bottle (empty) or to a clean sample bottle that is properly labeled, and
store it as appropriate. Analyze the solid phase by using only the soil,
sediment and paper pulp method. Take note of, and report, the estimated volume
of liquid before disposing of the liquid as a liquid waste.
6.6 Soil, Sediment, or Paper Sludge (Pulp) Percent Dry Weight
Determination - The percent dry weight of soil, sediment or paper pulp samples
showing detectable levels (see note below) of at least one 2,3,7,8-substituted
PCDD/PCDF congener is determined according to the following procedure. Weigh a
10 g portion of the soil or sediment sample (+ 0.5 g) to three significant
figures. Dry it to constant weight at 110°C in an adequately ventilated oven.
Allow the sample to cool in a desiccator. Weigh the dried solid to three
significant figures. Calculate and report the percent dry weight. Do not use
this solid portion of the sample for extraction, but instead dispose of it as
hazardous waste.
NOTE: Until detection limits have been established (Sec. 1.3), the lower
MCLs (Table 1) may be used to estimate the minimum detectable
levels.
% dry weight = q of dry sample x 100
g of sample
CAUTION: Finely divided soils and sediments contaminated with
PCDDs/PCDFs are hazardous because of the potential for
inhalation or ingestion of particles containing PCDDs/PCDFs
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(including 2,3,7,8-TCDD). Such samples should be handled in
a confined environment (i.e., a closed hood or a glove box).
6.7 Lipid Content Determination
6.7.1 Fish Tissue - To determine the lipid content of fish tissue,
concentrate 125 ml of the fish tissue extract (Sec. 7.2.2), in a tared 200
mL round bottom flask, on a rotary evaporator until a constant weight (W)
is achieved.
100 (W)
Percent lipid =
10
Dispose of the lipid residue as a hazardous waste if the results of
the analysis indicate the presence of PCDDs or PCDFs.
6.7.2 Adipose Tissue - Details for the determination of the adipose
tissue lipid content are provided in Sec. 7.3.3.
7.0 PROCEDURE
7.1 Internal standard addition
7.1.1 Use a portion of 1 g to 1000 g (± 5 percent) of the sample to
be analyzed. Typical sample size requirements for different matrices are
given in Sec. 7.4 and in Table 1. Transfer the sample portion to a tared
flask and determine its weight.
7.1.2 Except for adipose tissue, add an appropriate quantity of the
sample fortification mixture (Sec. 5.8) to the sample. All samples should
be spiked with 100 ^L of the sample fortification mixture to give internal
standard concentrations as indicated in Table 1. As an example, for 13C12-
2,3,7,8-TCDD, a 10 g soil sample requires the addition of 1000 pg of 13C12-
2,3,7,8-TCDD to give the required 100 ppt fortification level. The fish
tissue sample (20 g) must be spiked with 200 /iL of the internal standard
solution, because half of the extract will be used to determine the lipid
content (Sec. 6.7.1).
7.1.2.1 For the fortification of soil, sediment, fly ash,
water, fish tissue, paper pulp and wet sludge samples, mix the
sample fortification solution with 1.0 ml acetone.
7.1.2.2 Do not dilute the nonane solution for the other
matrices.
7.1.2.3 The fortification of adipose tissue is carried out
at the time of homogenization (Sec. 7.3.2.3).
7.2 Extraction and Purification of Fish and Paper Pulp Samples
7.2.1 Add 60 g anhydrous sodium sulfate to a 20 g portion of a
homogeneous fish sample (Sec. 6.3) and mix thoroughly with a stainless
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steel spatula. After breaking up any lumps, place the fish/sodium sulfate
mixture in the Soxhlet apparatus on top of a glass wool plug. Add 250 ml
methylene chloride or hexane/methylene chloride (1:1) to the Soxhlet
apparatus and reflux for 16 hours. The solvent must cycle completely
through the system five times per hour. Follow the same procedure for the
partially dewatered paper pulp sample (using a 10 g sample, 30 g of
anhydrous sodium sulfate and 200 ml of toluene).
NOTE: As an option, a Soxhlet/Dean Stark extractor system may be
used, with toluene as the solvent. No sodium sulfate is added
when using this option.
7.2.2 Transfer the fish extract from Sec. 7.2.1 to a 250 ml
volumetric flask and fill to the mark with methylene chloride. Mix well,
then remove 125 ml for the determination of the lipid content (Sec.
6.7.1). Transfer the remaining 125 ml of the extract, plus two 15 ml
hexane/methylene chloride rinses of the volumetric flask, to a KD
apparatus equipped with a Snyder column. Quantitatively transfer all of
the paper pulp extract to a KD apparatus equipped with a Snyder column.
NOTE: As an option, a rotary evaporator may be used in place of the
KD apparatus for the concentration of the extracts.
7.2.3 Add a Teflon™, or equivalent, boiling chip. Concentrate the
extract in a water bath to an apparent volume of 10 ml. Remove the
apparatus from the water bath and allow to cool for 5 minutes.
7.2.4 Add 50 mL hexane and a new boiling chip to the KD flask.
Concentrate in a water bath to an apparent volume of 5 mL. Remove the
apparatus from the water bath and allow to cool for 5 minutes.
NOTE: The methylene chloride must have been completely removed
before proceeding with the next step.
7.2.5 Remove and invert the Snyder column and rinse it into the KD
apparatus with two 1 ml portions of hexane. Decant the contents of the KD
apparatus and concentrator tube into a 125 mL separatory funnel. Rinse
the KD apparatus with two additional 5 mL portions of hexane and add the
rinses to the funnel. Proceed with the cleanup according to the
instructions starting in Sec. 7.5.1.1, but omit the procedures described
in Sees. 7.5.1.2 and 7.5.1.3.
7.3 Extraction and Purification of Human Adipose Tissue
7.3.1 Human adipose tissue samples must be stored at a temperature
of -20°C or lower from the time of collection until the time of analysis.
The use of chlorinated materials during the collection of the samples must
be avoided. Samples are handled with stainless steel forceps, spatulas,
or scissors. All sample bottles (glass) are cleaned as specified in the
note at the end of Sec. 4.3. Teflon™ lined caps should be used.
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NOTE: The specified storage temperature of -20°C is the maximum
storage temperature permissible for adipose tissue samples.
Lower storage temperatures are recommended.
7.3.2 Adipose Tissue Extraction
7.3.2.1 Weigh, to the nearest 0.01 g, a 10 g portion of a
frozen adipose tissue sample into a culture tube (2.2 x 15 cm).
NOTE: The sample size may be smaller, depending on
availability. In such a situation, the analyst is
required to adjust the volume of the internal standard
solution added to the sample to meet the fortification
level stipulated in Table 1.
7.3.2.2 Allow the adipose tissue specimen to reach room
temperature (up to 2 hours).
7.3.2.3 Add 10 ml methylene chloride and 100 /*L of the
sample fortification solution. Homogenize the mixture for
approximately 1 minute with a tissue homogenizer.
7.3.2.4 Allow the mixture to separate, then remove the
methylene chloride extract from the residual solid material with a
disposable pi pet. Percolate the methylene chloride through a filter
funnel containing a clean glass wool plug and 10 g anhydrous sodium
sulfate. Collect the dried extract in a graduated 100 ml volumetric
flask.
7.3.2.5 Add a second 10 ml portion of methylene chloride
to the sample and homogenize for 1 minute. Decant the solvent, dry
it, and transfer it to the 100 ml volumetric flask (Sec. 7.3.2.4).
7.3.2.6 Rinse the culture tube with at least two
additional portions of methylene chloride (10 ml each), and transfer
the entire contents to the filter funnel containing the anhydrous
sodium sulfate. Rinse the filter funnel and the anhydrous sodium
sulfate contents with additional methylene chloride (20 to 40 ml)
into the 100 ml flask. Discard the sodium sulfate.
7.3.2.7 Adjust the volume to the 100 ml mark with
methylerie chloride.
7.3.3 Adipose Tissue Lipid Content Determination
7.3.3.1 Preweigh a clean 1 dram (or metric equivalent)
glass vial to the nearest 0.0001 g on an analytical balance tared to
zero.
7.3.3.2 Accurately transfer 1.0 ml of the final extract
(100 mL) from Sec. 7.3.2.7 to the vial. Reduce the volume of the
extract on a water bath (50-60°C) by a gentle stream of purified
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nitrogen until an oily residue remains. Nitrogen blowdown is
continued until a constant weight is achieved.
NOTE: When the sample size of the adipose tissue is smaller
than 10 g, then the analyst may use a larger portion (up
to 10 percent) of the extract defined in Sec. 7.3.2.7
for the lipid determination.
7.3.3.3 Accurately weigh the vial with the residue to the
nearest 0.0001 g and calculate the weight of the lipid present in
the vial based on the difference of the weights.
7.3.3.4 Calculate the percent lipid content of the
original sample to the nearest 0.1 percent as shown below:
**lr X "ext
Lipid content, LC {%) = x 100
"""at X "al
where:
W,r = weight of the lipid residue to the nearest 0.0001
g calculated from Sec. 7.3.3.3,
Vext = total volume (100 ml) of the extract in ml from
Sec. 7.3.2.7,
Wat = weight of the original adipose tissue sample to
the nearest 0.01 g from Sec. 7.3.2.1, and
Val = volume of the aliquot of the final extract in mi
used for the quantitative measure of the lipid
residue (1.0 ml) from Sec. 7.3.3.2.
7.3.3.5 Record the lipid residue measured in Sec. 7.3.3.3
and the percent lipid content from Sec. 7.3.3.4.
7.3.4 Adipose Tissue Extract Concentration
7.3.4.1 Quantitatively transfer the remaining extract from
Sec. 7.3.3.2 (99.0 ml) to a 500 ml Erlenmeyer flask. Rinse the
volumetric flask with 20 to 30 ml of additional methylene chloride
to ensure quantitative transfer.
7.3.4.2 Concentrate the extract on a rotary evaporator and
a water bath at 40°C until an oily residue remains.
7.3.5 Adipose Tissue Extract Cleanup
7.3.5.1 Add 200 ml hexane to the lipid residue in the 500
ml Erlenmeyer flask and swirl the flask to dissolve the residue.
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7.3.5.2 Slowly add, with stirring, 100 g of 40 percent
(w/w) sulfuric acid-impregnated silica gel. Stir with a magnetic
stirrer for two hours at room temperature.
7.3.5.3 Allow the solid phase to settle, and decant the
liquid through a filter funnel containing 10 g anhydrous sodium
sulfate on a glass wool plug, into another 500 mL Erlenmeyer flask.
7.3.5.4 Rinse the solid phase with two 50 ml portions of
hexane. Stir each rinse for 15 minutes, decant, and dry as
described under Sec. 7.3.5.3. Combine the hexane extracts from Sec.
7.3.5.3 with the rinses.
7.3.5.5 Rinse the sodium sulfate in the filter funnel with
an additional 25 ml hexane and combine this rinse with the hexane
extracts from Sec. 7.3.5.4.
7.3.5.6 Prepare an acidic silica column as follows: Pack
a 2 cm x 10 cm chromatographic column with a glass wool plug, add
approximately 20 ml hexane, add 1 g silica gel and allow to settle,
then add 4 g of 40 percent (w/w) sulfuric acid-impregnated silica
gel and allow to settle. Elute the excess hexane from the column
until the solvent level reaches the top of the chromatographic
packing. Verify that the column does not have any air bubbles and
channels.
7.3.5.7 Quantitatively transfer the hexane extract from
the Erlenmeyer flask (Sees. 7.3.5.3 through 7.3.5.5) to the silica
gel column reservoir. Allow the hexane extract to percolate through
the column and collect the eluate in a 500 mL KD apparatus.
7.3.5.8 Complete the elution by percolating 50 ml hexane
through the column into the KD apparatus. Concentrate the eluate on
a steam bath to approximately 5 ml. Use nitrogen blowdown to bring
the final volume to about 100 juL.
NOTE: If the silica gel impregnated with 40 percent sulfuric
acid is highly discolored throughout the length of the
adsorbent bed, the cleaning procedure must be repeated
beginning with Sec. 7.3.5.1.
7.3.5.9 The extract is ready for the column cleanups
described in Sees. 7.5.2 through 7.5.3.6.
7.4 Extraction and Purification of Environmental and Waste Samples
7.4.1 Sludge/Wet Fuel Oil
7.4.1.1 Extract aqueous sludge or wet fuel oil samples by
refluxing a sample (e.g., 2 g) with 50 ml toluene in a 125 ml flask
fitted with a Dean-Stark water separator. Continue refluxing the
sample until all the water is removed.
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NOTE: If the sludge or fuel oil sample dissolves in toluene,
treat it according to the instructions in Sec. 7.4.2
below. If the labeled sludge sample originates from
pulp (paper mills), treat it according to the
instructions starting in Sec. 7.2, but without the
addition of sodium sulfate.
7.4.1.2 Cool the sample, filter the toluene extract
through a glass fiber filter, or equivalent, into a 100 mL round
bottom flask.
7.4.1.3 Rinse the filter with 10 ml toluene and combine
the extract with the rinse.
7.4.1.4 Concentrate the combined solutions to near dryness
on a rotary evaporator at 50°C. Use of an inert gas to concentrate
the extract is also permitted. Proceed with Sec. 7.4.4.
7.4.2 Still Bottom/Oil
7.4.2.1 Extract still bottom or oil samples by mixing a
sample portion (e.g., 1.0 g) with 10 ml toluene in a small beaker
and filtering the solution through a glass fiber filter (or
equivalent) into a 50 ml round bottom flask. Rinse the beaker and
filter with 10 ml toluene.
7.4.2.2 Concentrate the combined toluene solutions to near
dryness on a rotary evaporator at 50°C. Proceed with Sec. 7.4.4.
7.4.3 Fly Ash
NOTE: Because of the tendency of fly ash to "fly", all handling
steps should be performed in a hood in order to minimize
contamination.
7.4.3.1 Weigh about 10 g fly ash to two decimal places and
transfer to an extraction jar. Add 100 /xL sample fortification
solution (Sec. 5.8), diluted to 1 ml with acetone, to the sample.
Add 150 ml of 1 M HC1 to the fly ash sample. Seal the jar with the
Teflon™ lined screw cap and shake for 3 hours at room temperature.
7.4.3.2 Rinse a glass fiber filter with toluene, and
filter the sample through the filter paper, placed in a Buchner
funnel, into all flask. Wash the fly ash cake with approximately
500 ml organic-free reagent water and dry the filter cake overnight
at room temperature in a desiccator.
7.4.3.3 Add 10 g anhydrous powdered sodium sulfate, mix
thoroughly, let sit in a closed container for one hour, mix again,
let sit for another hour, and mix again.
7.4.3.4 Place the sample and the filter paper into an
extraction thimble, and extract in a Soxhlet extraction apparatus
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charged with 200 ml toluene for 16 hours using a five cycle/hour
schedule.
NOTE: As an option, a Soxhlet/Dean Stark extractor system may
be used, with toluene as the solvent. No sodium sulfate
is added when using this option.
7.4.3.5 Cool and filter the toluene extract through a
glass fiber filter into a 500 ml round bottom flask. Rinse the
filter with 10 ml toluene. Add the rinse to the extract and
concentrate the combined toluene solutions to near dryness on a
rotary evaporator at 50°C. Proceed with Sec. 7.4.4.
7.4.4 Transfer the concentrate to a 125 ml separatory funnel using
15 ml hexane. Rinse the flask with two 5 ml portions of hexane and add
the rinses to the funnel. Shake the combined solutions in the separatory
funnel for two minutes with 50 ml of 5 percent sodium chloride solution,
discard the aqueous layer, and proceed with Sec. 7.5.
7.4.5 Aqueous samples
7.4.5.1 Allow the sample to come to ambient temperature,
then mark the water meniscus on the side of the 1 L sample bottle
for later determination of the exact sample volume. Add the
required acetone diluted sample fortification solution (Sec. 5.8).
7.4.5.2 When the sample is judged to contain 1 percent or
more solids, the sample must be filtered through a glass fiber
filter that has been rinsed with toluene. If the suspended solids
content is too great to filter through the 0.45 /urn filter,
centrifuge the sample, decant, and then filter the aqueous phase.
NOTE: Paper mill effluent samples normally contain 0.02%-0.2%
solids, and would not require filtration. However, for
optimum analytical results, all paper mill effluent
samples should be filtered, the isolated solids and
filtrate extracted separately, and the extracts
recombined.
7.4.5.3 Combine the solids from the centrifuge bottle(s)
with the particulates on the filter and with the filter itself and
proceed with the Soxhlet extraction as specified in Sees. 7.4.6.1
through 7.4.6.4. Remove and invert the Snyder column and rinse it
down into the KD apparatus with two 1 ml portions of hexane.
7.4.5.4 Pour the aqueous filtrate into a 2 L separatory
funnel. Add 60 ml methylene chloride to the sample bottle, seal and
shake for 30 seconds to rinse the inner surface. Transfer the
solvent to the separatory funnel and extract the sample by shaking
the funnel for two minutes with periodic venting.
7..4.5.S Allow the organic layer to separate from the water
phase for a minimum of 10 minutes. If the emulsion interface
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between layers is more than one third the volume of the solvent
layer, the analyst must employ mechanical techniques to complete the
phase separation (e.g., glass stirring rod).
7.4.5.6 Collect the methylene chloride into a KD apparatus
(mounted with a 10 ml concentrator tube) by passing the sample
extracts through a filter funnel packed with a glass wool plug and
5 g anhydrous sodium sulfate.
NOTE: As an option, a rotary evaporator may be used in place
of the KD apparatus for the concentration of the
extracts.
7.4.5.7 Repeat the extraction twice with fresh 60 ml
portions of methylene chloride. After the third extraction, rinse
the sodium sulfate with an additional 30 ml methylene chloride to
ensure quantitative transfer. Combine all extracts and the rinse in
the KD apparatus.
NOTE: A continuous liquid-liquid extractor may be used in
place of a separatory funnel when experience with a
sample from a given source indicates that a serious
emulsion problem will result or an emulsion is
encountered when using a separatory funnel. Add 60 ml
methylene chloride to the sample bottle, seal, and shake
for 30 seconds to rinse the inner surface. Transfer the
solvent to the extractor. Repeat the rinse of the
sample bottle with an additional 50 to 100 ml portion of
methylene chloride and add the rinse to the extractor.
Add 200 to 500 ml methylene chloride to the distilling
flask, add sufficient organic-free reagent water (Sec.
5.1) to ensure proper operation, and extract for
24 hours. Allow to cool, then detach the distilling
flask. Dry and concentrate the extract as described in
Sees. 7.4.5.6 and 7.4.5.8 through 7.4.5.10. Proceed
with Sec. 7.4.5.11.
7.4.5.8 Attach a Snyder column and concentrate the extract
on a water bath until the apparent volume of the liquid is 5 ml.
Remove the KD apparatus and allow it to drain and cool for at least
10 minutes.
7.4.5.9 Remove the Snyder column, add 50 mL hexane, add
the concentrate obtained from the Soxhlet extraction of the
suspended solids (Sec. 7.4.5.3), if applicable, re-attach the Snyder
column, and concentrate to approximately 5 ml. Add a new boiling
chip to the KD apparatus before proceeding with the second
concentration step.
7.4.5.10 Rinse the flask and the lower joint with two 5 mL
portions of hexane and combine the rinses with the extract to give
a final volume of about 15 ml.
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7.4.5.11 Determine the original sample volume by filling
the sample bottle to the mark with water and transferring the water
to a 1000 ml graduated cylinder. Record the sample volume to the
nearest 5 ml. Proceed with Sec. 7.5.
7.4.6 Soil/Sediment
7.4.6.1 Add 10 g anhydrous powdered sodium sulfate to the
sample portion (e.g., 10 g) and mix thoroughly with a stainless
steel spatula. After breaking up any lumps, place the soil/sodium
sulfate mixture in the Soxhlet apparatus on top of a glass wool plug
(the use of an extraction thimble is optional).
NOTE: As an option, a Soxhlet/Dean Stark extractor system may
be used, with toluene as the solvent. No sodium sulfate
is added when using this option.
7.4.6.2 Add 200 to 250 ml toluene to the Soxhlet apparatus
and reflux for 16 hours. The solvent must cycle completely through
the system five times per hour.
NOTE: If the dried sample is not of free flowing consistency,
more sodium sulfate must be added.
7.4.6.3 Cool and filter the extract through a glass fiber
filter into a 500 ml round bottom flask for evaporation of the
toluene. Rinse the filter with 10 ml of toluene, and concentrate
the combined fractions to near dryness on a rotary evaporator at
50°C. Remove the flask from the water bath and allow to cool for
5 minutes.
7.4.6.4 Transfer the residue to a 125 mL separatory
funnel, using 15 mL of hexane. Rinse the flask with two additional
portions of hexane, and add the rinses to the funnel. Proceed with
Sec. 7.5.
7.5 Cleanup
7.5.1 Partition
7.5.1.1 Partition the hexane extract against 40 ml of
concentrated sulfuric acid. Shake for two minutes. Remove and
discard the sulfuric acid layer (bottom). Repeat the acid washing
until no color is visible in the acid layer (perform a maximum of
four acid washings).
7.5.1.2 Omit this step for the fish sample extract.
Partition the extract against 40 ml of 5 percent (w/v) aqueous
sodium chloride. Shake for two minutes. Remove and discard the
aqueous layer (bottom).
7,.5.1.3 Omit this step for the fish sample extract.
Partition the extract against 40 ml of 20 percent (w/v) aqueous
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potassium hydroxide (KOH). Shake for two minutes. Remove and
discard the aqueous layer (bottom). Repeat the base washing until
no color is visible in the bottom layer (perform a maximum of four
base washings). Strong base (KOH) is known to degrade certain
PCDDs/PCDFs, so contact time must be minimized.
7.5.1.4 Partition the extract against 40 ml of 5 percent
(w/v) aqueous sodium chloride. Shake for two minutes. Remove and
discard the aqueous layer (bottom). Dry the extract by pouring it
through a filter funnel containing anhydrous sodium sulfate on a
glass wool plug, and collect it in a 50 ml round bottom flask.
Rinse the funnel with the sodium sulfate with two 15 ml portions of
hexane, add the rinses to the 50 ml flask, and concentrate the
hexane solution to near dryness on a rotary evaporator (35°C water
bath), making sure all traces of toluene (when applicable) are
removed. (Use of blowdown with an inert gas to concentrate the
extract is also permitted.)
7.5.2 Silica/Alumina Column Cleanup
7.5.2.1 Pack a gravity column (glass, 30 cm x 10.5 mm),
fitted with a Teflon™ stopcock, with silica gel as follows: Insert
a glass wool plug into the bottom of the column. Place 1 g silica
gel in the column and tap the column gently to settle the silica
gel. Add 2 g sodium hydroxide-impregnated silica gel, 4 g sulfuric
acid-impregnated silica gel, and 2 g silica gel. Tap the column
gently after each addition. A small positive pressure (5 psi) of
clean nitrogen may be used if needed. Elute with 10 ml hexane and
close the stopcock just before exposure of the top layer of silica
gel to air. Discard the eluate. Check the column for channeling.
If channeling is observed, discard the column. Do not tap the
wetted column.
7.5.2.2 Pack a gravity column (glass, 300 mm x 10.5 mm),
fitted with a Teflon™ stopcock, with alumina as follows: Insert a
glass wool plug into the bottom of the column. Add a 4 g layer of
sodium sulfate. Add a 4 g layer of Woelm® Super 1 neutral alumina.
Tap the top of the column gently. Woelm® Super 1 neutral alumina
need not be activated or cleaned before use, but it should be stored
in a sealed desiccator. Add a 4 g layer of anhydrous sodium sulfate
to cover the alumina. Elute with 10 mL hexane and close the
stopcock just before exposure of the sodium sulfate layer to air.
Discard the eluate. Check the column for channeling. If channeling
is observed, discard the column. Do not tap a wetted column.
NOTE: Optionally, acidic alumina (Sec. 5.2.2) can be used in
place of neutral alumina.
7.5.2.3 Dissolve the residue from Sec. 7.5.1.4 in 2 ml
hexane and apply the hexane solution to the top of the silica gel
column. Rinse the flask with enough hexane (3-4 ml) to complete the
quantitative transfer of the sample to the surface of the silica
gel.
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7.5.2.4 Elute the silica gel column with 90 ml of hexane,
concentrate the eluate on a rotary evaporator (35°C water bath) to
approximately 1 ml, and apply the concentrate to the top of the
alumina column (Sec. 7.5.2.2). Rinse the rotary evaporator flask
twice with 2 ml of hexane, and add the rinses to the top of the
alumina column.
7.5.2.5 Add 20 ml hexane to the alumina column and elute
until the hexane level is just below the top of the sodium sulfate.
Do not discard the eluted hexane, but collect it in a separate flask
and store it for later use, as it may be useful in determining where
the labeled analytes are being lost if recoveries are not
satisfactory.
7.5.2.6 Add 15 ml of 60 percent methylene chloride in
hexane (v/v) to the alumina column and collect the eluate in a
conical shaped (15 ml) concentration tube. With a carefully
regulated stream of nitrogen, concentrate the 60 percent methylene
chloride/hexane fraction to about 2 ml.
7.5.3 Carbon Column Cleanup
7.5.3.1 Prepare an AX-21/Celite 545® column as follows:
Thoroughly mix 5.40 g active carbon AX-21 and 62.0 g Celite 545® to
produce an 8 percent (w/w) mixture. Activate the mixture at 130°C
for 6 hours and store it in a desiccator.
7.5.3.2 Cut off both ends of a 10 ml disposable
serological pipet to give a 10 cm long column. Fire polish both
ends and flare, if desired. Insert a glass wool plug at one end,
then pack the column with enough Celite 545® to form a 1 cm plug,
add 1 g of the AX-21/Celite 545® mixture, top with additional Celite
545® (enough for a 1 cm plug), and cap the packing with another
glass wool plug.
NOTE: Each new batch of AX-21/Celite 545® must be checked as
follows: Add 50 juL of the continuing calibration
solution to 950 juL hexane. Take this solution through
the carbon column cleanup step, concentrate to 50 inl-
and analyze. If the recovery of any of the analytes is
<80 percent, discard this batch of AX-21/Celite 545®.
7.5.3.3 Rinse the AX-21/Celite 545® column with 5 ml of
toluene, followed by 2 ml of 75:20:5 (v/v) methylene chloride/
methanol/toluene, 1 mL of 1:1 (v/v) cyclohexane/methylene chloride,
and 5 ml hexane. The flow rate should be less than 0.5 mL/min.
Discard the rinses. While the column is still wet with hexane, add
the sample concentrate (Sec. 7.5.2.6) to the top of the column.
Rinse the concentrator tube (which contained the sample concentrate)
twice with 1 ml hexane, and add the rinses to the top of the column.
7.5.3.4 Elute the column sequentially with two 2 mL
portions of hexane, 2 ml cyclohexane/methylene chloride (50:50,
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v/v), and 2 ml methylene chloride/methanol/toluene (75:20:5, v/v).
Combine these eluates; this combined fraction may be used as a check
on column efficiency.
7.5.3.5 Turn the column upside down and elute the
PCDD/PCDF fraction with 20 ml toluene. Verify that no carbon fines
are present in the eluate. If carbon fines are present in the
eluate, filter the eluate through a glass fiber filter (0.45 urn) and
rinse the filter with 2 ml toluene. Add the rinse to the eluate.
7.5.3.6 Concentrate the toluene fraction to about 1 ml on
a rotary evaporator by using a water bath at 50°C. Carefully
transfer the concentrate into a 1 ml minivial and, again at elevated
temperature (50°C), reduce the volume to about 100 pi using a stream
of nitrogen and a sand bath. Rinse the rotary evaporator flask
three times with 300 p.1 of a solution of 1 percent toluene in
methylene chloride, and add the rinses to the concentrate. Add
10 juL of the nonane recovery standard solution (Sec. 5.9) for soil,
sediment, water, fish, paper pulp and adipose tissue samples, or 50
juL of the recovery standard solution for sludge, still bottom and
fly ash samples. Store the sample at room temperature in the dark.
7.6 Chromatographic/Mass Spectrometric Conditions and Data Acquisition
Parameters
7.6.1 Gas Chromatograph
Column coating: DB-5
Film thickness: 0.25 urn
Column dimension: 60 m x 0.32 mm
Injector temperature: 270°C
Splitless valve time: 45 s
Interface temperature: Function of the final temperature
Temperature program:
Stage
Init.
Temp.
(°C)
Init.
Hold Time
(min)
Temp.
Ramp
(°C/min)
Final
Temp.
(°C)
Final
Hold
Time (min)
1 200 2 5 220 16
2 5 235 7
3 5 330 5
Total time: 60 min
7.6.2 Mass Spectrometer
7.6.2.1 The mass spectrometer must be operated in a
selected ion monitoring (SIM) mode with a total cycle time
(including the voltage reset time) of one second or less (Sec.
7.6.3.1). At a minimum, the ions listed in Table 6 for each of the
five SIM descriptors must be monitored. Note that with the
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exception of the last descriptor (OCDD/OCDF), all descriptors
contain 10 ions. The selection (Table 6) of the molecular ions M
and M+2 for 13C-HxCDF and 13C-HpCDF rather than M+2 and M+4 (for
consistency) was made to eliminate, even under high-resolution mass
spectrometric conditions, interferences occurring in these two ion
channels for samples containing high levels of native HxCDDs and
HpCDDs. It is important to maintain the same set of ions for both
calibration and sample extract analyses. The selection of the lock-
mass ion is left to the performing laboratory.
NOTE: At the option of the analyst, the tetra- and
pentachlorinated dioxins and furans can be
combined into a single descriptor.
7.6.2.2 The recommended mass spectrometer tuning
conditions are based on the groups of monitored ions shown in Table
6. By using a PFK molecular leak, tune the instrument to meet the
minimum required resolving power of 10,000 (10 percent valley) at
m/z 304.9824 (PFK) or any other reference signal close to m/z
303.9016 (from TCDF). By using peak matching conditions and the
aforementioned PFK reference peak, verify that the exact mass of m/z
380.9760 (PFK) is within 5 ppm of the required value. Note that the
selection of the low- and high-mass ions must be such that they
provide the largest voltage jump performed in any of the five mass
descriptors (Table 6).
7.6.3 Data Acquisition
7.6.3.1 The total cycle time for data acquisition must be
< 1 second. The total cycle time includes the sum of all the dwell
times and voltage reset times.
7.6.3.2 Acquire SIM data for all the ions listed in the
five descriptors of Table 6.
7.7 Calibration
7.7.1 Initial Calibration - Initial calibration is required before
any samples are analyzed for PCDDs and PCDFs. Initial calibration is also
required if any routine calibration (Sec. 7.7.3) does not meet the
required criteria listed in Sec. 7.7.2.
7.7.1.1 All five high-resolution concentration calibration
solutions listed in Table 5 must be used for the initial
calibration.
7.7.1.2 Tune the instrument with PFK as described in
Sec. 7.6.2.2.
7.7.1.3 Inject 2 fil of the GC column performance check
solution (Sec. 5.7) and acquire SIM mass spectral data as described
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earlier in Sec. 7.6.2. The total cycle time must be < 1 second. The
laboratory must not perform any further analysis until it is demon-
strated and documented that the criterion listed in Sec. 8.2.1 was
met.
7.7.1.4 By using the same GC (Sec. 7.6.1) and MS
(Sec. 7.6.2) conditions that produced acceptable results with the
column performance check solution, analyze a 2 /LtL portion of each
of the five concentration calibration solutions once with the
following mass spectrometer operating parameters.
7.7.1.4.1 The ratio of integrated ion current for the
ions appearing in Table 8 (homologous series quantitation
ions) must be within the indicated control limits (set for
each homologous series) for all unlabeled calibration
standards in Table 5.
7.7.1.4.2 The ratio of integrated ion current for the
ions belonging to the carbon-labeled internal and recovery
standards (Table 5) must be within the control limits
stipulated in Table 8.
NOTE: Sees. 7.7.1.4.1 and 7.7.1.4.2 require that 17 ion
ratios from Sec. 7.7.1.4.1 and 11 ion ratios from
Sec. 7.7.1.4.2 be within the specified control
limits simultaneously in one run. It is the
laboratory's responsibility to take corrective
action if the ion abundance ratios are outside
the limits.
7.7.1.4.3 For each selected ion current profile (SICP)
and for each GC signal corresponding to the elution of a
target analyte and of its labeled standards, the signal-to-
noise ratio (S/N) must be better than or equal to 2.5.
Measurement of S/N is required for any GC peak that has an
apparent S/N of less than 5:1. The result of the calculation
must appear on the SICP above the GC peak in question.
7.7.1.4.4 Referring to Table 9, calculate the 17
relative response factors (RF) for unlabeled target analytes
[RF(n); n = 1 to 17] relative to their appropriate internal
standards (Table 5) and the nine RFs for the labeled 13C12
internal standards [RF(m); m = 18 to 26)] relative to the two
recovery standards (Table 5) according to the following
formulae:
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Ax x Qjs Ais x Qre
RFn = RFm =
Qx * Ais Qi8 x Are
where:
Ax = sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for
unlabeled PCDDs/PCDFs,
Ais = sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for the
labeled internal standards,
Ars = sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for the
labeled recovery standards,
Qis = quantity of the internal standard injected
(pg),
Qr8 = quantity of the recovery standard injected
(pg), and
Qx = quantity of the unlabeled PCDD/PCDF analyte
injected (pg).
The RFn and RFm are dimensionless quantities; the units
used to express Qis, Qre and Qx must be the same.
7.7.1.4.5 Calculate the RF and their respective
percent relative standard deviations (%RSO) for the five
calibration solutions:
5
RFn - 1/5 Z RFnW
Where n represents a particular PCDD/PCDF (2,3,7,8-
substituted) congener (n = 1 to 17; Table 9), and j is the
injection number (or calibration solution number; j = 1 to
5).
7.7.1.4.6 The relative response factors to be used for
the determination of the concentration of total isomers in a
homologous series (Table 9) are calculated as follows:
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7.7.1.4.6.1 For congeners that belong to a
homologous series containing only one isomer (e.g., OCDD
and OCDF) or only one 2,3,7,8-substituted isomer
(Table 4; TCDD, PeCDD, HpCDD, and_JCDF), the mean RF
used will be the same as the mean RF determined in Sec.
7.7.1.4.5.
NOTE: The calibration solutions do not contain
13C12-OCDF as an internal standard. This is
because a minimum resolving power of 12,000
is required to resolve the [M+6]+ ion of
13C12-OCDF from the [M+2]+ ion of OCDD (and
[M+4]+ from 13C12lOCDF with [M]+ of OCDD).
Therefore, the RF for OCDF is calculated
relative to 13C12-OCDD.
7.7.1.4.6.2 For congeners that belong to a
homologous series containing more than_ one
2,3,7,8-substituted isomer (Table 4), the mean RF used
for those homologous series will be the mean of the RFs
calculated for all individual 2,3,7,8-substituted
congeners using the equation below:
1 t
RFU = - I RFn
k
t n=1
where:
k = 27 to 30 (Table 9), with 27 = PeCDF; 28 =
HxCDF; 29 = HxCDD; and 30 = HpCDF,
t = total number of 2,3,7,8-substituted isomers
present in the calibration solutions (Table
5) for each homologous series (e.g., two
for PeCDF, four for HxCDF, three for HxCDD,
two for HpCDF).
NOTE: Presumably, the HRGC/HRMS response factors
of different isomers within a homologous
series are different. However, this
analytical protocol will make the
assumption that the HRGC/HRMS responses of
all isomers in a homologous series that do
not have the 2,3,7,8-substitution pattern
are the same as the responses of one or
more of the 2,3,7,8-substituted isomer(s)
in that homologous series.
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7.7.1.4.7 Relative response factors [RFm] to be used
for the determination of the percent recoveries for the nine
internal standards are calculated as follows:
RF
Qism * Are
5
RFm = 1/5 I RFm(j)
m(j),
where:
m = 18 to 26 (congener type) and j = 1 to 5
(injection number),
Aism = sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for a
given internal standard (m - 18 to 26),
Ars = sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for the
appropriate recovery standard (see Table 5,
footnotes),
Qrs> QiSm = quantities of, respectively, the recovery
standard (rs) and a particular internal
standard (is = m) injected (pg),
RFm = relative response factor of a particular
internal standard (m) relative to an
appropriate recovery standard, as
determined from one injection, and
RFm = calculated mean relative response factor of
a particular internal standard (m) relative
to an appropriate recovery standard, as
determined from the five initial calibra-
tion injections (j).
7.7.2 Criteria for Acceptable Calibration - The criteria listed
below for acceptable calibration must be met before sample analyses are
performed.
7.7.2.1 The percent relative standard deviations for the
mean response factors [RFn and RFm] from the 17 unlabeled standards
must not exceed + 20 percent, and those for the nine labeled
reference compounds must not exceed + 30 percent.
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7.7.2.2 The S/N for the GC signals present in every SICP
(including the ones for the labeled standards) must be > 10.
7.7.2.3 The ion abundance ratios (Table 8) must be within
the specified control limits.
NOTE: If the criterion for acceptable calibration
listed in Sec. 7.7.2.1 is met, the analyte-
specific RF can then be considered independent of
the analyte quantity for the calibration concen-
tration range. The mean RFs will be used for all
calculations until the routine calibration
criteria (Sec. 7.7.4) are no longer met. At such
time, new mean RFs will be calculated from a new
set of injections of the calibration solutions.
7.7.3 Routine Calibration (Continuing Calibration Check) - Routine
calibrations must be performed at the beginning of a 12-hour period after
successful mass resolution and GC resolution performance checks. A
routine calibration is also required at the end of a 12-hour shift.
7.7.3.1 Inject 2 ^L of the concentration calibration
solution HRCC-3 standard (Table 5). By using the same HRGC/HRMS
conditions as used in Sees. 7.6.1 and 7.6.2, determine and document
an acceptable calibration as provided in Sec. 7.7.4.
7.7.4 Criteria for Acceptable Routine Calibration - The following
criteria must be met before further analysis is performed.
7.7.4.1 The measured RFs [RFn for the unlabeled standards]
obtained during the routine calibration runs must be within + 20
percent of the mean values established during the initial
calibration (Sec. 7.7.1.4.5).
1.1 A.2 The measured RFs [RFm for the labeled standards]
obtained during the routine calibration runs must be within
+ 30 percent of the mean values established during the initial
calibration (Sec. 7.7.1.4.7).
7.7.4.3 The ion abundance ratios (Table 8) must be within
the allowed control limits.
7.7.4.4 If either one of the criteria in Sees. 7.7.4.1 and
1.1 A.2 is not satisfied, repeat one more time. If these criteria
are still not satisfied, the entire routine calibration process
(Sec. 7.7.1) must be reviewed. It is realized that it may not
always be possible to achieve all RF criteria. For example, it has
occurred that the RF criteria for 13C12-HpCDD and 13C12-OCDD were not
met, however, the RF values for the corresponding unlabeled
compounds were routinely within the criteria established in the
method. In these cases, 24 of the 26 RF parameters have met the QC
criteria, and the data quality for the unlabeled HpCDD and OCDD
values were not compromised as a result of the calibration event.
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In these situations, the analyst must assess the effect on overall
data quality as required for the data quality objectives and decide
on appropriate action. Corrective action would be in order, for
example, if the compounds for which the RF criteria were not met
included both the unlabeled and the corresponding internal standard
compounds. If the ion abundance ratio criterion (Sec. 7.7.4.3) is
not satisfied, refer to the note in Sec. 7.7.1.4.2 for resolution.
NOTE: An initial calibration must be carried out
whenever the HRCC-3, the sample fortification, or
the recovery standard solution is replaced by a
new solution from a different lot.
7.8 Analysis
7.8.1 Remove the sample or blank extract (from Sec. 7.5.3.6) from
storage. With a stream of dry, purified nitrogen, reduce the extract
volume to 10 /ul_ to 50 juL.
NOTE: A final volume of 20 ML or more should be used whenever
possible. A 10 juL final volume is difficult to handle, and
injection of 2 /uL out of 10 /uL leaves little sample for
confirmations and repeat injections, and for archiving.
7.8.2 Inject a 2 /iL aliquot of the extract into the GC, operated
under the conditions that have been established to produce acceptable
results with the performance check solution (Sees. 7.6.1 and 7.6.2).
7.8.3 Acquire SIM data according to Sees. 7.6.2 and 7.6.3. Use the
same acquisition and mass spectrometer operating conditions previously
used to determine the relative response factors (Sees. 7.7.1.4.4 through
7.7.1.4.7). Ions characteristic of polychlorinated diphenyl ethers are
included in the descriptors listed in Table 6.
NOTE: The acquisition period must at least encompass the PCDD/PCDF
overall retention time window previously determined (Sec.
8.2.1.3). Selected ion current profiles (SICP) for the lock-
mass ions (one per mass descriptor) must also be recorded and
included in the data package. These SICPs must be true
representations of the evolution of the lock-mass ions
amplitudes during the HRGC/HRMS run (see Sec. 8.2.2 for the
proper level of reference compound to be metered into the ion
chamber.) The analyst may be required to monitor a PFK ion,
riot as a lock-mass, but as a regular ion, in order to meet
this requirement. It is recommended to examine the lock-mass
ion SICP for obvious basic sensitivity and stability changes
of the instrument during the GC/MS run that could affect the
measurements [Tondeur et al., 1984, 1987]. Report any
discrepancies in the case narrative.
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7.8.4 Identification Criteria - For a gas chromatographic peak to
be identified as a PCDD or PCDF, it must meet all of the following
criteria:
7.8.4.1 Retention Times
7.8.4.1.1 For 2,3,7,8-substituted congeners, which
have an isotopically labeled internal or recovery standard
present in the sample extract (this represents a total of 10
congeners including OCDD; Tables 2 and 3), the retention time
(RRT; at maximum peak height) of the sample components (i.e.,
the two ions used for quantitation purposes listed in Table
6) must be within -1 to +3 seconds of the isotopically
labeled standard.
7.8.4.1.2 For 2,3,7,8-substituted compounds that do
not have an isotopically labeled internal standard present in
the sample extract (this represents a total of six congeners;
Table 3), the retention time must fall within 0.005 retention
time units of the relative retention times measured in the
routine calibration. Identification of OCDF is based on its
retention time relative to 13C12-OCDD as determined from the
daily routine calibration results.
7,8.4.1.3 For non-2,3,7,8-substituted compounds (tetra
through octa; totaling 119 congeners), the retention time
must be within the corresponding homologous retention time
windows established by analyzing the column performance check
solution (Sec. 8.1.3).
7.8.4.1.4 The ion current responses for both ions used
for quantitative purposes (e.g., for TCDDs: m/z 319.8965 and
321.8936) must reach maximum simultaneously (+ 2 seconds).
7.8.4.1.5 The ion current responses for both ions used
for the labeled standards (e.g., for 13C12-TCDD: m/z 331.9368
and m/z 333.9339) must reach maximum simultaneously (+ 2
seconds).
NOTE: The analyst is required to verify the presence of
1,2,8,9-TCDD and 1,3,4,6,8-PeCDF (Sec. 8.1.3) in
the SICPs of the daily performance checks.
Should either one compound be missing, the
analyst is required to take corrective action as
it may indicate a potential problem with the
ability to detect all the PCDDs/PCDFs.
7.8.4.2 Ion Abundance Ratios
7.8.4.2.1 The integrated ion currents for the two ions
used for quantitation purposes must have a ratio between the
lower and upper limits established for the homologous series
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to which the peak is assigned. See Sees. 7.7.1.4.1 and
7.7.1.4.2 and Table 8 for details.
7.8.4.3 Signal-to-Noise Ratio
7.8.4.3.1 All ion current intensities must be > 2.5
times noise level for positive identification of a PCDD/PCDF
compound or a group of coeluting isomers. Figure 6 describes
the procedure to be followed for the determination of the
S/N.
7.8.4.4 Polychlorinated Diphenyl Ether Interferences
7.8.4.4.1 In addition to the above criteria, the
identification of a GC peak as a PCDF can only be made if no
signal having a S/N > 2.5 is detected at the same retention
time (+ 2 seconds) in the corresponding polychlorinated
diphenyl ether (PCDPE, Table 6) channel.
7.9 Calculations
7.9.1 For gas chromatographic peaks that have met the criteria
outlined in Sees. 7.8.4.1.1 through 7.8.4.3.1, calculate the concentration
of the PCDD or PCDF compounds using the formula:
Av n
x A *
Ax == sum of the integrated ion abundances of the quantitation
ions (Table 6) for unlabeled PCDDs/PCDFs,
Ajs = sum of the integrated ion abundances of the quantitation
ions (Table 6) for the labeled internal standards,
Qis = quantity, in pg, of the internal standard added to the
sample before extraction,
W = weight, in g, of the sample (solid or organic liquid),
or volume in ml of an aqueous sample, and
RFn = calculated mean relative response factor for the analyte
[RFn with n = 1 to 17; Sec. 7.7.1.4.5].
If the analyte is identified as one of the 2,3,7,8-substituted PCDDs
or PCDFs, RFn is the value calculated using the equation in Sec. 7.7.1.4.5.
However, if it is a non-2,3,7,8-substituted congener, the RF(k) value is
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the one calculated using the equation in Sec. 7.7.1.4.6.2. [RFk k = 27
to 30].
7.9.2 Calculate the percent recovery of the nine internal standards
measured in the sample extract, using the formula:
Ais x Qre
Internal standard percent recovery = 1=- x 100
Qis x Ars x RFm
where:
Ais = sum of the integrated ion abundances of the quantitation
ions (Table 6) for the labeled internal standard,
Ars = sum of the integrated ion abundances of the quantitation
ions (Table 6) for the labeled recovery standard; the
selection of the recovery standard depends on the type
of congeners (see Table 5, footnotes),
Qis = quantity, in pg, of the internal standard added to the
sample before extraction,
Qrs = quantity, in pg, of the recovery standard added to the
cleaned-up sample residue before HRGC/HRMS analysis, and
RFm = calculated mean relative response factor for the labeled
internal standard relative to the appropriate (see Table
5, footnotes) recovery standard. This represents the
mean obtained in Sec. 7.7.1.4.7 [RFm with m = 18 to 26].
NOTE: For human adipose tissue, adjust the percent recoveries by
adding 1 percent to the calculated value to compensate for
the 1 percent of the extract diverted for the lipid
determination.
7.9.3 If the concentration in the final extract of any of the
fifteen 2,3,7,8-substituted PCDD/PCDF compounds (Table 3) exceeds the
upper method calibration limits (MCL) listed in Table 1 (e.g., 200 pg//uL
for TCDD in soil), the linear range of response versus concentration may
have been exceeded, and a second analysis of the sample (using a one tenth
aliquot) should be undertaken. The volumes of the internal and recovery
standard solutions should remain the same as described for the sample
preparation (Sees. 7.1 to 7.9.3). For the other congeners (including
OCDD), however, report the measured concentration and indicate that the
value exceeds the MCL.
7.9.3.1 If a smaller sample size would not be
representative of the entire sample, one of the following options is
recommended:
(1) Re-extract an additional aliquot of sufficient size to insure
that it is representative of the entire sample. Spike it with a
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higher concentration of internal standard. Prior to GC/MS analysis,
dilute the sample so that it has a concentration of internal
standard equivalent to that present in the calibration standard.
Then, analyze the diluted extract.
(2) Re-extract an additional aliquot of sufficient size to insure
that it is representative of the entire sample. Spike it with a
higher concentration of internal standard. Immediately following
extraction, transfer the sample to a volumetric flask and dilute to
known volume. Remove an appropriate aliquot and proceed with
cleanup and analysis.
(3) Use the original analysis data to quantitate the internal
standard recoveries. Respike the original extract (note that no
additional cleanup is necessary) with 100 times the usual quantity
of internal standards. Dilute the re-spiked extract by a factor of
100. Reanalyze the diluted sample using the internal standard
recoveries calculated from the initial analysis to correct the
results for losses during isolation and cleanup.
7.9.4 The total concentration for each homologous series of PCDD and
PCDF is calculated by summing up the concentrations of all positively
identified isomers of each homologous series. Therefore, the total should
also include the 2,3,7,8-substituted congeners. The total number of GC
signals included in the homologous total concentration value must be
specified in the report. If an isomer is not detected, use zero (0) in
this calculation.
7.9.5 Sample Specific Estimated Detection Limit - The sample
specific estimated detection limit (EDL) is the concentration of a given
analyte required to produce a signal with a peak height of at least 2.5
times the background signal level. An EDL is calculated for each
2,3,7,8-substituted congener that is not identified, regardless of whether
or not other non-2,3,7,8-substituted isomers are present. Two methods of
calculation can be used, as follows, depending on the type of response
produced during the analysis of a particular sample.
7.9.5.1 Samples giving a response for both quantitation
ions (Tables 6 and 9) that is less than 2.5 times the background
level.
7.9.5.1.1 Use the expression for EDL (specific
2,3,7,8-substituted PCDD/PCDF) below to calculate an EDL for
each absent 2,3,7,8-substituted PCDD/PCDF (i.e., S/N < 2.5).
The background level is determined by measuring the range of
the noise (peak to peak) for the two quantitation ions (Table
6) of a particular 2,3,7,8-substituted isomer within an
homologous series, in the region of the SICP trace
corresponding to the elution of the internal standard (if the
congener possesses an internal standard) or in the region of
the SICP where the congener is expected to elute by
comparison with the routine calibration data (for those
congeners that do not have a 13C-labeled standard),
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multiplying that noise height by 2.5, and relating the
product to an estimated concentration that would produce that
peak height.
Use the formula:
2.5 x Hx x Qis
EDL (specific 2,3,7,8-subst. PCDD/PCDF) =
His x W x RFn
where:
EDL = estimated detection limit for homologous
2,3,7,8-substituted PCDDs/PCDFs.
Hx = sum of the height of the noise level for each
quantitation ion (Table 6) for the unlabeled
PCDDs/PCDFs, measured as shown in Figure 6.
His = sum of the height of the noise level for each
quantitation ion (Table 6) for the labeled
internal standard, measured as shown in Figure 6.
W, RFn, and Qis retain the same meanings as defined in
Sec. 7.9.1.
7.9.5.2 Samples characterized by a response above the
background level with a S/N of at least 2.5 for both quantitation
ions (Tables 6 and 9).
7.9.5.2.1 When the response of a signal having the
same retention time as a 2,3,7,8-substituted congener has a
S/N in excess of 2.5 and does not meet any of the other
qualitative identification criteria listed in Sec. 7.8.4,
calculate the "Estimated Maximum Possible Concentration"
(EMPC) according to the expression shown in Sec. 7.9.1,
except that Ax in Sec. 7.9.1 should represent the sum of the
area under the smaller peak and of the other peak area
calculated using the theoretical chlorine isotope ratio.
7.9.6 The relative percent difference (RPD) of any duplicate sample
results are calculated as follows:
I S, - S2 |
RPD = x 100
(S, + S2 ) / 2
S., and S2 represent sample and duplicate sample results.
7.9.7 The 2,3,7,8-TCDD toxicity equivalents (TE) of PCDDs and PCDFs
present in the sample are calculated, if requested by the data user,
according to the method recommended by the Chlorinated Dioxins Workgroup
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(CDWG) of the EPA and the Center for Disease Control (CDC). This method
assigns a 2,3,7,8-TCDD toxicity equivalency factor (TEF) to each of the
fifteen 2,3,7,8-substituted PCDDs and PCDFs (Table 3) and to OCDD and
OCDF, as shown in Table 10. The 2,3,7,8-TCDD equivalent of the PCDDs and
PCDFs present in the sample is calculated by summing the TEF times their
concentration for each of the compounds or groups of compounds listed in
Table 10. The exclusion of other homologous series such as mono-, di-,
and tri- chlorinated dibenzodioxins and dibenzofurans does not mean that
they are non-toxic. However, their toxicity, as known at this time, is
much lower than the toxicity of the compounds listed in Table 10. The
above procedure for calculating the 2,3,7,8-TCDD toxicity equivalents is
not claimed by the CDWG to be based on a thoroughly established scientific
foundation. The procedure, rather, represents a "consensus recommendation
on science policy". Since the procedure may be changed in the future,
reporting requirements for PCDD and PCDF data would still include the
reporting of the analyte concentrations of the PCDD/PCDF congener as
calculated in Sees. 7.9.1 and 7.9.4.
7.9.7.1 Two GC Column TEF Determination
7.9.7.1.1 The concentration of 2,3,7,8-TCDD (see note
below), is calculated from the analysis of the sample extract
on the 60 m DB-5 fused silica capillary column. The
experimental conditions remain the same as the conditions
described previously in Sec. 7.8, and the calculations are
performed as outlined in Sec. 7.9. The chromatographic
separation between the 2,3,7,8-TCDD and its close eluters
(1,2,3,7/1,2,3,8-TCDD and 1,2,3,9-TCDD) must be equal or less
than 25 percent valley.
7.9.7.1.2 The concentration of the 2,3,7,8-TCDF is
obtained from the analysis of the sample extract on the 30 m
DB-225 fused silica capillary column. However, the GC/MS
conditions must be altered so that: (1) only the first three
descriptors (i.e., tetra-, penta-, and hexachlorinated
congeners) of Table 6 are used; and (2) the switching time
between descriptor 2 (pentachlorinated congeners) and
descriptor 3 (hexachlorinated congeners) takes place
following the elution of 13C12-l,2,3,7,8-PeCDD. The
concentration calculations are performed as outlined in Sec.
7.9. The chromatographic separation between the 2,3,7,8-TCDF
and its close eluters (2,3,4,7-TCDF and 1,2,3,9-TCDF) must be
equal or less than 25 percent valley.
NOTE: The confirmation and quantitation of 2,3,7,8-TCDD
(Sec. 7.9.7.1.1) may be accomplished on the SP-
2330 GC column instead of the DB-5 column,
provided the criteria listed in Sec. 8.2.1 are
met and the requirements described in Sec.
8.3.2 are followed.
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7.9.7.1.3 For a gas chromatographic peak to be
identified as a 2,3,7,8-substituted PCDD/PCDF congener, it
must meet the ion abundance and signal-to-noise ratio
criteria listed in Sees. 7.8.4.2 and 7.8.4.3, respectively.
In addition, the retention time identification criterion
described in Sec. 7.8.4.1.1 applies here for congeners for
which a carbon-labeled analogue is available in the sample
extract. However, the relative retention time (RRT) of the
2,3,7,8-substituted congeners for which no carbon-labeled
analogues are available must fall within 0.006 units of the
carbon-labeled standard RRT. Experimentally, this is
accomplished by using the attributions described in Table 11
and the results from the routine calibration run on the
SP-2330 column.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control (QC) procedures.
Quality control to validate sample extraction is covered in Method 3500. If
extract cleanup was performed, follow the QC in Method 3600 and in the specific
cleanup method.
8.2 System Performance Criteria - System performance criteria are
presented below. The laboratory may use the recommended GC column described in
Sec. 4.2. It must be documented that all applicable system performance criteria
(specified in Sees. 8.2.1 and 8.2.2) were met before analysis of any sample is
performed. Sec. 7.6.1 provides recommended GC conditions that can be used to
satisfy the required criteria. Figure 3 provides a typical 12-hour analysis
sequence, whereby the response factors and mass spectrometer resolving power
checks must be performed at the beginning and the end of each 12-hour period of
operation. A GC column performance check is only required at the beginning of
each 12-hour period during which samples are analyzed. An HRGC/HRMS method blank
run is required between a calibration run and the first sample run. The same
method blank extract may thus be analyzed more than once if the number of samples
within a batch requires more than 12 hours of analyses.
8.2.1 GC Column Performance
8.2.1.1 Inject 2 /iL (Sec. 4.1.1) of the column performance
check solution (Sec. 5.7) and acquire selected ion monitoring (SIM)
data as described in Sec. 7.6.2 within a total cycle time of < 1
second (Sec. 7.6.3.1).
8.2.1.2 The chromatographic separation between 2,3,7,8-
TCDD and the peaks representing any other unlabeled TCDD isomers
must be resolved with a valley of < 25 percent (Figure 4), where:
Valley percent = (x/y) (100)
x = measured as in Figure 4 from the 2,3,7,8-closest TCDD
eluting isomer, and
y = the peak height of 2,3,7,8-TCDD.
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It is the responsibility of the laboratory to verify the
conditions suitable for the appropriate resolution of 2,3,7,8-TCDD
from all other TCDD isomers. The GC column performance check
solution also contains the known first and last PCDD/PCDF eluters
under the conditions specified in this protocol. Their retention
times are used to determine the eight homologue retention time
windows that are used for qualitative (Sec. 7.8.4.1) and
quantitative purposes. All peaks (that includes 13C12-2,3,7,8-TCDD)
should be labeled and identified on the chromatograms. Furthermore,
all first eluters of a homologous series should be labeled with the
letter F, and all last eluters of a homologous series should be
labeled with the letter L (Figure 4 shows an example of peak
labeling for TCDD isomers). Any individual selected ion current
profile'(SICP) (for the tetras, this would be the SICP for m/z 322
and m/z 304) or the reconstructed homologue ion current (for the
tetras, this would correspond to m/z 320 + m/z 322 + m/z 304 + m/z
306) constitutes an acceptable form of data presentation. An SICP
for the labeled compounds (e.g., m/z 334 for labeled TCDD) is also
required.
8.2.1.3 The retention times for the switching of SIM ions
characteristic of one homologous series to the next higher
homologous series must be indicated in the SICP. Accurate switching
at the appropriate times is absolutely necessary for accurate
monitoring of these compounds. Allowable tolerance on the daily
verification with the GC performance check solution should be better
than 10 seconds for the absolute retention times of all the
components of the mixture. Particular caution should be exercised
for the switching time between the last tetrachlorinated congener
(i.e., 1,2,8,9-TCDD) and the first pentachlorinated congener (i.e.,
1,3,4,6,8-PeCDF), as these two compounds elute within 15 seconds of
each other on the 60 m DB-5 column. A laboratory with a GC/MS
system that is not capable of detecting both congeners (1,2,8,9-TCDD
and 1,3,4,6,8-PeCDF) within one analysis must take corrective
action. If the recommended column is not used, then the first and
last eluting isomer of each homologue must be determined
experimentally on the column which is used, and the appropriate
isomers must then be used for window definition and switching times.
8.2.2 Mass Spectrometer Performance
8.2.2.1 The mass spectrometer must be operated in the
electron ionization mode. A static resolving power of at least
10,000 (10 percent valley definition) must be demonstrated at
appropriate masses before any analysis is performed (Sec. 7.8).
Static resolving power checks must be performed at the beginning and
at the end of each 12 hour period of operation. However, it is
recommended that a check of the static resolution be made and
documented before and after each analysis. Corrective action must
be implemented whenever the resolving power does not meet the
requirement.
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8.2.2.2 Chromatography time for PCDDs and PCDFs exceeds
the long term mass stability of the mass spectrometer. Because the
instrument is operated in the high-resolution mode, mass drifts of
a few ppm (e.g., 5 ppm in mass) can have serious adverse effects on
instrument performance. Therefore, a mass drift correction is
mandatory. To that effect, it is recommended to select a lock-mass
ion from the reference compound (PFK is recommended) used for tuning
the mass spectrometer. The selection of the lock-mass ion is
dependent on the masses of the ions monitored within each
descriptor. Table 6 offers some suggestions for the lock-mass ions.
However, an acceptable lock-mass ion at any mass between the
lightest and heaviest ion in each descriptor can be used to monitor
and correct mass drifts. The level of the reference compound (PFK)
metered into the ion chamber during HRGC/HRMS analyses should be
adjusted so that the amplitude of the most intense selected lock-
mass ion signal (regardless of the descriptor number) does not
exceed 10 percent of the full scale deflection for a given set of
detector parameters. Under those conditions, sensitivity changes
that might occur during the analysis can be more effectively
monitored.
NOTE: Excessive PFK (or any other reference substance) may cause
noise problems and contamination of the ion source resulting
in an increase in downtime for source cleaning.
8.2.2.3 Documentation of the instrument resolving power
must then be accomplished by recording the peak profile of the high-
mass reference signal (m/z 380.9760) obtained during the above peak
matching experiment by using the low-mass PFK ion at m/z 304.9824 as
a reference. The minimum resolving power of 10,000 must be
demonstrated on the high-mass ion while it is transmitted at a lower
accelerating voltage than the low-mass reference ion, which is
transmitted at full sensitivity. The format of the peak profile
representation (Figure 5) must allow manual determination of the
resolution, i.e., the horizontal axis must be a calibrated mass
scale (amu or ppm per division). The result of the peak width
measurement (performed at 5 percent of the maximum, which
corresponds to the 10 percent valley definition) must appear on the
hard copy and cannot exceed 100 ppm at m/z 380.9760 (or 0.038 amu at
that particular mass).
8.3 Quality Control Samples
8.3.1 Performance Evaluation Samples - Included among the samples
in all batches may be samples (blind or double blind) containing known
amounts of unlabeled 2,3,7,8-substituted PCDDs/PCDFs or other PCDD/PCDF
congeners.
8.3.2 Performance Check Solutions
8.3.2.1 At the beginning of each 12-hour period during
which samples are to be analyzed, an aliquot of the 1) GC column
performance check solution and 2) high-resolution concentration
calibration solution No. 3 (HRCC-3; see Table 5) shall be analyzed
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to demonstrate adequate GC resolution and sensitivity, response
factor reproducibility, and mass range calibration, and to establish
the PCDD/PCDF retention time windows. A mass resolution check shall
also be performed to demonstrate adequate mass resolution using an
appropriate reference compound (PFK is recommended). If the
required criteria are not met, remedial action must be taken before
any samples are analyzed.
8.3.2.2 To validate positive sample data, the routine or
continuing calibration (HRCC-3; Table 5) and the mass resolution
check must be performed also at the end of each 12-hour period
during which samples are analyzed. Furthermore, an HRGC/HRMS method
blank run must be recorded following a calibration run and the first
sample run.
8.3.2.2.1 If the laboratory operates only during one
period (shift) each day of 12 hours or less, the GC
performance check solution must be analyzed only once (at the
beginning of the period) to validate the data acquired during
the period. However, the mass resolution and continuing
calibration checks must be performed at the beginning as well
as at the end of the period.
8.3.2.2.2 If the laboratory operates during
consecutive 12-hour periods (shifts), analysis of the GC
performance check solution must be performed at the beginning
of each 12-hour period. The mass resolution and continuing
calibration checks from the previous period can be used for
the beginning of the next period.
8.3.2.3 Results of at least one analysis of the GC column
performance check solution and of two mass resolution and continuing
calibration checks must be reported with the sample data collected
during a 12 hour period.
8.3.2.4 Deviations from criteria specified for the GC
performance check or for the mass resolution check invalidate all
positive sample data collected between analyses of the performance
check solution, and the extracts from those positive samples shall
be reanalyzed.
If the routine calibration run fails at the beginning of a 12
hour shift, the instructions in Sec. 7.7.4.4 must be followed. If
the continuing calibration check performed at the end of a 12 hour
period fails by no more than 25 percent RPD for the 17 unlabeled
compounds and 35 percent RPD for the 9 labeled reference compounds,
use the mean RFs from the two daily routine calibration runs to
compute the analyte concentrations, instead of the RFs obtained from
the initial calibration. A new initial calibration (new RFs) is
required immediately (within two hours) following the analysis of
the samples, whenever the RPD from the end-of-shift routine
calibration exceeds 25 percent or 35 percent, respectively. Failure
to perform a new initial calibration immediately following the
analysis of the samples will automatically require reanalysis of all
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positive sample extracts analyzed before the failed end-of-shift
continuing calibration check.
8.3.3 The GC column performance check mixture, high-resolution
concentration calibration solutions, and the sample fortification
solutions may be obtained from the EMSL-CIN. However, if not available
from the EMSL-CIN, standards can be obtained from other sources, and
solutions can be prepared in the laboratory. Concentrations of all
solutions containing 2,3,7,8-substituted PCDDs/PCDFs, which are not
obtained from the EMSL-CIN, must be verified by comparison with the EPA
standard solutions that are available from the EMSL-CIN.
8.3.4 Field Blanks - Each batch of samples usually contains a field
blank sample of uncontaminated soil, sediment or water that is to be
fortified before analysis according to Sec. 8.3.4.1. In addition to this
field blank, a batch of samples may include a rinsate, which is a portion
of the solvent (usually trichloroethylene) that was used to rinse sampling
equipment. The rinsate is analyzed to assure that the samples were not
contaminated by the sampling equipment.
8.3.4.1 Fortified Field Blank
8.3.4.1.1 Weigh a 10 g portion or use 1 L (for aqueous
samples) of the specified field blank sample and add 100 juL
of the solution containing the nine internal standards
(Table 2) diluted with 1.0 mL acetone (Sec. 7.1).
8.3.4.1.2 Extract by using the procedures beginning
in Sees. 7.4.5 or 7.4.6, as applicable, add 10 juL of the
recovery standard solution (Sec. 7.5.3.6) and analyze a 2 /zL
aliquot of the concentrated extract.
8.3.4.1.3 Calculate the concentration (Sec. 7.9.1) of
2,3,7,8-substituted PCDDs/PCDFs and the percent recovery of
the internal standards (Sec. 7.9.2).
8.3.4.1.4 Extract and analyze a new simulated
fortified field blank whenever new lots of solvents or
reagents are used for sample extraction or for column
chromatographic procedures.
8.3.4.2 Rinsate Sample
8.3.4.2.1 The rinsate sample must be fortified like
a regular sample.
8.3.4.2.2 Take a 100 mL (+ 0.5 mL) portion of the
sampling equipment rinse solvent (rinsate sample), filter, if
necessary, and add 100 /uL of the solution containing the nine
internal standards (Table 2).
8.3.4.2.3 Using a KD apparatus, concentrate to
approximately 5 mL.
8290 - 41 Revision 0
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NOTE: As an option, a rotary evaporator may be used in
place of the KD apparatus for the concentration
of the rinsate.
8.3.4.2.4 Transfer the 5 ml concentrate from the KD
concentrator tube in 1 mL portions to a 1 ml minivial,
reducing the volume in the minivial as necessary with a
gentle stream of dry nitrogen.
8.3.4.2.5 Rinse the KD concentrator tube with two
0.5 ml portions of hexane and transfer the rinses to the 1 ml
minivial. Blow down with dry nitrogen as necessary.
8.3.4.2.6 Just before analysis, add 10 jitL recovery
standard solution (Table 2) and reduce the volume to its
final volume, as necessary (Sec. 7.8.1). No column
chromatography is required.
8.3.4.2.7 Analyze an aliquot following the same
procedures used to analyze samples.
8.3.4.2.8 Report percent recovery of the internal
standard and the presence of any PCDD/PCDF compounds in jug/L
of rinsate solvent.
8.3.5 Duplicate Analyses
8.3.5.1 In each batch of samples, locate the sample
specified for duplicate analysis, and analyze a second 10 g soil or
sediment sample portion or 1 L water sample, or an appropriate
amount of the type of matrix under consideration.
8.3.5.1.1 The results of the laboratory duplicates
(percent recovery and concentrations of 2,3,7,8-substituted
PCDD/PCDF compounds) should agree within 25 percent relative
difference (difference expressed as percentage of the mean).
Report all results.
8.3.5.1.2 Recommended actions to help locate problems:
8.3.5.1.2.1 Verify satisfactory instrument
performance (Sees. 8.2 and 8.3).
8.3.5.1.2.2 If possible, verify that no error was
made while weighing the sample portions.
8.3.5.1.2.3 Review the analytical procedures with
the performing laboratory personnel.
8.3.6 Matrix Spike and Matrix Spike Duplicate
8..3.6.1 Locate the sample for the MS and MSD analyses (the
sample may be labeled "double volume").
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8.3.6.2 Add an appropriate volume of the matrix spike
fortification solution (Sec. 5.10) and of the sample fortification
solution (Sec. 5.8), adjusting the fortification level as specified
in Table 1 under IS Spiking Levels.
8.3.6.3 Analyze the MS and MSD samples as described in
Sec. 7.
8.3.6.4 The results obtained from the MS and MSD samples
(concentrations of 2,3,7,8-substituted PCDDs/PCDFs) should agree
within 20 percent relative difference.
8.4 Percent Recovery of the Internal Standards - For each sample, method
blank and rinsate, calculate the percent recovery (Sec. 7.9.2). The percent
recovery should be between 40 percent and 135 percent for all 2,3,7,8-substituted
internal standards.
NOTE; A low or high percent recovery for a blank does not require
discarding the analytical data but it may indicate a
potential problem with future analytical data.
8.5 Identification Criteria
8.5.1 If either one of the identification criteria appearing in
Sees. 7.8.4.1.1 through 7.8.4.1.4 is not met for an homologous series, it
is reported that the sample does not contain unlabeled 2,3,7,8-substituted
PCDD/PCDF isomers for that homologous series at the calculated detection
limit (Sec. 7.9.5)
8.5.2 If the first initial identification criteria (Sees. 7.8.4.1.1
through 7.8.4.1.4) are met, but the criteria appearing in Sees. 7.8.4.1.5
and 7.8.4.2.1 are not met, that sample is presumed to contain interfering
contaminants. This must be noted on the analytical report form, and the
sample should be rerun or the extract reanalyzed.
8.6 Unused portions of samples and sample extracts should be preserved
for six months after sample receipt to allow further analyses.
8.7 Reuse of glassware is to be minimized to avoid the risk of
contamination.
9.0 METHOD PERFORMANCE
9.1 Data are currently not available.
10.0 REFERENCES
1. "Control of Interferences in the Analysis of Human Adipose Tissue for
2,3,7,8-Tetrachlorodibenzo-p-dioxin". D. G. Patterson, J.S. Holler, D.F.
Grote, L.R. Alexander, C.R. Lapeza, R.C. O'Connor and J.A. Liddle.
Environ. Toxicol. Chem. 5, 355-360 (1986).
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2. "Method 8290: Analytical Procedures and Quality Assurance for Multimedia
Analysis of Polychlorinated Dibenzo-p-Dioxins and Dibenzofurans by High-
Resolution Gas Chromatography/High-Resolution Mass Spectrometry". Y.
Tondeur and W.F. Beckert. U.S. Environmental Protection Agency,
Environmental Monitoring Systems Laboratory, Las Vegas, NV.
3. "Carcinogens - Working with Carcinogens", Department of Health, Education,
and Welfare, Public Health Service, Center for Disease Control. National
Institute for Occupational Safety and Health. Publication No. 77-206,
August 1977.
4. "OSHA Safety and Health Standards, General Industry", (29 CFR 1910),
Occupational Safety and Health Administration, OSHA 2206 (revised January
1976).
5. "Safety in Academic Chemistry Laboratories", American Chemical Society
Publication, Committee on Chemical Safety (3rd Edition, 1979.)
6. "Hybrid HRGC/MS/MS Method for the Characterization of Tetrachlorinated
Dibenzo-p-dioxins in Environmental Samples." Y. Tondeur, W.J. Niederhut,
S.R. Missler, and J.E. Campana, Mass Spectrom. 14, 449-456 (1987).
7. USEPA National Dioxin Study - Phase II, "Analytical Procedures and Quality
Assurance Plan for the Determination of PCDD/PCDF in Fish", EPA-Duluth,
October 26, 1987.
11.0 SAFETY
11.1 The following safety practices are excerpts from EPA Method 613,
Sec. 4 (July 1982 version) and amended for use in conjunction with this method.
The 2,3,7,8-TCDD isomer has been found to be acnegenic, carcinogenic, and
teratogenic in laboratory animal studies. Other PCDDs and PCDFs containing
chlorine atoms in positions 2,3,7,8 are known to have toxicities comparable to
that of 2,3,7,8-TCDD. The analyst should note that finely divided dry soils
contaminated with PCDDs and PCDFs are particularly hazardous because of the
potential for inhalation and ingestion. It is recommended that such samples be
processed in a confined environment, such as a hood or a glove box. Laboratory
personnel handling these types of samples should wear masks fitted with charcoal
filters to prevent inhalation of dust.
11.2 The toxicity or carcinogenicity of each reagent used in this method
is not precisely defined; however, each chemical compound should be treated as
a potential health hazard. From this viewpoint, exposure to these chemicals must
be kept to a minimum. The laboratory is responsible for maintaining a current
awareness file of OSHA regulations regarding the safe handling of the chemicals
specified in this method. A reference file of material safety data sheets should
be made available to all personnel involved in the chemical analysis of samples
suspected to contain PCDDs and/or PCDFs. Additional references to laboratory
safety are given in references 3, 4 and 5.
11.3 Each laboratory must develop a strict safety program for the handling
of PCDDs and PCDFs. The laboratory practices listed below are recommended.
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11.3.1 Contamination of the laboratory will be minimized by
conducting most of the manipulations in a hood.
11.3.2 The effluents of sample splitters for the gas
chromatograph and roughing pumps on the HRGC/HRMS system should pass
through either a column of activated charcoal or be bubbled through a trap
containing oil or high boiling alcohols.
11.3.3 Liquid waste should be dissolved in methanol or ethanol
and irradiated with ultraviolet light at a wavelength less than 290 nm for
several days (use F 40 BL lamps, or equivalent). Using this analytical
method, analyze the irradiated liquid wastes and dispose of the solutions
when 2,3,7,8-TCDD and -TCDF congeners can no longer be detected.
11.4 The following precautions were issued by Dow Chemical U.S.A. (revised
11/78) for safe handling of 2,3,7,8-TCDD in the laboratory and amended for use
in conjunction with this method.
11.4.1 The following statements on safe handling are as complete
as possible on the basis of available toxicological information. The
precautions for safe handling and use are necessarily general in nature
since detailed, specific recommendations can be made only for the
particular exposure and circumstances of each individual use. Assistance
in evaluating the health hazards of particular plant conditions may be
obtained from certain consulting laboratories and from State Departments
of Health or of Labor, many of which have an industrial health service.
The 2,3,7,8-TCDD isomer is extremely toxic to certain kinds of laboratory
animals. However, it has been handled for years without injury in
analytical and biological laboratories. Many techniques used in handling
radioactive and infectious materials are applicable to 2,3,7,8-TCDD.
11.4.1.1 Protective Equipment: Throw away plastic gloves,
apron or lab coat, safety glasses and laboratory hood adequate for
radioactive work. However, PVC gloves should not be used.
11.4.1.2 Training: Workers must be trained in the proper
method of removing contaminated gloves and clothing without
contacting the exterior surfaces.
11.4.1.3 Personal Hygiene: Thorough washing of hands and
forearms after each manipulation and before breaks (coffee, lunch,
and shift).
11.4.1.4 Confinement: Isolated work area, posted with
signs, segregated glassware and tools, plastic backed absorbent
paper on benchtops.
11.4.1.5 Waste: Good technique includes minimizing
contaminated waste. Plastic bag liners should be used in waste
cans.
11.4.1.6 Disposal of Hazardous Wastes: Refer to the
November 7, 1986 issue of the Federal Register on Land Ban Rulings
for details concerning the handling of dioxin containing wastes.
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11.4.1.7 Decontamination: Personnel - apply a mild soap
with plenty of scrubbing action. Glassware, tools and surfaces -
Chlorothene NU Solvent (Trademark of the Dow Chemical Company) is
the least toxic solvent shown to be effective. Satisfactory
cleaning may be accomplished by rinsing with Chlorothene, then
washing with a .detergent and water. Dish water may be disposed to
the sewer after percolation through a charcoal bed filter. It is
prudent to minimize solvent wastes because they require special
disposal through commercial services that are expensive.
11.4.1.8 Laundry: Clothing known to be contaminated should
be disposed with the precautions described under "Disposal of
Hazardous Wastes". Laboratory coats or other clothing worn in
2,3,7,8-TCDD work area may be laundered. Clothing should be
collected in plastic bags. Persons who convey the bags and launder
the clothing should be advised of the hazard and trained in proper
handling. The clothing may be put into a washer without contact if
the launderer knows the problem. The washer should be run through
one full cycle before being used again for other clothing.
11.4.1.9 Wipe Tests: A useful method for determining
cleanliness of work surfaces and tools is to wipe the surface with
a piece of filter paper, extract the filter paper and analyze the
extract.
NOTE: A procedure for the collection, handling,
analysis, and reporting requirements of wipe
tests performed within the laboratory is
described in Attachment A. The results and
decision making processes are based on the
presence of 2,3,7,8-substituted PCDDs/PCDFs.
11.4.1.10 Inhalation: Any procedure that may generate
airborne contamination must be carried out with good ventilation.
Gross losses to a ventilation system must not be allowed. Handling
of the dilute solutions normally used in analytical and animal work
presents no significant inhalation hazards except in case of an
accident.
11.4.1.11 Accidents: Remove contaminated clothing
immediately, taking precautions not to contaminate skin or other
articles. Wash exposed skin vigorously and repeatedly until medical
attention is obtained.
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Attachment A
PROCEDURES FOR THE COLLECTION, HANDLING, ANALYSIS, AND
REPORTING OF WIPE TESTS PERFORMED WITHIN THE LABORATORY
This procedure is designed for the periodic evaluation of potential con-
tamination by 2,3,7,8-substituted PCDD/PCDF congeners of the working areas inside
the laboratory.
A.I Perform the wipe tests on surface areas of two inches by one foot
with glass fiber paper saturated with distilled in glass acetone using a pair of
clean stainless steel forceps. Use one wiper for each of the designated areas.
Combine the wipers to one composite sample in an extraction jar containing 200
mL distilled in glass acetone. Place an equal number of unused wipers in 200 mL
acetone and use this as a control. Add 100 fj.1 of the sample fortification
solution to each jar containing used or unused wipers (Sec. 5.8).
A. 1.1 Close the jar containing the wipers and the acetone and
extract for 20 minutes using a wrist action shaker. Transfer the extract
into a KD apparatus fitted with a concentration tube and a three ball
Snyder column. Add two Teflon™ or Carborundum™ boiling chips and
concentrate the extract to an apparent volume of 1.0 mL on a steam bath.
Rinse the Snyder column and the KD assembly with two 1 mL portions of
hexane into the concentrator tube, and concentrate its contents to near
dryness with a gentle stream of nitrogen. Add 1.0 mL hexane to the
concentrator tube and swirl the solvent on the walls.
A.1.2 Prepare a neutral alumina column as described in Sec. 7.5.2.2
and follow the steps outlined in Sees. 7.5.2.3 through 7.5.2.5.
A. 1.3 Add 10 juL of the recovery standard solution as described in
Sec. 7.5.3.6.
A.2 Concentrate the contents of the vial to a final volume of 10 /xL
(either in a minivial or in a capillary tube). Inject 2 jtiL of each extract
(wipe and control) onto a capillary column and analyze for 2,3,7,8-substituted
PCDDs/PCDFs as specified in the analytical method in Sec. 7.8. Perform
calculations according to Sec. 7.9.
A.3 Report the presence of 2,3,7,8-substituted PCDDs and PCDFs as a
quantity (pg or ng) per wipe test experiment (WTE). Under the conditions out-
lined in this analytical protocol, a lower limit of calibration of 10 pg/WTE is
expected for 2,3,7,8-TCDD. A positive response for the blank (control) is
defined as a signal in the TCDD retention time window at any of the masses
monitored which is equivalent to or above 3 pg of 2,3,7,8-TCDD per WTE. For
other congeners, use the multiplication factors listed in Table 1, footnote (a)
(e.g., for OCDD, the lower MCL is 10 x 5 = 50 pg/WTE and the positive response
for the blank would be 3 x 5 = 15 pg). Also, report the recoveries of the
internal standards during the simplified cleanup procedure.
A.4 At a minimum, wipe tests should be performed when there is evidence
of contamination in the method blanks.
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A.5 An upper limit of 25 pg per TCDD isomer and per wipe test experiment
is allowed (use multiplication factors listed in footnote (a) from Table 1 for
other congeners). This value corresponds to 2? times the lower calibration limit
of the analytical method. Steps to correct the contamination must be taken
whenever these levels are exceeded. To that effect, first vacuum the working
places (hoods, benches, sink) using a vacuum cleaner equipped with a high
efficiency particulate absorbent (HEPA) filter and then wash with a detergent.
A new set of wipes should be analyzed before anyone is allowed to work in the
dioxin area of the laboratory after corrective action has been taken.
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Figure 1.
8
Dibenzodioxin
8
Dibenzofuran
General structures of dibenzo-p-dioxin and dibenzofuran.
8290 - 49
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Figure 2.
M/AM
5,600
5,600
8,550
Peak profile displays demonstrating the effect of the detector zero on the
measured resolving power. In this example, the true resolving power is 5,600.
A) The zero was set too high; no effect is observed upon the
measurement of the resolving power.
B) The zero was adjusted properly.
C) The zero was set too low; this results in overestimating the actual
resolving power because the peak-to-peak noise cannot be measured
accurately.
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Figure 3.
Analytical Procedure
8:00 AM
Mass Resolution
Mass Accuracy
Thaw Sample Extract
I
Concentrate to 10
I
9:00 AM
Initial or
Routine
Calibration
QC Column
Performance
11:00 AM
Method
Blank
8:00 PM
Mass
Resolution
Routine
Calibration
Typical 12 hour analysis sequence of events.
8290 - 51
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Figure 4.
00
>
_*-•
c
*-»
_c
0>
JH
0)
CC
oo
«n
—J—,—
24:00
Time
22:30
25:30
-, ,
27:00
Selected ion current profile for m/z 322 (TCDDs) produced by MS analysis of
the GC performance check solution on a 60 m DB-5 fused silica capillary column
under the conditions listed in Sec. 7.6.
8290 - 52
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Figure 5.
80-i
60-
40-
20-
Ref. mass 304.9824 Peak top
Span. 200 ppm
System file name
Data file name
Resolution
Group number
lonization mode
Switching
Ref. masses
YVES150
A:85Z567
10000
1
EI +
VOLTAGE
304.9824
380.9260
M/AM—10.500
Channel B 380.9260 Lock mass
Span 200 ppm
Peak profiles representing two PFK reference ions at m/z 305 and 381. The
resolution of the high-mass signal is 95 ppm at 5 percent of the peak height;
this corresponds to a resolving power M/dM of 10,500 (10 percent valley
definition).
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Figure 6.
20:00
22:00
24:00
26:00
28:00
30:00
Manual determination of S/N.
The peak height (S) is measured between the mean noise (lines C and D).
These mean signal values are obtained by tracing the line between the
baseline average noise extremes, El and E2, and between the apex average
noise extremes, E3 and E4, at the apex of the signal.
NOTE:
It is imperative that the instrument interface amplifier
electronic zero offset be set high enough so that negative
going baseline noise is recorded.
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Table 1.
Types of Matrices, Sample Sizes and 2,3,7,8-TCDD-Based
Method Calibration Limits (Parts per Trillion)
Lower MCLa
Upper MCLa
Weight (g)
IS Spiking
Levels (ppt)
Final Extr.
Vol. (jLtL)d
Water
0.
2
1000
1
10-50
Soil
Sediment
Paper Pulpb
01 1.0
200
10
100
10-50
Fly
Ash
1.0
200
10
100
50
Fish
Tissue
1.0
200
20
100
10-50
Human
Adipose
0 Tissue
1.0
200
10
100
10-50
Sludges,
Fuel Oil
5.0
1000
2
500
50
Still-
Bottom
10
2000
1
1000
50
a For other congeners multiply the values by 1 for TCDF/PeCDD/PeCDF, by 2.5
for HxCDD/HxCDF/HpCDD/HpCDF, and by 5 for OCDD/OCDF.
b Sample dewatered according to Sec. 6.5.
c One half of the extract from the 20 g sample is used for determination of
lipid content (Sec. 7.2.2).
d See Sec. 7.8.1, Note.
NOTE: Chemical reactor residues are treated as still bottoms if their
appearances so suggest.
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Table 2.
Composition of the Sample Fortification
and Recovery Standard Solutions8
Analyte
Sample Fortification
Solution
Concentration
(pg//LtL; Solvent:
Nonane)
Recovery Standard
Solution
Concentration
(pg//xL; Solvent:
Nonane)
13
C12-2,3,7,8-TCDD
13C12-2,3,7,8-TCDF
13C12-1,2,3,4-TCDD
13
13
13
13
13
C12-l,2,3,7,8-PeCDD
C12-l,2,3,7,8-PeCDF
C12-l,2,3,6,7,8-HxCDD
C12-l,2,3,4,7,8-HxCDF
C12-l,2,3,7,8,9-HxCDD
13C12-l,2,3,4,6,7,8-HpCDD
13C12-l,2,3,4,6,7,8-HpCDF
13C12-OCDD
10
10
10
10
25
25
25
25
50
50
50
(a) These solutions should be made freshly every day because of the possibility
of adsorptive losses to glassware. If these solutions are to be kept for more
than one day, then the sample fortification solution concentrations should be
increased ten fold, and the recovery standard solution concentrations should be
doubled. Corresponding adjustments of the spiking volumes must then be made.
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Table 3.
The Fifteen 2,3,7,8-Substituted PCDD and PCDF Congeners
PCDD PCDF
2,3,7,8-TCDD(*) 2,3,7,8-TCDF(*)
l,2,3,7,8-PeCDD(*) l,2,3,7,8-PeCDF(*)
l,2,3,6,7,8-HxCDD(*) 2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDD 1,2,3,6,7,8-HxCDF
l,2,3,7,8,9-HxCDD(+) 1,2,3,7,8,9-HxCDF
l,2,3,4,6,7,8-HpCDD(*) l,2,3,4,7,8-HxCDF(*)
2,3,4,6,7,8-HxCDF
l,2,3,4,6,7,8-HpCDF(*)
1,2,3,4,7,8,9-HpCDF
(*) The 13C-labeled analogue is used as an internal standard.
(+) The 13C-labeled analogue is used as a recovery standard.
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Table 4.
Isomers of Chlorinated Dioxins and Furans as a
Function of the Number of Chlorine Atoms
Number of
Chlorine
Atoms
1
2
3
4
5
6
7
8
Total
Number of
Dioxin
Isomers
2
10
14
22
14
10
2
1
75
Number of
2,3,7,8
Isomers
—
—
—
1
1
3
1
1
7
Number of
Furan
Isomers
4
16
28
38
28
16
4
1
135
Number of
2,3,7,8
Isomers
—
—
—
1
2
4
2
1
10
8290 - 58
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Table 5.
High-Resolution Concentration Calibration Solutions
Concentration (pq/uL, in Nonane)
Compound
HRCC
Unlabeled Analytes
2,3,7,8-TCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDD
1,2,3,7,8-PeCDF
2,3,
1,2,
1,2,
1,2,
1,2,
1,2,
1,2,
2,3,
1,2,
1,2,
1,2,
OCDD
OCDF
4,7,
3,4,
3,6,
3,7,
3,4,
3,6,
3,7,
4,6,
3,4,
3,4,
3,4,
8-PeCDF
7
7
,
,
8,
7
7
8
7
6
6
7
Internal
13C
13f12
u12
13C1?
13C
13/- 12
13p12
73C12
,, 12
13p
, L12
C
-2,3
-2,3
-1,2
-1,2
-1,2
-1,2
-1,2
-1,2
,
,
?
}
,
,
,
,
,
,
,
,
,
,
8-HxCDD
8-HxCDD
9-HxCDD
8-HxCDF
8-HxCDF
9-HxCDF
8-HxCDF
7,8-HpCDD
7,8-HpCDF
8,9-HpCDF
200
200
500
500
500
500
500
500
500
500
500
500
500
500
500
1,000
1,000
50
50
125
125
125
125
125
125
125
125
125
125
125
125
125
250
250
10
10
25
25
25
25
25
25
25
25
25
25
25
25
25
50
50
2
2
6
6
6
6
6
6
6
6
6
6
6
6
6
12
12
.5
.5
.25
.25
.25
.25
.25
.25
.25
.25
.25
.25
.25
.25
.25
.5
.5
1
1
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
Standards
7
7
3
3
3
3
3
3
,8-TCDD
,8-TCDF
,7,8-PeCDD
,7,8-PeCDF
,6,7,8-HxCDD
,4,7,8-HxCDF
,4,6,7,8-HpCDD
,4,6,7,8-HpCDF
-OCDD
Recovery
13f
V/12
13c
-1,2
-1,2
'
50
50
50
50
125
125
125
125
250
50
50
50
50
125
125
125
125
250
50
50
50
50
125
125
125
125
250
50
50
50
50
125
125
125
125
250
50
50
50
50
125
125
125
125
250
Standards
3
3
,4-TCDD(a|
,7,8,9-HxCDD(b|
50
125
50
125
50
125
50
125
50
125
(a)
Used for recovery determinations of TCDD, TCDF, PeCDD and PeCDF internal
standards.
(bl Used for recovery determinations of HxCDD, HxCDF, HpCDD, HpCDF and OCDD
internal standards.
8290 - 59
Revision 0
September 1994
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Table 6.
Ions Monitored for HRGC/HRMS Analysis of PCDDs/PCDFs
Descriptor
1
2
3
Accurate1"1
Mass
303.9016
305.8987
315.9419
317.9389
319.8965
321.8936
331.9368
333.9338
375.8364
[354.9792]
339.8597
341.8567
351.9000
353.8970
355.8546
357.8516
367.8949
369.8919
409.7974
[354.9792]
373.8208
375.8178
383.8639
385.8610
389.8156
391.8127
401.8559
403.8529
445.7555
[430.9728]
Ion
ID
M
M+2
M
M+2
M
M+2
M
M+2
M+2
LOCK
M+2
M+4
M+2
M+4
M+2
M+4
M+2
M+4
M+2
LOCK
M+2
M+4
M
M+2
M+2
M+4
M+2
M+4
M+4
LOCK
Elemental
Composition
C12H435C140
C12H435C1337C10
13C12H435C140
13C12H435C1337C10
C12H435C1402
C12H435C1337C102
13C12H435C1402
13C12H435C1337C102
C12H435C1537C10
£9^13
C12H335C1437C10
C12H335C1337C120
13C12H335C1437C10
Ci2H3 C 1 3 C 1 20
C12H335C1437C102
C12H335C1337C1202
13C12H335C1437C102
13C12H335C1337C1202
C12H335C1637C10
C9F13
C12H235C1537C10
C12H235C1437C120
13C12H235C160
13C12H235C1537C10
C12H235C1537C102
C12H235C1437C1202
13C12H235C1537C102
13C12H235C1437C1202
C12H235C1637C120
C9F17
Analyte
TCDF
TCDF
TCDF (S)
TCDF (S)
TCDD
TCDD
TCDD (S)
TCDD (S)
HxCDPE
PFK
PeCDF
PeCDF
PeCDF (S)
PeCDF (S)
PeCDD
PeCDD
PeCDD (S)
PeCDD (S)
HpCDPE
PFK
HxCDF
HxCDF
HxCDF (S)
HxCDF (S)
HxCDD
HxCDD
HxCDD (S)
HxCDD (S)
OCDPE
PFK
8290 - 60
Revision 0
September 1994
-------
Descriptor Accurate'8' Ion
Mass ID
4 407.7818 M+2
409.7788 M+4
417.8250 M
419.8220 M+2
423.7767 M+2
425.7737 M+4
435.8169 M+2
437.8140 M+4
479.7165 M+4
[430.9728] LOCK
5 441.7428 M+2
443.7399 M+4
457.7377 M+2
459.7348 M+4
469.7780 M+2
471.7750 M+4
513.6775 M+4
[442.9728] LOCK
Table 6.
Continued
Elemental
Composition
C12H35C1637C10
C12H35C1537C120
13C12H35C170
13C12H35C1637C10
C12H35C1637C102
C12H35C1537C1202
13C12H35C1637C102
13C12H35C1537C1202
C12H35C1737C120
CgF-i?
C1235C1737C10
C1235C1637C120
C1236C1737C102
C1235C1637C1202
13C1235C1737C102
13f 35f1 37f-i «
^12 ^'6 ^'2U2
r 35n 37fi o
^12 UI8 U ' 2U
^10^17
Analyte
HpCDF
HpCDF
HpCDF (S)
HpCDF
HpCDD
HpCDD
HpCDD (S)
HpCDD (S)
NCDPE
PFK
OCDF
OCDF
OCDD
OCDD
OCDD (S)
OCDD (S)
DCDPE
PFK
(8) The following nuclidic masses were used:
H = 1.007825
C = 12.000000
13C = 13.003355
F = 18.9984
0 = 15.994915
35C1 = 34.968853
37C1 = 36.965903
S = internal/recovery standard
8290 - 61
Revision 0
September 1994
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Table 7.
PCDD and PCDF Congeners Present in the GC Performance
Evaluation Solution and Used for Defining the
Homologous GC Retention Time Windows on a
60 m DB-5 Column
No. of
Chlorine
Atoms
4<°>
5
6
7
8
PCDD Positional
First
Eluter
1,3,6,8
1,2,4,6,8/
1,2,4,7,9
1,2,4,6,7,9/
1,2,4,6,8,9
1,2,3,4,6,7,9
Isomer
Last
Eluter
1,2,8,9
1,2,3,8,9
1,2,3,4,6,
1,2,3,4,6,
1,2,3,4,6,
PCDF Positional
First
Eluter
1,3,6,8
1,3,4,6,8
7 1,2,3,4,6,8
7,8 1,2,3,4,6,7,8
7,8,9
Isomer
Last
Eluter
1,2,8,9
1,2,3,8,9
1,2,3,4,8,9
1,2,3,4,7,8,9
1,2,3,4,6,7,8,9
In addition to these two TCDD isomers, the 1,2,3,4-, 1,2,3,7-, 1,2,3,8-, 2,3,7,8-,
13C12-2,3,7,8-, and 1,2,3,9-TCDD isomers must also be present as a check of column
resolution.
8290 - 62 Revision 0
September 1994
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Table 8.
Theoretical Ion Abundance Ratios and Their Control Limits
for PCDDs and PCDFs
Number of
Chlorine Ion
Atoms Type
4
5
6
6w
ylb)
7
8
M/M+2
M+2/M+4
M+2/M+4
M/M+2
M/M+2
M+2/M+4
M+2/M+4
Theoretical
Ratio
0.77
1.55
1.24
0.51
0.44
1.04
0.89
Control
lower
0.65
1.32
1.05
0.43
0.37
0.88
0.76
Limits
upper
0.89
1.78
1.43
0.59
0.51
1.20
1.02
M Used only for 13C-HxCDF (IS).
(bl Used only for 13C-HpCDF (IS).
8290 - 63
Revision 0
September 1994
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Table 9.
Relative Response Factor [RF (number)] Attributions
Number
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
2,3,7
2,3,7
1,2,3
1,2,3
2,3,4
1,2,3
1,2,3
1,2,3
1 ,?,3
1,2,3
1,2,3
2,3,4
1,2,3
1,2,3
1,2,3
OCDD
OCDF
13C12-
13p
13C12-
13C12-
13C12-
13C
13C12-
13C12-
13r -
L12-
Total
Total
Total
Total
Specific Congener Name
,8-TCDD (and total TCDDs)
,8-TCDF (and total TCDFs)
,7,8-PeCDD (and total PeCDDs)
,7,8-PeCDF
,7,8-PeCDF
,4,7,8-HxCDD
,6,7,8-HxCDD
,7,8,9-HxCDD
4,7,8-HxCDF
,6,7,8-HxCDF
,7,8,9-HxCDF
,6,7,8-HxCDF
,4,6,7,8-HpCDD (and total HpCDDs)
,4,6,7,8-HpCDF
,4,7,8,9-HpCDF
2,3,7,8-TCDD
2, 3, 7, 8-TCDF
1,2,3,7,8-PeCDD
1,2, 3, 7,8-PeCDF
1,2,3,6,7,8-HxCDD
1,2,3,4,7,8-HxCDF
1,2,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8-HpCDF
OCDD
PeCDFs
HxCDFs
HxCDDs
HpCDFs
8290 - 64 Revision 0
September 1994
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Table 10.
2,3,7,8-TCDD Toxicity Equivalency Factors (TEFs) for the
Polychlorinated Dibenzodioxins and Dibenzofurans
Number Compound(s) TEFa
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
2,3,7,8-TCDD
1,2,3,7,8-PeCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDD
1,2,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8,9-OCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
1,2,3,4,7,8-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
1,2,3,4,6,7,8,9-OCDF
1.00
0.50
0.10
0.10
0.10
0.01
0.001
0.1
0.05
0.5
0.1
0.1
0.1
0.1
0.01
0.01
0.001
Taken from "Interim Procedures for Estimating Risks Associated with Exposures
to Mixtures of Chlorinated Dibenzo-p-Dioxin and -Dibenzofurans (CDDs and CDFs)
and 1989 Update", (EPA/625/3-89/016, March 1989).
8290 - 65 Revision 0
September 1994
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Table 11.
Analyte Relative Retention Time Reference Attributions
(a)
Analyte Analyte RRT Reference
1,2,3,4,7,8-HxCDD 13C12-l,2,3,6,7,8-HxCDD
1,2,3,6,7,8-HxCDF 13C12-l,2,3,4,7,8-HxCDF
1,2,3,7,8,9-HxCDF 13C12-l,2,3,4,7,8-HxCDF
2,3,4,6,7,8-HxCDF 13C12-l,2,3,4,7,8-HxCDF
(al The retention time of 2,3,4,7,8-PeCDF on the DB-5 column is measured relative
13C
to 13C12-l,2,3,4,6,7,8-HpCDF.
to 13C12-l,2,3,7,8-PeCDF and the retention time of 1,2,3,4,7,8,9-HpCDF relative
8290 - 66 Revision 0
September 1994
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4.3 DETERMINATION OF ORGANIC ANALYTES
4.3.3 HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC METHODS
The following methods are included in this section:
Method 8310:
Method 8315:
Appendix A:
Method 8316:
Method 8318:
Method 8321:
Method 8330:
Method 8331:
Polynuclear Aromatic Hydrocarbons
Determination of Carbonyl Compounds by High
Performance Liquid Chromatography (HPLC)
Recrystallization of 2,4-
Dinitrophenylhydrazine (DNPH)
Acrylamide, Acrylonitrile and Acrolein by High
Performance Liquid Chromatography (HPLC)
N-Methylcarbamates by High Performance Liquid
Chromatography (HPLC)
Solvent Extractable Non-Volatile Compounds by
High Performance Liquid
Chromatography/Thermospray/Mass Spectrometry
(HPLC/TSP/MS) or Ultraviolet (UV) Detection
Nitroaromatics and Nitramines by High Performance
Liquid Chromatography (HPLC)
Tetrazene by Reverse Phase High Performance
Liquid Chromatography (HPLC)
FOUR - 12
Revision 2
September 1994
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METHOD 8310
POLYNUCLEAR AROMATIC HYDROCARBONS
1.0 SCOPE AND APPLICATION
1.1 Method 8310 is used to determine the concentration of certain poly-
nuclear aromatic hydrocarbons (PAH) in ground water and wastes. Specifically,
Method 8310 is used to detect the following substances:
Acenaphthene Chrysene
Acenaphthylene Di benzo(a,h)anthracene
Anthracene Fluoranthene
Benzo(a)anthracene Fluorene
Benzo(a)pyrene Indeno(1,2,3-cd)pyrene
Benzo(b)fluoranthene Naphthalene
Benzo(ghi)perylene Phenanthrene
Benzo(k)fluoranthene Pyrene
1.2 Use of Method 8310 presupposes a high expectation of finding the
specific compounds of interest. If the user is attempting to screen samples
for any or all of the compounds listed above, he must develop independent
protocols for the verification of identity.
1.3 The method detection limits for each compound in reagent water are
listed in Table 1. Table 2 lists the practical quantitation limit (PQL) for
other matrices. The sensitivity of this method usually depends on the level
of interferences rather than instrumental limitations. The limits of
detection listed in Table 1 for the liquid chromatographic approach represent
sensitivities that can be achieved in the absence of interferences. When
interferences are present, the level of sensitivity will be lower.
1.4 This method is recommended for use only by experienced residue
analysts or under the close supervision of such qualified persons.
2.0 SUMMARY OF METHOD
2.1 Method 8310 provides high performance liquid chromatographic (HPLC)
conditions for the detection of ppb levels of certain polynuclear aromatic
hydrocarbons. Prior to use of this method, appropriate sample extraction
techniques must be used. A 5- to 25-uL aliquot of the extract is injected
into an HPLC, and compounds in the effluent are detected by ultraviolet (UV)
and fluorescence detectors.
2.2 If interferences prevent proper detection of the analytes of
interest, the method may also be performed on extracts that have undergone
cleanup using silica gel column cleanup (Method 3630).
8310 - 1
Revision
Date September 1986
-------
TABLE 1. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY OF PAHsa
Compound
Naphthalene
Acenaphthylene
Acenaphthene
Fluorene
Phenanthrene
Anthracene
Fluoranthrene
Pyrene
Benzo (a) anthracene
Chrysene
Benzo (b) f 1 uoranthene
Benzo (k) f 1 uoranthene
Benzo(a)pyrene
D1 benzo (a , h) anthracene
Benzo (ghi)perylene
Indeno(l,2,3-cd)pyrene
Retention
time (mln)
16.6
18.5
20.5
21.2
22.1
23.4
24.5
25.4
28.5
29.3
31.6
32.9
33.9
35.7
36.3
37.4
Col umn
capacity
factor (k1)
12.2
13.7
15.2
15.8
16.6
17.6
18.5
19.1
21.6
22.2
24.0
25.1
25.9
27.4
27.8
28.7
Method Detection
limit (ug/L)
UV Fluorescence
1.8
2.3
1.8
0.21
0.64
0.66
0.21
0.27
0.013
0.15
0.018
0.017
0.023
0.030
0.076
0.043
a HPLC conditions: Reverse phase HC-ODS S11-X, 5 micron particle size,
1n a 250-mm x 2.6-mrn I.D. stainless steel column. Isocratlc elutlon for 5 mln
using acetom'trlle/water (4:6)(v/v), then linear gradient elutlon to 100%
acetonltrlle over 25 mln at 0.5 mL/min flow rate. If columns having other
Internal diameters are used, the flow rate should be adjusted to maintain a
linear velocity of 2 mm/sec.
TABLE 2. DETERMINATION OF PRACTICAL QUANTITATION LIMITS (PQL) FOR VARIOUS
MATRICES3
Matrix
Factor*3
Ground water
Low-level soil by sonication with GPC cleanup
High-level soil and sludges by sonlcatlon
Non-water mlsdble waste
10
670
10,000
100,000
aSample PQLs are highly matrix-dependent. The PQLs listed herein are
provided for guidance and may not always be achievable.
= [Method Detection Limit (Table 1) X [Factor (Table 2)]. For non-
aqueous samples, the factor is on a wet-weight basis.
8310 - 2
Revision 0
Date September 1986
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3.0 INTERFERENCES
3.1 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts and/or elevated baselines, causing misinterpreta-
tion of the chromatograms. All of these materials must be demonstrated to be
free from interferences, under the conditions of the analysis, by running
method blanks. Specific selection of reagents and purification of solvents by
distillation in all-glass systems may be required.
3.2 Interferences coextracted from the samples will vary considerably
from source to source. Although a general cleanup technique is provided as
part of this method, individual samples may require additional cleanup
approaches to achieve the sensitivities stated in Table 1.
3.3 The chromatographic conditions described allow for a unique
resolution of the specific PAH compounds covered by this method. Other PAH
compounds, in addition to matrix artifacts, may interfere.
4.0 APPARATUS AND MATERIALS
4.1 Kuderna-Danish (K-D) apparatus:
4.1.1 Concentrator tube: 10-ml, graduated (Kontes K-570050-1025 or
equivalent). Ground-glass stopper is used to prevent evaporation of
extracts.
4.1.2 Evaporation flask: 500-mL (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs.
4.1.3 Snyder column: Three-ball macro (Kontes K-503000-0121 or
equivalent).
4.1.4 Snyder column: Two-ball micro (Kontes K-569001-0219 or
equivalent).
4.2 Boiling chips; Solvent extracted, approximately 10/40 mesh (silicon
carbide or equivalent).
4.3 Water bath; Heated, with concentric ring cover, capable of
temperature control (+5'C). The bath should be used in a hood.
4.4 Syringe; 5-mL.
4.5 High pressure syringes.
4.6 HPLC apparatus;
4.6.1 Gradient pumping system: Constant flow.
4.6.2 Reverse phase column: HC-ODS Sil-X, 5-micron particle size
diameter, in a 250-mm x 2.6-mm I.D. stainless steel column (Perkin Elmer
No. 089-0716 or equivalent).
8310 - 3
Revision 0
Date September 1986
-------
4.6.3 Detectors: Fluorescence and/or UV detectors may be used.
4.6.3.1 Fluorescence detector: For excitation at 280-nm and
emission greater than 389-nm cutoff (Corning 3-75 or equivalent).
Fluorometers should have dispersive optics for excitation and can
utilize either filter or dispersive optics at the emission detector.
4.6.3.2 UV detector: 254-nm, coupled to the fluorescence
detector.
4.6.4 Strip-chart recorder: compatible with detectors. A data
system for measuring peak areas and retention times is recommended.
4.7 Volumetric flasks; 10-, 50-, and 100-mL.
5.0 REAGENTS
5.1 Reagent water: Reagent water is defined as water 1n which an
interferent is not observed at the method detection limit of the compounds of
interest.
5.2 Acetonitrile; HPLC quality, distilled in glass.
5.3 Stock standard solutions;
5.3.1 Prepare stock standard solutions at a concentration of 1.00
ug/uL by dissolving 0.0100 g of assayed reference material in aceto-
nitrile and diluting to volume in a 10-mL volumetric flask. Larger
volumes can be used at the convenience of the analyst. When compound
purity is assayed to be 96% or greater, the weight can be used without
correction to calculate the concentration of the stock standard.
Commercially prepared stock standards can be used at any concentration if
they are certified by the manufacturer or by an independent source.
5.3.2 Transfer the stock standard solutions into Teflon-sealed
screw-cap bottles. Store at 4*C and protect from light. Stock standards
should be checked frequently for signs of degradation or evaporation,
especially just prior to preparing calibration standards from them.
5.3.3 Stock standard solutions must be replaced after one year, or
sooner if comparison with check standards indicates a problem.
5.4 Calibration standards; Calibration standards at a minimum of five
concentration levels should be prepared through dilution of the stock
standards with acetonitrile. One of the concentration levels should be at a
concentration near, but above, the method detection limit. The remaining
concentration levels should correspond to the expected range of concentrations
found in real samples or should define the working range of the HPLC. Cali-
bration standards must be replaced after six months, or sooner if comparison
with check standards indicates a problem.
8310 - 4
Revision 0
Date September 1986
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5.5 Internal standards (1f Internal standard calibration 1s used); To
use this approach, the analyst must select one or more internal standards that
are similar in analytical behavior to the compounds of interest. The analyst
must further demonstrate that the measurement of the internal standard is not
affected by method or matrix interferences. Because of these limitations, no
internal standard can be suggested that is applicable to all samples.
5.5.1 Prepare calibration standards at a minimum of five
concentration levels for each analyte as described in Paragraph 5.4.
5.5.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with acetonitrlle.
5.5.3 Analyze each calibration standard according to Section 7.0.
5.6 Surrogate standards; The analyst should monitor the performance of
the extraction,cleanup(Tf necessary), and analytical system and the
effectiveness of the method in dealing with each sample matrix by spiking each
sample, standard, and reagent water blank with one or two surrogates (e.g.,
decafluorobiphenyl or other PAHs not expected to be present in the sample)
recommended to encompass the range of the temperature program used in this
method. Deuterated analogs of analytes should not be used as surrogates for
HPLC analysis due to coelutlon problems.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1. Extracts must be stored under refrigeration and must be analyzed
within 40 days of extraction.
7.0 PROCEDURE
7.1 Extraction;
7.1.1 Refer to Chapter Two for guidance on choosing the appropriate
extraction procedure. In general, water samples are extracted at a
neutral pH with methylene chloride, using either Method 3510 or 3520.
Solid samples are extracted using either Method 3540 or 3550. To achieve
maximum sensitivity with this method, the extract must be concentrated to
1 mL.
7.1.2 Prior to HPLC analysis, the extraction solvent must be
exchanged to acetonitrile. The exchange 1s performed during the K-D
procedures listed in all of the extraction methods. The exchange 1s
performed as follows.
7.1.2.1 Following K-D of the methylene chloride extract to
1 mL using the macro-Snyder column, allow the apparatus to cool and
drain for at least 10 min.
8310 - 5
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Date September 1986
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7.1.2.2 Increase the temperature of the hot water bath to 95-
100'C. Momentarily remove the Snyder column, add 4 ml of
acetonltrlle, a new boiling chip, and attach a two-ball mlcro-Snyder
column. Concentrate the extract using 1 ml of acetonltrlle to
prewet the Snyder column. Place the K-D apparatus on the water bath
so that the concentrator tube 1s partially Immersed 1n the hot
water. Adjust the vertical position of the apparatus and the water
temperature, as required, to complete concentration 1n 15-20 m1n.
At the proper rate of distillation the balls of the column will
actively chatter, but the chambers will not flood. When the
apparent volume of liquid reaches 0.5 ml, remove the K-D apparatus
and allow 1t to drain and cool for at least 10 m1n.
7.1.2.3 When the apparatus is cool, remove the micro-Snyder
column and rinse its lower joint Into the concentrator tube with
about 0.2 ml of acetonltrlle. A 5-mL syringe is recommended for
this operation. Adjust the extract volume to 1.0 ml. Stopper the
concentrator tube and store refrigerated at 4*C, if further
processing will not be performed immediately. If the extract will
be stored longer than two days, it should be transferred to a
Teflon-sealed screw-cap vial. Proceed with HPLC analysis 1f further
cleanup is not required.
7.2 HPLC conditions (Recommended);
7.2.1 Using the column described in Paragraph 4.6.2: Isocratlc
elution for 5 m1n using aceton1tr1le/water (4:6)(v/v), then linear
gradient elution to 100% acetonitrile over 25 min at 0.5 mL/min flow
rate. If columns having other Internal diameters are used, the flow rate
should be adjusted to maintain a linear velocity of 2 mm/sec.
7.3 Calibration:
7.3.1 Refer to Method 8000 for proper calibration procedures. The
procedure of internal or external standard calibration may be used. Use
Table 1 and especially Table 2 for guidance on selecting the lowest point
on the calibration curve.
7.3.2 Assemble the necessary HPLC apparatus and establish operating
parameters equivalent to those indicated in Section 7.2.1. By injecting
calibration standards, establish the sensitivity limit of the detectors
and the linear range of the analytical systems for each compound.
7.3.3 Before using any cleanup procedure, the analyst should
process a series of calibration standards through the procedure to
confirm elution patterns and the absence of Interferences from the
reagents.
7.4 HPLC analysis;
7.4.1 Table 1 summarizes the estimate retention times of PAHs
determinable by this method. Figure 1 1s an example of the separation
achievable using the conditions given in Paragraph 7.2.1.
8310 - 6
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Column: HC-ODSSIL-X
Mobile Phase: 40% to 100% Acetonitrile in Water
Dectector: Fluorescence
40
RETENTION TIME (MINUTES)
Figure 1. Liquid chromatogram of poly nuclear aromatics.
8310 - 7
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Date September 1986
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7.4.2 If Internal standard calibration Is to be performed, add
10 uL of Internal standard to the sample prior to Injection. Inject
2-5 uL of the sample extract with a high-pressure syringe or sample
injection loop. Record the volume Injected to the nearest 0.1 uL, and
the resulting peak size, in area units or peak heights. Re-equilibrate
the HPLC column at the initial gradient conditions for at least 10 min
between injections.
7.4.3 Using either the internal or external calibration procedure
(Method 8000), determine the identity and quantity of each component peak
in the sample chromatogram which corresponds to the compounds used for
calibration purposes. See Section 7.8 of Method 8000 for calculation
equations.
7.4.4 If the peak area exceeds the linear range of the system,
dilute the extract and .^analyze.
7.4.5 If the peak area measurement is prevented by the presence of
interferences, further cleanup is required.
7.5 Cleanup:
7.5.1 Cleanup of the acetonitrile extract takes place using Method
3630 (Silica Gel Cleanup). Specific instructions for cleanup of the
extract for PAHs is given in Section 7.1 of Method 3630.
7.5.2 Following cleanup, analyze the samples using HPLC as
described in Section 7.4.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method used. If extract cleanup was performed, follow the QC
in Method 3600 and in the specific cleanup method.
8.2 Mandatory quality control to validate the HPLC system operation is
found 1n Method 8000, Section 8.6.
8.2.1 The quality control check sample concentrate (Method 8000,
Section 8.6) should contain each analyte at the following concentrations
in acetonitrile: naphthalene, 100 ug/mL; acenaphthylene, 100 ug/mL;
acenaphthene, 100 ug/mL; fluorene, 100 ug/mL; phenanthrene, 100 ug/mL;
anthracene, 100 ug/mL; benzo(k)fluoranthene, 5 ug/mL; and any other PAH
at 10 ug/mL.
8.2.2 Table 3 indicates the calibration and QC acceptance criteria
for this method. Table 4 gives method accuracy and precision as
functions of concentration for the analytes of interest. The contents of
both Tables should be used to evaluate a laboratory's ability to perform
and generate acceptable data by this method.
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8.3 Calculate surrogate standard recovery on all samples, blanks, and
spikes. Determine if the recovery is within limits (limits established by
performing QC procedures outlined in Method 8000, Section 8.10).
8.3.1 If recovery is not within limits, the following procedures
are required.
• Check to be sure there are no errors in calculations,
surrogate solutions and internal standards. Also, check
instrument performance.
• Recalculate the data and/or reanalyze the extract if any of
the above checks reveal a problem.
• Reextract and reanalyze the sample if none of the above are
a problem or flag the data as "estimated concentration."
9.0 METHOD PERFORMANCE
9.1 The method was tested by 16 laboratories using reagent water,
drinking water, surface water, and three industrial wastewaters spiked at six
concentrations over the range 0.1 to 425 ug/L. Single operator precision,
overall precision, and method accuracy were found to be directly related to
the concentration of the analyte and essentially independent of the sample
matrix. Linear equations to describe these relationships are presented in
Table 4.
9.2 This method has been tested for linearity of spike recovery from
reagent water and has been demonstrated to be applicable over the
concentration range from 8 x MDL to 800 x MDL with the following exception:
benzo(ghi)perylene recovery at 80 x and 800 x MDL were low (35% and 45%,
respectively).
9.3 The accuracy and precision obtained will be determined by the sample
matrix, sample-preparation technique, and calibration procedures used.
10.0 REFERENCES
1. "Development and Application of Test Procedures for Specific Organic Toxic
Substances in Wastewaters, Category 9 - PAHs," Report for EPA Contract 68-03-
2624 (in preparation).
2. Sauter, A.D., L.D. Betowski, T.R. Smith, V.A. Strickler, R.6. Beimer, B.N.
Colby, and J.E. Wilkinson, "Fused Silica Capillary Column GC/MS for the
Analysis of Priority Pollutants," Journal of HRC&CC 4, 366-384, 1981.
3. "Determination of Polynuclear Aromatic Hydrocarbons in Industrial and
Municipal Wastewaters," EPA-600/4-82-025, U.S. Environmental Protection
Agency, Environmental Monitoring and Support Laboratory, Cincinnati, Ohio
45268, September 1982.
8310 - 9
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Date September 1986
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4. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects," Journal of the Association of Official Analytical
Chemists, 48, 1037, 1965.
5. "EPA Method Validation Study 20, Method 610 (Polynuclear Aromatic
Hydrocarbons)," Report for EPA Contract 68-03-2624 (in preparation).
6. U.S. EPA 40 CFR Part 136, "Guidelines Establishing Test Procedures for the
Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim Final
Rule and Proposed Rule," October 26, 1984.
7. Provost, L.P. and R.S. Elder, "Interpretation of Percent Recovery Data,"
American Laboratory, Ijj, pp. 58-63, 1983.
8310 - 10
Revision
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TABLE 3. QC ACCEPTANCE CRITERIA*
Parameter
Acenaphthene
Acenaphthylene
Anthracene
Benzo (a) anthracene
Benzo (a) pyrene
Benzo (b) f 1 uoranthene
Benzo (ghi)perylene
Benzo (k) f 1 uoranthene
Chrysene
01 benzo (a , h) anthracene
Fl uoranthene
Fluorene
Indeno (1 , 2 , 3-cd) pyrene
Naphthalene
Phenanthrene
Pyrene
Test
cone.
(ug/L)
100
100
100
10
10
10
10
5
10
10
10
100
10
100
100
10
Limit
for s
(ug/L)
40.3
45.1
28.7
4.0
4.0
3.1
2.3
2.5
4.2
2.0
3.0
43.0
3.0
40.7
37.7
3.4
Range
for X
(ug/L)
D-105.7
22.1-112.1
11.2-112.3
3.1-11.6
0.2-11.0
1.8-13.8
D-10.7
D-7.0
D-17.5
0.3-10.0
2.7-11.1
D-119
1.2-10.0
21.5-100.0
8.4-133.7
1.4-12.1
Range
P. Ps
(%)
D-124
D-139
D-126
12-135
D-128
6-150
D-116
D-159
D-199
D-110
14-123
D-142
D-116
D-122
D-155
D-140
s = Standard deviation of four recovery measurements, in ug/L.
7 = Average recovery for four recovery measurements, in ug/L.
p, ps = Percent recovery measured.
D = Detected; result must be greater than zero.
aCriteria from 40 CFR Part 136 for Method 610. These criteria are based
directly upon the method performance data in Table 3. Where necessary, the
limits for recovery have been broadened to assure applicability of the limits
to concentrations below those used to develop Table 3.
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TABLE 4. METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATIONa
Parameter
Acenaphthene
Acenaphthylene
Anthracene
Benzo (a) anthracene
Benzo(a)pyrene
Benzo (b) f 1 uoranthene
Benzo (ghi)perylene
Benzo (k) f 1 uoranthene
Chrysene
Dlbenzo (a, h) anthracene
Fl uoranthene
Fluorene
Indeno (1 , 2 , 3-cd)pyrene
Naphthalene
Phenanthrene
Pyrene
Accuracy, as
recovery, x1
(ug/L)
0.52C+0.54
0.69C-1.89
0.63C-1.26
0.73C+0.05
0.56C+0.01
0.78C+0.01
0.44C+0.30
0.59C+0.00
0.77C-0.18
0.41C-0.11
0.68C+0.07
0.56C-0.52
0.54C+0.06
0.57C-0.70
0.72C-0.95
0.69C-0.12
Single analyst
precision, sr'
(ug/L)
0.397+0.76
0.367+0.29
0.237+1.16
0.287+0.04
0.387-0.01
0.217+0.01
0.257+0.04
0.447-0.00
0.327-0.18
0.247+0.02
0.227+0.06
0.447-1.12
0.297+0.02
0.397-0.18
0.297+0.05
0.257+0.14
Overal 1
precision,
S1 (ug/L)
0.537+1.32
0.427+0.52
0.417+0.45
0.347+0.02
0.537-0.01
0.387-0.00
0.587+0.10
0.697+0.10
0.667-0.22
0.457+0.03
0.327+0.03
0.637-0.65
0.427+0.01
0.417+0.74
0.477-0.25
0.427-0.00
x' = Expected recovery for one or more measurements of a sample
containing a concentration of C, in ug/L.
sr' = Expected single analyst standard deviation of measurements at an
average concentration of 7, in ug/L.
S1 = Expected interlaboratory standard deviation of measurements at an
average concentration found of 7, in ug/L.
C = True value for the concentration, 1n ug/L.
7 = Average recovery found for measurements of samples containing a
concentration of C, in ug/L.
8310 - 12
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METHOD 8310
POLYNUCLEAR AROMATIC HYDROCARBONS
7.1.1
0
Choose
appropriate
•xtractlon
procedure
(see Chapter 2)
7. l.Z
7.3.3
Process
a series of
calibration
standards
Exchange
extract-
ion solvent to
acetonltrlle
durlne K-D
procedures
7.2
7.4
Perform
HPLC
analysis (see
Method 6000
for calculation
equations
Set HPLC
conditions
7.3
Refer to
Method 8OOO
for proper
calibration
techniques
Cleanup using
Method 3630
7.3.2
HPLC apparatus:
establish
operating
parameters
o
8310 - 13
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Date September 1986
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METHOD 8315
DETERMINATION OF CARBONYL COMPOUNDS
BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
1.0 SCOPE AND APPLICATION
1.1 This method provides procedures for the determination of free
carbonyl compounds in various matrices by derivatization with
2,4-dinitrophenylhydrazine (DNPH). The method utilizes high performance liquid
chromatography (HPLC) with ultraviolet/visible (UV/vis) detection to identify and
quantitate the target analytes using two different sets of conditions: Option 1
and Option 2. Option 1 has been shown to perform well for one set of target
analytes for aqueous samples, soil or waste samples, and stack samples collected
by Method 0011. Option 2 has been shown to work well for another set of target
analytes in indoor air samples collected by Method 0100. The two sets of target
analytes overlap for some compounds. Refer to the Analysis Option listed in the
following table to determine which analytes may be analyzed by which option. The
following compounds may be determined by this method:
Compound Name CAS No.a Analysis Optionb
Acetaldehyde
Acetone
Acrolein
Benzaldehyde
Butanal (butyraldehyde)
Crotonaldehyde
Cyclohexanone
Decanal
2,5-Dimethylbenzaldehyde
Formaldehyde
Heptanal
Hexanal (hexaldehyde)
Isovaleraldehyde
Nonanal
Octanal
Pentanal (valeraldehyde)
Propanal (propionaldehyde)
m-Tolualdehyde
o-Tolualdehyde
p-Tolualdehyde
a Chemical Abstract Services
" Tk-io 1 -i o +• rv-P + •» v»n r\4- •!» m •* 1
75-07-0
67-64-1
107-02-8
100-52-7
123-72-8
123-73-9
108-94-1
112-31-2
5779-94-2
50-00-0
111-71-7
66-25-1
590-86-3
124-19-6
124-13-0
110-62-3
123-38-6
620-23-5
529-20-4
104-87-0
Registry Number.
\/4- n o /->/MTt-^-ir\r* •% rv n
1,2
2
2
2
1,2
1,2
1
1
2
1,2
1
1,2
2
1
1
1,2
1,2
2
2
2
klf^\V*l^l"Vr\TM/'t 1 T f» 41 f\ f
compounds that have been evaluated using modifications of the
8315 - 1 Revision 0
September 1994
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analysis. Refer to the respective option number when choosing the
appropriate analysis technique for a particular compound.
1.2 The Option 1 method detection limits (MDL) are listed in Tables 1 and
2. The sensitivity data for sampling and analysis using Method 0100 (Option 2)
are given in Table 3. The MDL for a specific sample may differ from that listed,
depending upon the nature of interferences in the sample matrix and the amount
of sample used in the procedure.
1.3 The extraction procedure for solid samples is similar to that
specified in Method 1311. Thus, a single sample may be extracted to measure the
analytes included in the scope of other appropriate methods. The analyst is
allowed the flexibility to select chromatographic conditions appropriate for the
simultaneous measurement of combinations of these analytes.
1.4 When this method is used to analyze unfamiliar sample matrices,
compound identification should be supported by at least one additional
qualitative technique. A gas chromatograph/mass spectrometer (GC/MS) may be used
for the qualitative confirmation of results for the target analytes, using the
extract produced by this method.
1.5 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of chromatography and in the interpretation of
chromatograms. Each analyst must demonstrate the ability to generate acceptable
results with this method, using the procedure described in Sec. 7.0.
2.0 SUMMARY OF METHOD
2.1 Liquid and Solid Samples (Option 1)
2.1.1 For wastes comprised of solids, or for aqueous wastes
containing significant amounts of solid material, the aqueous phase, if
any, is separated from the solid phase and stored for later analysis. If
necessary, the particle size of the solids in the waste is reduced. The
solid phase is extracted with an amount of extraction fluid equal to 20
times the weight of the solid phase. The extraction fluid employed is a
function of the alkalinity of the solid phase of the waste. A special
extractor vessel is used when testing for volatiles. Following extraction,
the aqueous extract is separated from the solid phase by filtration
employing 0.6 to 0.8 jum glass fiber filter.
2.1.2 If compatible (i.e., multiple phases will not form on
combination), the initial aqueous phase of the waste is added to the
aqueous extract, and these liquids are analyzed together. If
incompatible, the liquids are analyzed separately and the results are
mathematically combined to yield a volume-weighted average concentration.
2.1.3 A measured volume of aqueous sample (approx. 100 mL) or an
appropriate amount of solids extract (approx. 25 g), is buffered to pH 3
and derivatized with 2,4-dinitrophenylhydrazine (DNPH), using either the
liquid-solid or a liquid-liquid extraction option. If the liquid-solid
8315 - 2 Revision 0
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option is used, the derivative is extracted using solid sorbent
cartridges, followed by elution with ethanol. If the liquid-liquid option
is used, the derivative is extracted from the sample with three (3)
portions of methylene chloride. The methylene chloride extracts are
concentrated using the Kuderna-Danish (K-D) procedure and exchanged with
acetonitrile prior to HPLC analysis. Liquid chromatographic conditions
are described which permit the separation and measurement of various
carbonyl compounds in the extract by absorbance detection at 360 nm.
2.1.4 If formaldehyde is the only analyte of interest, the aqueous
sample or solids extract should be buffered to pH 5.0 to minimize artifact
formaldehyde formation.
2.2 Stack Gas Samples Collected by Method 0011 (Option 1) - The entire
sample returned to the laboratory is extracted with methylene chloride and the
methylene chloride extract is brought up to a known volume. An aliquot of the
methylene chloride extract is solvent exchanged and concentrated or diluted as
necessary. Liquid chromatographic conditions are described that permit the
separation and measurement of various carbonyl compounds in the extract by
absorbance detection at 360 nm.
2.3 Indoor Air Samples by Method 0100 (Option 2) - The sample cartridges
are returned to the laboratory and backflushed with acetonitrile into a 5 mL
volumetric flask. The eluate is brought up to volume with more acetonitrile.
Two (2) aliquots of the acetonitrile extract are pipetted into two (2) sample
vials having Teflon-lined septa. Liquid chromatographic conditions are described
that permit the separation and measurement of the various carbonyl compounds in
the extract by absorbance detection at 360 nm.
3.0 INTERFERENCES
3.1 Method interferences may be caused by contaminants in solvents,
reagents, glassware, and other sample processing hardware that lead to discrete
artifacts and/or elevated baselines in the chromatograms. All of these materials
must be routinely demonstrated to be free from interferences under the conditions
of the analysis by analyzing laboratory reagent blanks as described in Sec. 8.5.
3.1.1 Glassware must be scrupulously cleaned. Clean all glassware
as soon as possible after use by rinsing with the last solvent used. This
should be followed by detergent washing with hot water, and rinses with
tap water and organic-free reagent water. It should then be drained,
dried, and heated in a laboratory oven at 130°C for several hours before
use. Solvent rinses with acetonitrile may be substituted for the oven
heating. After drying and cooling, glassware should be stored in a clean
environment to prevent any accumulation of dust or other contaminants.
NOTE: Do not use acetone or methanol. These solvents react with
DNPH to form interfering compounds.
8315 - 3 Revision 0
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3.1.2 The use of high purity reagents and solvents helps to minimize
interference problems. Purification of solvents by distillation in all
glass systems may be required.
3.1.3 Polyethylene gloves must be worn when handling the silica gel
cartridges to reduce the possibility of contamination.
3.2 Formaldehyde contamination of the DNPH reagent is a frequently
encountered problem due to its widespread occurrence in the environment. The
DNPH reagent in Option 2 must be purified by multiple recrystallizations in UV-
grade acetonitrile. Recrystallization is accomplished, at 40-60°C, by slow
evaporation of the solvent to maximize crystal size. The purified DNPH crystals
are stored under UV-grade acetonitrile until use. Impurity levels of carbonyl
compounds in the DNPH are determined prior to the analysis of the samples and
should be less than 25 mg/L. Refer to Appendix A for the recrystallization
procedure.
3.3 Matrix interferences may be caused by contaminants that are
coextracted from the sample. The extent of matrix interferences will vary
considerably from source to source, depending upon the nature and diversity of
the matrix being sampled. Although the HPLC conditions described allow for a
resolution of the specific compounds covered by this method, other matrix
components may interfere. If interferences occur in subsequent samples,
modification of the mobile phase or some additional cleanup may be necessary.
3.4 In Option 1, acetaldehyde is generated during the derivatization step
if ethanol is present in the sample. This background will impair the measurement
of acetaldehyde at levels below 0.5 ppm (500 ppb).
3.5 For Option 2, at the stated two column analysis conditions, the
identification and quantitation of butyraldehyde may be difficult due to
coelution with isobutyraldehyde and methyl ethyl ketone. Precautions should be
taken and adjustment of the analysis conditions should be done, if necessary, to
avoid potential problems.
4.0 APPARATUS AND MATERIALS
4.1 High performance liquid chromatograph (modular)
4.1.1 Pumping system - Gradient, with constant flow control capable
of 1.50 mL/min.
4.1.2 High pressure injection valve with 20 /zL loop.
4.1.3 Column - 250 mm x 4.6 mm ID, 5 /zm particle size, CIS (Zorbax
or equivalent).
4.1.4 Absorbance detector - 360 nm.
4.1.5 Strip-chart recorder compatible with detector - Use of a data
system for measuring peak areas and retention times is recommended.
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4.1.6 Helium - for degassing mobile phase solvents. (Options
1 and 2)
4.1.7 Mobile Phase Reservoirs and Suction Filtration Apparatus - For
holding and filtering HPLC mobile phase. Filtering system should be all
glass and Teflon and use a 0.22 /urn polyester membrane filter. (Option 2)
4.1.8 Syringes - for HPLC injection loop loading, with capacity at
least four times the loop volume.
4.2 Apparatus and Materials for Option 1
4.2.1 Reaction vessel - 250 ml Florence flask.
4.2.2 Separatory funnel - 250 ml, with Teflon stopcock.
4.2.3 Kuderna-Danish (K-D) apparatus.
4.2.3.1 Concentrator tube - 10 ml graduated (Kontes
K-570050-1025 or equivalent). A ground glass stopper is used to
prevent evaporation of extracts.
4.2.3.2 Evaporation flask - 500 ml (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.2.3.3 Snyder column - Three ball macro (Kontes
K-503000-0121 or equivalent).
4.2.3.4 Snyder column - Two ball micro (Kontes
K-569001-0219 or equivalent).
4.2.3.5 Springs - 1/2 inch (Kontes K-662750 or
equivalent).
4.2.4 Boiling chips - Solvent extracted with methylene chloride,
approximately 10/40 mesh (silicon carbide or equivalent).
4.2.5 pH meter - Capable of measuring to the nearest 0.01 units.
4.2.6 Glass fiber filter paper - 1.2 ^m pore size (Fisher Grade G4
or equivalent).
4.2.7 Solid sorbent cartridges - Packed with 2 g C18 (Baker or
equivalent).
4.2.8 Vacuum manifold - Capable of simultaneous extraction of up to
12 samples (Supelco or equivalent).
4.2.9 Sample reservoirs - 60 ml capacity (Supelco or equivalent).
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4.2.10 Pipet - Capable of accurately delivering 0.10 ml
solution (Pipetman or equivalent).
4.2.11 Water bath - Heated, with concentric ring cover, capable
of temperature control (+ 2°C). The bath should be used under a hood.
4.2.12 Sample shaker - Controlled temperature incubator (+ 2°C)
with orbital shaking (Lab-Line Orbit Environ-Shaker Model 3527 or
equivalent).
4.2.13 Syringes - 5 mL, 500 juL, 100 /jL, (Luer-Lok or
equivalent).
4.2.14 Syringe Filters - 0.45 /urn filtration disks (Gelman
Acrodisc 4438 or equivalent).
4.3 Apparatus and Materials for Option 2
4.3.1 Syringes - 10 ml, with Luer-Lok type adapter, used to
backflush the sample cartridges by gravity feed.
4.3.2 Syringe Rack - made of an aluminum plate with adjustable legs
on all four corners. Circular holes of a diameter slightly larger than
the diameter of the 10 mL syringes are drilled through the plate to allow
batch processing of cartridges for cleaning, coating, and sample elution.
A plate (0.16 x 36 x 53 cm) with 45 holes in a 5x9 matrix is recommended.
See Figure 2 in Method 0100.
4.4 Volumetric Flasks - 5 mL, 10 mL, and 250 or 500 mL.
4.5 Vials - 10 or 25 mL, glass with Teflon-lined screw caps or crimp
tops.
4.6 Balance - Analytical, capable of accurately weighing to 0.0001 g.
4.7 Glass Funnels
4.8 Polyethylene Gloves - used to handle silica gel cartridges.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first, ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water - Water in which an interferant is not
observed at the method detection limit for the compounds of interest.
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5.3 Formalin - Solution of formaldehyde (CH20) in organic-free reagent
water, nominally 37.6 percent (w/w). Exact concentration will be determined for
the stock solution in Sec. 5.7.1.1.
5.4 Aldehydes and ketones - analytical grade, used for preparation of
DNPH derivative standards of target analytes other than formaldehyde. Refer to
the target analyte list.
5.5 Option 1 Reagents
5.5.1 Methylene chloride, CH2C12 - HPLC grade or equivalent.
5.5.2 Acetonitrile, CH3CN - HPLC grade or equivalent.
5.5.3 Sodium hydroxide solutions, NaOH, 1.0 N and 5 N.
5.5.4 Sodium chloride, NaCl, saturated solution - Prepare by
dissolving an excess of the reagent grade solid in organic-free reagent
water.
5.5.5 Sodium sulfite solution, Na2S03, 0.1 M.
5.5.6 Sodium sulfate, Na2S04 - granular, anhydrous.
5.5.7 Citric Acid, C8H807, 1.0 M solution.
5.5.8 Sodium Citrate, C6H5Na307.2H20, 1.0 M trisodium salt dihydrate
solution.
5.5.9 Acetic acid (glacial), CH3C02H.
5.5.10 Sodium acetate, CH3C02Na.
5.5.11 Hydrochloric Acid, HC1, 0.1 N.
5.5.12 Citrate buffer, 1 M, pH 3 - Prepare by adding 80 ml of 1 M
citric acid solution to 20 ml of 1 M sodium citrate solution. Mix
thoroughly. Adjust pH with NaOH or HC1 as needed.
5.5.13 pH 5.0 Acetate buffer (5M) - Formaldehyde analysis only.
Prepared by adding 40 ml 5M acetic acid solution to 60 ml 5M sodium
acetate solution. Mix thoroughly. Adjust pH with NaOH or HC1 as needed.
5.5.14 2,4-Dinitrophenylhydrazine, 2,4-(02N)2C6H3]NHNH2, (DNPH), 70%
in organic-free reagent water (w/w).
5.5.14.1 DNPH (3.00 mg/mL) - Dissolve 428.7 mg of 70% (w/w)
DNPH solution in 100 ml acetonitrile.
5.5.15 Extraction fluid for Option 1 - Dilute 64.3 ml of 1.0 N NaOH
and 5.7 ml glacial acetic acid to 900 ml with organic-free reagent water.
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Dilute to 1 liter with organic-free reagent water. The pH should be 4.93
± 0.02.
5.6 Option 2 Reagents
5.6.1 Acetonitrile, CH3CN - UV grade.
5.6.2 2,4-Dinitrophenylhydrazine, C6H6N404, (DNPH) - recrystallize
at least twice with UV grade acetonitrile using the procedure in Appendix
A.
5.7 Stock Standard Solutions for Option 1
5.7.1 Stock formaldehyde (approximately 1000 mg/L) - Prepare by
diluting an appropriate amount of the certified or standardized
formaldehyde (approximately 265 jxL) to 100 ml with organic-free reagent
water. If a certified formaldehyde solution is not available or there is
any question regarding the quality of a certified solution, the solution
may be standardized using the procedure in Sec. 5.7.1.1.
5.7.1.1 Standardization of formaldehyde stock solution -
Transfer a 25 ml aliquot of a 0.1 M Na2S03 solution to a beaker and
record the pH. Add a 25.0 ml aliquot of the formaldehyde stock
solution (Sec. 5.18.1) and record the pH. Titrate this mixture back
to the original pH using 0.1 N HC1. The formaldehyde concentration
is calculated using the following equation:
Concentration (mg/L) =
where:
(30.03)(N HCl)(mL HC1)
25.0 ml
N HC1
mL HC1
30.03
Normality of HC1 solution used (in milli-
equivalents/mL) (1 mmole of HC1 = 1 mini-
equivalent of HC1)
mL of standardized HC1 solution used
Molecular of weight of formaldehyde (in
mg/mmole)
5.7.2 Stock aldehyde(s) and ketone(s) - Prepare by adding an
appropriate amount of the pure material to 90 mL of acetonitrile and
dilute to 100 mL, to give a final concentration of 1000 mg/L.
5.8 Stock Standard Solutions for Option 2
5.8.1 Preparation of the DNPH Derivatives for HPLC analysis
5.8.1.1 To a portion of the recrystallized DNPH, add
sufficient 2N HC1 to obtain an approximately saturated solution.
Add to this solution the target analyte in molar excess of the DNPH.
Filter the DNPH derivative precipitate, wash it with 2N HC1, wash it
again with water, and allow it to dry in air.
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5.8.1.2 Check the purity of the DNPH derivative by melting
point determination or HPLC analysis. If the impurity level is not
acceptable, recrystallize the derivative in acetonitrile. Repeat
the purity check and recrystallization as necessary until 99% purity
is achieved.
5.8.2 Preparation of DNPH Derivative Standards and Calibration
Standards for HPLC analysis
5.8.2.1 Stock Standard Solutions - Prepare individual
stock standard solutions for each of the target analyte DNPH
derivatives by dissolving accurately weighed amounts in
acetonitrile. Individual stock solutions of approximately 100 mg/L
may be prepared by dissolving 0.010 g of the solid derivative in
100 ml of acetonitrile.
5.8.2.2 Secondary Dilution Standard(s) - Using the
individual stock standard solutions, prepare secondary dilution
standards in acetonitrile containing the DNPH derivatives from the
target analytes mixed together. Solutions of 100 ^g/i may be
prepared by placing 100 jiL of a 100 mg/L solution in a 100 mL
volumetric flask and diluting to the mark with acetonitrile.
5.8.2.3 Calibration Standards - Prepare a working
calibration standard mix from the secondary dilution standard, using
the mixture of DNPH derivatives at concentrations of 0.5-2.0 /ig/L
(which spans the concentration of interest for most indoor air
work). The concentration of the DNPH derivative in the standard mix
solutions may need to be adjusted to reflect relative concentration
distribution in a real sample.
5.9 Standard Storage - Store all standard solutions at 4°C in a glass
vial with a Teflon-lined cap, with minimum headspace, and in the dark. They
should be stable for about 6 weeks. All standards should be checked frequently
for signs of degradation or evaporation, especially just prior to preparing
calibration standards from them.
5.10 Calibration Standards
5.10.1 Prepare calibration solutions at a minimum of 5
concentrations for each analyte of interest in organic-free reagent water
(or acetonitrile for Option 2) from the stock standard solution. The
lowest concentration of each analyte should be at, or just above, the MDLs
listed in Tables 1 or 2. The other concentrations of the calibration
curve should correspond to the expected range of concentrations found in
real samples.
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes, Sec.
4.1.
6.2 Samples must be refrigerated at 4°C. Aqueous samples must be
derivatized and extracted within 3 days of sample collection. The holding times
of leachates of solid samples should be kept at a minimum. All derivatized
sample extracts should be analyzed within 3 days after preparation.
6.3 Sample;; collected by Methods 0011 or 0100 must be refrigerated at
4°C. It is recommended that samples be extracted and analyzed within 30 days of
collection.
7.0 PROCEDURE
7.1 Extraction of Solid Samples (Option 1)
7.1.1 All solid samples should be made as homogeneous as possible
by stirring and removal of sticks, rocks, and other extraneous material.
When the sample is not dry, determine the dry weight of the sample, using
a representative aliquot. If particle size reduction is necessary,
proceed as per Method 1311.
7.1.1.1 Determination of dry weight - In certain cases,
sample results are desired based on a dry weight basis. When such
data are desired or required, a portion of sample for dry weight
determination should be weighed out at the same time as the portion
used for analytical determination.
WARNING: The drying oven should be contained in a hood or
vented. Significant laboratory contamination may
result from drying a heavily contaminated
hazardous waste sample.
7.1.1.2 Immediately after weighing the sample for
extraction, weigh 5-10 g of the sample into a tared crucible.
Determine the % dry weight of the sample by drying overnight at
105°C. Allow to cool in a desiccator before weighing:
g of dry sample
% dry weight = x 100
g of sample
7.1.2 Measure 25 g of solid into a 500 mL bottle with a Teflon
lined screw cap or crimp top, and add 500 mL of extraction fluid (Sec.
5.5.15). Extract the solid by rotating the bottle at approximately 30 rpm
for 18 hours. Filter the extract through glass fiber filter paper and
store in sealed bottles at 4°C. Each mL of extract represents 0.050 g
solid. Smaller quantities of solid sample may be used with
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correspondingly reduced volumes of extraction fluid maintaining the 1:20
mass to volume ratio.
7.2 Cleanup and Separation (Option 1)
7.2.1 Cleanup procedures may not be necessary for a relatively
clean sample matrix. The cleanup procedures recommended in this method
have been used for the analysis of various sample types. If particular
samples demand the use of an alternative cleanup procedure, the analyst
must determine the elution profile and demonstrate that the recovery of
formaldehyde from a spiked sample is greater than 85%. Recovery may be
lower for samples which form emulsions.
7.2.2 If the sample is not clear, or the complexity is unknown, the
entire sample should be centrifuged at 2500 rpm for 10 minutes. Decant
the supernatant liquid from the centrifuge bottle, and filter through
glass fiber filter paper into a container which can be tightly sealed.
7.3 Derivatization (Option 1)
7.3.1 For aqueous samples, measure an aliquot of sample which is
appropriate to the anticipated analyte concentration range (nominally
100 ml). Quantitatively transfer the sample aliquot to the reaction
vessel (Sec. 4.2).
7.3.2 For solid samples, 1 to 10 ml of extract (Sec. 7.1) will
usually be required. The amount used for a particular sample must be
determined through preliminary experiments.
NOTE: In cases where the selected sample or extract volume is less
than 100 ml, the total volume of the aqueous layer should be
adjusted to 100 ml with organic-free reagent water. Record
original sample volume prior to dilution.
7.3.3 Derivatization and extraction of the target analytes may be
accomplished using the liquid-solid (Sec. 7.3.4) or liquid-liquid (Sec.
7.3.5) procedures.
7.3.4 Liquid-Solid Derivatization and Extraction
7.3.4.1 For analytes other than formaldehyde, add 4 ml of
citrate buffer and adjust the pH to 3.0 + 0.1 with 6M HC1 or 6M
NaOH. Add 6 ml of DNPH reagent, seal the container, and place in a
heated (40°C), orbital shaker (Sec. 4.2.12) for 1 hour. Adjust the
agitation to produce a gentle swirling of the reaction solution.
7.3.4.2 If formaldehyde is the only analyte of interest,
add 4 mL acetate buffer and adjust pH to 5.0 + 0.1 with 6M HC1 or 6M
NaOH. Add 6 ml of DNPH reagent, seal the container, and place in a
heated (40°C), orbital shaker (Sec. 4.2.12) for 1 hour. Adjust the
agitation to produce a gentle swirling of the reaction solution.
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7.3.4.3 Assemble the vacuum manifold and connect to a
water aspirator or vacuum pump. Attach a 2 g sorbent cartridge to
the vacuum manifold. Condition each cartridge by passing 10 ml
dilute citrate buffer (10 ml of 1 M citrate buffer dissolved in 250
ml of organic-free reagent water) through each sorbent cartridge.
7.3.4.4 Remove the reaction vessel from the shaker
immediately at the end of the one hour reaction period and add 10 ml
saturated NaCl solution to the vessel.
7.3.4.5 Quantitatively transfer the reaction solution to
the sorbent cartridge and apply a vacuum so that the solution is
drawn through the cartridge at a rate of 3 to 5 mL/min. Continue
applying the vacuum for about 1 minute after the liquid sample has
passed through the cartridge.
7.3.4.6 While maintaining the vacuum conditions described
in Sec. 7.3.4.4, elute each cartridge train with approximately 9 ml
of acetonitrile directly into a 10 ml volumetric flask. Dilute the
solution to volume with acetonitrile, mix thoroughly, and place in
a tightly sealed vial until analyzed.
NOTE:
Because this method uses an excess of DNPH, the
cartridges will remain a yellow color after
completion of Sec. 7.3.4.5. The presence of this
color is not indicative of the loss of the
analyte derivatives.
7.3.5 Liquid-Liquid Derivatization and Extraction
7.3.5.1 For analytes other than formaldehyde, add 4 mL of
citrate buffer and adjust the pH to 3.0 + 0.1 with 6M HC1 or 6M
NaOH. Add 6 mL of DNPH reagent, seal the container, and place in a
heated (40°C), orbital shaker for 1 hour. Adjust the agitation to
produce a gentle swirling of the reaction solution.
7.3.5.2 If formaldehyde is the only analyte of interest,
add 4 mL acetate buffer and adjust pH to 5.0 ± 0.1 with 6M HC1 or 6M
NaOH. Add 6 mL of DNPH reagent, seal the container, and place in a
heated (40°C), orbital shaker for 1 hour. Adjust the agitation to
produce a gentle swirling of the reaction solution.
7.3.5.3 Serially extract the solution with three 20 mL
portions of methylene chloride using a 250 mL separatory funnel. If
an emulsion forms upon extraction, remove the entire emulsion and
centrifuge at 2000 rpm for 10 minutes. Separate the layers and
proceed with the next extraction. Combine the methylene chloride
layers in a 125 mL Erlenmeyer flask containing 5.0 grams of
anhydrous sodium sulfate. Swirl contents to complete the extract
drying process.
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7.3.5.4 Assemble a Kuderna-Danish (K-D) concentrator by
attaching a 10 ml concentrator tube to a 500 ml evaporator flask.
Pour the extract into the evaporator flask being careful to minimize
transfer of sodium sulfate granules. Wash the Erlenmeyer flask with
30 ml of methylene chloride and add wash to the evaporator flask to
complete quantitative transfer.
7.3.5.5 Add one to two clean boiling chips to the
evaporative flask and attach a three ball Snyder column. Prewet the
Snyder column by adding about 1 ml methylene chloride to the top.
Place the K-D apparatus on a hot water bath (80-90°C) so that the
concentrator tube is partially immersed in the hot water and the
entire lower rounded surface of the flask is bathed with hot vapor.
Adjust the vertical position of the apparatus and the water
temperature, as required, to complete the concentration in 10-15
min. At the proper rate of distillation the balls of the column
will actively chatter, but the chambers will not flood with
condensed solvent. When the apparent volume of liquid reaches 5 ml,
remove the K-D apparatus and allow it to drain and cool for at least
10 min.
7.3.5.6 Prior to liquid chromatographic analysis, the
extract solvent must be exchanged to acetonitrile. The analyst must
ensure quantitative transfer of the extract concentrate. The
exchange is performed as follows:
7.3.5.6.1 Remove the three-ball Snyder column and
evaporator flask. Add 5 ml of acetonitrile , a new glass
bead or boiling chip, and attach the micro-Snyder column to
the concentrator tube. Concentrate the extract using 1 ml
of acetonitrile to prewet the Snyder column. Place the K-D
apparatus on the water bath so that the concentrator tube is
partially immersed in the hot water. Adjust the vertical
position of the apparatus and the water temperature, as
required, to complete concentration. At the proper rate of
distillation the balls of the column will actively chatter,
but the chambers will not flood. When the apparent volume
of liquid reaches less than 5 ml, remove the K-D apparatus
and allow it to drain and cool for at least 10 minutes.
7.3.5.6.2 Remove the Snyder column and rinse the flask
and its lower joint with 1-2 ml of acetonitrile and add to
concentrator tube. Quantitatively transfer the sample to a
10 mL volumetric flask using a 5 ml syringe with an attached
Acrodisc 0.45 /im filter cassette. Adjust the extract volume
to 10 ml. Stopper the flask and store refrigerated at 4°C
if further processing will not be performed immediately. If
the extract will be stored longer than two (2) days, it
should be transferred to a vial with a Teflon lined screw
cap or crimp top. Proceed with HPLC chromatographic
analysis if further cleanup is not required.
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7.4 Extraction of Samples from Methods 0011 and 0100 (Options 1 and 2)
7.4.1 Stack gas samples collected by Method 0011 (Option 1)
7.4.1.1 Measure the volume of the aqueous phase of the
sample prior to extraction (for moisture determination in case the
volume was not measured in the field). Pour the sample into a
separatory funnel and drain the methylene chloride into a volumetric
flask.
7.4.1.2 Extract the aqueous solution with two or three
aliquots of methylene chloride. Add the methylene chloride extracts
to the volumetric flask.
7.4.1.3 Fill the volumetric flask to the line with
methylene chloride. Mix well and remove an aliquot.
7.4.1.4 If high concentrations of formaldehyde are
present, the extract can be diluted with mobile phase, otherwise the
extract solvent must be exchanged as described in Sec. 7.3.5.5. If
low concentrations of formaldehyde are present, the sample should be
concentrated during the solvent exchange procedure.
7.4.1.5 Store the sample at 4°C. If the extract will be
stored longer than two days, it should be transferred to a vial with
a Teflon-lined screw cap, or a crimp top with a Teflon-lined septum.
Proceed with HPLC chromatographic analysis if further cleanup is not
required.
7.4.2 Ambient air samples collected by Method 0100 (Option 2)
7.4.2.1 The samples will be received by the laboratory in
a friction-top can containing 2 to 5 cm of granular charcoal, and
should be stored in this can, in a refrigerator, until analysis.
Alternatively, the samples may also be stored alone in their
individual glass containers. The time between sampling and analysis
should not exceed 30 days.
7.4.2.2 Remove the sample cartridge from the labeled
culture tube. Connect the sample cartridge (outlet or long end
during sampling) to a clean syringe.
NOTE: The liquid flow during desorption should be in
the opposite direction from the air flow during
sample collection (i.e, backflush the cartridge).
7.4.2.3 Place the cartridge/syringe in the syringe rack.
7.4.2.4 Backflush the cartridge (gravity feed) by passing
6 ml of acetonitrile from the syringe through the cartridge to a
graduated test tube, or to a 5 ml volumetric flask.
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NOTE: A dry cartridge has an acetonitrile holdup volume
slightly greater than 1 ml. The eluate flow may
stop before the acetonitrile in the syringe is
completely drained into the cartridge because of
air trapped between the cartridge filter and the
syringe Luer-Lok tip. If this happens, displace
the trapped air with the acetonitrile in the
syringe using a long-tip disposable Pasteur
pipet.
7.4.2.5 Dilute to the 5 ml mark with acetonitrile. Label
the flask with sample identification. Pipet two aliquots into
sample vials having Teflon-lined septa.
7.4.2.6 Store the sample at 4°C. Proceed with HPLC
chromatographic analysis of the first aliquot if further cleanup is
not required. Store the second aliquot in the refrigerator until
the results of the analysis of the first aliquot are complete and
validated. The second aliquot can be used for confirmatory
analysis, if necessary.
7.5 Chromatographic Conditions (Recommended):
7.5.1 Option 1 - For aqueous samples, soil or waste samples, and
stack gas samples collected by Method 0011.
Column: CIS, 4.6 mm x 250 mm ID, 5 /zm particle size
Mobile Phase Gradient: 70%/30% acetonitrile/water (v/v), hold for
20 min.
70%/30% acetonitrile/water to 100%
acetonitrile in 15 min.
100% acetonitrile for 15 min.
Flow Rate: 1.2 mL/min
Detector: Ultraviolet, operated at 360 nm
Injection Volume: 20 fj,l
7.5.2 Option 2 - For ambient air samples collected by Method 0100.
Column: Two HPLC columns, 4.6 mm x 250 mm ID,
(Zorbax ODS, or equivalent) in series
Mobile Phase Gradient: 60%/40% CH3CN/H20, hold for 0 min.
60%/40% to 75%/25% CH3CN/H20, linearly in 30
min.
75%/25% to 100%/0% CH3CN/H20, linearly in 20
min.
100% CH3CN for 5 minutes.
100%/0% to 60%/40% CH3CN/H20, linearly in 1
min.
60%/40% CH3CN/H20 for 15 minutes.
Detector: Ultraviolet, operated at 360 nm
Flow Rate: 1.0 mL/min
Sample Injection volume:25 ^l (suggested)
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NOTE: For Options 1 and 2, analysts are advised to adjust their
HPLC systems to optimize chromatographic conditions for
their particular analytical needs. The separation of
acrolein, acetone, and propionaldehyde should be a minimum
criterion of the optimization in Option 2.
7.5.3 Filter and degas the mobile phase to remove dissolved gasses,
using the following procedure:
7.5.3.1 Filter each solvent (water and acetonitrile)
through a 0.22 pm polyester membrane filter, in an all glass and
Teflon suction filtration apparatus.
7.5.3.2 Degas each filtered solution by purging with
helium for 10-15 minutes (100 mL/min) or by heating to 60°C for 5-10
minutes in an Erlenmeyer flask covered with a watch glass. A
constant back pressure restrictor (350 kPa) or 15-30 cm of 0.25 mm
ID Teflon tubing should be placed after the detector to eliminate
further mobile phase outgassing.
7.5.3.3 Place the mobile phase components in their
respective HPLC solvent reservoirs, and program the gradient system
according to the conditions listed in Sec. 7.5.2. Allow the system
to pump for 20-30 minutes at a flow rate of 1.0 mL/min with the
initial solvent mixture ratio (60%/40% CH3CN/H20). Display the
detector output on a strip chart recorder or similar output device
to establish a stable baseline.
7.6 Calibration
7.6.1 Establish liquid chromatographic operating conditions to
produce a retention time similar to that indicated in Table 1 for the
liquid-solid derivatization and extraction or in Table 2 for liquid-liquid
derivatization and extraction. For determination of retention time
windows, see Sec. 7.5 of Method 8000. Suggested chromatographic
conditions are provided in Sec. 7.5.
7.6.2 Process each calibration standard solution through
derivatization and extraction, using the same procedure employed for
sample processing (Sees. 7.3.4 or 7.3.5).
7.6.3 Analyze a solvent blank to ensure that the system is clean
and interference free.
NOTE: The samples and standards must be allowed to come to ambient
temperature before analysis.
7.6.4 Analyze each processed calibration standard using the
chromatographic conditions listed in Sec. 7.5, and tabulate peak area
against calibration solution concentration in /jg/L.
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7.6.5 Tabulate the peak area along with standard concentration
injected to determine the response factor (RF) for the analyte at each
concentration (see Sec. 7.8.1 for equations). The percent relative
standard deviation (%RSD) of the mean RF of the calibration standards
should be no greater than ± 20 percent or a system check will have to be
performed. If a calibration check after the system check does not meet
the criteria, a recalibration will have to be performed. If the
recalibration does not meet the established criteria, new calibration
standards must be made.
7.6.6 The working calibration curve must be verified each day,
before and after analyses are performed, by analyzing one or more
calibration standards. The response obtained should fall within + 15
percent of the initially established response or a system check will have
to be performed. If a calibration check after the system check does not
meet the criteria, the system must be recalibrated.
7.6.7 After 10 sample runs, or less, one of the calibration
standards must be reanalyzed to ensure that the DNPH derivative response
factors remain within +15% of the original calibration response factors.
7.7 Sample Analysis
7.7.1 Analyze samples by HPLC, using conditions established in Sec.
7.5. For analytes to be analyzed by Option 1, Tables 1 and 2 list the
retention times and MDLs that were obtained under these conditions. For
Option 2 analytes, refer to Figure 3 for the sample chromatogram.
7.7.2 If the peak area exceeds the linear range of the calibration
curve, a smaller sample injection volume should be used. Alternatively,
the final solution may be diluted with acetonitrile and reanalyzed.
7.7.3 After elution of the target analytes, calculate the
concentration of analytes found in the samples using the equations found
in Sec. 7.8 or the specific sampling method used.
7.7.4 If the peak area measurement is prevented by the presence of
observed interferences, further cleanup is required.
7.8 Calculations
7.8.1 Calculate each response factor, mean response factor, and
percent relative standard deviation as follows:
Concentration of standard injected,
Area of signal
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_ IRF(
Mean RF = RF =
! I (RF, - RF)2 /N-l
%RSD = — x 100%
RF
where:
RF = Mean response factor or mean of the response factors
using the 5 calibration concentrations.
RFj = Response factor for calibration standard i (i = 1-5).
%RSD = Percent relative standard deviation of the response
factors.
N = Number of calibration standards.
7.8.2 Calculate the analyte concentrations in liquid samples as
follows:
Concentration of aldehydes in fj.g/1 - (RF)(Area of signal)(100/V8)
where:
RF = Mean response factor for a particular analyte.
Vs = Number of ml of sample (unitless).
7.8.3 Calculate the analyte concentration in solid samples as
follows:
Concentration of aldehydes in ng/g = (RF)(Area of signal)(20/ Vex)
where:
RF = Mean response factor for a particular analyte.
Vex = Number of ml extraction fluid aliquot (unitless).
7.8.4 Calculate the concentration of formaldehyde in stack gas
samples (Method 0011) as follows: (Option 1)
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7.8.4.1 Calculation of Total Formaldehyde: To determine
the total formaldehyde in mg, use the following equation:
[g/mole formaldehyde]
Total mg formaldehyde = Cd x V x DF x x 10~3 mg/Mg
[g/mole DNPH derivative]
where:
Cd = measured concentration of DNPH-formaldehyde
derivative, mg/L
V = organic extract volume, ml
DF = dilution factor
7.8.4.2 Formaldehyde concentration in stack gas: Determine
the formaldehyde concentration in the stack gas using the following
equation:
Cf = K [total formaldehyde, mg] / Vm(8td)
where:
K = 35.31 ft3/m3, if Vm(8td) is expressed in
English units
1.00 m3/m3, if Vm(std) is expressed in metric
units
Vm(std) = volume of gas sample as measured by dry gas
meter, corrected to standard conditions,
dscm (dscf)
7.8.5 Calculation of the Concentration of Formaldehyde and Other
Carbonyls from Indoor Air Sampling by Method 0100. (Option 2)
7.8.5.1 The concentration of target analyte "a" in air at
standard conditions (25°C and 101.3 kPa), Concastd in ng/L, may be
calculated using the following equation:
(AreaJ (RF) (VolJ (MWJ (1000 ng/Mg)
Conca = x DF
(MWd)(VTotStd)(1000 ml/I)
where:
Areaa = Area of the sample peak for analyte "a"
RF = Mean response factor for analyte "a" from
the calibration in M9/L- (See Sec, 7.8.1)
Vola = Total volume of the sample cartridge eluate
(ml)
MWa = Molecular weight of analyte "a" in g/mole
MWd = Molecular weight of the DNPH derivative of
analyte "a" in g/mole
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"TolStd
DF
Total volume of air sampled converted to
standard conditions in liters (L). (To
calculate the concentration at sampling
conditions use Vtot.)(See Sec. 9.1.3 of
Method 0100)
Dilution Factor for the sample cartridge
eluate, if any. If there is no dilution,
DF = 1
7.8.5.2 The target analyte "a" concentration at standard
conditions may be converted to parts per billion by volume, Cone, in
ppbv, using the following equation:
Conca in ppbv
where:
Conca
22.4
MW,
(Cone.) (22.4)
(MWJ
Concentration of analyte "a" in ng/L
Ideal gas law volume (22.4 nl_ of gas = 1
nmole at standard conditions)
Molecular weight of analyte "a" in g/mole
(or ng/nmole)
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures. Refer to Table 4 for QC acceptance limits derived from the
interlaboratory method validation study on Method 8315.
9.0 METHOD PERFORMANCE
9.1 The MDLs for Option 1 listed in Table 1 were obtained using organic-
free reagent water and liquid-solid extraction. The MDLs for Option 1 listed in
Table 2 were obtained using organic-free reagent water and methylene chloride
extraction. Results reported in Tables 1 and 2 were achieved using fortified
reagent water volumes of 100 mL. Lower detection limits may be obtained using
larger sample volumes.
9.1.1 Option 1 of this method has been tested for linearity of
recovery from spiked organic-free reagent water and has been demonstrated
to be applicable over the range 50-1000 jug/L .
9.1.2 To generate the MDL and precision and accuracy data reported
in this section, analytes were segregated into two spiking groups, A and
B. Representative chromatograms using liquid-solid and liquid-liquid
extraction are presented in Figures 1 (a and b) and 2 (a and b),
respectively.
8315 - 20
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September 1994
-------
9.2 The Sensitivity of Option 2 sampling (Method 0100) and analysis is
listed in Table 3.
9.3 Method 8315, Option 1, was tested by 12 laboratories using reagent
water and ground waters spiked at six concentration levels over the range 30-2200
jug/L. Method accuracy and precision were found to be directly related to the
concentration of the analyte and independent of the sample matrix. Mean recovery
weighted linear regression equations, calculated as a function of spike
concentration, as well as overall and single-analyst precision regression
equations, calculated as functions of mean recovery, are presented in Table 5.
These equations can be used to estimate mean recovery and precision at any
concentration value within the range tested.
10.0 REFERENCES
1. "OSHA Safety and Health Standards, General Industry", (29CRF1910).
Occupational Safety and Health Administration, OSHA 2206, (Revised,
January 1976).
11.0 SAFETY
11.1 The toxicity or carcinogenicity of each reagent used in this method
has not been precisely defined; however, each chemical compound should be treated
as a potential health hazard. From this viewpoint, exposure to these chemicals
must be reduced to the lowest possible level by whatever means available. The
laboratory is responsible for maintaining a current awareness file of OSHA
regulations regarding the safe handling of the chemicals specified in this
method. A reference file of material safety data sheets should also be made
available to all personnel involved in the chemical analysis. Additional
references to laboratory safety are available.
11.2 Formaldehyde has been tentatively classified as a known or suspected,
human or mammalian carcinogen.
8315 - 21 Revision 0
September 1994
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TABLE 1.
OPTION 1 - METHOD DETECTION LIMITS8 USING
LIQUID-SOLID EXTRACTION
Analyte Retention Time MDL
(minutes)
Formaldehyde
Acetaldehyde
Propanal
Crotonaldehyde
Butanal
Cyclohexanone
Pentanal
Hexanal
Heptanal
Octanal
Nonanal
Decanal
5.3
7.4
11.7
16.1
18.1
27.6
28.4
34.1
35.0
40.1
40.4
44.1
6.2
43. 7b
11.0
5.9
6.3
5.8
15.3
10.7
10.0
6.9
13.6
4.4
The method detection limit (MDL) is defined as the minimum
concentration that can be measured with 99% confidence that the
value is above background level. With the exception of
acetaldehyde, all reported MDLs are based upon analyses of 6 to 8
replicate blanks spiked at 25 ng/i. The MDL was computed as
fol1ows:
MDL = t(N.1i0.01)(Std. Dev.)
where:
t(N-i,o.oi> = Tne upper first percentile point of the
t-distribution with n-1 degrees of freedom.
Std. Dev. = Standard deviation, calculated using n-1
degrees of freedom.
The reported MDL is based upon analyses of 3 replicate, fortified
blanks at 250
8315 - 22 Revision 0
September 1994
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TABLE 2.
OPTION 1 - METHOD DETECTION LIMITS8 USING
LIQUID-LIQUID EXTRACTION
Analyte Retention Time
(minutes)
Formaldehyde
Acetaldehyde
Propanal
Crotonaldehyde
Butanal
Cyclohexanone
Pentanal
Hexanal
Heptanal
Octanal
Nonanal
Decanal
5.3
7.4
11.7
16.1
18.1
27.6
28.4
34.1
35.0
40.1
40.4
44.1
MDL
(M9/L)a
23.2
110. 2b
8.4
5.9
7.8
6.9
13.4
12.4
6.6
9.9
7.4
13.1
The method detection limit (MDL) is defined as the minimum
concentration that can be measured with 99% confidence that the value
is above background level. With the exception of acetaldehyde, all
reported MDLs are based upon analyses of 6 to 8 replicate blanks
spiked at 25 /ag/L. The MDL was computed as follows:
= t(N.1i0.ol)(Std. Dev.)
where:
t
-------
TABLE 3.
OPTION 2 - SENSITIVITY (ppb, v/v) OF SAMPLING AND ANALYSIS FOR
CARBONYL COMPOUNDS IN AMBIENT AIR USING AN ADSORBENT CARTRIDGE
FOLLOWED BY GRADIENT HPLC"
Compound
10
Sample Volume (L)b
20 30 40 50 100 200 300 400 500
Acetaldehyde
Acetone
Acrolein
Benzaldehyde
Butyraldehyde
Crotonaldehyde
2,5-Dimethyl-
benzaldehyde
Formaldehyde
Hexanal
Isovaleraldehyde
Propionaldehyde
m-Tolualdehyde
o-Tolualdehyde
p-Tolualdehyde
Valeraldehyde
1
1
1
1
1
1
0
1
1
1
1
1
1
1
1
.36
.28
.29
.07
.21
.22
.97
.45
.09
.15
.28
.02
.02
.02
.15
0.68
0.64
0.65
0.53
0.61
0.61
0.49
0.73
0.55
0.57
0.64
0.51
0.51
0.51
0.57
0.45
0.43
0.43
0.36
0.40
0.41
0.32
0.48
0.36
0.38
0.43
0.34
0.34
0.34
0.38
0.34
0.32
0.32
0.27
0.30
0.31
0.24
0.36
0.27
0.29
0.32
0.25
0.25
0.25
0.29
0.27
0.26
0.26
0.21
0.24
0.24
0.19
0.29
0.22
0.23
0.26
0.20
0.20
0.20
0.23
0.14
0.13
0.13
0.11
0.12
0.12
0.10
0.15
0.11
0.11
0.13
0.10
0.10
0.10
0.11
0.07
0.06
0.06
0.05
0.06
0.06
0.05
0.07
0.05
0.06
0.06
0.05
0.05
0.05
0.06
0.05
0.04
0.04
0.04
0.04
0.04
0.03
0.05
0.04
0.04
0.04
0.03
0.03
0.03
0.04
0.03
0.03
0.03
0.03
0.03
0.03
0.02
0.04
0.03
0.03
0.03
0.03
0.03
0.03
0.03
0.03
0.03
0.03
0.02
0.02
0.02
0.02
0.03
0.02
0.02
0.03
0.02
0.02
0.02
0.02
a The ppb values are measured at 1 atm and 25°C. The sample cartridge is
eluted with 5 mL acetonitrile and 25 /iL is injected into the HPLC. The
maximum sampling flow through a DNPH-coated Sep-Pak is about 1.5 L/minute.
b A sample volume of 1000 L was also analyzed. The results show a
sensitivity of 0.01 ppb for all the target analytes.
8315 - 24
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September 1994
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TABLE 4.
PERFORMANCE-BASED QC ACCEPTANCE LIMITS CALCULATED
USING THE COLLABORATIVE STUDY DATA
Spike
Analyte Concentration"
Formaldehyde
Propanal
Crotonaldehyde
Butanal
Cyclohexanone
Hexanal
Octanal
Decanal
160
160
160
160
160
160
160
160
X"
154
148
160
151
169
151
145
153
s»°
30.5
22.4
34.8
22.7
39.2
34.6
40.1
40.0
Acceptance
Limits, %d
39-153
50-134
35-165
52-137
32-179
30-159
15-166
21-171
Spike concentration, M9/L-
Mean recovery calculated using the reagent water, mean recovery, linear
regression equation, ng/l.
Overall standard deviation calculated using the reagent water, overall
standard deviation linear regression equation, M9/L.
Acceptance limits calculated as (X + 3sR)100/spike concentration.
8315 - 25 Revision 0
September 1994
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TABLE 5.
WEIGHTED LINEAR REGRESSION EQUATIONS FOR MEAN RECOVERY AND PRECISION
Analyte
Formaldehyde
Propanal
Crotonaldehyde
Butanal
Cyclohexanone
Hexanal
Octanal
Decanal
Appl icable
Cone. Range
39.2-2450
31.9-2000
32.4-2030
35.4-2220
31.6-1970
34.1-2130
32.9-2050
33.2-2080
Reagent Water
X 0.909C + 8.79
SR 0.185X + 1.98a
sr 0.093X + 5.79
X 0.858C + 10.49
SR 0.140X + 1.63
sr 0.056X + 2.76
X 0.975C + 4.36
SR 0.185X + 5.15
sr 0.096X + 1.85
X 0.902C + 6.65
SR 0.149X + 0.21
s, 0.086X - 0.71
X 0.962C + 14.97
SR 0.204X + 4.73a
sr 0.187X + 3.46
X 0.844C + 15.81
SR 0.169X + 9.07
sr 0.098X + 0.37a
X 0.856C + 7.88
SR 0.200X + 11.17
sr 0.092X + 1.71a
X 0.883C + 12.00
SR 0.225X + 5.52
sr 0.088X + 2.28a
a Variance is not constant over concentration range.
X Mean recovery, M9/U exclusive of outliers.
SR Overall standard deviation, M9/L> exclusive of outl
sr Single-analyst standard deviation, M9/U exclusive
Ground Water
0.870C +14.84
0.177X + 13.85
0.108X + 6.24
0.892C + 22.22
0.180X + 12.37
0.146X + 2.08a
0.971C + 2.94
0.157X + 6.09
0.119X - 2.27
0.925C + 12.71
0.140X + 6.89
0.108X - 1.63a
0.946C + 28.95
0.345X + 5.02
0.123X + 7.64
0.926C + 9.16
0.132X + 8.31
0.074X - 0.40a
0.914C + 13.09
0.097X + 12.41
0.039X + 1.14
0.908C + 6.46
0.153X + 2.23
0.052X + 0.37
iers.
of outliers.
8315 - 26
Revision 0
September 1994
-------
FIGURE la.
OPTION 2 - LIQUID-SOLID PROCEDURAL STANDARD OF GROUP A ANALYTES AT 625 M9/L
-0.80-
-1.00-
•-1.20-
V*
X
-1.40-
-1.80-
-1.80-'
-2.M
8
1.00
a.oo 3.00
x 10* Minute*
4.00
Retention Time
(minutes)
5.33
11.68
18.13
27.93
36.60
42.99
Analyte
Derivative
Formaldehyde
Propanal
Butanal
Cyclohexanone
Heptanal
Nonanal
8315 - 27
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September 1994
-------
FIGURE Ib.
OPTION 1 - LIQUID-SOLID PROCEDURAL STANDARD OF GROUP B ANALYTES AT 625 M9/L
-0.00-
-0.80-
^
Li
1.00
2.00
3.00
4.00
10*
Retention Time
(minutes)
7.50
16.68
26.88
32.53
40.36
45.49
Analyte
Derivative
Acetaldehyde
Crotonaldehyde
Pentanal
Hexanal
Octanal
Decanal
8315 - 28
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September 1994
-------
FIGURE 2a.
OPTION 1 - LIQUID-LIQUID PROCEDURAL STANDARD OF GROUP A ANALYTES AT 625 M9/L
-1.40-
1.00
a.oo
3.00
> 10* BinutM
4.00
Retention Time
(minutes)
5.82
13.23
20.83
29.95
37.77
43.80
Analyte
Derivative
Formaldehyde
Propanal
Butanal
Cyclohexanone
Heptanal
Nonanal
8315 - 29
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September 1994
-------
FIGURE 2b.
OPTION 1 - LIQUID-LIQUID PROCEDURAL STANDARD OF GROUP B ANALYTES AT 625
-8.00-
1.00
f.OO
3.00
a 10* •inutM
4.00
Retention Time
(minutes)
7.79
17.38
27.22
32.76
40.51
45.62
Analyte
Derivative
Acetaldehyde
Crotonaldehyde
Pentanal
Hexanal
Octanal
Decanal
8315 - 30
Revision 0
September 1994
-------
FIGURE 3.
OPTION 2 - CHROMATOGRAPHIC SEPARATION OF THE DNPH DERIVATIVES
OF 15 CARBONYL COMPOUNDS
ONPH
JuJ
14
15
10
20
TIME, mm
Peak Identification
30
40
Number Compound
Concentration^/ L)
1 Formaldehyde 1.140
2 Acetaldehyde 1.000
3 Acrolein 1.000
4 Acetone 1.000
5 Propanal 1.000
6 Crotonaldehyde 1.000
7 Butanal 0.905
8 Benzaldehyde 1.000
9 Isovaleraldehyde 0.450
10 Pentanal 0.485
11 o-Tolualdehyde 0.515
12 m-Tolualdehyde 0.505
13 p-Tolualdehyde 0.510
14 Hexanal 1.000
15 2,4-Dimethylbenzaldehyde 0.510
8315 - 31
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September 1994
-------
METHOD 8315
DETERMINATION OF CARBONYL COMPOUNDS
BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
Start
0
Ambient Air (Option 2)
7.0 What is
the sample
marix?
stack Gas (Option i
(Media (Option 1)
7.1.1-7.1.1.1
Homogenize sample
and determine dry
weight
Solid
7.1.2 Extract
sample tor 18
hours; filter and
store extract
7.0 Is media
solid or
aqueous?
I Aqueous
Is sample
dear or sample
complexity
known?
Yes
7.2.2 Centrifuge sample
at 2500 rpm for 10
minutes; decant
and filter
7.3.2 Measure 1-10
ml extract; adjust
volume to 100 mL
with water
Solid
7.3.1 Is
media solid
Aqueous
7.3.1 Measure
aliquot of sample;
adjust volume to
100 ml with water
7.3.S.S Exchange
solvent to methanol
O
8315 - 32
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September 1994
-------
METHOD 8315
continued
7.4.1.1 Measure volume
of aqueous phase of
sample: pour sample Into
separatory funnel and
drain methytene chloride
(from Method 0011) Into
volumetric flask
I
7.4.1.2 Extract aqueous
solution with methvtene
chloride; add methytene
chloride extracts to
volumetric task
I
7.4.1.3 Mute to volume
w«h metiytene chloride;
mix wed; remove aliquot
7.4.1.5 Store
sample at 40
7.4.1.4
sample have
a high concentration
of formaldehyde?
7.4.1.4 Dilute
extract with mobile
phase
7.4.1.4 Exchange
solvent with methanol
as in 7.3.5.5
7.4.1.4 Concentrate
extract during
solvent exchange
process
8315 - 33
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September 1994
-------
METHOD 8315
continued
O
7.4.2.2 - 7.4.2.3
Connect sample cartridge
to dean syringe and
place in syringe tack
1
7.4.2.4 Backflush
carfrldgewith
acetonitrite
^7.4.2,4 N.
Doesekiate N. Yes __
ttowb
bkx
.
No
eoome /
tod? /
7.4.24 Displace
trapped air will
it ni*tit*mtltmUit !•>
acetannmein
syringe using a long-tip
disposable Pasteur pjpet
7.4.2.5 DflutB to 5
mLwtthacetonitrile;
label flask; pipat 2
aNquoteinto
sample viato
I
7.4.2.6 Store
sample at 4C
8315 - 34
Revision 0
September 1994
-------
METHOD 8315
continued
7.5.2 Set LC conditions
to produce appropriate
retention times
I
7.5.1 Option
1W2LC
conditions?
7.5.1 Set LC
conditions to produce
appropriate retention
7.5.2.1 Filter and
degas mobile phase
7.6.2 Process calibration
standards through same
processing steps as samples
7.6.3 - 7.6.4
Analyze solvent blank
and calibration standards:
tabulate peak areas
7.6.5 Determine response
factor at each concentration
7.6.5
Does
calibration
check meet
criteria?
7.6.5 Recalibrate
Yes
Yes
7.6.5
Does
__ IJik^MiJMM
caiioraoon
check meet
criteria?
0
7.6.5 Prepare new
calibration
standards
8315 - 35
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September 1994
-------
METHOD 8315
continued
O
1
f
7.6.6 - 7.6.7 Verify
calibration curve every day;
reanalyze 1 calibration
standard after 10
sample runs or less
7.7 Analyze samples
byHPLC
7.7.2 Inject a smaller
volume or dilute sample
7.7.4 Further
cleanup Is required
7.7.2
Does peak
area exceed
calibration
curve?
7.7.4 Are
interferences
present?
781 Calculate each
response factor, mean
response factor, and
percent BSD
i
i
7.8.2 - 7.8.5
Calculate analyte
concentrations
i
Stop
831S - 36
Revision 0
September 1994
-------
APPENDIX A
RECRYSTALLIZATION OF 2,4-DINITROPHENYLHYDRAZINE (DNPH)
NOTE: This procedure should be performed under a properly ventilated hood.
Inhalation of acetonitrile can result in nose and throat irritation (brief
exposure at 500 ppm) or more serious effects at higher concentration
and/or longer exposures.
A.I Prepare a saturated solution of DNPH by boiling excess DNPH in 200 ml
of acetonitrile for approximately 1 hour.
A.2 After 1 hour, remove and transfer the supernatant to a covered beaker
on a hot plate and allow gradual cooling to 40 to 60°C. Maintain this
temperature range until 95% of the solvent has evaporated, leaving crystals.
A.3 Decant the solution to waste and rinse the remaining crystals twice
with three times their apparent volume of acetonitrile.
A.4 Transfer the crystals to a clean beaker, add 200 ml of acetonitrile,
heat to boiling, and again let the crystals grow slowly at 40 to 60°C until 95%
of the solvent has evaporated. Repeat the rinsing process as in Sec. A.3.
A.5 Take an aliquot of the second rinse, dilute 10 times with
acetonitrile, acidify with 1 ml of 3.8 M perchloric acid per 100 mL of DNPH
solution, and analyze with HPLC as in Sec. 7.0 for Option 2. An acceptable
impurity level is less than 0.025 ng/jiL of formaldehyde in recrystallized DNPH
reagent or below the sensitivity (ppb, v/v) level indicated in Table 3 for the
anticipated sample volume.
A.6 If the impurity level is not satisfactory, pipet off the solution to
waste, repeat the recrystallization as in Sec. A.4 but rinse with two 25 ml
portions of acetonitrile. Prep and analyze the second rinse as in Sec. A.5.
A.7 When the impurity level j_s satisfactory, place the crystals in an
all-glass reagent bottle, add another 25 ml of acetonitrile, stopper, and shake
the bottle. Use clean pipets when removing the saturated DNPH stock solution to
reduce the possibility of contamination of the solution. Maintain only a minimum
volume of the saturated solution adequate for day to day operation to minimize
waste of the purified reagent.
8315 - 37 Revision 0
September 1994
-------
METHOD 8316
ACRYLAMIDE, ACRYLONITRILE AND ACROLEIN BY HIGH PERFORMANCE
LIQUID CHROMATOGRAPHY (HPLC)
1.0 SCOPE AND APPLICATION
1.1 The following compounds can be determined by this method:
Compound Name CAS No.a
Acrylamide 79-06-1
Acrylonitrile 107-13-1
Acrolein (Propenal) 107-02-8
a Chemical Abstract Services Registry Number.
1.2 The method detection limits (MDLs) for the target analytes in
organic-free reagent water are listed in Table 1. The method may be applicable
to other matrices.
1.3 This method is restricted to use by or under the supervision of
analysts experienced in the use of high performance liquid chromatographs and
skilled in the interpretation of high performance liquid chromatograms. Each
analyst must demonstrate the ability to generate acceptable results with this
method.
2.0 SUMMARY OF METHOD
2.1 Water samples are analyzed by high performance liquid chromatography
(HPLC). A 200 juL aliquot is injected onto a C-18 reverse-phase column, and
compounds in the effluent are detected with an ultraviolet (UV) detector.
3.0 INTERFERENCES
3.1 Contamination by carryover can occur whenever high-concentration and
low-concentration samples are sequentially analyzed. To reduce carryover, the
sample syringe must be rinsed out between samples with solvent. Whenever an
unusually concentrated sample is encountered, it should be followed by the
analysis of solvent to check for cross contamination.
8316 - 1 Revision 0
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4.0 APPARATUS AND MATERIALS
4.1 HPLC system
4.1.1 One high pressure pump.
4.1.2 Octadecyl Si lane (ODS, C-18) reverse phase HPLC column,
25 cm x 4.6 mm, 10 /xm, (Zorbax, or equivalent).
4.1.3 Variable wavelength UV detector.
4.1.4 Data system.
4.2 Other apparatus
4.2.1 Water degassing unit - 1 liter filter flask with stopper and
pressure tubing.
4.2.2 Analytical balance - + 0.0001 g.
4.2.3 Magnetic stirrer and magnetic stirring bar.
4.2.4 Sample filtration unit - syringe filter with 0.45 /xm filter
membrane, or equivalent disposable filter unit.
4.3 Materials
4.3.1 Syringes - 10, 25, 50 and 250 /iL and 10 mL.
4.3.2 Volumetric pipettes, Class A, glass -1,5 and 10 mL.
4.3.3 Volumetric flasks - 5, 10, 50 and 100 mL.
4.3.4 Vials - 25 mL, glass with Teflon lined screw caps or crimp
tops.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Acrylamide, CH2:CHCONH2, 99+% purity, electrophoresis reagent grade.
5.3 Acrylonitrile, H2C:CHCN, 99+% purity.
5.4 Acrolein, CH2:CHCHO, 99+7. purity.
8316 - 2 Revision 0
September 1994
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5.5 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One. Sparge with He
to eliminate 02 to prevent significant absorption interference from 02 at the 195
nm wavelength.
5.6 Stock standard solutions - Can be prepared from pure standard
materials or can be purchased as certified solutions. Commercially prepared
stock standards can be used if they are certified by the manufacturer and
verified against a standard made from pure material.
5.6.1 Aery1 amide
5.6.1.1 Weigh 0.0100 g of acrylamide neat standard into a
100 ml volumetric flask, and dilute to the mark with organic-free
reagent water. Calculate the concentration of the standard solution
from the actual weight used. When compound purity is assayed to be
96% or greater, the weight can be used without correction to
calculate the concentration of the stock standard.
5.6.1.2 Transfer the stock solution into vials with Teflon
lined screw caps or crimp tops. Store at 4°C, protected from light.
5.6.1.3 Stock solutions must be replaced after six months,
or sooner if comparison with the check standards indicates a
problem.
5.6.2 Acrylonitrile and Acrolein - Prepare separate stock solutions
for acrylonitrile and acrolein.
5.6.2.1 Place about 9.8 ml of organic-free reagent water
into a 10 ml volumetric flask before weighing the flask and stopper.
Weigh the flask and record the weight to the nearest 0.0001 g. Add
two drops of neat standard, using a 50 juL syringe, to the flask.
The liquid must fall directly into the water, without contacting the
inside wall of the flask.
CAUTION: Acrylonitrile and acrolein are toxic. Standard
preparation should be performed in an laboratory
fume hood.
5.6.2.2 Stopper the flask and then reweigh. Dilute to
volume with organic-free reagent water. Calculate the concentration
from the net gain in weight. When compound purity is assayed to be
96% or greater, the weight can be used without correction to
calculate the concentration of the stock standard.
5.6.2.3 Stock solutions must be replaced after six months,
or sooner if comparison with the check standards indicates a
problem.
8316 - 3 Revision 0
September 1994
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5.7 Calibration standards
5.7.1 Prepare calibration standards at a minimum of five
concentrations by diluting the stock solutions with organic-free reagent
water.
5.7.2 One calibration standard should be prepared at a concentration
near, but above, the method detection limit; the remaining standards should
correspond to the range of concentrations found in real samples, but should not
exceed the working range of the HPLC system (1 mg/L to 10 mg/L).
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Sec. 4.1.
7.0
8.0
PROCEDURE
7.1 HPLC Conditions
Mobile Phase:
Injection Volume:
Flow Rate:
Pressure:
Temperature:
Detector UV wavelength:
7.2 Calibration:
Degassed organic-free reagent water
200 ML
2.0 mL/min
38 atm
25°C
195 nm
7.2.1 Prepare standard solutions of acrylamide as described in Sec.
5.7.1. Inject 200 ,uL aliquots of each solution into the chromatograph.
See Method 8000 for additional guidance on calibration by the external
standard method.
7.3 Chromatographic analysis:
7.3.1 Analyze the samples using the same chromatographic conditions
used to prepare the standard curve. Suggested chromatographic conditions
are given in Sec. 7.1. Table 1 provides the retention times that were
obtained under these conditions during method development.
QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Before processing any samples, the analyst must demonstrate, through
the analysis of a method blank, that all glassware and reagents are interference
free.
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9.0 METHOD PERFORMANCE
9.1 Method performance data are not available.
10.0 REFERENCES
1. Hayes, Sam; "Aery!amide, Acrylonitrile, and Acrolein Determination in
Water by High Pressure Liquid Chromatography," USEPA.
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TABLE 1
ANALYTE RETENTION TIMES AND METHOD DETECTION LIMITS
Retention MDL
Compound Time (min) (M9/L)
Acrylamide 3.5 10
Acrylonitrile 8.9 20
Acrolein (Propenal) 10.1 30
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METHOD 8316
ACRYLAMIDE. ACRYLONITRILE AND ACROLEIN BY HIGH PERFORMANCE
LIQUID CHRQMATOGRAPHY (HPLC)
( Start J
7.1 Set by
HPLC
Conditions.
7.2 Calibrate
Chromatograph.
7.3
Chromatographic
analysis.
( Stop J
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METHOD 8318
N-METHYLCARBAMATES BY HIGH PERFORMANCE LIQUID
CHROMATOGRAPHY (HPLC)
1.0 SCOPE AND APPLICATION
1.1 Method 8318 is used to determine the concentration of
N-methylcarbamates in soil, water and waste matrices. The following compounds can
be determined by this method:
Compound Name CAS No.a
Aldicarb (Temik) 116-06-3
Aldicarb Sulfone 1646-88-4
Carbaryl (Sevin) 63-25-2
Carbofuran (Furadan) 1563-66-2
Dioxacarb 6988-21-2
3-Hydroxycarbofuran 16655-82-6
Methiocarb (Mesurol) 2032-65-7
Methomyl (Lannate) 16752-77-5
Promecarb 2631-37-0
Propoxur (Baygon) 114-26-1
a Chemical Abstract Services Registry Number.
1.2 The method detection limits (MDLs) of Method 8318 for determining the
target analytes in organic-free reagent water and in soil are listed in Table 1.
1.3 This method is restricted to use by, or under the supervision of,
analysts experienced in the use of high performance liquid chromatography (HPLC)
and skilled in the interpretation of chromatograms. Each analyst must
demonstrate the ability to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 N-methylcarbamates are extracted from aqueous samples with methylene
chloride, and from soils, oily solid waste and oils with acetonitrile. The
extract solvent is exchanged to methanol/ethylene glycol, and then the extract
is cleaned up on a C-18 cartridge, filtered, and eluted on a C-18 analytical
column. After separation, the target analytes are hydrolyzed and derivatized
post-column, then quantitated fluorometrically.
2.2 Due to the specific nature of this analysis, confirmation by a
secondary method is not essential. However, fluorescence due to post-column
derivatization may be confirmed by substituting the NaOH and o-phthalaldehyde
solutions with organic-free reagent water and reanalyzing the sample. If
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fluorescence is still detected, then a positive interference is present and care
should be taken in the interpretation of the results.
2.3 The sensitivity of the method usually depends on the level of
interferences present, rather than on the instrumental conditions. Waste samples
with a high level of extractable fluorescent compounds are expected to yield
significantly higher detection limits.
3.0 INTERFERENCES
3.1 Fluorescent compounds, primarily alkyl amines and compounds which
yield primary alkyl amines on base hydrolysis, are potential sources of
interferences.
3.2 Coeluting compounds that are fluorescence quenchers may result in
negative interferences.
3.3 Impurities in solvents and reagents are additional sources of
interferences. Before processing any samples, the analyst must demonstrate
daily, through the analysis of solvent blanks, that the entire analytical system
is interference free.
4.0 APPARATUS AND MATERIALS
4.1 HPLC system
4.1.1 An HPLC system capable of injecting 20 juL aliquots and
performing multilinear gradients at a constant flow. The system must also
be equipped with a data system to measure the peak areas.
4.1.2 C-18 reverse phase HPLC column, 25 cm x 4.6 mm (5 /zm).
4.1.3 Post Column Reactor with two solvent delivery systems (Kratos
PCRS 520 with two Kratos Spectroflow 400 Solvent Delivery Systems, or
equivalent).
4.1.4 Fluorescence detector (Kratos Spectroflow 980, or equivalent).
4.2 Other apparatus
4.2.1 Centrifuge.
4.2.2 Analytical balance - + 0.0001 g.
4.2.3 Top loading balance - + 0.01 g.
4.2.4 Platform shaker.
4.2.5 Heating block, or equivalent apparatus, that can accommodate
10 mL graduated vials (Sec. 4.3.11).
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4.3 Materials
4.3.1 HPLC injection syringe - 50 ,uL.
4.3.2 Filter paper, (Whatman #113 or #114, or equivalent).
4.3.3 Volumetric pipettes, Class A, glass, assorted sizes.
p
4.3.4 Reverse phase cartridges, (C-18 Sep-Pak [Waters Associates],
or equivalent).
4.3.5 Glass syringes - 5 ml.
4.3.6 Volumetric flasks, Class A - Sizes as appropriate.
4.3.7 Erlenmeyer flasks with teflon-lined screw caps, 250 ml.
4.3.8 Assorted glass funnels.
4.3.9 Separatory funnels, with ground glass stoppers and teflon
stopcocks - 250 ml.
4.3.10 Graduated cylinders - 100 ml.
4.3.11 Graduated glass vials - 10 ml, 20 ml_.
4.3.12 Centrifuge tubes - 250 ml.
4.3.13 Vials - 25 ml, glass with Teflon lined screw caps or
crimp tops.
4.3.14 Positive displacement micro-pipettor, 3 to 25 /zL
displacement, (Gilson Microman [Rainin #M-25] with tips, [Rainin #CP-25],
or equivalent).
4.3.15 Nylon filter unit, 25 mm diameter, 0.45 /zm pore size,
disposable (Alltech Associates, #2047, or equivalent).
5.0 REAGENTS
5.1 HPLC grade chemicals shall be used in all tests. It is intended that
all reagents shall conform to the specifications of the Committee on Analytical
Reagents of the American Chemical Society, where such specifications are
available. Other grades may be used, provided it is first ascertained that the
reagent is of sufficiently high purity to permit its use without lowering the
accuracy of the determination.
5.2 General
5.2.1 Acetonitrile, CH,CN - HPLC grade - minimum UV cutoff at 203 nm
(EM Omnisolv #AX0142-1, or equivalent).
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5.2.2 Methanol, CH3OH - HPLC grade - minimum UV cutoff at 230 nm (EM
Omni solv #MX0488-1, or equivalent).
5.2.3 Methylene chloride, CH?C1? - HPLC grade - minimum UV cutoff at
230 nm (EM Omnisolv #0X0831-1, or Equivalent).
5.2.4 Hexane, CgH,, - pesticide grade - (EM Omnisolv #HX0298-1, or
equivalent).
5.2.5 Ethylene glycol, HOCH2CH2OH - Reagent grade - (EM Science, or
equivalent).
5.2.6 Organic-free reagent water - All references to water in this
method refer to organic-free reagent water, as defined in Chapter One.
5.2.7 Sodium hydroxide, NaOH - reagent grade - 0.05N NaOH solution.
5.2.8 Phosphoric acid, H3P04 - reagent grade.
5.2.9 pH 10 borate buffer (J.T. Baker #5609-1, or equivalent).
5.2.10 o-Phthalaldehyde, o-CcMCHO),, - reagent grade (Fisher
#0-4241, or equivalent). ° H c
5.2.11 2-Mercaptoethanol, HSCH2CH2OH - reagent grade (Fisher
#0-3446, or equivalent).
5.2.12 N-methylcarbamate neat standards (equivalence to EPA
standards must be demonstrated for purchased solutions).
5.2.13 Chloroacetic acid, C1CH2COOH, 0.1 N.
5.3 Reaction solution
5.3.1 Dissolve 0.500 g of o-phthalaldehyde in 10 ml of methanol, in
a 1 L volumetric flask. To this solution, add 900 ml of organic-free
reagent water, followed by 50 mL of the borate buffer (pH 10). After
mixing well, add 1 ml of 2-mercaptoethanol, and dilute to the mark with
organic-free reagent water. Mix the solution thoroughly. Prepare fresh
solutions on a weekly basis, as needed. Protect from light and store
under refrigeration.
5.4 Standard solutions
5.4.1 Stock standard solutions: prepare individual 1000 mg/L
solutions by adding 0.025 g of carbamate to a 25 mL volumetric flask, and
diluting to the mark with methanol. Store solutions, under refrigeration,
in glass vials with Teflon lined screw caps or crimp tops. Replace every
six months.
5.4.2 Intermediate standard solution: prepare a mixed 50.0 mg/L
solution by adding 2.5 mL of each stock solution to a 50 mL volumetric
flask, and diluting to the mark with methanol. Store solutions, under
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refrigeration, in glass vials with Teflon lined screw caps or crimp tops.
Replace every three months.
5.4.3 Working standard solutions: prepare 0.5, 1.0, 2.0, 3.0 and 5.0
mg/L solutions by adding 0.25, 0.5, 1.0, 1.5 and 2.5 mL of the
intermediate mixed standard to respective 25 ml volumetric flasks, and
diluting each to the mark with methanol. Store solutions, under
refrigeration, in glass vials with Teflon lined screw caps or crimp tops.
Replace every two months, or sooner if necessary.
5.4.4 Mixed QC standard solution: prepare a 40.0 mg/L solution from
another set of stock standard solutions, prepared similarly to those
described in Sec. 5.4.1. Add 2.0 ml of each stock solution to a 50 ml
volumetric flask and dilute to the mark with methanol. Store the
solution, under refrigeration, in a glass vial with a Teflon lined screw
cap or crimp top. Replace every three months.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Due to the extreme instability of N-methylcarbamates in alkaline
media, water, waste water and leachates should be preserved immediately after
collection by acidifying to pH 4-5 with 0.1 N chloroacetic acid.
6.2 Store samples at 4°C and out of direct sunlight, from the time of
collection through analysis. N-methylcarbamates are sensitive to alkaline
hydrolysis and heat.
6.3 All samples must be extracted within seven days of collection, and
analyzed within 40 days of extraction.
7.0 PROCEDURE
7.1 Extraction
7.1.1 Water, domestic wastewater, aqueous industrial wastes, and
leachates
7.1.1.1 Measure 100 mL of sample into a 250 mL separatory
funnel and extract by shaking vigorously for about 2 minutes with 30
mL of methylene chloride. Repeat the extraction two more times.
Combine all three extracts in a 100 mL volumetric flask and dilute
to volume with methylene chloride. If cleanup is required, go to
Sec. 7.2. If cleanup is not required, proceed directly to Sec.
7.3.1.
7.1.2 Soils, solids, sludges, and heavy aqueous suspensions
7.1.2.1 Determination of sample % dry weight - In certain
cases, sample results are desired based on dry-weight basis. When
such data is desired, a portion of sample for this determination
should be weighed out at the same time as the portion used for
analytical determination.
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WARNING: The drying oven should be contained in a hood or
vented. Significant laboratory contamination may
result from a heavily contaminated hazardous
waste sample.
7.1.2.1.1 Immediately after weighing the sample for
extraction, weigh 5-10 g of the sample into a tared crucible.
Determine the % dry weight of the sample by drying overnight
at 105°C. Allow to cool in a desiccator before weighing:
% dry weight = q of dry sample x 100
g of sample
7.1.2.2 Extraction - Weigh out 20 + 0.1 g of sample into
a 250 ml Erlenmeyer flask with a Teflon-lined screw cap. Add 50 ml
of acetonitrile and shake for 2 hours on a platform shaker. Allow
the mixture to settle (5-10 min), then decant the extract into a 250
ml centrifuge tube. Repeat the extraction two more times with 20 ml
of acetonitrile and 1 hour shaking each time. Decant and combine
all three extracts. Centrifuge the combined extract at 200 rpm for
10 min. Carefully decant the supernatant into a 100 mL volumetric
flask and dilute to volume with acetonitrile. (Dilution factor = 5)
Proceed to Sec. 7.3.2.
7.1.3 Soils heavily contaminated with non-aqueous substances, such
as oils
7.1.3.1 Determination of sample % dry weight - Follow
Sees. 7.1.2.1 through 7.1.2.1.1.
7.1.3.2 Extraction - Weigh out 20 + 0.1 g of sample into
a 250 ml Erlenmeyer flask with a Teflon-lined screw cap. Add 60 ml
of hexane and shake for 1 hour on a platform shaker. Add 50 ml of
acetonitrile and shake for an additional 3 hours. Allow the mixture
to settle (5-10 min), then decant the solvent layers into a 250 mL
separatory funnel. Drain the acetonitrile (bottom layer) through
filter paper into a 100 ml volumetric flask. Add 60 mL of hexane and
50 mL of acetonitrile to the sample extraction flask and shake for
1 hour. Allow the mixture to settle, then decant the mixture into
the separatory funnel containing the hexane from the first
extraction. Shake the separatory funnel for 2 minutes, allow the
phases to separate, drain the acetonitrile layer through filter
paper into the volumetric flask, and dilute to volume with
acetonitrile. (Dilution factor = 5) Proceed to Sec. 7.3.2.
7.1.4 Non-aqueous liquids such as oils
7.1.4.1 Extraction - Weigh out 20 + 0.1 g of sample into
a 125 rnL separatory funnel. Add 40 mL of hexane and 25 mL of
acetonitrile and vigorously shake the sample mixture for 2 minutes.
Allow the phases to separate, then drain the acetonitrile (bottom
layer) into a 100 mL volumetric flask. Add 25 mL of acetonitrile to
the sample funnel, shake for 2 minutes, allow the phases to
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Repeat the extraction with another 25 ml portion of acetonitrile,
combining the extracts. Dilute to volume with acetonitrile.
(Dilution factor = 5). Proceed to Sec. 7.3.2.
7.2 Cleanup - Pipet 20.0 ml of the extract into a 20 ml glass vial
containing 100 /uL of ethylene glycol. Place the vial in a heating block set at
50° C, and gently evaporate the extract under a stream of nitrogen (in a fume
hood) until only the ethylene glycol keeper remains. Dissolve the ethylene
glycol residue in 2 ml of methanol, pass the extract through a pre-washed C-18
reverse phase cartridge, and collect the eluate in a 5 ml volumetric flask.
Elute the cartridge with methanol, and collect the eluate until the final volume
of 5.0 mL is obtained. (Dilution factor = 0.25) Using a disposable 0.45 /urn
filter, filter an aliquot of the clean extract directly into a properly labelled
autosampler vial. The extract is now ready for analysis. Proceed to Sec. 7.4.
7.3 Solvent Exchange
7.3.1 Water, domestic wastewater, aqueous industrial wastes, and
leachates:
Pipet 10.0 ml of the extract into a 10 ml graduated glass vial
containing 100 /uL of ethylene glycol. Place the vial in a heating block
set at 50 C, and gently evaporate the extract under a stream of nitrogen
(in a fume hood) until only the ethylene glycol keeper remains. Add
methanol to the ethylene glycol residue, dropwise, until the total volume
is 1.0 ml. (Dilution factor = 0.1). Using a disposable 0.45 jum filter,
filter this extract directly into a properly labelled autosampler vial.
The extract is now ready for analysis. Proceed to Sec. 7.4.
7.3.2 Soils, solids, sludges, heavy aqueous suspensions, and non-
aqueous liquids:
Elute 15 ml of the acetonitrile extract through a C-18 reverse phase
cartridge, prewashed with 5 ml of acetonitrile. Discard the first 2 ml of
eluate and collect the remainder. Pipet 10.0 ml of the clean extract into
a 10 ml graduated glass vial containing 100 juL of ethylene glycol. Place
the vial in a heating block set at 50° C, and gently evaporate the extract
under a stream of nitrogen (in a fume hood) until only the ethylene glycol
keeper remains. Add methanol to the ethylene glycol residue, dropwise,
until the total volume is 1.0 ml. (Additional dilution factor = 0.1;
overall dilution factor = 0.5). Using a disposable 0.45 jum filter, filter
this extract directly into a properly labelled autosampler vial. The
extract is now ready for analysis. Proceed to Sec. 7.4.
7.4 Sample Analysis
7.4.1 Analyze the samples using the chromatographic conditions,
post-column reaction parameters and instrument parameters given in Sees.
7.4.1.1, 7.4.1.2, 7.4.1.3 and 7.4.1.4. Table 2 provides the retention
times that were obtained under these conditions during method development.
A chromatogram of the separation is shown in Figure 1.
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7.4.1.1
Chromatographic Conditions (Recommended)
Solvent A: Organic-free reagent water, acidified with
0.4 mL of phosphoric acid per liter of
water
Solvent B: Methanol/acetonitrile (1:1, v/v)
Flow rate: 1.0 mL/min
Injection Volume: 20 juL
Solvent delivery system program:
36.02
Function
FR
B%
B%
B%
B%
B%
B%
ALARM
Duration
(min)
20
5
5
3
7
0.01
7.4.1.2
Post-column Hydrolysis Parameters (Recommended)
Solution:
Flow Rate:
Temperature:
Residence Time:
0.05 N aqueous sodium hydroxide
0.7 mL/min
95° C
7.4.1.3
(Recommended)
Solution:
35 seconds (1 mL reaction coil)
Post-column Derivatization Parameters
Flow Rate:
Temperature:
Residence time:
o-phthalaldehyde/2-mercaptoethanol
5.3.1)
0.7 mL/min
40° C
25 seconds (1 mL reaction coil)
(Sec.
7.4.1.4
Fluorometer Parameters (Recommended)
Cell: 10 ML
Excitation wavelength: 340 nm
Emission wavelength: 418 nm cutoff filter
Sensitivity wavelength: 0.5 juA
PMT voltage: -800 V
Time constant: 2 sec
7.4.2 If the peak areas of the sample signals exceed the calibration
range of the system, dilute the extract as necessary and reanalyze the
diluted extract.
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7.5 Calibration:
7.5.1 Analyze a solvent blank (20 juL of methanol) to ensure that the
system is clean. Analyze the calibration standards (Sec. 5.4.3), starting
with the 0.5 mg/L standards and ending with the 5.0 mg/L standard. If the
percent relative standard deviation (%RSD) of the mean response factor
(RF) for each analyte does not exceed 20%, the system is calibrated and
the analysis of samples may proceed. If the %RSD for any analyte exceeds
20%, recheck the system and/or recalibrate with freshly prepared
calibration solutions.
7.5.2 Using the established calibration mean response factors, check
the calibration of the instrument at the beginning of each day by
analyzing the 2.0 mg/L mixed standard. If the concentration of each
analyte falls within the range of 1.70 to 2.30 mg/L (i.e., within + 15% of
the true value), the instrument is considered to be calibrated and the
analysis of samples may proceed. If the observed value of any analyte
exceeds its true value by more than ± 15%, the instrument must be
recalibrated (Sec. 7.5.1).
7.5.3 After 10 sample runs, or less, the 2.0 mg/L standards must be
analyzed to ensure that the retention times and response factors are still
within acceptable limits. Significant variations (i.e., observed
concentrations exceeding the true concentrations by more than + 15%) may
require a re-analysis of the samples.
7.6 Calculations
7.6.1 Calculate each response factor as follows (mean value based on
5 points):
concentration of standard
RF =
area of the signal
5
(I RF.)
mean RF = RF =
[(I RFi - RF)2]1/2 / 4
i
%RSD of RF = — X 100%
RF
7.6.2 Calculate the concentration of each N-methylcarbamate as
follows:
jug/g or mg/L = (RF) (area of signal) (dilution factor)
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8.0 QUALITY CONTROL
8.1 Before processing any samples, the analyst must demonstrate, through
the analysis of a method blank for each matrix type, that all glassware and
reagents are interference free. Each time there is a change of reagents, a
method blank must be processed as a safeguard against laboratory contamination.
8.2 A QC check solution must be prepared and analyzed with each sample
batch that is processed. Prepare this solution, at a concentration of 2.0 mg/L
of each analyte, from the 40.0 mg/L mixed QC standard solution (Sec. 5.4.4). The
acceptable response range is 1.7 to 2.3 mg/L for each analyte.
8.3 Negative interference due to quenching may be examined by spiking the
extract with the appropriate standard, at an appropriate concentration, and
examining the observed response against the expected response.
8.4 Confirm any detected analytes by substituting the NaOH and OPA
reagents in the post column reaction system with deionized water, and reanalyze
the suspected extract. Continued fluorescence response will indicate that a
positive interference is present (since the fluorescence response is not due to
the post column derivatization). Exercise caution in the interpretation of the
chromatogram.
9.0 METHOD PERFORMANCE
9.1 Table 1 lists the single operator method detection limit (MDL) for
each compound in organic-free reagent water and soil. Seven/ten replicate
samples were analyzed, as indicated in the table. See reference 7 for more
details.
9.2 Tables 2, 3 and 4 list the single operator average recoveries and
standard deviations for organic-free reagent water, wastewater and soil. Ten
replicate samples were analyzed at each indicated spike concentration for each
matrix type.
9.3 The method detection limit, accuracy and precision obtained will be
determined by the sample matrix.
10.0 REFERENCES
1. California Department of Health Services, Hazardous Materials Laboratory,
"N-Methylcarbamates by HPLC", Revision No. 1.0, September 14, 1989.
2. Krause, R.T. Journal of Chromatographic Science, 1978, vol. 16, pg 281.
3. Klotter, Kevin, and Robert Cunico, "HPLC Post Column Detection of
Carbamate Pesticides", Varian Instrument Group, Walnut Creek, CA 94598.
4. USEPA, "Method 531. Measurement of N-Methylcarbomyloximes and N-
Methylcarbamates in Drinking Water by Direct Aqueous Injection HPLC with
Post Column Derivatization", EPA 600/4-85-054, Environmental Monitoring
and Support Laboratory, Cincinnati, OH 45268.
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5. USEPA, "Method 632. The Determination of Carbamate and Urea Pesticides in
Industrial and Municipal Wastewater", EPA 600/4-21-014, Environmental
Monitoring and Support Laboratory, Cincinnati, OH 45268.
6. Federal Register, "Appendix B to Part 136 - Definition and Procedure for
the Determination of the Method Detection Limit - Revision 1.11", Friday,
October 26, 1984, 49, No. 209, 198-199.
7. Okamoto, H.S., D. Wijekoon, C. Esperanza, J. Cheng, S. Park, J. Garcha, S.
Gill, K. Perera "Analysis for N-Methylcarbamate Pesticides by HPLC in
Environmental Samples", Proceedings of the Fifth Annual USEPA Symposium on
Waste Testing and Quality Assurance, July 24-28, 1989, Vol. II, 57-71.
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TABLE 1
ELUTION ORDER, RETENTION TIMESa AND
SINGLE OPERATOR METHOD DETECTION LIMITS
Method Detection Limits
Compound
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
cr-Naphthor
Methiocarb (Mesurol)
Promecarb
Retention
Time
(min)
9.59
9.59
12.70
13.50
16.05
18.06
18.28
19.13
20.30
22.56
23.02
Organic-free
Reagent Water
(M9/L)
1.9C
1.7
2.6
2.2
9.4C
2.4
2.0
1.7
-
3.1
2.5
Soil
(M9/kg)
44C
12c
10c
>50C
12C
17
22
31
-
32
17
a
b
c
d
See Sec. 7.4 for chromatographic conditions
MDL for organic-free reagent water, sand, soil were determined by
analyzing 10 low concentration spike replicate for each matrix type
(except where noted). See reference 7 for more details.
MDL determined by analyzing 7 spiked replicates.
Breakdown product of Carbaryl.
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TABLE 2
SINGLE OPERATOR AVERAGE RECOVERY AND
PRECISION DATA3 FOR ORGANIC-FREE REAGENT WATER
Compound
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
Methiocarb (Mesurol)
Promecarb
Recovered
225
244
210
241
224
232
239
242
231
227
% Recovery
75.0
81.3
70.0
80.3
74.7
77.3
79.6
80.7
77.0
75.7
SD
7.28
8.34
7.85
8.53
13.5
10.6
9.23
8.56
8.09
9.43
%RSD
3.24
3.42
3.74
3.54
6.03
4.57
3.86
3.54
3.50
4.1
Spike Concentration = 300 jug/L of each compound, n = 10
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TABLE 3
SINGLE OPERATOR AVERAGE RECOVERY AND
PRECISION DATAa FOR WASTEWATER
Compound
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
Methiocarb (Mesurol)
Promecarb
a Spike Concentration
KI/% w«ns*s*t*«*\vttf
Recovered
235
247
2§J
D
258
263
262
262
254
263
= 300 jug/L of each
% Recovery
78.3
82.3
83.7
-
86.0
87.7
87.3
87.3
84.7
87.7
compound, n = 10
SD
17.6
29.9
25.4
-
16.4
16.7
15.7
17.2
19.9
15.1
%RSD
7.49
12.10
10.11
-
6.36
6.47
5.99
6.56
7.83
5.74
recovery
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TABLE 4
SINGLE OPERATOR AVERAGE RECOVERY AND
PRECISION DATA3 FOR SOIL
Compound
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
Methiocarb (Mesurol)
Promecarb
Recovered
1.57
1.48
1.60
1.51
1.29
1.33
1.46
1.53
1.45
1.29
% Recovery
78.5
74.0
80.0
75.5
64.5
66.5
73.0
76.5
72.5
64.7
SD
0.069
0.086
0.071
0.073
0.142
0.126
0.092
0.076
0.071
0.124
%RSD
4.39
5.81
4.44
4.83
11.0
9.47
6.30
4.90
4.90
9.61
Spike Concentration = 2.00 mg/kg of each compound, n = 10
8318 - 15
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FIGURE 1
100-
R
E
S
P
0
N
S
E
TIME (MIN)
1.00 ]ug/mL EACH OF:
1. ALDICARB SULFONE
2. METHOMYL
3. 3-HYDROXYCARBOFURAN
4. DIOXACARB
5. ALDICARB
6. PROPOXUR
7. CARBOFURAN
8. CARBARYL
9. METHIOCARB
10. PROMECARB
8318 - 16
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METHOD 8321
SOLVENT EXTRACTABLE NON-VOLATILE COMPOUNDS BY
HIGH PERFORMANCE LIQUID CHRQMATOGRAPHY/THERMOSPRAY/MASS SPECTROMETRY
(HPLC/TSP/MS) OR ULTRAVIOLET (UV) DETECTION
1.0 SCOPE AND APPLICATION
1.1 This method covers the use of high performance liquid chromatography
(HPLC), coupled with either thermospray-mass spectrometry (TSP-MS), and/or
ultraviolet (UV), for the determination of disperse azo dyes, organophosphorus
compounds, and Tris-(2,3-dibromopropyl)phosphate in wastewater, ground water,
sludge, and soil/sediment matrices, and chlorinated phenoxyacid compounds and
their esters in wastewater, ground water, and soil/sediment matrices. Data are
also provided for chlorophenoxy acid herbicides in fly ash (Table 15), however,
recoveries for most compounds are very poor indicating poor extraction efficiency
for these analytes using the extraction procedure included in this method.
Additionally, this method may apply to other non-volatile compounds that are
solvent extractable, are amenable to HPLC, and are ionizable under thermospray
introduction for mass spectrometric detection. The following compounds can be
determined by this method:
Compound Name
Fluorescent Brightener 61
Fluorescent Brightener 236
Alkaloids
Caffeine
Strychnine
CAS No.'
Azo Dyes
Disperse Red 1
Disperse Red 5
Disperse Red 13
Disperse Yellow 5
Disperse Orange 3
Disperse Orange 30
Disperse Brown 1
Solvent Red 3
Solvent Red 23
Anthraquinone Dyes
Disperse Blue 3
Disperse Blue 14
Disperse Red 60
Coumarin Dyes
(Fluorescent Briqhteners)
2872-52-8
3180-81-2
2832-40-8
6439-53-8
730-40-5
5261-31-4
17464-91-4
6535-42-8
85-86-9
2475-46-9
2475-44-7
17418-58-5
8066-05-5
63590-17-0
58-08-2
57-24-9
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Compound Name CAS No.'
Organophosphorus Compounds
Methomyl 16752-77-5
Thiofanox 39196-18-4
Famphur 52-85-7
Asulam 3337-71-1
Dichlorvos 62-73-7
Dimethoate 60-51-5
Disulfoton 298-04-4
Fensulfothion 115-90-2
Merphos 150-50-5
Methyl parathion 298-00-0
Monocrotophos 919-44-8
Naled 300-76-5
Phorate 298-02-2
Trichlorfon 52-68-6
Tris-(2,3-Dibromopropyl) phosphate, (Tris-BP) 126-72-7
Chlorinated Phenox.yacid Compounds
Dalapon 75-99-0
Dicamba 1918-00-9
2,4-D 94-75-7
MCPA 94-74-6
MCPP 7085-19-0
Dichlorprop 120-36-5
2,4,5-T 93-76-5
Silvex (2,4,5-TP) 93-72-1
Dinoseb 88-85-7
2,4-DB 94-82-6
2,4-D, butoxyethanol ester 1929-73-3
2,4-D, ethylhexyl ester 1928-43-4
2,4,5-T, butyl ester 93-79-8
2,4,5-T, butoxyethanol ester 2545-59-7
a Chemical Abstract Services Registry Number.
1.2 This method may be applicable to the analysis of other non-volatile
or semi volatile compounds.
1.3 Tris-BP has been classified as a carcinogen. Purified standard
material and stock standard solutions should be handled in a hood.
1.4 Method 8321 is designed to detect the chlorinated phenoxyacid
compounds (free acid form) and their esters without the use of hydrolysis and
esterification in the extraction procedure.
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1.5 The compounds were chosen for analysis by HPLC/MS because they have
been designated as problem compounds that are hard to analyze by traditional
chromatographic methods (e.g. gas chromatography). The sensitivity of this
method is dependent upon the level of interferants within a given matrix, and
varies with compound class and even with compounds within that class.
Additionally, the limit of detection (LOD) is dependent upon the mode of
operation of the mass spectrometer. For example, the LOD for caffeine in the
selected reaction monitoring (SRM) mode is 45 pg of standard injected (10 /xL
injection), while for Disperse Red 1 the LOD is 180 pg. The LOD for caffeine
under single quadrupole scanning is 84 pg and is 600 pg for Disperse Red 1 under
similar scanning conditions.
1.6 The experimentally determined limits of detection (LOD) for the
target analytes are presented in Tables 3, 10, 13, and 14. For further compound
identification, MS/MS (CAD - collision activated dissociation) can be used as an
optional extension of this method.
1.7 This method is restricted to use by or under the supervision of
analysts experienced in the use of high performance liquid chromatographs/mass
spectrometers and skilled in the interpretation of liquid chromatograms and mass
spectra. Each analyst must demonstrate the ability to generate acceptable
results with this method.
2.0 SUMMARY OF METHOD
2.1 This method provides reverse phase high performance liquid
chromatographic (RP/HPLC) and thermospray (TSP) mass spectrometric (MS)
conditions for the detection of the target analytes. Quantitative analysis is
performed by TSP/MS, using an external standard approach. Sample extracts can
be analyzed by direct injection into the thermospray or onto a liquid
chromatographic-thermospray interface. A gradient elution program is used on the
chromatograph to separate the compounds. Detection is achieved both by negative
ionization (discharge electrode) and positive ionization, with a single
quadrupole mass spectrometer. Since this method is based on an HPLC technique,
the use of ultraviolet (UV) detection is optional on routine samples.
2.2 Prior to the use of this method, appropriate sample preparation
techniques must be used.
2.2.1 Samples for analysis of chlorinated phenoxyacid compounds are
prepared by a modification of Method 8151 (see Sec. 7.1.2). In general,
one liter of aqueous sample or fifty grams of solid sample are pH
adjusted, extracted with diethyl ether, concentrated and solvent exchanged
to acetonitrile.
2.2.2 Samples for analysis of the other target analytes are prepared
by established extraction techniques. In general, water samples are
extracted at a neutral pH with methylene chloride, using a separatory
funnel (Method 3510) or a continuous liquid-liquid extractor (Method
3520). Soxhlet (Methods 3540/3541) or ultrasonic (Method 3550) extraction
using methylene chloride/acetone (1:1) is used for solid samples. A
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micro-extraction technique is included for the extraction of Tris-BP from
aqueous and non-aqueous matrices.
2.3 An optional thermospray-mass spectrometry/mass spectrometry
(TS-MS/MS) confirmatory method is provided. Confirmation is obtained by using
MS/MS collision activated dissociation (CAD) or wire-repeller CAD.
3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, 8000 and 8150/8151.
3.2 The use of Florisil Column Cleanup (Method 3620) has been
demonstrated to yield recoveries less than 85% for some of the compounds in this
method, and is therefore not recommended for all compounds. Refer to Table 2 of
Method 3620 for recoveries of organophosphorus compounds as a function of
Florisil fractions.
3.3 Compounds with high proton affinity may mask some of the target
analytes. Therefore, an HPLC must be used as a chromatographic separator, for
quantitative analysis.
3.4 Analytical difficulties encountered with specific organophosphorus
compounds, as applied in this method, may include (but are not limited to) the
following:
3.4.1 Methyl parathion shows some minor degradation upon analysis.
3.4.2 Naled can undergo debromination to form dichlorvos.
3.4.3 Merphos often contains contamination from merphos oxide.
Oxidation of merphos can occur during storage, and possibly upon
introduction into the mass spectrometer.
Refer to Method 8141 for other compound problems as related to the
various extraction methods.
3.5 The chlorinated phenoxy acid compounds, being strong organic acids,
react readily with alkaline substances and may be lost during analysis.
Therefore, glassware and glass wool must be acid-rinsed, and sodium sulfate must
be acidified with sulfuric acid prior to use to avoid this possibility.
3.6 Due to the reactivity of the chlorinated herbicides, the standards
must be prepared in acetonitrile. Methylation will occur if prepared in
methanol.
3.7 Solvents, reagents, glassware, and other sample processing hardware
may yield discrete artifacts or elevated baselines, or both, causing
misinterpretation of chromatograms or spectra. All of these materials must be
demonstrated to be free from interferences under the conditions of the analysis
by running reagent blanks. Specific selection of reagents and purification of
solvents by distillation in all-glass systems may be required.
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3.8 Interferants co-extracted from the sample will vary considerably
from source to source. Retention times of target analytes must be verified by
using reference standards.
3.9 The optional use of HPLC/MS/MS methods aids in the confirmation of
specific analytes. These methods are less subject to chemical noise than other
mass spectrometric methods.
4.0 APPARATUS AND MATERIALS
4.1 HPLC/MS
4.1.1 High Performance Liquid Chromatograph (HPLC) - An analytical
system with programmable solvent delivery system and all required
accessories including 10 /xL injection loop, analytical columns, purging
gases, etc. The solvent delivery system must be capable, at a minimum, of
a binary solvent system. The chromatographic system must be capable of
interfacing with a Mass Spectrometer (MS).
4.1.1.1 HPLC Post-Column Addition Pump - A pump for post-
column addition should be used. Ideally, this pump should be a
syringe pump, and does not have to be capable of solvent
programming.
4.1.1.2 Recommended HPLC Columns - A guard column and an
analytical column are required.
4.1.1.2.1 Guard Column - C18 reverse phase guard
column, 10 mm x 2.6 mm ID, 0.5 /zm frit, or equivalent.
4.1.1.2.2 Analytical Column - C18 reverse phase
column, 100 mm x 2 mm ID, 5 /u,m particle size of ODS-Hypersil;
or Ca reversed phase column, 100 mm x 2 mm ID, 3 urn particle
size of MOS2-Hypersil, or equivalent.
4.1.2 HPLC/MS interface(s)
4.1.2.1 Micromixer - 10 /nL, interfaces HPLC column system
with HPLC post-column addition solvent system.
4.1.2.2 Interface - Thermospray ionization interface and
source that will give acceptable calibration response for each
analyte of interest at the concentration required. The source must
be capable of generating both positive and negative ions, and have
a discharge electrode or filament.
4.1.3 Mass spectrometer system - A single quadrupole mass
spectrometer capable of scanning from 1 to 1000 amu. The spectrometer
must also be capable of scanning from 150 to 450 amu in 1.5 sec or less,
using 70 volts (nominal) electron energy in the positive or negative
electron impact modes. In addition, the mass spectrometer must be capable
8321 - 5 Revision 0
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of producing a calibrated mass spectrum for PEG 400, 600, or 800 (see Sec.
5.14).
4.1.3.1 Optional triple quadrupole mass spectrometer -
capable of generating daughter ion spectra with a collision gas in
the second quadrupole and operation in the single quadrupole mode.
4.1.4 Data System - A computer system that allows the continuous
acquisition and storage on machine-readable media of all mass spectra
obtained throughout the duration of the chromatographic program must be
interfaced to the mass spectrometer. The computer must have software that
allows any MS data file to be searched for ions of a specified mass, and
such ion abundances to be plotted versus time or scan number. This type
of plot is defined as an Extracted Ion Current Profile (EICP). Software
must also be available that allows integration of the abundances in any
EICP between specified time or scan-number limits. There must be computer
software available to operate the specific modes of the mass spectrometer.
4.2 HPLC with UV detector - An analytical system with solvent
programmable pumping system for at least a binary solvent system, and all
required accessories including syringes, 10 /uL injection loop, analytical
columns, purging gases, etc. An automatic injector is optional, but is useful
for multiple samples. The columns specified in Sec. 4.1.1.2 are also used with
this system.
4.2.1 If the UV detector is to be used in tandem with the
thermospray interface, then the detector cell must be capable of
withstanding high pressures (up to 6000 psi). However, the UV detector
may be attached to an HPLC independent of the HPLC/TS/MS and, in that
case, standard HPLC pressures are acceptable.
4.3 Purification Equipment for Azo Dye Standards
4.3.1 Soxhlet extraction apparatus.
4.3.2 Extraction thimbles, single thickness, 43 x 123 mm.
4.3.3 Filter paper, 9.0 cm (Whatman qualitative No. 1 or
equivalent).
4.3.4 Silica-gel column - 3 in. x 8 in., packed with Silica gel
(Type 60, EM reagent 70/230 mesh).
4.4 Extraction equipment for Chlorinated Phenoxyacid Compounds
4.4.1 Erlenmeyer flasks - 500-mL wide-mouth Pyrex, 500-mL Pyrex,
with 24/40 ground glass joint, 1000-mL pyrex.
4.4.2 Separatory funnel - 2000 mL.
4.4.3 Graduated cylinder - 1000 mL.
4.4.4 Funnel - 75 mm diameter.
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4.4.5 Wrist shaker - Burrell Model 75 or equivalent.
4.4.6 pH meter.
4.5 Kuderna-Danish (K-D) apparatus (optional).
4.5.1 Concentrator tube - 10 mi graduated (Kontes K-570050-1025 or
equivalent). A ground glass stopper is used to prevent evaporation of
extracts.
4.5.2 Evaporation flask - 500 mi (Kontes K-570001-500 or
equivalent). Attach to concentrator tube with springs, clamps, or
equivalent.
4.5.3 Snyder column - Two ball micro (Kontes K-569001-0219 or
equivalent).
4.5.4 Springs - 1/2 in. (Kontes K-662750 or equivalent).
4.6 Disposable serological pipets - 5 ml x 1/10, 5.5 mm ID.
4.7 Collection tube - 15 mi conical, graduated (Kimble No. 45165 or
equivalent).
4.8 Vials - 5 ml conical, glass, with Teflon lined screw-caps or crimp
tops.
4.9 Glass wool - Supelco No. 2-0411 or equivalent.
4.10 Microsyringes - 100 jil, 50 jzL, 10 p.i (Hamilton 701 N or equivalent),
and 50 ni (Blunted, Hamilton 705SNR or equivalent).
4.11 Rotary evaporator - Equipped with 1000 ml receiving flask.
4.12 Balances - Analytical, 0.0001 g, Top-loading, 0.01 g.
4.13 Volumetric flasks, Class A - 10 ml to 1000 ml.
4.14 Graduated cylinder - 100 ml.
4.15 Separatory funnel - 250 ml.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests.
Unless otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
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5.2 Organic free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Sodium sulfate (granular, anhydrous), Na2S04. Purify by heating at
400°C for 4 hours in a shallow tray, or by precleaning the sodium sulfate with
methylene chloride..
5.4 Ammonium acetate, NH4OOCCH3, solution (0.1 M). Filter through a 0.45
micron membrane filter (Millipore HA or equivalent).
5.5 Acetic acid, CH3C02H
5.6 Sulfuric acid solution
5.6.1 ((1:1) (v/v)) - Slowly add 50 ml H2S04 (sp. gr. 1.84) to 50
mL of water.
5.6.2 ((1:3) (v/v)) - slowly add 25 ml H2S04 (sp. gr. 1.84) to 75
ml of water.
5.7 Argon gas, 99+% pure.
5.8 Solvents
5.8.1 Methylene chloride, CH2C12 - Pesticide quality or equivalent.
5.8.2 Toluene, C6H5CH3 - Pesticide quality or equivalent.
5.8.3 Acetone, CH3COCH3 - Pesticide quality or equivalent.
5.8.4 Diethyl Ether, C2H5OC2H5 - Pesticide quality or equivalent.
Must be free of peroxides as indicated by test strips (EM Quant, or
equivalent). Procedures for removal of peroxides are provided with the
test strips. After cleanup, 20 ml of ethyl alcohol preservative must be
added to each liter of ether.
5.8.5 Methanol, CH3OH - HPLC quality or equivalent.
5.8.6 Acetonitrile, CH3CN - HPLC quality or equivalent.
5.8.7 Ethyl acetate CH3C02C2H5 - Pesticide quality or equivalent.
5.9 Standard Materials - pure standard materials or certified solutions
of each analyte targeted for analysis. Disperse azo dyes must be purified before
use according to Sec. 5.10.
5.10 Disperse Azo Dye Purification
5.10.1 Two procedures are involved. The first step is the
Soxhlet extraction of the dye for 24 hours with toluene and evaporation of
the liquid extract to dryness, using a rotary evaporator. The solid is
then recrystallized from toluene, and dried in an oven at approximately
100°C. If this step does not give the required purity, column
8321 - 8 Revision 0
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chromatography should be employed. Load the solid onto a 3 x 8 inch
silica gel column (Sec. 4.3.4), and elute with diethyl ether. Separate
impurities chromatographically, and collect the major dye fraction.
5.11 Stock standard solutions - Can be prepared from pure standard
materials or can be purchased as certified solutions.
5.11.1 Prepare stock standard solutions by accurately weighing
0.0100 g of pure material. Dissolve the material in methanol or other
suitable solvent (e.g. prepare Tris-BP in ethyl acetate), and dilute to
known volume in a volumetric flask.
NOTE: Due to the reactivity of the chlorinated herbicides, the
standards must be prepared in acetonitrile. Methylation will
occur if prepared in methanol.
If compound purity is certified at 96% or greater, the weight can
be used without correction to calculate the concentration of the stock
standard. Commercially prepared stock standards can be used at any
concentration if they are certified by the manufacturer or by an
independent source.
5.11.2 Transfer the stock standard solutions into glass vials
with Teflon lined screw-caps or crimp-tops. Store at 4°C and protect from
light. Stock standard solutions should be checked frequently for signs of
degradation or evaporation, especially just prior to preparing calibration
standards.
5.12 Calibration standards - A minimum of five concentrations for each
parameter of interest should be prepared through dilution of the stock standards
with methanol (or other suitable solvent). One of these concentrations should
be near, but above, the MDL. The remaining concentrations should correspond to
the expected range of concentrations found in real samples, or should define the
working range of the HPLC-UV/VIS or HPLC-TSP/MS. Calibration standards must be
replaced after one or two months, or sooner if comparison with check standards
indicates a problem.
5.13 Surrogate standards - The analyst should monitor the performance of
the extraction, cleanup (when used), and analytical system, along with the
effectiveness of the method in dealing with each sample matrix, by spiking each
sample, standard, and blank with one or two surrogates (e.g., organophosphorus
or chlorinated phenoxyacid compounds not expected to be present in the sample).
5.14 HPLC/MS tuning standard - Polyethylene glycol 400 (PEG-400), PEG-600
or PEG-800. Dilute to 10 percent (v/v) in methanol. Dependent upon analyte
molecular weight range: m.w. < 500 amu, use PEG-400; m.w. > 500 amu, use PEG-600,
or PEG-800.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Sec. 4.1.
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7.0 PROCEDURE
7.1 Sample preparation - Samples for analysis of disperse azo dyes and
organophosphorus compounds must be prepared by one of the following methods prior
to HPLC/MS analysis:
Matrix Methods
Water 3510, 3520
Soil/sediment 3540, 3541, 3550
Waste 3540, 3541, 3550, 3580
Samples for the analysis of Tris-(2,3-dibromopropyl )phosphate in wastewater
must be prepared according to Sec. 7.1.1 prior to HPLC/MS analysis. Samples for
the analysis of chlorinated phenoxyacid compounds and their esters should be
prepared according to Sec. 7.1.2 prior to HPLC/MS analysis.
7.1.1 Microextraction for Tris-BP:
7.1.1.1 Solid Samples
7.1.1.1.1 Weigh a 1 gram portion of the sample into
a tared beaker. If the sample appears moist, add an
equivalent amount of anhydrous sodium sulfate and mix well.
Add 100 /iL of Tris-BP (approximate concentration 1000 mg/L)
to the sample selected for spiking; the amount added should
result in a final concentration of 100 ng//iL in the 1 mL
extract.
7.1.1.1.2 Remove the glass wool plug from a disposable
serological pipet. Insert a 1 cm plug of clean silane
treated glass wool to the bottom (narrow end) of the pipet.
Pack 2 cm of anhydrous sodium sulfate onto the top of the
glass wool. Wash pipet and contents with 3 - 5 mL of
methanol.
7.1.1.1.3 Pack the sample into the pipet prepared
according to Sec. 7.1.1.1.2. If packing material has dried,
wet with a few mL of methanol first, then pack sample into
the pipet.
7.1.1.1.4 Extract the sample with 3 mL of methanol
followed by 4 mL of 50% (v/v) methanol/methylene chloride
(rinse the sample beaker with each volume of extraction
solvent prior to adding it to the pipet containing the
sample). Collect the extract in a 15 mL graduated glass
tube.
7.1.1.1.5 Evaporate the extract to 1 mL using the
nitrogen blowdown technique (Sec. 7.1.1.1.6). Record the
volume. It may not be possible to evaporate some sludge
samples to a reasonable concentration.
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7.1.1.1.6 Nitrogen Slowdown Technique
7.1.1.1.6.1 Place the concentrator tube in
a warm water bath (approximately 35°C) and evaporate the
solvent volume to the required level using a gentle
stream' of clean, dry nitrogen (filtered through a
column of activated carbon).
CAUTION: Do not use plasticized tubing
between the carbon trap and the
sample.
7.1.1.1.6.2 The internal wall of the tube
must be rinsed down several times with methylene
chloride during the operation. During evaporation, the
solvent level in the tube must be positioned to prevent
water from condensing into the sample (i.e., the
solvent level should be below the level of the water
bath). Under normal operating conditions, the
extract should not be allowed to become dry. Proceed
to Sec. 7.1.1.1.7.
7.1.1.1.7 Transfer the extract to a glass vial with
a Teflon lined screw-cap or crimp-top and store refrigerated
at 4°C. Proceed with HPLC analysis.
7.1.1.1.8 Determination of percent dry weight - In
certain cases, sample results are desired based on a dry
weight basis. When such data are desired, or required, a
portion of sample for this determination should be weighed
out at the same time as the portion used for analytical
determination.
WARNING: The drying oven should be contained in a
hood or vented. Significant laboratory
contamination may result from drying a
heavily contaminated hazardous waste
sample.
7.1.1.1.9 Immediately after weighing the sample for
extraction, weigh 5-10 g of the sample into a tared crucible.
Determine the % dry weight of the sample by drying overnight
at 105°C. Allow to cool in a desiccator before weighing:
% dry weight = q of dry sample x 100
g of sample
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7.1.1.2 Aqueous Samples
7.1.1.2.1 Using a 100 ml graduated cylinder, measure
100 ml of sample and transfer it to a 250 ml separatory
funnel. Add 200 juL of Tris-BP (approximate concentration
1000 mg/L) to the sample selected for spiking; the amount
added should result in a final concentration of 200 ng/juL in
the 1 ml extract.
7.1.1.2.2 Add 10 ml of methylene chloride to the
separatory funnel. Seal and shake the separatory funnel
three times, approximately 30 seconds each time, with
periodic venting to release excess pressure. NOTE: Methylene
chloride creates excessive pressure rapidly; therefore,
initial venting should be done immediately after the
separatory funnel has been sealed and shaken once. Methylene
chloride is a suspected carcinogen, use necessary safety
precautions.
7.1.1.2.3 Allow the organic layer to separate from the
water phase for a minimum of 10 minutes. If the emulsion
interface between layers is more than one-third the size of
the solvent layer, the analyst must employ mechanical
techniques to complete phase separation. See Sec. 7.5,
Method 3510.
7.1.1.2.4 Collect the extract in a 15 ml graduated
glass tube. Proceed as in Sec. 7.1.1.1.5.
7.1.2 Extraction for chlorinated phenoxyacid compounds - Preparation
of soil, sediment, and other solid samples must follow Method 8151, with
the exception of no hydrolysis or esterification. Sec. 7.1.2.1 presents
an outline of the procedure with the appropriate changes necessary for
determination by Method 8321. Sec. 7.1.2.2 describes the extraction
procedure for aqueous samples.
7.1.2.1 Extraction of solid samples
7.1.2.1.1 Add 50 g of soil/sediment sample to a 500
ml, wide mouth Erlenmeyer. Add spiking solutions if
required, mix well and allow to stand for 15 minutes. Add 50
ml of organic-free reagent water and stir for 30 minutes.
Determine the pH of the sample with a glass electrode and pH
meter, while stirring. Adjust the pH to 2 with cold H2S04
(1:1) and monitor the pH for 15 minutes, with stirring. If
necessary, add additional H2S04 until the pH remains at 2.
7.1.2.1.2 Add 20 mL of acetone to the flask, and mix
the contents with the wrist shaker for 20 minutes. Add 80 ml
of diethyl ether to the same flask, and shake again for 20
minutes. Decant the extract and measure the volume of
solvent recovered.
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7.1.2.1.3 Extract the sample twice more using 20 ml
of acetone followed by 80 ml of diethyl ether. After
addition of each solvent, the mixture should be shaken with
the wrist shaker for 10 minutes and the acetone-ether extract
decanted.
7.1.2.1.4 After the third extraction, the volume of
extract recovered should be at least 75% of the volume of
added solvent. If this is not the case, additional
extractions may be necessary. Combine the extracts in a 2000
ml separatory funnel containing 250 ml of reagent water. If
an emulsion forms, slowly add 5 g of acidified sodium sulfate
(anhydrous) until the solvent-water mixture separates. A
quantity of acidified sodium sulfate equal to the weight of
the sample may be added, if necessary.
7.1.2.1.5 Check the pH of the extract. If it is not
at or below pH 2, add more concentrated HC1 until the extract
is stabilized at the desired pH. Gently mix the contents of
the separatory funnel for 1 minute and allow the layers to
separate. Collect the aqueous phase in a clean beaker, and
the extract phase (top layer) in a 500 ml ground-glass
Erlenmeyer flask. Place the aqueous phase back into the
separatory funnel and re-extract using 25 ml of diethyl
ether. Allow the layers to separate and discard the aqueous
layer. Combine the ether extracts in the 500 ml Erlenmeyer
flask.
7.1.2.1.6 Add 45 - 50 g acidified anhydrous sodium
sulfate to the combined ether extracts. Allow the extract to
remain in contact with the sodium sulfate for approximately
2 hours.
NOTE: The drying step is very critical. Any moisture
remaining in the ether will result in low
recoveries. The amount of sodium sulfate used is
adequate if some free flowing crystals are
visible when swirling the flask. If all of the
sodium sulfate solidifies in a cake, add a few
additional grams of acidified sodium sulfate and
again test by swirling. The 2 hour drying time is
a minimum; however, the extracts may be held
overnight in contact with the sodium sulfate.
7.1.2.1.7 Transfer the ether extract, through a funnel
plugged with acid-washed glass wool, into a 500 ml K-D flask
equipped with a 10 ml concentrator tube. Use a glass rod to
crush caked sodium sulfate during the transfer. Rinse the
Erlenmeyer flask and column with 20-30 ml of diethyl ether to
complete the quantitative transfer. Reduce the volume of the
extract using the macro K-D technique (Sec. 7.1.2.1.8).
7.1.2.1.8 Add one or two clean boiling chips to the
flask and attach a three ball macro-Snyder column. Prewet
8321 - 13 Revision 0
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the Snyder column by adding about 1 ml of diethyl ether to
the top. Place the apparatus on a hot water bath (60°-65°C)
so that the concentrator tube is partially immersed in the
hot water and the entire lower rounded surface of the flask
is bathed in vapor. Adjust the vertical position of the
apparatus and the water temperature, as required, to complete
the concentration in 15-20 minutes. At the proper rate of
distillation the balls of the column will actively chatter,
but the chambers will not flood. When the apparent volume of
liquid reaches 5 ml, remove the K-D apparatus from the water
bath and allow it to drain and cool for at least 10 minutes.
7.1.2.1.9 Exchange the solvent of the extract to
acetonitrile by quantitatively transferring the extract with
acetonitrile to a blow-down apparatus. Add a total of 5 ml
acetonitrile. Reduce the extract volume according to Sec.
7.1.1.1.6, and adjust the final volume to 1 ml.
7.1.2.2 Preparation of aqueous samples
7.1.2.2.1 Using a 1000 mL graduated cylinder, measure
1 liter (nominal) of sample, record the sample volume to the
nearest 5 mL, and transfer it to a separatory funnel. If
high concentrations are anticipated, a smaller volume may be
used and then diluted with organic-free reagent water to 1
liter. Adjust the pH to less than 2 with sulfuric acid (1:1).
7.1.2.2.2 Add 150 mL of diethyl ether to the sample
bottle, seal, and shake for 30 seconds to rinse the walls.
Transfer the solvent wash to the separatory funnel and
extract the sample by shaking the funnel for 2 minutes with
periodic venting to release excess pressure. Allow the
organic layer to separate from the water layer for a minimum
of 10 minutes. If the emulsion interface between layers is
more than one-third the size of the solvent layer, the
analyst must employ mechanical techniques to complete the
phase separation. The optimum technique depends upon the
sample, and may include stirring, filtration of the emulsion
through glass wool, centrifugation, or other physical
methods. Drain the aqueous phase into a 1000 ml Erlenmeyer
flask.
7.1.2.2.3 Repeat the extraction two more times using
100 mL of diethyl ether each time. Combine the extracts in
a 500 mL Erlenmeyer flask. (Rinse the 1000 mL flask with
each additional aliquot of extracting solvent to make a
quantitative transfer.)
7.1.2.2.4 Proceed to Sec. 7.1.2.1.6 (drying, K-D
concentration, solvent exchange, and final volume
adjustment).
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7.2 Prior to HPLC analysis, the extraction solvent must be exchanged to
methanol or acetonitrile (Sec. 7.1.2.1.9). The exchange is performed using the
K-D procedures listed in all of the extraction methods.
7.3 HPLC Chromatographic Conditions:
7.3.1 Analyte-specific Chromatographic conditions are shown in
Table 1. Chromatographic conditions which are not analyte-specific are as
follows:
Flow rate: 0.4 mL/min
Post-column mobile phase: 0.1 M ammonium acetate (1% methanol)
(0.1 M ammonium acetate for
phenoxyacid compounds)
Post-column flow rate: 0.8 mL/min
7.3.2 If there is a Chromatographic problem from compound retention
when analyzing for disperse azo dyes, organophosphorus compounds, or
Tris-(2,3-dibromopropyl)phosphate, a 2% constant flow of methylene
chloride may be applied as needed. Methylene chloride/aqueous methanol
solutions must be used with caution as HPLC eluants. Acetic acid (1%),
another mobile phase modifier, can be used with compounds with acid
functional groups.
7.3.3 A total flow rate of 1.0 to 1.5 mL/min is necessary to
maintain thermospray ionization.
7.3.4 Retention times for organophosphorus compounds on the
specified analytical column are presented in Table 9.
7.4 Recommended HPLC/Thermospray/MS operating conditions:
7.4.1 Positive Ionization mode
Repeller (wire or plate, optional): 170 to 250 v (sensitivity
optimized). See Figure 2 for schematic of source with wire repeller.
Mass range: 150 to 450 amu (compound dependent, expect 1 to 18 amu
higher than molecular weight of the compound).
Scan time: 1.50 sec/scan.
7.4.2 Negative Ionization mode
Discharge electrode: on
Filament: off
Mass Range: 135 to 450 amu
Scan time: 1.50 sec/scan.
7.4.3 Thermospray temperatures:
Vaporizer control 110°C to 130°C.
Vaporizer tip 200°C to 215°C.
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Jet 210°C to 220°C.
Source block 230°C to 265°C. (Some compounds may degrade in
the source block at higher temperatures, the
operator should use knowledge of chemical
properties to estimate proper source
temperature).
7.4.4 Sample injection volume: 20 /xL is necessary in order to
overfill the 10 pi injection loop. If solids are present in the extract,
allow them to settle or centrifuge the extract and withdraw the injection
volume from the clear layer.
7.5 Calibration:
7.5.1 Thermospray/MS system - Must be hardware-tuned on quadrupole
1 (and quadrupole 3 for triple quadrupoles) for accurate mass assignment,
sensitivity, and resolution. This is accomplished using polyethylene
glycol (PEG) 400, 600, or 800 (see Sec. 5.14) which have average molecular
weights of 400, 600, and 800, respectively. A mixture of these PEGs can
be made such that it will approximate the expected working mass range for
the analyses. Use PEG 400 for analysis of chlorinated phenoxyacid
compounds. The PEG is introduced via the thermospray interface,
circumventing the HPLC.
7.5.1.1 The mass calibration parameters are as follows:
for PEG 400 and 600 for PEG 800
Mass range: 15 to 765 amu Mass range: 15 to 900 amu
Scan time: 5.00 sec/scan Scan time: 5.00 sec/scan
Approximately 100 scans should be acquired, with 2 to 3
injections made. The scan with the best fit to the accurate mass
table (see Tables 7 and 8) should be used as the calibration table.
7.5.1.2 The low mass range from 15 to 100 amu is covered
by the ions from the ammonium acetate buffer used in the thermospray
process: NH4+ (18 amu), NH4+H20 (36), CH3OHNH4 + (50) (methanol),
or CH3CNNH4+ (59) (acetonitrile), and CH3COOHNH4+ (78) (acetic
acid). The appearance of the m/z 50 or 59 ion depends upon the use
of methanol or acetonitrile as the organic modifier. The higher
mass range is covered by the ammonium ion adducts of the various
ethylene glycols (e.g. H(OCH2CH2)nOH where n=4, gives the
H(OCH2CH2)4OH-NH4+ ion at m/z 212).
7.5.2 Liquid Chromatograph
7.5.2.1 Prepare calibration standards as outlined in Sec.
5.12.
7.5.2.2 Choose the proper ionization conditions, as
outlined in Sec. 7.4. Inject each calibration standard onto the
HPLC, using the chromatographic conditions outlined in Table 1.
Calculate the area under the curve for the mass chromatogram of each
8321 - 16 Revision 0
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quantitation ion. For example, Table 9 lists the retention times
and the major ions (>5%) present in the positive ionization
thermospray single quadrupole spectra of the organophosphorus
compounds. In most cases the (M+H)+ and (M+NH4)+ adduct ions are
the only ions of significant abundance. Plot these ions as area
response versus the amount injected. The points should fall on a
straight line, with a correlation coefficient of at least 0.99 (0.97
for chlorinated phenoxyacid analytes).
7.5.2.3 If HPLC-UV detection is also being used,
calibrate the instrument by preparing calibration standards as
outlined in Sec. 5.12, and injecting each calibration standard onto
the HPLC using the chromatographic conditions outlined in Table 1.
Integrate the area under the full chromatographic peak for each
concentration. Quantitation by HPLC-UV may be preferred if it is
known that sample interference and/or analyte coelution are not a
problem.
7.5.2.4 For the methods specified in Sec. 7.5.2.2 and
7.5.2.3, the retention time of the chromatographic peak is an
important variable in analyte identification. Therefore, the ratio
of the retention time of the sample analyte to the standard analyte
should be 1.0 - 0.1.
7.5.2.5 The concentration of the sample analyte will be
determined by using the calibration curves determined in Sees.
7.5.2.2 and 7.5.2.3. These calibration curves must be generated on
the same day as each sample is analyzed. At least duplicate
determinations should be made for each sample extract. Samples
whose concentrations exceed the standard calibration range should
be diluted to fall within the range.
7.5.2.6 Refer to Method 8000 for further information on
calculations.
7.5.2.7 Precision can also be calculated from the ratio
of response (area) to the amount injected; this is defined as the
calibration factor (CF) for each standard concentration. If the
percent relative standard deviation (%RSD) of the CF is less than
20 percent over the working range, linearity through the origin can
be assumed, and the average calibration factor can be used in place
of a calibration curve. The CF and %RSD can be calculated as
follows:
CF = Total Area of Peak/Mass injected (ng)
%RSD = SD/CF x 100
where:
SD = Standard deviation between CFs
CF = Average CF
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7.6 Sample Analysis
7.6.1 Once the LC/MS system has been calibrated as outlined in Sec.
7.5, it is ready for sample analysis. It is recommended that the samples
initially be analyzed in the negative ionization mode. If low levels of
compounds are suspected, then the samples should also be screened in the
positive ionization mode.
7.6.1.1 A blank 20 p,L injection (methanol) must be
analyzed after the standard(s) analyses, in order to determine any
residual contamination of the Thermospray/HPLC/MS system.
7.6.1.2 Take a 20 /xL aliquot of the sample extract from
Sec. 7.4.4. Start the HPLC gradient elution, load and inject the
sample aliquot, and start the mass spectrometer data system
analysis.
7.7 Calculations
7.7.1 Using the external standard calibration procedure (Method
8000), determine the identity and quantity of each component peak in the
sample reconstructed ion chromatogram which corresponds to the compounds
used for calibration processes. See Method 8000 for calculation
equations.
7.7.2 The retention time of the chromatographic peak is an important
parameter for the identity of the analyte. However, because matrix
interferences can change chromatographic column conditions, the retention
times are not as significant, and the mass spectra confirmations are
important criteria for analyte identification.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
8.2 Tables 4, 5, 6, 11, 12, and 15 indicate the single operator accuracy
and precision for this method. Compare the results obtained with the results in
the tables to determine if the data quality is acceptable. Tables 4, 5, and 6
provide single lab data for Disperse Red 1, Table 11 with organophoshorus
pesticides, Table 12 with Tris-BP and Table 15 with chlorophenoxyacid herbicides.
8.2.1 If recovery is not acceptable, check the following:
8.2.1.1 Check to be sure that there are no errors in the
calculations, surrogate solutions or internal standards. If errors
are found, recalculate the data accordingly.
8.2.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and
re-analyze the extract.
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8.2.1.3 If no problem is found, re-extract and re-analyze
the sample.
8.2.1.4 If, upon re-analysis, the recovery is again not
within limits, flag the data as "estimated concentration".
8.3 Instrument performance - Check the performance of the entire
analytical system daily using data gathered from analyses of blanks, standards,
and replicate samples.
8.3.1 See Sec. 7.5.2.7 for required HPLC/MS parameters for standard
calibration curve %RSD limits.
8.3.2 See Sec. 7.5.2.4 regarding retention time window QC limits.
8.3.3 If any of the chromatographic QC limits are not met, the
analyst should examine the LC system for:
• Leaks,
• Proper pressure delivery,
• A dirty guard column; may need replacing or repacking, and
• Possible partial thermospray plugging.
Any of the above items will necessitate shutting down the HPLC/TSP
system, making repairs and/or replacements, and then restarting the
analyses. The calibration standard should be reanalyzed before any sample
analyses, as described in Sec. 7.5.
8.3.4 The experience of the analyst performing liquid
chromatography is invaluable to the success of the method. Each day that
analysis is performed, the daily calibration standard should be evaluated
to determine if the chromatographic system is operating properly. If any
changes are made to the system (e.g. column change), the system must be
recalibrated.
8.4 Optional Thermospray HPLC/MS/MS confirmation
8.4.1 With respect to this method, MS/MS shall be defined as the
daughter ion collision activated dissociation acquisition with quadrupole
one set on one mass (parent ion), quadrupole two pressurized with argon
and with a higher offset voltage than normal, and quadrupole three set to
scan desired mass range.
8.4.2 Since the thermospray process often generates only one or two
ions per compound, the use of MS/MS is a more specific mode of operation,
yielding molecular structural information. In this mode, fast screening
of samples can be accomplished through direct injection of the sample into
the thermospray.
8.4.3 For MS/MS experiments, the first quadrupole should be set to
the protonated molecule or ammoniated adduct of the analyte of interest.
The third quadrupole should be set to scan from 30 amu to just above the
mass region of the protonated molecule.
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8.4.4 The collision gas pressure (Ar) should be set at about 1.0
mTorr and the collision energy at 20 eV. If these parameters fail to give
considerable fragmentation, they may be raised above these settings to
create more and stronger collisions.
8.4.5 For analytical determinations, the base peak of the collision
spectrum shall be taken as the quantification ion. For extra specificity,
a second ion should be chosen as a backup quantification ion.
8.4.6 Generate a calibration curve as outlined in Sec. 7.5.2.
8.4.7 For analytical determinations, calibration blanks must be run
in the MS/MS mode to determine specific ion interferences. If no
calibration blanks are available, chromatographic separation must be
performed to assure no interferences at specific masses. For fast
screening, the MS/MS spectra of the standard and the analyte could be
compared and the ratios of the three major (most intense) ions examined.
These ratios should be approximately the same, unless there is an
interference. If an interference appears, chromatography must be
utilized.
8.4.8 For unknown concentrations, the total area of the quantitation
ion(s) is calculated and the calibration curves generated as in Sec. 7.5
are used to attain an injected weight number.
8.4.9 MS/MS techniques can also be used to perform structural
analysis on ions represented by unassigned m/z ratios. The procedure for
compounds of unknown structures is to set up a CAD experiment on the ion
of interest. The spectrum generated from this experiment will reflect the
structure of the compound by its fragmentation pattern. A trained mass
spectroscopist and some history of the sample are usually needed to
interpret the spectrum. (CAD experiments on actual standards of the
expected compound are necessary for confirmation or denial of that
substance.)
8.5 Optional wire-repeller CAD confirmation
8.5.1 See Figure 3 for the correct position of the wire-repeller in
the thermospray source block.
8.5.2 Once the wire-repeller is inserted into the thermospray flow,
the voltage can be increased to approximately 500 - 700 v. Enough voltage
is necessary to create fragment ions, but not so much that shorting
occurs.
8.5.3 Continue as outlined in Sec. 7.6.
9.0 METHOD PERFORMANCE
9.1 Single operator accuracy and precision studies have been conducted
using spiked sediment, wastewater, sludge, and water samples. The results are
presented in Tables 4, 5, 6, 11, 12, and 15. Tables 4, 5, and 6 provide single
8321 - 20 Revision 0
September 1994
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lab data for Disperse Red 1, Table 11 for organophoshorus pesticides, Table 12
for Tris-BP and Table 15 with chlorophenoxyacid herbicides.
9.2 LODs should be calculated for the known analytes, on each instrument
to be used. Tables 3, 10, and 13 list limits of detection (LOD) and/or estimated
quantitation limits (EQL) that are typical with this method.
9.2.1 The LODs presented in this method were calculated by analyzing
three replicates of four standard concentrations, with the lowest
concentration being near the instrument detection limit. A linear
regression was performed on the data set to calculate the slope and
intercept. Three times the standard deviation (3a) of the lowest standard
amount, along with the calculated slope and intercept, were used to find
the LOD. The LOD was not calculated using the specifications in Chapter
One, but according to the ACS guidelines specified in Reference 4.
9.2.2 Table 17 presents a comparison of the LODs from Method 8151
and Method 8321 for the chlorinated phenoxyacid compounds.
9.3 Table 16 presents multilaboratory accuracy and precision data for
the chlorinated phenoxyacid herbicides. The data summary is based on data from
three laboratories that analyzed duplicate solvent solutions at each
concentration specified in the Table.
10.0 REFERENCES
1. Voyksner, R.D.; Haney, C.A. "Optimization and Application of Thermospray
High-Performance Liquid Chromatography/Mass Spectrometry"; Anal. Chem.
1985, 57, 991-996.
2. Blakley, C.R.; Vestal, M.L. "Thermospray Interface for Liquid
Chromatography/Mass Spectrometry"; Anal. Chem. 1983, 55, 750-754.
3. Taylor, V.; Mickey, D. M., Marsden, P. J. "Single Laboratory Validation of
EPA Method 8140"; EPA-600/4-87/009, U.S. Environmental Protection Agency,
Las Vegas, NV, 1987, 144 pp.
4. "Guidelines for Data Acquisition and Data Quality Evaluation in
Environmental Chemistry"; Anal. Chem. 1980, 5_2, 2242-2249.
5. Betowski, L. D.; Jones, T. L. "The Analysis of Organophosphorus Pesticide
Samples by HPLC/MS and HPLC/MS/MS"; Environmental Science and Technology.
1988,
8. EPA: 2nd Annual Report on Carcinogens, NTP 81-43, Dec. 1981, pp. 236-237.
9. Blum, A.; Ames, B. N. Science 195, 1977, 17.
10. Zweidinger, R. A.; Cooper, S. D.; Pellazari, E. D., Measurements of
Organic Pollutants in Water and Wastewater, ASTM 686.
11. Cremlyn, R. Pesticides: Preparation and mode of Action; John Wiley and
Sons: Chichester, 1978; p. 142.
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12. Cotterill, E. 6.; Byast, T. H. "HPLC of Pesticide Residues in
Environmental Samples." in Liquid Chromatography in Environmental
Analysis; Laurence, J. F., Ed.; Humana Press: Clifton, NJ, 1984.
13. Voyksner, R. I). "Thermospray HPLC/MS for Monitoring the Environment." In
Applications of New Mass Spectrometry Techniques in Pesticide Chemistry;
Rosen, J. D., Ed., John Wiley and Sons: New York, 1987.
14. Yinon, J.; Jones, T. L.; Betowski, L. D. Rap. Comm. Mass Spectrom. 1989,
3, 38.
15. Shore, F. L.; Amick, E. N., Pan, S. T., Gurka, D. F. "Single Laboratory
Validation of EPA Method 8150 for the Analysis of Chlorinated Herbicides
in Hazardous Waste"; EPA/600/4-85/060, U.S. Environmental Protection
Agency, Las Vegas, NV, 1985.
16. "Development and Evaluations of an LC/MS/MS Protocol", EPA/600/X-86/328,
Dec. 1986.
17. "An LC/MS Performance Evaluation Study of Organophosphorus Pesticides",
EPA/600/X-89/006, Jan. 1989.
18. "A Performance Evaluation Study of a Liquid Chromatography/Mass
Spectrometry Method for Tris-(2,3-Dibromopropyl) Phosphate",
EPA/600/X-89/135, June 1989.
19. "Liquid Chromatography/Mass Spectrometry Performance Evaluation of
Chlorinated Phenoxyacid Herbicides and Their Esters", EPA/600/X-89/176,
July 1989.
20. "An Inter!aboratory Comparison of an SW-846 Method for the Analysis of the
Chlorinated Phenoxyacid Herbicides by LC/MS", EPA/600/X-90/133, June 1990.
8321 - 22 Revision 0
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TABLE 2.
COMPOUNDS AMENABLE TO THERMOSPRAY MASS SPECTROMETRY
Disperse Azo Dyes Alkaloids
Methine Dyes Aromatic ureas
Arylmethane Dyes Amides
Coumarin Dyes Amines
Anthraquinone Dyes Amino acids
Xanthene Dyes Organophosphorus Compounds
Flame retardants Chlorinated Phenoxyacid Compounds
TABLE 3.
LIMITS OF DETECTION (LOD) AND METHOD SENSITIVITIES
FOR DISPERSE RED 1 AND CAFFEINE
Compound
Disperse Red 1
Caffeine
Mode
SRM
Single Quad
CAD
SRM
Single Quad
CAD
LOD
(P9)
180
600
2,000
45
84
240
EQL(7s)
(P9)
420
1400
4700
115
200
560
EQL(lOs)
(pg)
600
2000
6700
150
280
800
EQL = Estimated Quantitation Limit
Data from Reference 16.
8321 - 24 Revision 0
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TABLE 4.
PRECISION AND ACCURACY COMPARISONS OF MS AND MS/MS WITH
HPLC/UV FOR ORGANIC-FREE REAGENT WATER SPIKED WITH DISPERSE RED 1
Sample
Spike 1
Spike 2
RPD
HPLC/UV
82.2 ± 0.2
87.4 ± 0.6
6.1%
Percent
MS
92.5 ± 3.7
90.2 ± 4.7
2.5%
Recovery
CAD
87.6 ± 4.6
90.4 ± 9.9
3 . 2%
SRM
95.5 ± 17.1
90.0 ± 5.9
5.9%
Data from Reference 16.
TABLE 5.
PRECISION AND ACCURACY COMPARISONS OF MS AND MS/MS WITH
HPLC/UV FOR MUNICIPAL WASTEWATER SPIKED WITH DISPERSE RED 1
Percent Recovery
Sample
Spike 1
Spike 2
RPD
HPLC/UV
93.4 ± 0.3
96.2 + 0.1
3.0%
MS
102.0 + 31
79.7 ± 15
25%
CAD
82.7 + 13
83.7 + 5.2
1 . 2%
Data from Reference 16.
8321 - 25
Revision 0
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TABLE 6.
RESULTS FROM ANALYSES OF ACTIVATED SLUDGE PROCESS WASTEWATER
Sample
5 mg/L Spiking
Concentration
1
1-D
2
3
RPD
Unspiked
Sample
1
1-D
2
3
RPD
Recovery
HPLC/UV
0.721 + 0.003
0.731 + 0.021
0.279 + 0.000
0.482 ± 0.001
1.3%
0.000
0.000
0.000
0.000
--
of Disperse Red 1
MS
0.664 + 0.030
0.600 + 0.068
0.253 + 0.052
0.449 + 0.016
10.1%
0.005 ± 0.0007
0.006 ± 0.001
0.002 ± 0.0003
0.003 ± 0.0004
18.2%
(mq/L)
CAD
0.796 + 0.008
0.768 + 0.093
0.301 ± 0.042
0.510 + 0.091
3 . 6%
<0.001
<0.001
<0.001
<0.001
--
Data from Reference 16.
8321 - 26 Revision 0
September 1994
-------
TABLE 7.
CALIBRATION MASSES AND % RELATIVE ABUNDANCES
OF PEG 400
Mass
18.0
35.06
36.04
50.06
77.04
168.12
212.14
256.17
300.20
344.22
388.25
432.28
476.30
520.33
564.35
608.38
652.41
653.41
696.43
697.44
% Relative
Abundances8
32.3
13.5
40.5
94.6
27.0
5.4
10.3
17.6
27.0
45.9
64.9
100
94.6
81.1
67.6
32.4
16.2
4.1
8.1
2.7
Intensity is normalized to mass 432,
8321 - 27 Revision 0
September 1994
-------
TABLE 8.
CALIBRATION MASSES AND % RELATIVE ABUNDANCES
OF PEG 600
Mass
18.0
36.04
50.06
77.04
168.12
212.14
256.17
300.20
344.22
388.25
432.28
476.30
520.33
564.35
608.38
652.41
653.41
696.43
% Relative
Abundances8
4.7
11.4
64.9
17.5
9.3
43.9
56.1
22.8
28.1
38.6
54.4
64.9
86.0
100
63.2
17.5
5.6
1.8
Intensity is normalized to mass 564.
8321 - 28 Revision 0
September 1994
-------
TABLE 9.
RETENTION TIMES AND THERMOSPRAY MASS SPECTRA
OF ORGANOPHOSPHORUS COMPOUNDS
Compound
Monocrotophos
Trichlorfon
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
Retention Time
(minutes)
1:09
1:22
1:28
4:40
9:16
9:52
10:52
13:30
13:55
18:51
Mass Spectra
(% Relative Abundance)8
241 (100), 224 (14)
274 (100), 257 (19), 238 (19)
230 (100), 247 (20)
238 (100), 221 (40)
398 (100), 381 (23), 238 (5),
221 (2)
326 (10), 309 (100)
281 (100), 264 (8), 251 (21),
234 (48)
278 (4), 261 (100)
292 (10), 275 (100)
315 (100), 299 (15)
a For molecules containing Cl, Br and S, only the base peak of the isotopic
cluster is listed.
Data from Reference 17.
8321 - 29
Revision 0
September 1994
-------
TABLE 10.
PRECISION AND METHOD DETECTION LIMITS (MDLs) FOR
ORGANOPHOSPHORUS COMPOUND STANDARDS
Compound
Dichlorvos
Dimethoate
Phorate
Disulfoton
Fensulfothion
Naled
Merphos
Methyl
parathion
Ion
238
230
261
275
309
398
299
281
Standard
Quantitation
Concentration
(ng/juL)
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
2
12.5
25
50
%RSD
16
13
5.7
4.2
2.2
4.2
13
7.3
0.84
14
7.1
4.0
2.2
14
6.7
3.0
4.1
9.2
9.8
2.5
9.5
9.6
5.2
6.3
5.5
17
3.9
5.3
7.1
4.8
1.5
MDL (ng)
4
2
2
1
0.4
0.2
1
30
Data from Reference 17.
8321 - 30
Revision 0
September 1994
-------
TABLE 11.
SINGLE OPERATOR ACCURACY AND PRECISION FOR LOW CONCENTRATION DRINKING
WATER (A), LOW CONCENTRATION SOIL (B), MEDIUM CONCENTRATION DRINKING
WATER (C), MEDIUM CONCENTRATION SEDIMENT (D)
Average
Recovery
Compound (%)
A
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
B
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
C
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
D
Dimethoate
Dichlorvos
Naled
Fensulfothion
Methyl parathion
Phorate
Disulfoton
Merphos
70
40
0.5
112
50
16
3.5
237
16
ND
ND
45
ND
78
36
118
52
146
4
65
85
10
2
101
74
166
ND
72
84
58
56
78
Standard
Deviation
7.7
12
1.0
3.3
28
35
8
25
4
5
--
15
7
19
4
29
3
7
24
15
1
13
8.5
25
--
8.6
9
6
5
4
Spike
Amount
ug/L
5
5
5
5
10
5
5
5
uq/kq
50
50
50
50
100
50
50
50
uq/L
50
50
50
50
100
50
50
50
mg/kq
2
2
2
2
3
2
2
2
Range of
Recovery
(%)
54
14
0
106
0
0
0
187
7
-
-
34
-
48
22
81
43
89
0
51
37
0
0
75
57
115
-
55
66
46
47
70
- 85
- 64
- 2
- 119
- 105
- 86
- 19
-287
- 24
-
-
- 56
-
- 109
- 49
- 155
- 61
- 204
- 9
- 79
- 133
- 41
- 4
- 126
- 91
- 216
-
- 90
- 102
- 70
- 66
- 86
Number
of
Analyses
15
15
15
15
15
15
15
15
15
15
15
15
15
15
15
15
12
12
12
12
12
12
12
12
15
15
15
15
15
15
15
12
Data from Reference 17.
8321 - 31
Revision 0
September 1994
-------
TABLE 12
SINGLE OPERATOR ACCURACY AND PRECISION FOR MUNICIPAL WASTE
WATER (A), DRINKING WATER (B), CHEMICAL SLUDGE WASTE (C)
Average
Recovery
Compound (%)
Tris-BP (A) 25
(B) 40
(C) 63
Spike Range
Standard Amount of % Number of
Deviation (ng/juL) Recovery Analyses
8.0 2 41 - 9.0 15
5.0 2 50-30 12
11 100 84-42 8
Data from Reference 18.
8321 - 32
Revision 0
September 1994
-------
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-------
TABLE 14
LIMITS OF DETECTION (LOD) IN THE POSITIVE AND NEGATIVE ION MODES
FOR THE CHLORINATED PHENOXYACID HERBICIDES AND FOUR ESTERS
Compound
Dalapon
Dicamba
2,4-D
MCPA
Dichlorprop
MCPP
2,4,5-T
2,4,5-TP (Silvex)
Dinoseb
2,4-DB
2,4-D,Butoxy
ethanol ester
2,4,5-T,Butoxy
ethanol ester
2,4,5-T, Butyl
ester
2,4-D,ethyl-
hexyl ester
Positive Mode
Quantitation
Ion
Not detected
238 (M+NH4)+
238 (M+NH4)+
218 (M+NH4)+
252 (M+NH4}+
232 (M+NHJ +
272 (M+NH4)+
286 (M+NH4)+
228 (M+NH4-NO)+
266 (M+NH4)+
321 (M+H)+
372 (M+NH4)+
328 (M+NH4)+
350 (M+NHJ +
LOD
(ng)
13
2.9
120
2.7
5.0
170
160
24
3.4
1.4
0.6
8.6
1.2
Negative Mode
Quantitation
Ion
141 (M-H)'
184 (M-HC1)'
184 (M-HC1)-
199 (M-l)-
235 (M-l)-
213 (M-l)-
218 (M-HC1)'
269 (M-l)'
240 (M)-
247 (M-l)-
185 (M-CeH^OJ-
195 (M-C8H1503)-
195 (M-C6Hn02)-
161 (M-C10H1903)-
LOD
(ng)
11
3.0
50
28
25
12
6.5
43
19
110
Data from Reference 19.
8321 - 34
Revision 0
September 1994
-------
TABLE 15
SINGLE LABORATORY OPERATOR ACCURACY AND PRECISION
FOR THE CHLORINATED PHENOXYACID HERBICIDES
Compound
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Si 1 vex
2,4-DB
Dinoseb
Dalapon
2,4-D, ester
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Silvex
2,4-DB
Dinoseb
Dalapon
2,4-D, ester
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Silvex
2,4-DB
Dinoseb
Dalapon
2,4-D, ester
(a)
Average Standard
Recovery(%) Deviation
LOW LEVEL
63
26
60
78
43
72
62
29
73
ND
73
HIGH LEVEL
54
60
67
66
66
61
74
83
91
43
97
LOW
117
147
167
142
ND
134
121
199
76
ND
180
DRINKING WATER
22
13
23
21
18
31
14
24
11
ND
17
DRINKING WATER
30
35
41
33
33
23
35
25
10
9.6
19
LEVEL SAND
26
23
79
39
ND
27
23
86
74
ND
58
Spike
Amount
M9/L
5
5
5
5
5
5
5
5
5
5
5
M9/L
50
50
50
50
50
50
50
50
50
50
50
M9/9
.1
.1
.1
.1
.1
.1
.1
.1
.1
.1
.1
Range of
Recovery
(%)
33
0
37
54
0
43
46
0
49
48
26
35
32
35
27
44
45
52
76
31
76
82
118
78
81
99
85
0
6
59
- 86
- 37
- 92
- 116
- 61
- 138
- 88
- 62
- 85
ND
- 104
- 103
- 119
- 128
- 122
- 116
- 99
- 132
- 120
- 102
- 56
- 130
- 147
- 180
- 280
- 192
ND
- 171
- 154
- 245
- 210
ND
- 239
Number
of
Analyses
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
9
6
9
10
10
10
10
10
10
10
10
10
10
7
la)All recoveries are in negative ionization mode, except for 2,4-D,ester.
ND = Not Detected.
8321 - 35
Revision 0
September 1994
-------
TABLE 15 (cont.)
SINGLE LABORATORY OPERATOR ACCURACY'AND PRECISION
FOR THE CHLORINATED PHENOXYACID HERBICIDES
Compound
(a)
Average
Recovery(%)
Standard
Deviation
Spike
Amount
Range of
Recovery
(%)
Number
of
Analyses
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Si 1 vex
2,4-DB
Dinoseb
Dalapon
2,4-D,ester
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Si 1 vex
2,4-DB
Dinoseb
Dalapon
2,4-D,ester
Dicamba
2,4-D
MCPA
MCPP
Dichlorprop
2,4,5-T
Si 1 vex
2,4-DB
Dinoseb
Dalapon
2,4-D,ester
HIGH LEVEL SAND
153
218
143
158
92
160
176
145
114
287
20
33
27
30
34
37
29
34
22
28
86
3.6
LOW LEVEL MUNICIPAL ASH
83
IND
ND
ND
ND
27
68
ND
44
ND
29
22
ND
ND
ND
ND
25
38
ND
13
ND
23
66
8.7
3.2
10
ND
2.9
6.0
ND
16
ND
1.9
21
4.8
4.8
4.3
ND
1.2
3.1
ND
6.8
ND
1.7
M9/9
1
1
1
1
1
1
1
1
1
1
1
.1
.1
.1
.1
.1
.1
.1
.1
.1
.1
.1
HIGH LEVEL MUNICIPAL ASH M9/9
119
187
111
115
51
131
141
110
65
166
17
- 209
- 276
- 205
- 226
- 161
- 204
- 225
- 192
- 140
- 418
- 25
48 - 104
ND
ND
ND
ND
0 - 60
22 - 128
ND
26 - 65
ND
0 - 53
41 - 96
5 - 21
0 - 10
4.7 - 16
ND
0 - 3.6
2.8 - 12
ND
0 - 23
ND
0 - 6.7
9
9
9
9
9
9
9
9
9
9
7
9
9
9
9
9
9
9
9
9
9
6
9
9
9
9
9
9
9
9
9
9
6
lalAll recoveries are in negative ionization mode, except for 2,4-D,ester.
ND = Not Detected.
8321 - 36
Revision 0
September 1994
-------
TABLE 16
MULTI LABORATORY ACCURACY AND PRECISION DATA
FOR THE CHLORINATED PHENOXYACID HERBICIDES
Compounds
Spiking
Concentration
Mean
(% Recovery)8
Data from Reference 20.
a Mean of duplicate data from 3 laboratories.
b % RSD of duplicate data from 3 laboratories.
% Relative
Standard Deviation6
500 mg/L
2,4,5-T
2,4,5-T,butoxy
2,4-D
2,4-DB
Dalapon
Dicamba
Dichlorprop
Dinoseb
MCPA
MCPP
Si 1 vex
50 mg/L
2,4,5-T
2,4,5-T,butoxy
2,4-D
2,4-DB
Dalapon
Dicamba
Dichlorprop
Dinoseb
MCPA
MCPP
Silvex
5 mq/L
2,4,5-T
2,4,5-T,butoxy
2,4-D
2,4-DB
Dalapon
Dicamba
Dichlorprop
Dinoseb
MCPA
MCPP
Silvex
90
90
86
95
83
77
84
78
89
86
96
62
85
64
104
121
90
96
86
96
76
65
90
99
103
96
150
105
102
108
94
98
87
23
29
17
22
13
25
20
15
11
12
27
68
9
80
28
99
23
15
57
20
74
71
28
17
31
21
4
12
22
30
18
15
15
8321 - 37
Revision 0
September 1994
-------
TABLE 17
COMPARISON OF LODs: METHOD 8151 vs. METHOD 8321
Compound
Method 8151
LOD(Mg/L)
Method 8321
LOD (Mg/L)
lonization
Mode
Dalapon
Dicamba
2,4-D
MCPA
Dichloroprop
MCPP
2,4,5-T
2,4,5-TP (Silvex)
2,4-DB
Dinoseb
1.3
0.8
0.2
0.06
0.26
0.09
0.08
0.17
0.8
0.19
1.1
0.3
0.29
2.8
0.27
0.50
0.65
4.3
0.34
1.9
8321 - 38
Revision 0
September 1994
-------
FIGURE 1.
SCHEMATIC OF THE THERMOSPRAY PROBE AND ION SOURCE
r
&
Flange
Source
Mounting
Ion Sampling
Cone
Ions
Electron Vaporizer
Beam .x Probe
T,
\
— 1C
i
Vapor || Heater Vaporizer
Temperature | Coupling
T4 Block
Temperature
T.
8321 - 39
Revision 0
September 1994
-------
FIGURE 2.
THERMOSPRAY SOURCE WITH WIRE-REPELLER
(High sensitivity configuration)
CERAMIC INSULATOR
WIRE REPELLER
8321 - 40
Revision 0
September 1994
-------
FIGURE 3.
THERMOSPRAY SOURCE WITH WIRE-REPELLER
(CAD configuration)
CERAMIC INSULATOR
WIRE REPELLER
8321 - 41
Revision 0
September 1994
-------
METHOD 8321
SOLVENT EXTRACTABLE NON-VOLATILE COMPOUNDS BY
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY/THERMOSPRAY/MASS SPECTROMETRY
(HPLC/TSP/MS) OR ULTRAVIOLET (UV) DETECTION
8321 - 42
Revision 0
September 1994
-------
METHOD 8330
NITROAROMATICS AND NITRAMINES BY HIGH
PERFORMANCE LIQUID CHROMAT06RAPHY (HPLC)
1.0 SCOPE AND APPLICATION
1.1 Method 8330 is intended for the trace analysis of explosives residues
by high performance liquid chromatography using a UV detector. This method is
used to determine the concentration of the following compounds in a water, soil,
or sediment matrix:
Compound
Abbreviation
CAS No8
Octahydro-l,3,5,7-tetranitro-l,3,5,7-tetrazocine HMX
Hexahydro-l,3,5-trinitro-l,3,5-triazine RDX
1,3,5-Trinitrobenzene 1,3,5-TNB
1,3-Dinitrobenzene 1,3-DNB
Methyl-2,4,6-tri ni trophenylni trami ne Tetryl
Nitrobenzene NB
2,4,6-Trinitrotoluene 2,4,6-TNT
4-Amino-2,6-dinitrotoluene 4-Am-DNT
2-Amino-4, 6-dinitrotoluene 2-Am-DNT
2,4-Dinitrotoluene 2,4-DNT
2,6-Dinitrotoluene 2,6-DNT
2-Nitrotoluene 2-NT
3-Nitrotoluene 3-NT
4-Nitrotoluene 4-NT
2691-41-0
121-82-4
99-35-4
99-65-0
479-45-8
98-95-3
118-96-7
1946-51-0
355-72-78-2
121-14-2
606-20-2
88-72-2
99-08-1
99-99-0
a Chemical Abstracts Service Registry number
1.2 Method 8330 provides a salting-out extraction procedure for low
concentration (parts per trillion, or nanograms per liter) of explosives residues
in surface or ground water. Direct injection of diluted and filtered water
samples can be used for water samples of higher concentration (See Table 1).
1.3 All of these compounds are either used in the manufacture of
explosives or are the degradation products of compounds used for that purpose.
When making stock solutions for calibration, treat each explosive compound with
caution. See NOTE in Sec. 5.3.1 and Sec. 11 on Safety.
1.4 The estimated quantitation limits (EQLs) of target analytes
determined by Method 8330 in water and soil are presented in Table 1.
1.5 This method is restricted to use by or under the supervision of
analysts experienced in the use of HPLC, skilled in the interpretation of
chromatograms, and experienced in handling explosive materials. (See Sec. 11.0
8330 - 1
Revision 0
September 1994
-------
on SAFETY.) Each analyst must demonstrate the ability to generate acceptable
results with this method.
2.0 SUMMARY OF METHOD
2.1 Method 8330 provides high performance liquid chromatographic (HPLC)
conditions for the detection of ppb levels of certain explosives residues in
water, soil and sediment matrix. Prior to use of this method, appropriate sample
preparation techniques must be used.
2.2 Low-Level Salting-out Method With No Evaporation: Aqueous samples
of low concentration are extracted by a salting-out extraction procedure with
acetonitrile and sodium chloride. The small volume of acetonitrile that remains
undissolved above the salt water is drawn off and transferred to a smaller
volumetric flask. It is back-extracted by vigorous stirring with a specific
volume of salt water. After equilibration, the phases are allowed to separate
and the small volume of acetonitrile residing in the narrow neck of the
volumetric flask is removed using a Pasteur pi pet. The concentrated extract is
diluted 1:1 with reagent grade water. An aliquot is separated on a C-18 reverse
phase column, determined at 254 nm, and confirmed on a CN reverse phase column.
2.3 High-level Direct Injection Method: Aqueous samples of higher
concentration can be diluted 1/1 (v/v) with methanol or acetonitrile, filtered,
separated on a C-18 reverse phase column, determine at 254 nm, and confirmed on
a CN reverse phase column. If HMX is an important target analyte, methanol is
preferred.
2.4 Soil and sediment samples are extracted using acetonitrile in an
ultrasonic bath, filtered and chromatographed as in Sec. 2.3.
3.0 INTERFERENCES
3.1 Solvents, reagents, glassware and other sample processing hardware
may yield discrete artifacts and/or elevated basel ines, causing misinterpretation
of the chromatograms. All of these materials must be demonstrated to be free
from interferences.
3.2 2,4-DNT and 2,6-DNT elute at similar retention times (retention time
difference of 0.2 minutes). A large concentration of one isomer may mask the
response of the other isomer. If it is not apparent that both isomers are
present (or are not detected), an isomeric mixture should be reported.
3.3 Tetryl decomposes rapidly in methanol/water solutions, as well as
with heat. All aqueous samples expected to contain tetryl should be diluted with
acetonitrile prior to filtration and acidified to pH <3. All samples expected
to contain tetryl should not be exposed to temperatures above room temperature.
3.4 Degradation products of tetryl appear as a shoulder on the 2,4,6-TNT
peak. Peak heights rather than peak areas should be used when tetryl is present
in concentrations that are significant relative to the concentration of
2,4,6-TNT.
8330 - 2 Revision 0
September 1994
-------
4.0 APPARATUS AND MATERIALS
4.1 HPLC system
4.1.1 HPLC - equipped with a pump capable of achieving 4000 psi, a
100 /zl loop injector and a 254 nm UV detector (Perkin Elmer Series 3, or
equivalent). For the low concentration option, the detector must be
capable of a stable baseline at 0.001 absorbance units full scale.
4.1.2 Recommended Columns:
4.1.2.1 Primary column: C-18 Reverse phase HPLC column,
25 cm x 4.6 mm (5 jum), (Supelco LC-18, or equivalent).
4.1.2.2 Secondary column: CN Reverse phase HPLC column,
25 cm x 4.6 mm (5 /^m), (Supelco LC-CN, or equivalent).
4.1.3 Strip chart recorder.
4.1.4 Digital integrator (optional).
4.1.5 Autosampler (optional).
4.2 Other Equipment
4.2.1 Temperature controlled ultrasonic bath.
4.2.2 Vortex mixer.
4.2.3 Balance, + 0.0001 g.
4.2.4 Magnetic stirrer with stirring pellets.
4.2.5 Water bath - Heated, with concentric ring cover, capable of
temperature control (± 5°C). The bath should be used in a hood.
4.2.6 Oven - Forced air, without heating.
4.3 Materials
4.3.1 High pressure injection syringe - 500 p,L, (Hamilton liquid
syringe or equivalent).
4.3.2 Disposable cartridge filters - 0.45 /im Teflon filter.
4.3.3 Pipets - Class A, glass, Appropriate sizes.
4.3.4 Pasteur pipets.
4.3.5 Scintillation Vials - 20 mL, glass.
4.3.6 Vials - 15 mL, glass, Teflon-lined cap.
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4.3.7 Vials- 40 ml, glass, Teflon-lined cap.
4.3.8 Disposable syringes - Plastipak, 3 ml and 10 ml or equivalent.
4.3.9 Volumetric flasks - Appropriate sizes with ground glass
stoppers, Class A.
NOTE: The 100 ml and 1 L volumetric flasks used for magnetic stirrer
extraction must be round.
4.3.10 Vacuum desiccator - Glass.
4.3.11 Mortar and pestle - Steel.
4.3.12 Sieve - 30 mesh.
4.3.13 Graduated cylinders - Appropriate sizes.
4.4 Preparation of Materials
4.4.1 Prepare all materials to be used as described in Chapter 4 for
semivolatile organics.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first, ascertained that the reagent is of sufficiently high purity
to permit its use without lowering the accuracy of the determination.
5.1.1 Acetonitrile, CH3CN - HPLC grade.
5.1.2 Methanol, CH3OH - HPLC grade.
5.1.3 Calcium chloride, CaCl2 - Reagent grade. Prepare an aqueous
solution of 5 g/L.
5.1.4 Sodium chloride, NaCl, shipped in glass bottles - reagent
grade.
5.2 Organic-free reagent water - All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Stock Standard Solutions
5.3.1 Dry each solid analyte standard to constant weight in a vacuum
desiccator in the dark. Place about 0.100 g (weighed to 0.0001 g) of a
single analyte into a 100 ml volumetric flask and dilute to volume with
acetonitrile. Invert flask several times until dissolved. Store in
8330 - 4 Revision 0
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refrigerator at 4°C in the dark. Calculate the concentration of the stock
solution from the actual weight used (nominal concentration = 1,000 mg/L).
Stock solutions may be used for up to one year.
NOTE: The HMX, RDX, Tetryl, and 2,4,6-TNT are explosives and the
neat material should be handled carefully. See SAFETY in Sec.
11 for guidance. HMX, RDX, and Tetryl reference materials
are shipped under water. Drying at ambient temperature
requires several days. DO NOT DRY AT HEATED TEMPERATURES!
5.4 Intermediate Standards Solutions
5.4.1 If both 2,4-DNT and 2,6-DNT are to be determined, prepare two
separate intermediate stock solutions containing (1) HMX, RDX, 1,3,5-TNB,
1,3-DNB, NB, 2,4,6-TNT, and 2,4-DNT and (2) Tetryl, 2,6-DNT, 2-NT, 3-NT,
and 4-NT. Intermediate stock standard solutions should be prepared at
1,000 M9/L, in acetonitrile when analyzing soil samples, and in methanol
when analyzing aqueous samples.
5.4.2 Dilute the two concentrated intermediate stock solutions, with
the appropriate solvent, to prepare intermediate standard solutions that
cover the range of 2.5 - 1,000 jug/L. These solutions should be
refrigerated on preparation, and may be used for 30 days.
5.4.3 For the low-level method, the analyst must conduct a detection
limit study and devise dilution series appropriate to the desired range.
Standards for the low level method must be prepared immediately prior to
use.
5.5 Working standards
5.5.1 Calibration standards at a minimum of five concentration
levels should be prepared through dilution of the intermediate standards
solutions by 50% (v/v) with 5 g/L calcium chloride solution (Sec. 5.1.3).
These solutions must be refrigerated and stored in the dark, and prepared
fresh on the day of calibration.
5.6 Surrogate Spiking Solution
5.6.1 The analyst should monitor the performance of the extraction
and analytical system as well as the effectiveness of the method in
dealing with each sample matrix by spiking each sample, standard and
reagent water blank with one or two surrogates (e.g., analytes not
expected to be present in the sample).
5.7 Matrix Spiking Solutions
5.7.1 Prepare matrix spiking solutions in methanol such that the
concentration in the sample is five times the Estimated Quantitation Limit
(Table 1). All target analytes should be included.
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5.8 HPLC Mobile Phase
5.8.1 To prepare 1 liter of mobile phase, add 500 ml of methanol to
500 ml of organic-free reagent water.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Follow conventional sampling and sample handling procedures as
specified for semivolatile organics in Chapter Four.
6.2 Samples and sample extracts must be stored in the dark at 4"C.
Holding times are the same as for semivolatile organics.
7.0 PROCEDURE
7.1 Sample Preparation
7.1.1 Aqueous Samples: It is highly recommended that process waste
samples be screened with the high-level method to determine if the low
level method (1-50 /ig/L) is required. Most groundwater samples will fall
into the low level method.
7.1.1.1 Low-Level Method (salting-out extraction)
7.1.1.1.1 Add 251.3 g of sodium chloride to a 1 L
volumetric flask (round). Measure out 770 mL of a water
sample (using a 1 L graduated cylinder) and transfer it to the
volumetric flask containing the salt. Add a stir bar and mix
the contents at maximum speed on a magnetic stirrer until the
salt is completely dissolved.
7.1.1.1.2 Add 164 mL of acetonitrile (measured with a
250 mL graduated cylinder) while the solution is being stirred
and stir for an additional 15 minutes. Turn off the stirrer
and allow the phases to separate for 10 minutes.
7.1.1.1.3 Remove the acetonitrile (upper) layer (about
8 mL) with a Pasteur pipet and transfer it to a 100 mL
volumetric flask (round). Add 10 mL of fresh acetonitrile to
the water sample in the 1 L flask. Again stir the contents of
the flask for 15 minutes followed by 10 minutes for phase
separation. Combine the second acetonitrile portion with the
initial extract. The inclusion of a few drops of salt water
at this point is unimportant.
7.1.1.1.4 Add 84 mL of salt water (325 g Nad per 1000
ml. of reagent water) to the acetonitrile extract in the 100 mL
volumetric flask. Add a stir bar and stir the contents on a
magnetic stirrer for 15 minutes, followed by 10 minutes for
phase separation. Carefully transfer the acetonitrile phase
8330 - 6 Revision 0
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to a 10 ml graduated cylinder using a Pasteur pipet. At this
stage, the amount of water transferred with the acetonitrile
must be minimized. The water contains a high concentration of
NaCl that produces a large peak at the beginning of the
chromatogram, where it could interfere with the HMX
determination.
7.1.1.1.5 Add an additional 1.0 ml of acetonitrile to
the 100 ml volumetric flask. Again stir the contents of the
flask for 15 minutes, followed by 10 minutes for phase
separation. Combine the second acetonitrile portion with the
initial extract in the 10 ml graduated cylinder (transfer to
a 25 ml graduated cylinder if the volume exceeds 5 ml).
Record the total volume of acetonitrile extract to the nearest
0.1 ml. (Use this as the volume of total extract [Vt] in the
calculation of concentration after converting to juL). The
resulting extract, about 5 - 6 ml, is then diluted 1:1 with
organic-free reagent water (with pH <3 if tetryl is a
suspected analyte) prior to analysis.
7.1.1.1.6 If the diluted extract is turbid, filter it
through a 0.45 - jum Teflon filter using a disposable syringe.
Discard the first 0.5 ml of filtrate, and retain the remainder
in a Teflon-capped vial for RP-HPLC analysis as in Sec. 7.4.
7.1.1.2 High-level Method
7.1.1.2.1 Sample filtration: Place a 5 ml aliquot of
each water sample in a scintillation vial, add 5 ml of
acetonitrile, shake thoroughly, and filter through a 0.45-jum
Teflon filter using a disposable syringe. Discard the first
3 ml of filtrate, and retain the remainder in a Teflon-capped
vial for RP-HPLC analysis as in Sec. 7.4. HMX quantitation
can be improved with the use of methanol rather than
acetonitrile for dilution before filtration.
7.1.2 Soil and Sediment Samples
7.1.2.1 Sample homogenization: Dry soil samples in air at
room temperature or colder to a constant weight, being careful not
to expose the samples to direct sunlight. Grind and homogenize the
dried sample thoroughly in an acetonitrile-rinsed mortar to pass a
30 mesh sieve.
NOTE: Soil samples should be screened by Method 8515 prior to
grinding in a mortar and pestle (See Safety Sec. 11.2).
7.1.2.2 Sample extraction
7.1.2.2.1 Place a 2.0 g subsample of each soil sample
in a 15 ml glass vial. Add 10.0 ml of acetonitrile, cap with
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Teflon-lined cap, vortex swirl for one minute, and place in a
cooled ultrasonic bath for 18 hours.
7.1.2.2.2 After sonication, allow sample to settle for
30 minutes. Remove 5.0 ml of supernatant, and combine with
5.0 ml of calcium chloride solution (Sec. 5.1.3) in a 20 ml
vial. Shake, and let stand for 15 minutes.
7.1.2.2.3 Place supernatant in a disposable syringe
and filter through a 0.45-jum Teflon filter. Discard first 3
mL and retain remainder in a Teflon-capped vial for RP-HPLC
analysis as in Sec. 7.4.
7.2 Chromatographic Conditions (Recommended)
Primary Column: C-18 reverse phase HPLC column, 25-cm
x 4.6-mm, 5 /urn, (Supelco LC-18 or equivalent).
Secondary Column: CN reverse phase HPLC column, 25-cm x
4.6-mm, 5 /j,m, (Supelco LC-CN or
equivalent).
Mobile Phase: 50/50 (v/v) methanol/organic-free
reagent water.
Flow Rate: 1.5 mL/min
Injection volume: 100-^L
UV Detector: 254 nm
7.3 Calibration of HPLC
7.3.1 All electronic equipment is allowed to warm up for 30 minutes.
During this period, at least 15 void volumes of mobile phase are passed
through the column (approximately 20 min at 1.5 mL/min) and continued
until the baseline is level at the UV detector's greatest sensitivity.
7.3.2 Initial Calibration. Injections of each calibration standard
over the concentration range of interest are made sequentially into the
HPLC in random order. Peak heights or peak areas are obtained for each
analyte. Experience indicates that a linear calibration curve with zero
intercept is appropriate for each analyte. Therefore, a response factor
for each analyte can be taken as the slope of the best-fit regression
line.
7.3.3 Daily Calibration. Analyze midpoint calibration standards, at
a minimum, at the beginning of the day, singly at the midpoint of the run,
and singly after the last sample of the day (assuming a sample group of 10
samples or less). Obtain the response factor for each analyte from the
mean peak heights or peak areas and compare it with the response factor
obtained for the initial calibration. The mean response factor for the
8330 - 8 Revision 0
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daily calibration must agree within ±15% of the response factor of the
initial calibration. The same criteria is required for subsequent
standard responses compared to the mean response of the triplicate
standards beginning the day. If this criterion is not met, a new initial
calibration must be obtained.
7.4 HPLC Analysis
7.4.1 Analyze the samples using the chromatographic conditions given
in Sec. 7.2. All positive measurements observed on the C-18 column must
be confirmed by injection onto the CN column.
7.4.2 Follow Sec. 7.0 in Method 8000 for instructions on the
analysis sequence, appropriate dilutions, establishing daily retention
time windows, and identification criteria. Include a mid-level standard
after each group of 10 samples in the analysis sequence. If column
temperature control is not employed, special care must be taken to ensure
that temperature shifts do not cause peak misidentification.
7.4.3 Table 2 summarizes the estimated retention times on both C-18
and CN columns for a number of analytes analyzable using this method. An
example of the separation achieved by Column 1 is shown in Figure 1.
7.4.4 Record the resulting peak sizes in peak heights or area units.
The use of peak heights is recommended to improve reproducibility of low
level samples.
7.4.5 Calculation of concentration is covered in Sec. 7.0 of Method
8000.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500.
8.2 Quality control required to validate the HPLC system operation is
found in Method 8000, Sec. 8.0.
8.3 Prior to preparation of stock solutions, acetonitrile, methanol, and
water blanks should be run to determine possible interferences with analyte
peaks. If the acetonitrile, methanol, or water blanks show contamination, a
different batch should be used.
9.0 METHOD PERFORMANCE
9.1 Table 3 presents the single laboratory precision based on data from
the analysis of blind duplicates of four spiked soil samples and four field
contaminated samples analyzed by seven laboratories.
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9.2 Table 4 presents the multilaboratory error based on data from the
analysis of blind duplicates of four spiked soil samples and four field
contaminated samples analyzed by seven laboratories.
9.3 Table 5 presents the multilaboratory variance of the high
concentration method for water based on data from nine laboratories.
9.4 Table 6 presents multi laboratory recovery data from the analysis of
spiked soil samples by seven laboratories.
9.5 Table 7 presents a comparison of method accuracy for soil and aqueous
samples (high concentration method).
9.6 Table 8 contains precision and accuracy data for the salting-out
extraction method.
10.0 REFERENCES
1. Bauer, C.F., T.F. Jenkins, S.M. Koza, P.M. Schumacher, P.H. Miyares and
M.E. Walsh (1989). Development of an analytical method for the
determination of explosive residues in soil. Part 3. Collaborative test
results and final performance evaluation. USA Cold Regions Research and
Engineering Laboratory, CRREL Report 89-9.
2. Grant, C.L., A.D. Hewitt and T.F. Jenkins (1989) Comparison of low
concentration measurement capability estimates in trace analysis: Method
Detection Limits and Certified Reporting Limits. USA Cold Regions
Research and Engineering Laboratory, Special Report 89-20.
3. Jenkins, T.F., C.F. Bauer, D.C. Leggett and C.L. Grant (1984)
Reversed-phased HPLC method for analysis of TNT, RDX, HMX and 2,4-DNT in
munitions wastewater. USA Cold Regions Research and Engineering
Laboratory, CRREL Report 84-29.
4. Jenkins, T.F. and M.E. Walsh (1987) Development of an analytical method
for explosive residues in soil. USA Cold Regions Research and Engineering
Laboratory, CRREL Report 87-7.
5. Jenkins, T.F., P.H. Miyares and ME. Walsh (1988a) An improved RP-HPLC
method for determining nitroaromatics and nitramines in water. USA Cold
Regions Research and Engineering Laboratory, Special Report 88-23.
6. Jenkins, T.F. and P.H. Miyares (1992) Comparison of Cartridge and
Membrane Solid-Phase Extraction with Salting-out Solvent Extraction for
Preconcentration of Nitroaromatic and Nitramine Explosives from Water.
USA Cold Regions Research and Engineering Laboratory, Draft CRREL Special
Report.
7. Jenkins, T.F., P.W. Schumacher, M.E. Walsh and C.F. Bauer (1988b)
Development of an analytical method for the determination of explosive
residues in soil. Part II: Further development and ruggedness testing.
USA Cold Regions Research and Engineering Laboratory, CRREL Report 88-8.
8330 - 10 Revision 0
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8. Leggett, D.C., T.F. Jenkins and P.M. Miyares (1990) Salting-out solvent
extraction for preconcentration of neutral polar organic solutes from
water. Analytical Chemistry, 62: 1355-1356.
9. Miyares, P.H. and T.F. Jenkins (1990) Salting-out solvent extraction for
determining low levels of nitroaromatics and nitramines in water. USA
Cold Regions Research and Engineering Laboratory, Special Report 90-30.
11.0 SAFETY
11.1 Standard precautionary measures used for handling other organic
compounds should be sufficient for the safe handling of the analytes targeted by
Method 8330. The only extra caution that should be taken is when handling the
analytical standard neat material for the explosives themselves and in rare cases
where soil or waste samples are highly contaminated with the explosives. Follow
the note for drying the neat materials at ambient temperatures.
11.2 It is advisable to screen soil or waste samples using Method 8515 to
determine whether high concentrations of explosives are present. Soil samples
as high as 2% 2,4,6-TNT have been safely ground. Samples containing higher
concentrations should not be ground in the mortar and pestle. Method 8515 is for
2,4,6-TNT, however, the other nitroaromatics will also cause a color to be
developed and provide a rough estimation of their concentrations. 2,4,6-TNT is
the analyte most often detected in high concentrations in soil samples. Visual
observation of a soil sample is also important when the sample is taken from a
site expected to contain explosives. Lumps of material that have a chemical
appearance should be suspect and not ground. Explosives are generally a very
finely ground grayish-white material.
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TABLE 1
ESTIMATED QUANTITATION LIMITS
Compounds
HMX
RDX
1,3,5-TNB
1,3-DNB
Tetryl
NB
2,4,6-TNT
4-Am-DNT
2-Am-DNT
2,6-DNT
2,4-DNT
2-NT
4-NT
3-NT
Water
Low-Level
-
0.84
0.26
0.11
-
-
0.11
0.060
0.035
0.31
0.020
-
-
-
(UQ/U
High-Level
13.0
14.0
7.3
4.0
4.0
6.4
6.9
-
-
9.4
5.7
12.0
8.5
7.9
Soil (mg/kg)
2.2
1.0
0.25
0.25
0.65
0.26
0.25
-
-
0.26
0.25
0.25
0.25
0.25
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TABLE 2
RETENTION TIMES AND CAPACITY FACTORS ON LC-18 AND LC-CN COLUMNS
Retention time
(min)
Compound
HMX
RDX
1,3,5-TNB
1,3-DNB
Tetry]
NB
2,4,6-TNT
4-Am-DNT
2-Am-DNT
2,6-DNT
2,4-DNT
2-NT
4-NT
3-NT
LC-18
2.44
3.73
5.11
6.16
6.93
7.23
8.42
8.88
9.12
9.82
10.05
12.26
13.26
14.23
LC-CN
8.35
6.15
4.05
4.18
7.36
3.81
5.00
5.10
5.65
4.61
4.87
4.37
4.41
4.45
Capacity
(k)
LC-18
0.49
1.27
2.12
2.76
3.23
3.41
4.13
4.41
4.56
4.99
5.13
6.48
7.09
7.68
factor
*
LC-CN
2.52
1.59
0.71
0.76
2.11
0.61
1.11
1.15
1.38
0.95
1.05
0.84
0.86
0.88
* Capacity factors are based on an unretained peak for nitrate at 1.71 min on
LC-18 and at 2.00 min on LC-CN.
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TABLE 3
SINGLE LABORATORY PRECISION OF METHOD FOR SOIL SAMPLES
Spiked Soils
RDX
1,3,5-TNB
1,3-DNB
Tetryl
2,4,6-TNT
2,4-DNT
Mean Cone.
(mg/kg) SD
%RSD
Field-Contaminated Soils
Mean Cone.
(mg/kg) SD %RSD
HMX
46
1.7
3.7
14
153
1.8
21.6
12.8
14.1
60
8.6
46
3.5
17
40
5.0
1.4
0.4
1.9
0.14
3.1
1.4
0.17
2.3
4.6
4.1
4.0
17.9
3.5
3.4
104
877
2.8
72
1.1
2.3
7.0
669
1.0
12
29.6
0.2
6.0
0.11
0.41
0.61
55
0.44
11.5
3.4
7.1
8.3
9.8
18.0
9.0
8.2
42.3
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TABLE 4
MULTILABORATORY ERROR OF METHOD FOR SOIL SAMPLES
Spiked Soils
HMX
RDX
1,3,5-TNB
1,3-DNB
Tetryl
2,4,6-TNT
2,4-DNT
Mean
(nig/ kg)
46
60
8.6
46
3.5
17
40
5.0
Cone.
SO
2.6
2.6
0.61
2.97
0.24
5.22
1.88
0.22
%RSD
5.7
4.4
7.1
6.5
6.9
30.7
4.7
4.4
Field-Contaminated Soils
(mg/kg)
14
153
104
877
2.8
72
1.1
2.3
7.0
669
1.0
Mean Cone.
SD °/.RSD
3.7
37.3
17.4
67.3
0.23
8.8
0.16
0.49
1.27
63.4
0.74
26.0
24.0
17.0
7.7
8.2
12.2
14.5
21.3
18.0
9.5
74.0
TABLE 5
MULTI LABORATORY VARIANCE OF METHOD FOR WATER SAMPLES8
Compounds
HMX
RDX
2,4-DNT
2,4,6-TNT
Mean Cone.
(MgA)
203
274
107
107
SD
14.8
20.8
7.7
11.1
%RSD
7.3
7.6
7.2
10.4
a Nine Laboratories
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TABLE 6
MULTILABORATORY RECOVERY DATA FOR SPIKED SOIL SAMPLES
Laboratory
1
3
4
5
6
7
8
True Cone
Mean
Std Dev
% RSD
% Diff*
Mean %
Recovery
HMX
44.97
50.25
42.40
46.50
56.20
41.50
52.70
50.35
47.79
5.46
11.42
5.08
95
Concentration (/ig/g)
1,3,5- 1,3-
RDX TNB DNB
48.78
48.50
44.00
48.40
55.00
41.50
52.20
50.20
48.34
4.57
9.45
3.71
96
48.99
45.85
43.40
46.90
41.60
38.00
48.00
50.15
44.68
3.91
8.75
10.91
89
49.94
45.96
49.50
48.80
46.30
44.50
48.30
50.05
47.67
2.09
4.39
4.76
95
Tetryl
32.48
47.91
31.60
32.10
13.20
2.60
44.80
50.35
29.24
16.24
55.53
41.93
58
2,4,6-
TNT
49.73
46.25
53.50
55.80
56.80
36.00
51.30
50.65
49.91
7.11
14.26
1.46
98
2,4-
DNT
51.05
48.37
50.90
49.60
45.70
43.50
49.10
50.05
48.32
2.78
5.76
3.46
96
* Between true value and mean determined value.
8330 - 16
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Analyte
TABLE 7
COMPARISON OF METHOD ACCURACY FOR SOIL AND AQUEOUS SAMPLES
(HIGH CONCENTRATION METHOD)
Recovery (%)
Soil Method*
Aqueous Method**
2,4-DNT
2,4,6-TNT
RDX
HMX
96.0
96.8
96.8
95.4
98.6
94.4
99.6
95.5
* Taken from Bauer et al. (1989), Reference 1.
** Taken from Jenkins et al. (1984), Reference 3.
8330 - 17
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TABLE 8
PRECISION AND ACCURACY DATA FOR THE SALTING-OUT EXTRACTION METHOD
Analyte
HMX
RDX
1,3,5-TNB
1,3-DNB
Tetryl
2,4,6-TNT
2-Am-DNT
2,4-DNT
1,2-NT
1,4-NT
1,3-NT
No. of Samples1
20
20
20
20
20
20
20
20
20
20
20
Precision
(% RSD)
10.5
8.7
7.6
6.6
16.4
7.6
9.1
5.8
9.1
18.1
12.4
Ave. Recovery
(%)
106
106
119
102
93
105
102
101
102
96
97
Cone. Range
(M9A)
0-1.14
0-1.04
0-0.82
0-1.04
0-0.93
0-0.98
0-1.04
0-1.01
0-1.07
0-1.06
0-1.23
1Reagent water
8330 - 18
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EXPLOSIVES ON A
C18 COLUMN
x
5
I
'(5
k 4
1
6
i
a
id
u
U
EXPLOSIVES ON A
CN COLUMN
1
A
12 I 4
FIGURE 1
CHROMATOGRAMS FOR COLUMNS DESCRIBED IN Sec. 4.1.2.
COURTESY OF U.S. ARMY CORPS OF ENGINEERS, OMAHA, NE.
8330 - 19
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METHOD 8330
NITROAROMATICS AND NITRAMINES BY HIGH
PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
Low
Sailing Out
71 1 1 1 Add 251 3 got salt
and 1 L ot water sample to a
1 L vol flask Mix the contents
i >
71 1 12Add164mLof
acetonitnte (ACN) and stir
toe 1 5 mirts
i
i
71113 Transfer ACN layer
to 100 mL vol flask. Add 10 mU
of fresh ACN to 1 L flask and
stir Transfer 2nd portion and
combine with 1st in 100 mL flask.
I
i
71 1 1 4 Add 84 mL of sal
water to the ACN extract and stir
Transfer ACN extract to 10 mL
grad cylinder
i
7 1 1 1 5 Add 1 rnL of ACN to
1 00 mL vol flask Stir and
transfer to the 10 mL grad.
cylinder Record volume
Dilute 1 1 with reagent water
i
71116 Filter if turbid
Transfer to a vial lor
RP-HPLC analysis
7111 Sample Filtration
Place 5 mL sample in
scintiltaion vtal. AddSmL
methanol: shake, filler
through 0 5 um filter Discard
first 3 mL Retain remainder
for use
8330 - 20
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METHOD 8330
(continued)
7121 Sample riomogemzation
Air dry sample at room Temp
or below Avoid exposure to
direct sunlight Grind sample
in an acetonitrite nnsed mortar
7122 Sample Extraction
71221
Place 2 g soil subsample.
10 mLs acetonitnle in 15 mL
glass vial Cap. vortex swirl.
place in room Temp or below
ultrasonic bath for 18 hrs
71222
Let sdn settle Add 5 ml
supernatant to 5 ml calcium
chloride soln in 20 mL vial
Shake, let stand 15 mms
71223
Filter supernatant through
0 5 urn filter. Discard initial
3 mL. retain remainder
for analysis
7 2 Set Chromatographic Conditions
7 3 Calibration of HPLC
732
Run working stds in triplicate
Plot ng vs peak area or ht
Curve should be linear with
zero intercept
733
Analyze midrange calibration
std at beginning, middle.
and end of sample analyses
Redo Section 7 3 1 if peak
areas or hts do not agree
to w/m w- 20% of initial
calibration values
7 4 Sample Analysis
741
Analyze samples Confirm
measurment w/injection onto
CN column
743
Refer to Table 2 for typical
analyte retention times
8330 - 21
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September 1994
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METHOD 8331
TETRAZENE BY REVERSE PHASE
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
1.0 SCOPE AND APPLICATION
1.1 This method is intended for the analysis of tetrazene, an explosive
residue, in soil and water. This method is limited to use by analysts
experienced in handling and analyzing explosive materials. The following
compounds can be determined by this method:
Compound CAS No8
Tetrazene 31330-63-9
a Chemical Abstracts Service Registry number
1.2 Tetrazene degrades rapidly in water and methanol at room temperature.
Special care must be taken to refrigerate or cool all solutions throughout the
analytical process.
1.3 Tetrazene, in its dry form, is extremely explosive. Caution must be
taken during preparation of standards.
1.4 The estimated quantitation limit (EQL) of Method 8331 for determining
the concentration of tetrazene is approximately 7 jug/L in water and
approximately 1 mg/kg in soil.
1.5 This method is restricted to use by or under the supervision of
analysts experienced in the use of HPLC, skilled in the interpretation of
chromatograms, and experienced in handling explosive materials. Each analyst
must demonstrate the ability to generate acceptable results with this method.
2.0 SUMMARY OF METHOD
2.1 A 10 mL water sample is filtered, eluted on a C-18 column using ion
pairing reverse phase HPLC, and quantitated at 280 nm.
2.2 2 g of soil are extracted with 55:45 v/v methanol-water and
1-decanesulfonic acid on a platform shaker, filtered, and eluted on a C-18 column
using ion pairing reverse phase HPLC, and quantitated at 280 nm.
8331 - 1 Revision 0
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3.0 INTERFERENCES
3.1 No interferences are known. Tetrazene elutes early, however, and if
a computing integrator is used for peak quantification, the baseline setting may
have to be set to exclude baseline aberrations. Baseline setting is particularly
important at low concentrations of analyte.
4.0 APPARATUS AND MATERIALS
4.1 HPLC system
4.1.1 HPLC - Pump capable of achieving 4000 psi.
4.1.2 100 jiL loop injector.
4.1.3 Variable or fixed wavelength detector capable of reading
280 nm.
4.1.4 C-18 reverse phase HPLC column, 25 cm x 4.6 mm (5 nm)
(Supelco LC-18, or equivalent).
4.1.5 Digital integrator - HP 3390A (or equivalent)
4.1.6 Strip chart recorder.
4.2 Other apparatus
4.2.1 Platform orbital shaker.
4.2.2 Analytical balance - + 0.0001 g.
4.2.3 Desiccator.
4.3 Materials
4.3.1 Injection syringe - 500 /uL.
4.3.2 Filters - 0.5 nm Millex-SR and 0.5 /itm Millex-HV, disposable,
or equivalent..
4.3.3 Pipets - volumetric, glass, Class A.
4.3.4 Scintillation vials - 20 mL, glass.
4.3.5 Syringes - 10 mL.
4.3.6 Volumetric flasks, Class A - 100 mL, 200 mL.
4.3.7 Erlenmeyer flasks with ground glass stoppers - 125 mL.
8331 - 2 Revision 0
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4.4 Preparation
4.4.1 Prepare all materials as described in Chapter 4 for volatile
organics.
5.0 REAGENTS
5.1 HPLC grade chemicals shall be used in all tests. It is intended that
all reagents shall conform to the specifications of the Committee on Analytical
Reagents of the American Chemical Society, where such specifications are
available. Other grades may be used, provided it is first ascertained that the
reagent is of sufficiently high purity to permit its use without lowering the
accuracy of the determination.
5.2 General
5.2.1 Methanol, CH3OH - HPLC grade.
5.2.2 Organic-free reagent water - All references to water in this
method refer to organic-free reagent water, as defined in Chapter One.
5.2.3 1-Decanesulfonic acid, sodium salt, C10H21S03Na - HPLC grade.
5.2.4 Acetic acid (glacial), CH3COOH - reagent grade.
5.3 Standard Solutions
5.3.1 Tetrazene - Standard Analytical Reference Material.
5.3.2 Stock standard solution - Dry tetrazene to constant weight
in a vacuum desiccator in the dark. (Tetrazene is extremely explosive in
the dry state. Do not dry more reagent than is necessary to prepare stock
solutions.) Place about 0.0010 g (weighed to 0.0001 g) into a 100-ml
volumetric flask and dilute to volume with methanol. Invert flask several
times until tetrazene is dissolved. Store in freezer at -10°C. Stock
solution is about 100 mg/L. Replace stock standard solution every week.
5.3.3 Intermediate standard solutions
5.3.3.1 Prepare a 4 mg/L standard by diluting the stock
solution 1/25 v/v with methanol.
5.3.3.2 Pipet 0.5, 1.0, 2.0, 5.0, 10.0, and 20.0 mL of the
4 mg/L standard solution into 6 separate 100 mL volumetric flasks,
and make up to volume with methanol. Pipet 25.0 mL of the 4 mg/L
standard solution into a 50 mL volumetric flask, and make up to
volume with methanol. This results in intermediate standards of
about 0.02, 0.04, 0.08, 0.2, 0.4, 0.8, 2 and 4 mg/L.
5.3.3.3 Cool immediately on preparation in refrigerator or
ice bath.
8331 - 3 Revision 0
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5.3.4 Working standard solutions
5.3.4.1 Inject 4 mL of each of the intermediate standard
solutions into 6.0 ml of water. This results in concentrations of
about 0.008, 0.016, 0.032, 0.08, 0.16, 0.3, 0.8 and 1.6 mg/L.
5.3.4.2 Cool immediately on preparation in refrigerator or
ice bath.
5.5 QC spike concentrate solution
5.5.1 Dry tetrazene to constant weight in a vacuum desiccator in
the dark. (Tetrazene is extremely explosive in the dry state. Do not dry
any more than necessary to prepare standards.) Place about 0.0011 g
(weighed to 0.0001 g) into a 200-ml volumetric flask and dilute to volume
with methanol. Invert flask several times until tetrazene is dissolved.
Store in freezer at -10°C. QC spike concentrate solution is about 55
mg/L. Replace stock standard solution every week.
5.5.2 Prepare spiking solutions, at concentrations appropriate to
the concentration range of the samples being analyzed, by diluting the QC
spike concentrate solution with methanol. Cool on preparation in
refrigerator or ice bath.
5.6 Eluent
5.6.1 To make about 1 liter of eluent, add 2.44 g of
1-decanesulfonic acid, sodium salt to 400/600 v/v methanol/water, and add
2.0 ml of glacial acetic acid.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Section 4.1.
6.2 Samples must be collected and stored in glass containers. Follow
conventional sampling procedures.
6.3 Samples must be kept below 4°C from the time of collection through
analysis.
7.0 PROCEDURE
7.1 Sample Preparation
7.1.1 Filtration of Water Samples
7.1.1.1 Place a 10 mL portion of each water sample in a
syringe and filter through a 0.5 /urn Millex-HV filter unit. Discard
first 5 mL of filtrate, and retain 5 mL for analysis.
8331 - 4 Revision 0
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7.1.2 Extraction and Filtration of Soil Samples
7.1.2.1 Determination of sample % dry weight - In certain
cases, sample results are desired based on dry-weight basis. When
such data is desired, a portion of sample for this determination
should be weighed out at the same time as the portion used for
analytical determination.
WARNING: The drying oven should be contained in a hood or
vented. Significant laboratory contamination may
result from a heavily contaminated hazardous
waste sample.
7.1.2.1.1 Immediately after weighing the sample for
extraction, weigh 5-10 g of the sample into a tared
crucible. Determine the % dry weight of the sample by
drying overnight at 105°C. Allow to cool in a desiccator
before weighing:
% dry weight = q of dry sample x 100
g of sample
7.1.2.2 Weigh 2 g soil subsamples into 125 ml Erlenmeyer
flasks with ground glass stoppers.
7.1.2.3 Add 50 ml of 55/45 v/v methanol-water with
1-decanesulfonic acid, sodium salt added to make a 0.1 M solution.
7.1.2.4 Vortex for 15 seconds.
7.1.2.5 Shake for 5 hr at 2000 rpm on platform shaker.
7.1.2.6 Place a 10 ml portion of each soil sample extract
in a syringe and filter through a 0.5 /jm Millex-SR filter unit.
Discard first 5 ml of filtrate, and retain 5 ml for analysis.
7.2 Sample Analysis
7.2.1 Analyze the samples using the chromatographic conditions
given in Section 7.2.1.1. Under these conditions, the retention time of
tetrazene is 2.8 min. A sample chromatogram, including other compounds
likely to be present in samples containing tetrazene, is shown in
Figure 1.
7.2.1.1 Chromatographic Conditions
Solvent: 0.01 M 1-decanesulfonic acid, in
acidic methanol/water (Section 5.5)
Flow rate: 1.5 mL/min
Injection volume: 100 /xL
UV Detector: 280 nm
8331 - 5 Revision 0
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7.3 Calibration of HPLC
7.3.1 Initial Calibration - Analyze the working standards
(Section 5.3.4), starting with the 0.008 mg/L standards and ending with
the 0.30 mg/L standard. If the percent relative standard deviation (%RSD)
of the mean response factor (RF) for each analyte does not exceed 20%, the
system is calibrated and the analysis of samples may proceed. If the %RSD
for any analyte exceeds 20%, recheck the system and/or recalibrate with
freshly prepared calibration solutions.
7.3.2 Continuing Calibration - On a daily basis, inject 250 /j,l of
stock standard into 20 ml water. Keep solution in refrigerator until
analysis. Analyze in triplicate (by overfilling loop) at the beginning of
the day, singly after each five samples, and singly after the last sample
of the day. Compare response factors from the mean peak area or peak
height obtained over the day with the response factor at initial
calibration. If these values do not agree within 10%, recalibrate.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Prior to preparation of stock solutions, methanol should be analyzed
to determine possible interferences with the tetrazene peak. If the methanol
shows contamination, a different batch of methanol should be used.
8.3 Method Blanks
8.3.1 Method blanks for the analysis of water samples should be
organic-free reagent water carried through all sample storage and handling
procedures.
8.3.2 Method blanks for the analysis of soil samples should be
uncontaminated soil carried through all sample storage, extraction, and
handling procedures.
9.0 METHOD PERFORMANCE
9.1 Method 8331 was tested in a laboratory over a period of four days.
Spiked organic-free reagent water and standard soil were analyzed in duplicate
each day for four days. The HPLC was calibrated daily according to the
procedures given in Section 7.1. Method performance data are presented in Tables
1 and 2.
10.0 REFERENCES
1. Walsh, M.E., and T.F. Jenkins, "Analytical Method for Determining
Tetrazene in Water," U.S. Army Corps of Engineers, Cold Regions Research
& Engineering Laboratory, Special Report 87-25, 1987.
8331 - 6 Revision 0
September 1994
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2. Walsh, M.E., and T.F. Jenkins, "Analytical Method for Determining
Tetrazene in Soil," U.S. Army Corps of Engineers, Cold Regions Research &
Engineering Laboratory, Special Report 88-15, 1988.
11.0 SAFETY
11.1 Standard precautionary measures used for handling other organic
compounds should be sufficient for safe handling of the analytes targeted by
Method 8331.
8331 - 7
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FIGURE 1
16
12
^m
8 8
TNT
HMX
Tttroztnt
O1-
0.064
Absorbonct Units
8331 - 8
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September 1994
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TABLE 1.
METHOD PERFORMANCE, WATER MATRIX
Spike
Cone.
(M9A)
0.00
7.25
14.5
29
72.5
145
290
725
OVERALL
Avq % Recovery
Replicate
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Day 1
0.0
NA
0.0
NA
8.9
122
6.6
91
14.6
101
14.8
102
31.8
110
29.5
102
71.1
98
71.2
98
140.6
97
138.5
96
289.4
100
282.0
97
737.6
102
700.2
97
Day 2
0.0
NA
0.0
NA
7.8
108
9.9
' 137
14.6
101
14.1
97
30.0
103
29.7
102
73.6
102
71.3
98
143.8
99
140.8
97
288.5
99
284.2
98
707.2
98
695.8
96
Day 3
0.0
NA
0.0
NA
7.4
102
8.5
117
13.8
95
14.1
98
30.8
106
30.4
105
75.7
104
70.7
98
144.7
100
140.9
97
291.0
100
281.9
97
714.3
99
714.2
99
Average
Day 4
0.0
NA
0.0
NA
9.4
130
6.7
92
14.6
101
15.2
105
28.7
99
30.7
106
73.9
102
71.6
99
142.1
98
136.9
94
289.8
100
282.5
97
722.0
100
716.3
99
% Recovery
NA
NA
116
109
99
100
105
104
101
98
98
96
100
97
99
97
102
8331 - 9
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September 1994
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TABLE 2
METHOD PERFORMANCE, SOIL MATRIX
Spike
Cone.
(M9/L)
0.00
1.28
2.56
5.12
12.8
25.6
OVERALL
Avq % Recovery
Replicate
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Repl icate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Replicate 1
% Recovery
Replicate 2
% Recovery
Day 1
0.0
NA
0.0
NA
0.6
49
1.2
92
1.4
56
1.5
59
2.9
57
3.0
58
7.8
61
8.0
62
17.2
67
16.7
65
Day 2
0.0
NA
0.0
NA
0.9
73
0.7
56
1.5
58
2.0
79
3.0
58
3.0
59
7.6
59
8.4-
66
16.7
65
16.8
66
Day 3
0.0
NA
0.0
NA
0.6
48
0.8
63
1.6
61
1.4
56
2.9
56
3.5
69
7.8
61
7.7
60
17.4
68
17.6
69
Average
Day 4 %
0.0
NA
0.0
NA
1.0
74
0.7
56
1.6
61
1.3
50
2.9
56
3.1
60
8.1
63
8.2
64
17.3
68
17.2
67
Recovery
NA
NA
61
67
59
61
57
61
61
63
67
67
62
8331 - 10
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September 1994
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METHOD 8331
TETRAZENE BY REVERSE PHASE
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
Start
7.1 .1 Filter 10 ml
water sample; discard
first 5 mL; analyze last 5.
7.1.2.1 Determine
% dry weight.
7.1.2.2 - 7.1.2.5
Extract 2 g soil
with 50 mL solvent.
7.1.2.6 Filter 10 mL
extract; discard 5 mL;
analyze last 5 mL.
7.2 Analyze samples
using chromatographic
conditions in
Section 7.2.1.1.
7.3.1 Initial calibration:
Analyze working
standards
(Section 5.3.3).
7.3.1 Recheck system/
recalibrate with new
calibration solution.
7.3.2
Continuing
Calibration.
Stop
8331 - 11
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September 1994
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4.3 DETERMINATION OF ORGANIC ANALYTES
4.3.4 FOURIER TRANSFORM INFRARED METHODS
The following method is included in this section:
Method 8410: Gas Chromatography/Fourier Transform Infrared
(GC/FT-IR) Spectrometry for Semi volatile
Organics: Capillary Column
FOUR - 13 RevisionZ
September 1994
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METHOD 8410
GAS CHROMATOGRAPHY/FOURIER TRANSFORM INFRARED
(GC/FT-IR) SPECTROMETRY FOR SEMIVOLATILE ORGANICS:
CAPILLARY COLUMN
1.0 SCOPE AND APPLICATION
1.1 This method covers the automated identification, or compound class
assignment of unidentifiable compounds, of solvent extractable semivolatile
organic compounds which are amenable to gas chromatography, by GC/FT-IR.
GC/FT-IR can be a useful complement to GC/MS analysis (Method 8270). It is
particularly well suited for the identification of specific isomers that are not
differentiated using GC/MS. Compound class assignments are made using infrared
group absorption frequencies. The presence of an infrared band in the
appropriate group frequency region may be taken as evidence of the possible
presence of a particular compound class, while its absence may be construed as
evidence that the compound class in question is not present. This evidence will
be further strengthened by the presence of confirmatory group frequency bands.
Identification limits of the following compounds have been demonstrated by this
method.
Compound Name
CAS No.'
Acenaphthene
Acenaphthylene
Anthracene
Benzo( a) anthracene
Benzo(a)pyrene
Benzoic acid
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl } ether
Bis(2-ethylhexyl) phthalate
4-Bromophenyl phenyl ether
Butyl benzyl phthalate
4-Chloroanil ine
4 -Chloro- 3 -methyl phenol
2-Chloronaphthalene
2-Chlorophenol
4-Chlorophenol
4-Chlorophenyl phenyl ether
Chrysene
Dibenzofuran
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
2,4-Dichlorophenol
83-32-9
208-96-8
120-12-7
56-55-3
50-32-8
65-85-0
111-91-1
111-44-4
39638-32-9
117-81-7
101-55-3
85-68-7
106-47-8
59-50-7
91-58-7
95-57-8
106-48-9
7005-72-3
218-01-9
132-64-9
84-74-2
95-50-1
541-73-1
106-46-7
120-83-2
8410 - 1
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September 1994
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Compound Name CAS No.a
Dimethyl phthalate 131-11-3
Diethyl phthalate 84-66-2
4,6-Dinitro-2-methylphenol 534-52-1
2,4-Dinitrophenol 51-28-5
2,4-Dinitrotoluene 121-14-2
2,6-Dinitrotoluene 606-20-2
Di-n-octyl phthalate 117-84-0
Di-n-propyl phthalate 131-16-8
Fluoranthene 206-44-0
Fluorene 86-73-7
Hexachloroberizene 118-74-1
1,3-Hexachlorobutadiene 87-68-3
Hexachlorocyclopentadiene 77-47-4
Hexachloroethane 67-72-1
Isophorone 78-59-1
2-Methylnaphthalene 91-57-6
2-Methylphenol 95-48-7
4-Methylphenol 106-44-5
Naphthalene 91-20-3
2-Nitroaniline 88-74-4
3-Nitroaniline 99-09-2
4-Nitroaniline 100-01-6
Nitrobenzene 98-95-3
2-Nitrophenol 88-75-5
4-Nitrophenol 100-02-7
N-Nitrosodimethylamine 62-75-9
N-Nitrosodiphenylamine 86-30-9
N-Nitroso-di-n-propylamine 621-64-7
Pentachlorophenol 87-86-5
Phenanthrene 85-01-8
Phenol 108-95-2
Pyrene 129-00-0
1,2,4-Trichlorobenzene 120-82-1
2,4,5-Trichlorophenol 95-95-4
2,4,6-Trichlorophenol 88-06-2
a Chemical Abstract Services Registry Number.
1.2 This method is applicable to the determination of most extractable,
semivolatile-organic compounds in wastewater, soils and sediments, and solid
wastes. Benzidine can be subject to losses during solvent concentration and GC
analysis; a-BHC, /3-BHC, Endosulfan I and II, and Endrin are subject to
decomposition under the alkaline conditions of the extraction step; Endrin is
subject to decomposition during GC analysis; and Hexachlorocyclopentadiene and
N-Nitrosodiphenylamine may decompose during extraction and GC analysis. Other
extraction and/or instrumentation procedures should be considered for unstable
analytes.
8410 - 2 Revision 0
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1.3 The identification limit of this method may depend strongly upon the
level and type of gas chromatographable (GC) semivolatile extractants. The
values listed in Tables 1 and 2 represent the minimum quantities of semivolatile
organic compounds which have been identified by the specified GC/FT-IR system,
using this method and under routine environmental analysis conditions. Capillary
GC/FT-IR wastewater identification limits of 25 M9/L may be achieved for weak
infrared absorbers with this method, while the corresponding identification
limits for strong infrared absorbers is 2 M9/L. Identification limits for other
sample matrices can be calculated from the wastewater values after choice of the
proper sample workup procedure (see Sec. 7.1).
2.0 SUMMARY OF METHOD
2.1 Prior to using this method, the samples should be prepared for
chromatography using the appropriate sample preparation and cleanup methods.
This method describes chromatographic conditions that will allow for the
separation of the compounds in the extract and uses FT-IR for detection and
quantitation of the target analytes.
3.0 INTERFERENCES
3.1 Glassware and other sample processing hardware must be thoroughly
cleaned to prevent contamination and misinterpretation. All of these materials
must be demonstrated to be free from interferences under the conditions of the
analysis by running method blanks. Specific selection of reagents or
purification of solvents by distillation in all-glass systems may be required.
3.2 Matrix interference will vary considerably from source to source,
depending upon the diversity of the residual waste being sampled. While general
cleanup techniques are provided as part of this method, unique samples may
require additional cleanup to isolate the analytes of interest from interferences
in order to achieve maximum sensitivity.
3.3 4-Chlorophenol and 2-nitrophenol are subject to interference from co-
eluting compounds.
3.4 Clean all glassware as soon as possible after use by rinsing with the
last solvent used. Glassware should be sealed/stored in a clean environment
immediately after drying to prevent any accumulation of dust or other
contaminants.
4.0 APPARATUS AND MATERIALS
4.1 Gas Chromatographic/Fourier Transform Infrared Spectrometric
Equipment
4.1.1 Fourier Transform-Infrared Spectrometer - A spectrometer
capable of collecting at least one scan set per second at 8 cm"1 resolution
is required. In general, a spectrometer purchased after 1985, or
retrofitted to meet post-1985 FT-IR improvements, will be necessary to
8410 - 3 Revision 0
September 1994
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meet the detection limits of this protocol. A state-of-the-art A/D
converter is required, since it has been shown that the signal-to-
noise ratio of single beam GC/FT-IR systems is A/D converter
1imited.
4.1.2 GC/FT-IR Interface - The interface should be lightpipe volume-
optimized for the selected chromatographic conditions (lightpipe volume of
100-200 fj,L for capillary columns). The shortest possible inert transfer
line (preferably fused silica) should be used to interface the end of the
chromatographic column to the lightpipe. If fused silica capillary
columns are employed, the end of the GC column can serve as the transfer
line if it is adequately heated. It has been demonstrated that the
optimum lightpipe volume is equal to the full width at half height of the
GC eluate peak.
4.1.3 Capillary Column - A fused silica DB-5 30 m x 0.32 mm
capillary column with 1.0 jum film thickness (or equivalent).
4.1.4 Data Acquisition - A computer system dedicated to the GC/FT-IR
system to allow the continuous acquisition of scan sets for a full
chromatographic run. Peripheral data storage systems should be available
(magnetic tape and/or disk) for the storage of all acquired data.
Software should be available to allow the acquisition and storage of every
scan set to locate the file numbers and transform high S/N scan sets, and
to provide a real time reconstructed chromatogram.
4.1.5 Detector - A cryoscopic, medium-band HgCdTe (MCT) detector
with the smallest practical focal area. Typical narrow-band MCT detectors
operate from 3800-800 cm"1, but medium-band MCT detectors can reach
650 cm"1. A 750 cm"1 cutoff (or lower) is desirable since it allows the
detection of typical carbon-chlorine stretch and aromatic out-of-plane
carbon-hydrogen vibrations of environmentally important organo-chlorine
and polynuclear aromatic compounds. The MCT detector sensitivity (D)*
should be > 1 x 1010 cm.
4.1.6 Lightpipe - Constructed of inert materials, gold coated, and
volume-optimized for the desired chromatographic conditions (see Sec.
7.3).
4.1.7 Gas Chromatograph - The FT-IR spectrometer should be
interfaced to a temperature programmable gas chromatograph equipped with
a Grob-type (or equivalent) purged splitless injection system suitable for
capillary glass columns or an on-column injector system.
A short, inert transfer line should interface the gas chromatograph
to the FT-IR lightpipe and, if applicable, to the GC detector. Fused
silica GC columns may be directly interfaced to the lightpipe inlet and
outlet.
4.2 Dry Purge Gas - If the spectrometer is the purge-type, provisions
should be made to provide a suitable continuous source of dry purge-gas to the
FT-IR spectrometer.
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4.3 Dry Carrier Gas - The carrier gas should be passed through an
efficient cartridge-type drier.
4.4 Syringes - 1-jj.l, 10-juL.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available. Other grades may be used,
provided it is first ascertained that the reagent is of sufficiently high purity
to permit its use without lessening the accuracy of the determination.
5.2 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
5.3 Solvents
5.3.1 Acetone, CH3COCH3 - Pesticide quality, or equivalent.
5.3.2 Methylene chloride, CH2C12 - Pesticide quality, or equivalent.
5.4 Stock Standard Solutions (1000 mg/L) - Standard solutions can be
prepared from pure standard materials or purchased as a certified solution.
5.4.1 Prepare stock standard solutions by accurately weighing 0.1000
+ 0.0010 g of pure material. Dissolve the material in pesticide quality
acetone or other suitable solvent and dilute to volume in a 100 ml
volumetric flask. Larger volumes can be used at the convenience of the
analyst. When compound purity is assayed to be 96 percent or greater, the
weight may be used without correction to calculate the concentration of
the stock standard. Commercially prepared stock standards may be used at
any concentration if they are certified by the manufacturer or by an
independent source.
5.4.2 Transfer the stock standard solutions into bottles with Teflon
lined screw-caps. Store at 4°C and protect from light. Stock standard
solutions should be checked frequently for signs of degradation or
evaporation, especially just prior to preparing calibration standards from
them.
5.4.3 Stock standard solutions must be replaced after 6 months or
sooner if comparison with quality control reference samples indicates a
problem.
5.5 Calibration Standards and Internal Standards - For use in situations
where GC/FT-IR will be used for primary quantitation of analytes rather than
confirmation of GC/MS identification.
5.5.1 Prepare calibration standards that contain the compounds of
interest, either singly or mixed together. The standards should be
8410 - 5 Revision 0
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prepared at concentrations that will completely bracket the working range
of the chromatographic system (at least one order of magnitude is
suggested).
5.5.2 Prepare internal standard solutions. Suggested internal
standards are 1-Fluoronaphthalene, Terphenyl, 2-Chlorophenol, Phenol,
Bis(2-chloroethoxy)methane, 2,4-Dichlorophenol, Phenanthrene, Anthracene,
and Butyl benzyl phthalate. Determine the internal standard concentration
levels from the minimum identifiable quantities. See Tables 1 and 2.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes, Sec.
4.1.
7.0 PROCEDURE
7.1 Sample Preparation - Samples must be prepared by one of the following
methods prior to GC/FT-IR analysis.
Matrix Methods
Water 3510, 3520
Soil/sediment 3540, 3541, 3550
Waste 3540, 3541, 3550, 3580
7.2 Extracts may be cleaned up by Method 3640, Gel-Permeation Cleanup.
7.3 Initial Calibration - Recommended GC/FT-IR conditions:
Scan time: At least 2 scan/sec.
Initial column temperature and hold time: 40°C for 1 minute.
Column temperature program: 40-280°C at 10°C/min.
Final column temperature hold: 280°C.
Injector temperature: 280-300°C.
Transfer line temperature: 270°C.
Lightpipe: 280°C.
Injector: Grob-type, splitless or on-
column.
Sample volume: 2-3 yl.
Carrier gas: Dry helium at about 1 mL/min.
7.4 With an oscilloscope, check the detector centerburst intensity versus
the manufacturer's specifications. Increase the source voltage, if necessary,
to meet these specifications. For reference purposes, laboratories should
prepare a plot of time versus detector voltage over at least a 5 day period.
7.5 Capillary Column Interface Sensitivity Test - Install a 30 m x
0.32 mm fused silica capillary column coated with 1.0 jum of DB-5 (or
equivalent). Set the lightpipe and transfer lines at 280°C, the injector at
225°C and the GC detector at 280°C (if used). Under splitless Grob-type or on-
8410 - 6 Revision 0
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column injection conditions, inject 25 ng of nitrobenzene, dissolved in 1 jul_ of
methylene chloride. The nitrobenzene should be identified by the on-line library
software search within the first five hits (nitrobenzene should be contained
within the search library).
7.6 Interferometer - If the interferometer is air-driven, adjust the
interferometer drive air pressure to manufacturer's specifications.
7.7 MCT Detector Check - If the centerburst intensity is 75 percent or
less of the mean intensity of the plot maximum obtained by the procedure of Sec.
7.4, install a new source and check the MCT centerburst with an oscilloscope
versus the manufacturer's specifications (if available). Allow at least five
hours of new source operation before data acquisition.
7.8 Frequency Calibration - At the present time, no consensus exists
within the spectroscopic community on a suitable frequency reference standard for
vapor-phase FT-IR. One reviewer has suggested the use of indene as an on-the-fly
standard.
7.9 Minimum Identifiable Quantities - Using the GC/FT-IR operating
parameters specified in Sec. 7.3, determine the minimum identifiable quantities
for the compounds of interest.
7.9.1 Prepare a plot of lightpipe temperature versus MCT centerburst
intensity (in volts or other vertical height units). This plot should
span the temperature range between ambient and the lightpipe thermal limit
in increments of about 20°C. Use this plot for daily QA/QC (see Sec. 8.4).
Note that modern GC/FT-IR interfaces (1985 and later) may have eliminated
most of this temperature effect.
7.10 GC/FT-IR Extract Analysis
7.10.1 Analysis - Analyze the dried methylene chloride extract
using the chromatographic conditions specified in Sec. 7.3 for capillary
column interfaces.
7.10.2 GC/FT-IR Identification - Visually compare the analyte
infrared (IR) spectrum versus the search library spectrum of the most
promising on-line library search hits. Report, as identified, those
analytes with IR frequencies for the five (maximum number) most intense IR
bands (S/N > 5) which are within + 5.0 cm"1 of the corresponding bands in
the library spectrum. Choose IR bands which are sharp and well resolved.
The software used to locate spectral peaks should employ the peak "center
of gravity" technique. In addition, the IR frequencies of the analyte and
library spectra should be determined with the same computer software.
7.10.3 Retention Time Confirmation - After visual comparison of
the analyte and library spectrum as described in Sec. 7.10.2, compare the
relative retention times (RRT) of the analyte and an authentic standard of
the most promising library search hit. The standard and analyte RRT
should agree within + 0.01 RRT units when both are determined at the same
chromatographic conditions.
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7.10.4 Compound Class or Functionality Assignment - If the
analyte cannot be unequivocally identified, report its compound class or
functionality. See Table 3 for gas-phase group frequencies to be used as
an aid for compound class assignment. It should be noted that FT-IR gas-
phase group stretching frequencies are 0-30 cm"1 higher in frequency than
those of the condensed phase.
7.10.5 Quantitation - This protocol can be used to confirm GC/MS
identifications, with subsequent quantitation. Two FT-IR quantitation and
a supplemental GC detector technique are also provided.
7.10.5.1 Integrated Absorbance Technique - After analyte
identification, construct a standard calibration curve of
concentration versus integrated infrared absorbance. For this
purpose, choose for integration only those FT-IR scans which are at
or above the peak half-height. The calibration curve should span at
least one order of magnitude and the working range should bracket
the analyte concentration.
7.10.5.2 Maximum Absorbance Infrared Band Technique -
Following analyte identification, construct a standard calibration
curve of concentration versus maximum infrared band intensity. For
this purpose, choose an intense, symmetrical and well resolved IR
absorbance band.
(Note that IR transmission is not proportional to concentra-
tion). Select the FT-IR scan with the highest absorbance to plot
against concentration. The calibration curve should span at least
one order of magnitude and the working range should bracket the
analyte concentration. This method is most practical for
repetitive, target compound analyses. It is more sensitive than the
integrated absorbance technique.
7.10.5.3 Supplemental GC Detector Technique - If a GC
detector is used in tandem with the FT-IR detector, the following
technique may be used: following analyte identification, construct
a standard calibration curve of concentration versus integrated peak
area. The calibration curve should span at least one order of
magnitude and the working range should bracket the analyte
concentration. This method is most practical for repetitive, target
compound analyses.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
Quality control to validate sample extraction is covered in Method 3500 and in
the extraction method utilized. If extract cleanup was performed, follow the QC
in Method 3600 and in the specific cleanup method.
8.2 One Hundred Percent Line Test - Set the GC/FT-IR operating conditions
to those employed for the Sensitivity Test (see Sec. 7.5). Collect 16 scans over
the entire detector spectral range. Plot the test and measure the peak-to-peak
8410 - 8 Revision 0
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noise between 1800 and 2000 cm"1. This noise should be < 0.15%. Store this plot
for future reference.
8.3 Single Beam Test - With the GC/FT-IR at analysis conditions, collect
16 scans in the single beam mode. Plot the co-added file and compare with a
subsequent file acquired in the same fashion several minutes later. Note if the
spectrometer is at purge equilibrium. Also check the plot for signs of
deterioration of the lightpipe potassium bromide windows. Store this plot for
future reference.
8.4 Align Test - With the lightpipe and MCT detector at thermal
equilibrium, check the intensity of the centerburst versus the signal temperature
calibration curve. Signal intensity deviation from the predicted intensity may
mean thermal equilibrium has not yet been achieved, loss of detector coolant,
decrease in source output, or a loss in signal throughput resulting from
lightpipe deterioration.
8.5 Mirror Alignment - Adjust the interferometer mirrors to attain the
most intense signal. Data collection should not be initiated until the
interferogram is stable. If necessary, align the mirrors prior to each GC/FT-IR
run.
8.6 Lightpipe - The lightpipe and lightpipe windows should be protected
from moisture and other corrosive substances at all times. For this purpose,
maintain the lightpipe temperature above the maximum GC program temperature but
below its thermal degradation limit. When not in use, maintain the lightpipe
temperature slightly above ambient. At all times, maintain a flow of dry, inert,
carrier gas through the lightpipe.
8.7 Beamsplitter - If the spectrometer is thermostated, maintain the
beamsplitter at a temperature slightly above ambient at all times. If the
spectrometer is not thermostated, minimize exposure of the beamsplitter to
atmospheric water vapor.
9.0 METHOD PERFORMANCE
9.1 Method 8410 has been in use at the U.S. Environmental Protection
Agency Environmental Monitoring Systems Laboratory for more than two years.
Portions of it have been reviewed by key members of the FT-IR spectroscopic
community (9). Side-by-side comparisons with GC/MS sample analyses indicate
similar demands upon analytical personnel for the two techniques. Extracts
previously subjected to GC/MS analysis are generally compatible with GC/FT-IR.
However, it should be kept in mind that lightpipe windows are typically water
soluble. Thus, extracts must be vigorously dried prior to analysis.
9.2 Table 4 provides performance data for this method.
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10.0 REFERENCES
1. Handbook for Analytical Quality Control in Water and Wastewater
Laboratories; U.S. Environmental Protection Agency. Environmental
Monitoring and Support Laboratory. ORD Publication Offices of Center for
Environmental Research Information: Cincinnati, OH, March 1979; Sec. 4,
EPA-600/4-79-019.
2. Freeman, R.R. Hewlett Packard Application Note: Quantitative Analysis
Using a Purged Splitless Injection Technique; ANGC 7-76.
3. Cole, R.H. Tables of Wavenumbers for the Calibration of Infrared
Spectrometers; Pergamon: New York, 1977.
4. Grasselli, J.G.; Griffiths, P.R.; Hannah, R.W. "Criteria for Presentation
of Spectra from Computerized IR Instruments"; Appl. Spectrosc. 1982, 36,
87.
5. Nyquist, R.A. The Interpretation of Vapor-Phase Infrared Spectra. Group
Frequency Data; Volume I. Sadtler Laboratories: Philadelphia, PA, 1984.
6. Socrates, G. Infrared Characteristic Group Frequencies; John Wiley and
Sons: New York, NY, 1980.
7. Bellamy, L.J. The Infrared Spectra of Complex Organic Molecules; 2nd ed.;
John Wiley and Sons: New York, NY, 1958.
8. Szymanski, H.A. Infrared Band Handbook, Volumes I and II; Plenum: New
York, NY, 1965.
9. Gurka, D.F. "Interim Protocol for the Automated Analysis of Semivolatile
Organic Compounds by Gas Chromatography/Fourier Transform-Infrared
Spectrometry"; Appl. Spectrosc. 1985, 39, 826.
10. Griffiths, P.R.; de Haseth, J.A.; Azarraga, L.V. "Capillary GC/FT-IR";
Anal. Chem. 1983, 55, 1361A.
11. Griffiths, P.P.; de Haseth, J.A. Fourier Transform-Infrared Spectrometry;
Wiley-Interscience: New York, NY, 1986.
12. Gurka, D. F.; Farnham, I.; Potter, B. B.; Pyle, S.; Titus, R. and Duncan,
W. "Quantitation Capability of a Directly Linked Gas
Chromatography/Fourier Transform Infrared/Mass Spectrometry System"; Anal.
Chem., 1989, 61, 1584.
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TABLE 1.
FUSED SILICA CAPILLARY COLUMN GAS CHROMATOGRAPHIC/FOURIER TRANSFORM
INFRARED IDENTIFICATION LIMITS FOR BASE/NEUTRAL EXTRACTABLES
Compound
Acenaphthene
Acenaphthylene
Anthracene
Benzo(a)anthracene
Benzo(a)pyrene
Bis(Z-cnloroethyl) ether
Bis(2-chloroethoxy)methane
Bis(2-chloroisopropyl) ether
Butyl benzyl phthalate
4-Bromophenyl phenyl ether
2-Chloronaphthalene
4-Chloroaniline
4-Chlorophenyl phenyl ether
Chrysene
Di-n-butyl phthalate
Dibenzofuran
Diethyl phthalate
Dimethyl phthalate
Di-n-octyl phthalate
Di-n-propyl phthalate
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1 ,4-Dichlorobenzene
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Bis-(2-ethylhexyl) phthalate
Fluoranthene
Fluorene
Hexachlorobenzene
Hexachl orocycl opentadi ene
Hexachloroethane
1 , 3 -Hexachl orobutadi ene
Isophorone
2-Methylnaphthalene
Naphthalene
Nitrobenzene
N-Nitrosodi methyl ami ne
N-Nitrosodi-n-propylamine
N-Nitrosodi phenyl ami ned
2-Nitroaniline
3-Nitroaniline
Identification
ng injected8
40(25)
50(50)
40(50)
(50)
(100)
70(10)
50(10)
50(10)
25(10)
40(5)
110
40
20(5)
(100)
20(5)
40
20(5)
20(5)
25(10)
25(5)
50
50
50
20
20
25(10)
100(50)
40(50)
40
120
50
120
40
110
40(25)
25
20(5)
50(5)
40
40
40
Limit
Mg/L"
20(12.5)
25(25)
20(25)
(25)
(50)
35(5)
25(5)
25(5)
12.5(5)
20(2.5)
55
20
10(2.5)
(50)
10(2.5)
20
10(2.5)
10(2.5)
12.5(5)
12.5(2.5)
25
25
25
10
10
12.5(5)
50(25)
20(25)
20
60
25
60
20
55
20(12.5)
12.5
10(2.5)
25(2.5)
20
20
20
»max, cm"
799
799
874
745
756
1115
1084
1088
1748
1238
851
1543
1242
757
1748
1192
1748
1751
1748
1748
1458
779
1474
1547
1551
1748
773
737
1346
814
783
853
1690
3069
779
1539
1483
1485
1501
1564
1583
8410 - 11
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TABLE 1.
(Continued)
Identification Limit
Compound ng injected8 M9/Lb i>max, cm"1c
4-Nitroanil ine
Phenanthrene
Pyrene
1,2,4-Trichlorobenzene
40
50(50)
100(50)
50(25)
20
25(25)
50(25)
25(12.5)
1362
729
820
750
a Determined using on-column injection and the conditions of Sec. 7.3. A medium
band HgCdTe detector [3800-700 cm'1; D'value Upeak 1000 Hz, 1) 4.5 x 1010 cm
Hz1/2W"1] type with a 0.25 mm2 focal chip was used. The GC/FT-IR system is a
1976 retrofitted model. Values in parentheses were determined with a new
(1986) GC/FT-IR system. A narrow band HgCdTe detector [3800-750cm"1; D'value
(/Jpeak 1000 Hz, 1) 4 x 1010 cm Hz1'2^1] was used. Chromatographic conditions
are those of Sec. 7.3.
b Based on a 2 juL injection of a one liter sample that has been extracted and
concentrated to a volume of 1.0 mL. Values in parentheses were determined
with a new (1986) GC/FT-IR system. A narrow band HgCdTe detector [3800-750cm~
1; D'value (/Ipeak 1000 Hz, 1) 4 x 1010 cm Hz1/2W"1] was used. Chromatographic
conditions are those of Sec. 7.3.
0 Most intense IR peak and suggested quantitation peak.
d Detected as diphenylamine.
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TABLE 2.
FUSED SILICA CAPILLARY COLUMN GAS CHROMATOGRAPHIC/FOURIER TRANSFORM
INFRARED ON-LINE AUTOMATED IDENTIFICATION LIMITS FOR ACIDIC EXTRACTABLES
Identification Limit
Compound
ng injected8
j>max, cm"
Benzoic acid
2-Chlorophenol
4-Chlorophenold
4-Chloro-3 -methyl phenol
2-Methyl phenol
4-Methyl phenol
2,4-Dichlorophenol
2,4-Dinitrophenol
4, 6-Dinitro-2-methyl phenol
2-NitrophenoTd
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
2,4,5-Trichlorophenol
70
50
100
25
50
50
50
60
60
40
50
50
70
120
120
35
25
50
12.5
25
25
25
30
30
20
25
25
35
60
60
1751
1485
1500
1177
748
1177
1481
1346
1346
1335
1350
1381
1184
1470
1458
* Operating conditions are the same as those cited in Sec. 7.3.
b Based on a 2 /LtL injection of a one liter sample that has been extracted and
concentrated to a volume of 1.0 mL.
0 Most intense IR peak and suggested quantitation peak.
d Subject to interference from co-eluting compounds.
8410 - 13
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TABLE 3.
GAS-PHASE GROUP FREQUENCIES
Number of
Functionality Class Compounds
Ether
Ester
Nitro
Nitrile
Ketone
Amide
AT kyne
Acid
Phenol
Aryl, Alkyl
Benzyl , Al kyl
Di aryl
Dial kyl
Alkyl, Vinyl
Unsubstituted Aliphatic
Aromatic
Monosubstituted Acetate
Aliphatic
Aromatic
Aliphatic
Aromatic
Aliphatic (acyclic)
(a,|8 unsaturated)
Aromatic
Substituted Acetamides
Aliphatic
Aliphatic
Dimerized-Aliphatic
Aromatic
1,4-Disubstituted
1,3-Disubstituted
1,2-Disubstituted
14
3
5
12
3
29
11
34
5
18
9
9
13
2
16
8
8
24
22
2
10
10
15
15
15
10
10
10
6
Frequency
Range, i/cm"1
1215-1275
1103-1117
1238-1250
1084-1130
1204-1207
1128-1142
1748-1761
1703-1759
1753-1788
1566-1594
1548-1589
1377-1408
1327-1381
1535-1566
1335-1358
2240-2265
2234-2245
1726-1732
1638-1699
1701-1722
1710-1724
3323-3329
3574-3580
1770-1782
3586-3595
3574-3586
1757-1774
3645-3657
1233-1269
1171-1190
3643-3655
1256-1315
1157-1198
3582-3595
1255-1274
(continued)
8410 - 14
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TABLE 3.
(Continued)
Functional ity
Alcohol
Amine
Alkane
Aldehyde
Benzene
Class
Primary Aliphatic
Secondary Aliphatic
Tertiary Aliphatic
Primary Aromatic
Secondary Aromatic
Al iphatic
Aromatic
Aliphatic
Monosubstituted
Number of
Compounds
20
11
16
17
10
10
6
15
5
10
14
12
12
12
6
6
6
7
24
24
11
23
25
Frequency
Range, j>cm~1
3630-3680
1206-1270
1026-1094
3604-3665
1231-1270
3640-3670
1213-1245
3480-3532
3387-3480
760- 785
2930-2970
2851-2884
1450-1475
1355-1389
1703-1749
2820-2866
2720-2760
1742-1744
2802-2877
2698-2712
1707-1737
1582-1630
1470-1510
831- 893
735- 790
675- 698
8410 - 15
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TABLE 4. FUSED SILICA CAPILLARY COLUMN GC/FT-IR QUANTITATION RESULTS
Compound
Concentration
Range, and
Identification
Limit, nga
Maximum
Absorbanceb
Correlation
Coefficient"1
Integrated
Absorbance0
Correlation
Coefficient11
Acenaphthene
Acenaphthylene
Anthracene
Benzo(a)anthracene
Benzoic acid
Benzo(a)pyrene
Bis(2-chloroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
4-Bromophenyl phenyl ether
Butyl benzyl phthalate
4-Chloroaniline
4-Chloro-3 -methyl phenol
2-Chloronaphthalene
2-Chlorophenol
4-Chlorophenole
4-Chlorophenyl phenyl ether
Chrysene
Dibenzofuran
Di-n-butyl phthalate
1,2-Dichlorobenzene
1,3'Dichlorobenzene
1,4-Dichlorobenzene
2,4-Dichlorophenol
Dimethyl phthalate
Dimethyl phthalate
Dinitro-2-methyl phenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Bis(2-ethylhexyl) phthalate
Fluoranthene
Fluorene
Hexachl orobenzene
1,3-Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachl oroethane
Isophorone
2-Methylnaphthalene
25-250
25-250
50-250
50-250
50-250
100-250
25-250
25-250
50-250
25-250
25-250
25-250
25-250
100-250
25-250
25-250
100-250
25-250
25-250
25-250
25-250
25-250
25-250
25-250
25-250
50-250
50-250
25-250
25-250
25-250
25-250
25-250
25-250
50-250
50-250
100-250
25-250
25-250
50-250
0.9995
0.9959
0.9969
0.9918
0.9864
0.9966
0.9992
0.9955
0.9981
0.9995
0.9999
0.9991
0.9975
0.9897
0.9976
0.9999
0.9985
0.9697
0.9998
0.9937
0.9985
0.9994
0.9964
0.9998
0.9998
0.9936
0.9920
0.9966
0.9947
0.9983
0.9991
0.9983
0.9987
0.9981
0.9960
0.9862
0.9986
0.9984
0.9981
0.9985
0.9985
0.9971
0.9921
0.9892
0.9074
0.9991
0.9992
0.9998
0.9996
0.9994
0.9965
0.9946
0.9988
0.9965
0.9997
0.9984
0.8579
0.9996
0.9947
0.9950
0.9994
0.9969
0.9996
0.9997
0.9967
0.9916
0.9928
0.9966
0.9991
0.9993
0.9966
0.9989
0.9995
0.9979
0.9845
0.9992
0.9990
0.9950
(continued)
8410 - 16
Revision 0
September 1994
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TABLE 4. (Continued)
Compound
Concentration
Range, and
Identification
Limit, nga
Maximum
Absorbanceb
Correlation
Coefficient11
Integrated
Absorbance0
Correlation
Coefficient11
2-Methyl phenol
4-Methyl phenol
Naphthalene
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Nitrobenzene
2-Nitrophenole
4-Nitrophenol
N-Nitrosodi methyl ami ne
N-Nitrosodiphenylamine
N-Nitrosodi -n-propylamine
Pentachlorophenol
Phenanthrene
Phenol
Pyrene
1 , 2 , 4-Trichl orobenzene
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
25-250
25-250
25-250
25-250
25-250
25-250
25-250
50-250
25-250
25-250
25-250
50-250
25-250
25-250
50-250
50-250
25-250
25-250
0.9972
0.9972
0.9956
0.9996
0.9985
0.9936
0.9997
0.9951
0.9982
0.9994
0.9991
0.9859
0.9941
0.9978
0.9971
0.9969
0.9952
0.9969
0.9964
0.9959
0.9954
0.9994
0.9990
0.9992
0.9979
0.9953
0.9993
0.9971
0.9995
0.9883
0.9989
0.9966
0.9977
0.991
0.9966
0.9965
a Lower end of range is at or near the identification limit.
b FT-IR scan with highest absorbance plotted against concentration.
c Integrated absorbance of combined FT-IR scans which occur at or above the
chromatogram peak half-height.
d Regression analysis carried out at four concentration levels. Each level
analyzed in duplicate. Chromatographic conditions are stated in Sec. 7.3.
e Subject to interference from co-eluting compounds.
8410 - 17
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September 1994
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METHOD 8410
GAS CHROMATOGRAPHY/FOURIER TRANSFORM INFRARED (GC/FT-IR)
SPECTROMETRY FOR SEMIVOLATILE ORGANICS: CAPILLARY COLUMN
( Start j
7.1 Sample
preparation
prior to
GC/FT-IR
analysis.
7.2 Optional
Gel
Permeation
Cleanup of
extracts.
7.3 Initial
Calibration;
recommended
GC/FT-IR
conditions.
7.4 Check
detector
centerburst
intensity.
7.5 Column
Interface
Sensitivity.
7.6 Adjust
interferometer
drive air
pressure.
7.10.1 Analyze
extracts using
conditions of
Section 7.3.
7.10.2 GC/FT-IR
Identification;
compare analyte
IR spectrum;
report.
7.7 MC
Detector;
centerburst
intensity <75%
plot max of
Section
7.4
7.10.3
Retention Time;
compare RRT of
analyte with
standard.
7.7 Replace
Source.
7.10.4 Report
compound class
if no library
match is found.
7.8 Frequency
Calibration.
7.10.5
Quantitation
desired.
7.9 Determine
min. identifiable
quantities of
analyte of
interest.
7.10.6 Standard
calibration curve
of cone. vs.
integrated IR
absorbance.
7.10.6
Quantitation
by integrated
absorbance?
7.9.1 Prepare
plot of
lightpipe T vs.
MCT centerburst
intensity.
7.10.8 Is
GC Detector
used in tandem
with FT-IR
detector?
7.10.7 Standard
calibration
curve of cone.
vs. max. IR band
intensity.
\ Yes
/
L*
7.10.8
Supplemental
GC Detector
Technique.
8410 - 18
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September 1994
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4.4 MISCELLANEOUS SCREENING METHODS
The following methods are included in this section:
Method 3810: Headspace
Method 3820: Hexadecane Extraction and Screening of Purgeable
Organics
Method 4010: Screening for Pentachlorophenol by Immunoassay
Method 8275: Thermal Chromatography/Mass Spectrometry (TC/MS)
for Screening Semi volatile Organic Compounds
FOUR - 14 Revision 2
September 1994
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METHOD 3810
HEADSPACE
1.0 SCOPE AND APPLICATION
1.1 Method 3810 was formerly Method 5020 in the second edition of this
manual.
1.2 Method 3810 is a static headspace technique for extracting volatile
organic compounds from samples. It is a simple method that allows large
numbers of samples to be screened in a relatively short period of time. It is
ideal for screening samples prior to using the purge-and-trap method.
Detection limits for this method may vary widely among samples because of the
large variability and complicated matrices of waste samples. The method works
best for compounds with boiling points of less than 125*C. The sensitivity of
this method will depend on the equilibria of the various compounds between the
vapor and dissolved phases.
1.3 Due to the variability of this method, this procedure is recommended
for use only as a screening procedure for other, more accurate determinative
methods (Methods 8010, 8015, 8020, 8030, and 8240).
2.0 SUMMARY OF METHOD
2.1 The sample is collected in sealed glass containers and allowed to
equilibrate at 90*C. A sample of the headspace gas is withdrawn with a gas-
tight syringe for screening analysis using the conditions specified in one of
the GC or GC/MS determinative methods (8010, 8015, 8020, 8030, or 8240).
3.0 INTERFERENCES
3.1 Samples can be contaminated by diffusion of volatile organics
(particularly chlorofluorocarbons and methylene chloride) through the sample
container septum during shipment and storage. A field sample blank prepared
from reagent water and carried through sampling and subsequent storage and
handling can serve as a check on such contamination.
3.2 Contamination by carryover can occur whenever high-level and low-
level samples are sequentially analyzed. To reduce carryover, the sample
syringe must be rinsed out between samples with reagent water. Whenever an
unusually concentrated sample is encountered, it should be followed by an
analysis of reagent water. It may be necessary to wash out the syringe with
detergent, rinse with distilled water, and dry in a 105*C oven between
analyses.
3.3 Before processing any samples, the analyst should demonstrate daily
through the analysis of an organic-free water or solvent blank that the entire
analytical system is interference-free.
3810 - 1
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Date September 1986
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4.0 APPARATUS AND MATERIALS
4.1 Refer to the specific determinative method for appropriate apparatus
and materials.
4.2 Vials; 125-mL Hypo-Vials (Pierce Chemical Co., #12995, or
equivalent), four each.
4.3 Septa: Tuf-Bond (Pierce #12720 or equivalent).
4'4 Seals; Aluminum (Pierce #132141 or equivalent).
4.5 Crimper; Hand (Pierce #13212 or equivalent).
4«6 Syringe; 5-mL, gas-tight with shutoff valve and chromatographic
needles.
4'7 Microsyringe: 250- or 500-uL.
4.8 Water bath; Heated, with concentric ring cover, capable of
temperature controT~[+5*C). The bath should be used in a hood.
5.0 REAGENTS
5.1 Refer to the specific determinative method and Method 8000 for
preparation of calibration standards.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Refer to the introductory material to this chapter, Organic
Analytes, Section 4.1.
7.0 PROCEDURE
7.1 Gas chromatographic conditions and Calibration; Refer to the
specific determinative method for GC operating conditions and to Method 8000,
Section 7.4, for calibration procedures.
7.2 Sample preparation;
7.2.1 Place 10.0 g of a well-mixed waste sample into each of two
separate 125-mL septum-seal vials.
7.2.2 Dose one sample vial through the septum with 200 uL of a
50 ng/uL calibration standard containing the compounds of interest.
Label this "1-ppm spike."
3810 - 2
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Date September 1986
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7.2.3 Dose a separate (empty) 125-mL septum seal vial with 200 uL
of the same 50 ng/uL calibration standard. Label this "1-ppm standard."
7.2.4 Place the sample, 1-ppm-spike, and 1-ppm-standard vials into
a 90*C water bath for 1 hr. Store the remaining sample vial at 4.0*C for
possible future analysis.
7.3 Sample analysis:
7.3.1 While maintaining the vials at 90°C, withdraw 2 ml of the
headspace gas with a gas-tight syringe and analyze by direct injection
into a GC. The GC should be operated using the same GC conditions listed
in the method being screened (8010, 8015, 8020, 8030, or 8240).
7.3.2 Analyze the 1-ppm standard and adjust instrument sensitivity
to give a minimum response of at least 2 times the background. Record
retention times (RT) and peak areas of compounds of interest.
7.3.3 Analyze the 1-ppm spiked sample in the same manner. Record
RTs and peak areas.
7.3.4 Analyze the undosed sample as in Paragraph 7.3.3.
7.3.5 Use the results obtained to determine if the sample requires
dilution or methanolic extraction as indicated in Method 5030.
8.0 QUALITY CONTROL
8.1 Before processing any samples, the analyst should demonstrate
through the analysis of a distilled water method blank that all glassware and
reagents are interference-free. Each time a set of samples is extracted or
there is a change in reagents, a method blank should be processed as a
safeguard against chronic laboratory contamination. The blank samples should
be carried through all stages of the sample preparation and measurement.
8.2 Standard quality assurance practices should be used with this
method. Fortified samples should be carried through all stages of sample
preparation and measurement; they should be analyzed to validate the
sensitivity and accuracy of the analysis. If the fortified waste samples do
not indicate sufficient sensitivity to detect less than or equal to 1 ug/g of
sample, then the sensitivity of the instrument should be increased.
9.0 METHOD PERFORMANCE
9.1 No data provided.
3810 - 3
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Date September 1986
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10.0 REFERENCES
1. Hachenberg, H. and A. Schmidt, Gas Chromatographlc Headspace Analysis,
Philadelphia: Hayden & Sons Inc., 1979.
2. Friant, S.L. and I.H. Suffet, "Interactive Effects of Temperature, Salt
Concentration and pH on Headspace Analysis for Isolating Volatile Trace
Organlcs in Aqueous Environmental Samples," Anal. Chem. 51, 2167-2172, 1979.
3810 - 4
Revision
Date September 1986
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METHOO 3B1O
HEAOSPACE METHOD
C
Start
7. 1
Set GC
opcrat ing
cono1tIons
7.2
Prepare sample
7.3
Analyzc
by direct
inj ect ion
into a GC
7
3.5
di
fl
c
Determine
if cample
required
lution or
letnanol ic
xtractlon
f Stop J
3810 - 5
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Date September 1986
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METHOD 3820
HEXADECANE EXTRACTION AND SCREENING OF PUR6EABLE ORGANICS
1.0 SCOPE AND APPLICATION
1.1 This method is a screening procedure for use with purge-and-trap GC
or GC/MS. The results of this analysis are purely qualitative and should not
be used as an alternative to more detailed and accurate quantitation methods.
2.0 SUMMARY OF METHOD
2.1 An aliquot of sample is extracted with hexadecane and then analyzed
by GC/FID. The results of this analysis will indicate whether the sample
requires dilution or methanolic extraction prior to purge-and-trap GC or GC/MS
analysis.
3.0 INTERFERENCES
3.1 Method interferences may be caused by contaminants in solvents,
reagents, and glassware. All these materials must be routinely demonstrated
to be free from contaminants by running laboratory reagent blanks. Matrix
interferences may be caused by contaminants that are coextracted from the
sample. The extent of matrix interferences will vary considerably from sample
to sample depending upon the nature and diversity of the water being sampled.
3.2 The flame ionization detector varies considerably in sensitivity
when comparing aromatics and halogenated methanes and ethanes. Halomethanes
are approximately 20x less sensitive than aromatics and haloethanes
approximately lOx less sensitive. Low-molecular-weight, water-soluble
solvents (e.g., alcohols and ketones) will not extract from the water, and
therefore will not be detected by GC/FID.
4.0 APPARATUS AND MATERIALS
4.1 Balance; Analytical, capable of accurately weighing 0.0001 gm.
4.2 Gas Chromatograph; An analytical system complete with gas
chromatograph suitable for on-column injection and all required accessories
including syringes, analytical columns, gases, detector, and strip-chart
recorder (or equivalent). A data system is recommended for measuring peak
heights and/or peak areas.
4.2.1 Detector: Flame ionization (FID).
4.2.2 GC column: 3-m x 2-mm I.D. glass column packed with
10% OV-101 on 100/120 mesh Chromosorb W-HP (or equivalent). The
column temperature should be programmed from 80*C to 280'C at 16*C/min
and held at 280*C for 10 min.
3820 - 1
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Date September 1986
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4.3 Centrifuge; Capable of accommodating 50-mL glass tubes.
4.4 Vials and caps; 2-mL for GC autosampler.
4.5 Volumetric flasks; 10- and 50-mL with ground-glass stopper or
Teflon-lined screw-cap.
4.6 Centrifuge tubes; 50-mL with ground-glass stopper or Teflon-lined
screw-cap.
4.7 Pasteur plpets: Disposable.
4.8 Bottles; Teflon-sealed screw-cap.
5.0 REAGENTS
5.1 Hexadecane and methanol; Pesticide quality or equivalent.
5.2 Reagent water; Reagent water is defined as water in which an
Interference is not observed at the method detection limit of each parameter
of Interest.
5.3 Stock standard solutions (1.00 ug/uL): Stock standard solutions can
be purchased as certifiedsolutions or can be prepared from pure standard
materials.
5.3.1 Prepare stock standard solutions by accurately weighing about
0.0100 grams of pure material. Dissolve the material 1n methanol 1n a
10-mL volumetric flask and dilute to volume (larger volumes may be used
at the convenience of the analyst). If compound purity is certified at
96% or greater, the weight can be used without correction to calculate
the concentration of the stock standard. Commercially available stock
standards may be used if they are certified by the manufacturer.
5.3.2 Transfer the stock standard solutions into Teflon-sealed
screw-cap bottles. Store at 4*C and protect from light. These standards
should be checked frequently for signs of degradation or evaporation.
5.4 Standard mixture II; Standard mixture #1 should contain benzene,
toluene, ethyl benzene, and xylene. Prepare a stock solution containing these
compounds as described In Paragraph 5.3 and then prepare a working standard
(through dilution) in which the concentration of each compound in the standard
1s 100 ng/uL 1n methanol.
5.5 Standard mixture 12; Standard mixture #2 should contain n-nonane
and n-dodecane. Prepare a stock solution containing these compounds as
described 1n Paragraph 5.3. Dilute the stock standard with methanol so that
the concentration of each compound is 100 ng/uL.
3820 - 2
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Date September 1986
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6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Sample preparation;
7.1.1 Water;
7.1.1.1 Allow the contents of the 40-mL sample vial to come to
room temperature. Quickly transfer the contents of the 40-mL vial
to a 50-mL volumetric flask. Immediately add 2.0 mL of hexadecane,
cap the flask, and shake the contents vigorously for 1 min. Let
phases separate. Open the flask and add sufficient reagent water to
bring the hexadecane layer into the neck of the flask.
7.1.1.2 Transfer approximately 1 mL of the hexadecane layer to
a 2.0-mL GC vial. If an emulsion is present after shaking the
sample, break it by:
1. pulling the emulsion through a small plug of Pyrex
glass wool packed in a pipet, or
2. transferring the emulsion to a centrifuge tube and
centrifuging for several min.
7.1.2 Standards;
7.1.2.1 Add 200 uL of the working standard mixtures #1 and #2
to separate 40-mL portions of reagent water. Follow the
instructions in Sections 7.1.1.1 and 7.1.1.2 'with the immediate
addition of 2.0 mL of hexadecane.
7.1.3 Sediment/Soil;
7.1.3.1 Add approximately 10 g of sample (wet weight) to 40 mL
of reagent water in a 50-mL centrifuge tube. Cap and shake
vigorously for 1 min. Centrifuge the sample briefly. Quickly
transfer the supernatant water to a 50-mL volumetric flask.
7.1.3.2 Follow the instructions given in Sections 7.1.1.1 and
7.1.1.2, starting with the addition of 2.0 mL of hexadecane.
7.2 Analysis;
7.2.1 Calibration:
7.2.1.1 External standard calibration; The GC/FID must be
calibrated each 12-hour shiftforhalf of full-scale response when
injecting 1-5 uL of each extracted standard mixture #1 and #2
(Paragraphs 5.4 and 5.5).
3820 - 3
Revision 0
Date September 1986
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7.2.2 GC/FID analysis: Inject the same volume of hexadecane
extract for the sample under Investigation as was used to perform the
external standard calibration. The GC conditions used for the standards
analysis must also be the same as those used to analyze the samples.
7.2.3 Interpretation of the GC/FID chromatograms: There are two
options for interpretation of the GC/FID results.
7.2.3.1 Option A; The standard mixture #1 is used to
calculate an approximate concentration of the aromatics in the
sample. Use this information to determine the proper dilution for
purge-and-trap if the sample is a water. If the sample is a
sediment/soil, use this information to determine which GC/MS purge-
and-trap method (low- or high-level) should be used. If aromatics
are absent from the sample or obscured by higher concentrations of
other purgeables, use Option B.
7.2.3.2 Option B; The response of standard mixture #2 is used
to determine which purge-and-trap method should be used for
analyzing a sample. All purgeables of interest have retention times
less than the n-dodecane retention time. A dilution factor
(Paragraph 7.2.4.1.3) may be calculated for water samples, and an X
factor (Paragraph 7.2.4.2.3) for soil/sediment samples, to determine
whether the low- or high-level purge-and-trap procedure should be
used.
7.2.4 Analytical decision point;
7.2.4.1 Water samples; Compare the hexadecane sample extract
chromatograms against an extracted standard chromatogram.
7.2.4.1.1 If no peaks are noted, analyze a 5-mL water
sample by the purge-and-trap method.
7.2.4.1.2 If peaks are present prior to the n-dodecane
peak and aromatics are distinguishable, follow Option A
(Paragraph 7.2.3.1).
7.2.4.1.3 If peaks are present prior to the n-dodecane
but the aromatics are absent or indistinguishable, Option B
should be used as follows: If all peaks (prior to n-dodecane)
are <3% of the n-nonane, analyze 5 ml of water sample by the
purge-and-trap method. If any peak is >3% of the n-nonane,
measure the area of the major peak and calculate the necessary
dilution factor as follows:
dilution factor = 50 x area of major peak in sample
peak area of n-nonane
The water sample should be diluted using the calculated factor
just prior to purge-and-trap GC or GC/MS analysis.
3820 - 4
Revision 0
Date September 1986
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7.2.4.2 Soil/sediment samples: Compare the hexadecane sample
extract chromatograms against an extracted standard chromatogram.
7.2.4.2.1 If no peaks are noted, analyze a 5-g sample by
the low-level purge-and-trap procedure.
7.2.4.2.2 If peaks are present prior to the n-dodecane
and aromatics are distinguishable, follow Option A using the
concentration information given in Table 1 to determine whether
to analyze the sample by a low- or high-level purge-and-trap
technique.
7.2.4.2.3 If peaks are present prior to n-dodecane but
aromatics are absent or indistinguishable, use Option B.
Calculate an X factor for the sample using the following
equation:
X factor = area of major peak in sample
area of n-nonane
Use the information provided in Table 1 to determine how the
sample should be handled for GC/MS analysis.
7.2.4.2.4 If a high-level method is indicated, the
information provided in Table 2 can be used to determine the
volume of methanol extract to add to 5 ml of reagent water for
analysis (see Methods 5030 and 8240 for methanolic extraction
procedure).
8.0 QUALITY CONTROL
8.1 It is recommended that a reagent blank be analyzed by this screening
procedure to ensure that no laboratory contamination exists. A blank should
be performed for each set of samples undergoing extraction and screening.
9.0 METHOD PERFORMANCE
9.1 No data available.
10.0 REFERENCES
1. U.S. EPA Contract Laboratory Program, Statement of Work for Organic
Analysis, July 1985, Revision.
3820 - 5
Revision 0
Date September 1986
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TABLE 1. DETERMINATION OF GC/MS PURGE-AND-TRAP METHOD
Approximate
X Factor Concentration Range a Analyze by
0-1.0 0-1,000 ug/kg Low-level method
>1.0 >1,000 ug/kg High-level method
a This concentration range 1s based upon the response of aromatics to
GC/FID. The concentration for halomethanes 1s 20x higher, and haloethanes
lOx higher, when comparing GC/FID responses.
TABLE 2. QUANTITY OF METHANOL EXTRACT REQUIRED FOR ANALYSIS OF HIGH-LEVEL
SOIL/SEDIMENTS
Approximate Volume of
X Factor Concentration Range a Methanol Extract b
0.25-5.0 500-10,000 ug/kg 100 ul
0.5-10.0 1,000-20,000 ug/kg 50 ul
2.5-50.0 5,000-100,000 ug/kg 10 ul
12.5-250 25,000-500,000 ug/kg 100 uL of
1/50 dilution c
a Actual concentration ranges could be 10 to 20 times higher than this
1f the compounds are halogenated and the estimates are from GC/FID.
b The volume of methanol added to 5 mL of water being purged should be
100 uL. Therefore 1f the amount of methanol extract required is less than 100
uL, additional methanol should be added to maintain the constant 100-uL
volume.
c Dilute an aliquot of the methanol extract and then take 100 uL for
analysis.
3820 - 6
Revision 0
Date September 1986
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METHOD 3820
HEXAOECANE EXTRACTION AND SCREENING OF PURGEABLE ORGANICS
7. 1
Prepare sample
7.3.1
Calibrate
6C/FIO each
13-hour shift
7.2.3
Perform
GC/FIO analysis
o
3820 - 7
Revision 0
Date September 1986
-------
METHOD 3620
HEXAOECANE EXTRACTION AND SCREENING OF PURGEABLE ORGANICS
(Continued!
7 .2 .A
Compare chroma togr-ams
of nexadecane sample
extract and extracted
standard
Are aromatic
distinguishable?
7.2.4
Compare chromatograms
of hexadecane sample
extract and extracted
standard
7.2.4.2
Use low-level
purge-ana—trap
procedure
Use standard
mixture
-------
METHOD 4010
SCREENING FOR PENTACHLOROPHENOL BY IMMUNOASSAY
1.0 SCOPE AND APPLICATION
1.1 Method 4010 is a procedure for screening solids such as soils,
sludges, and aqueous media such as waste water and leachates for
pentachlorophenol (PCP) (CAS Registry 87-86-5).
1.2 Method 4010 is recommended for screening samples to determine whether
PCP is likely to be present at concentrations above 0.5 mg/Kg for solids or
0.005 mg/L for aqueous samples. Method 4010 provides an estimate for the
concentration of PCP by comparison with a standard.
1.3 Using the test kits from which this method was developed, 95 % of
aqueous samples containing 2 ppb or less of PCPs will produce a negative result
in the 5 ppb test configuration. Also, 95 % of soil samples containing 125 ppb
or less of PCBs will produce a negative result in the 500 ppb test configuration.
1.4 In cases where the exact concentration of PCP is required, additional
techniques (i.e., gas chromatography (Method 8040) or gas chromatography/mass
spectrometry (Method 8270)) should be used.
2.0 SUMMARY OF METHOD
2.1 Test kits are commercially available for this method. The
manufacturer's directions should be followed. In general, the method is
performed using a water sample or an extract of a soil sample. Sample and an
enzyme conjugate reagent are added to immobilized antibody. The enzyme conjugate
"competes" with PCP present in the sample for binding to immobilized anti-PCP
antibody. The test is interpreted by comparing the response produced by testing
a sample to the response produced by testing standard(s) simultaneously.
3.0 INTERFERENCES
3.1 Compounds that are chemically similar may cause a positive test (false
positive) for PCP. The test kit used in preparation of this method was evaluated
for interferences. Table 1 provides the concentration of compounds found to give
a false positive test at the indicated concentration.
3.2 Other compounds have been tested for cross reactivity with PCP, and
have been demonstrated to not interfere with the specific kit tested. Consult
the information provided by the manufacturer of the kit used for additional
information regarding cross reactivity with other compounds.
3.3 Storage and use temperatures may modify the method performance. Follow
the manufacturer's directions for storage and use.
4010-1 Revision 0
August 1993
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4.0 APPARATUS AND MATERIALS
4.1 PENTA RISc Test Kits (EnSys, Inc.), or equivalent. Each commercially
available test kit will supply or specify the apparatus and materials necessary
for successful completion of the test.
5.0 REAGENTS
5.1 Each commercially available test kit will supply or specify the
reagents necessary for successful completion of the test.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this chapter, Organic Analytes,
Section 4.1.
7.0 PROCEDURE
7.1 Follow the manufacturer's instructions for the test kit being used.
Those test kits used must meet or exceed the performance indicated in Tables 2-3.
8.0 QUALITY CONTROL
8.1 Follow the manufacturer's instructions for the test kit being used for
quality control procedures specific to the test kit used. Additionally, guidance
provided in Chapter One should be followed.
8.2 Use of replicate analyses, particularly when results indicate
concentrations near the action level, is recommended to refine information
gathered with the kit.
8.3 Do not use test kits past their expiration date.
8.4 Do not use tubes or reagents designated for use with other kits.
8.5 Use the test kits within their specified storage temperature and
operating temperature limits.
8.6 Method 4010 is intended for field or laboratory use. The appropriate
level of quality assurance should accompany the application of this method to
document data quality.
9.0 METHOD PERFORMANCE
9.1 This method has been applied to a series of groundwater, process
water, and wastewater samples from industries which use PCP, and the results
compared with GC/MS determination of PCP (Method 8270). These results are
provided in Table 2. These results represent determinations by two laboratories.
4010-2 Revision 0
August 1993
-------
9.2 This method has been applied to a series of soils from industries
which use PCP and the results compared with GC/MS determination of PCP via Method
8270. These results are provided in Table 3. These results represent
determinations by two laboratories.
10.0 REFERENCES
1. J.P. Mapes, K.D. McKenzie, L.R. McClelland, S. Movassaghi, R.A. Reddy, R.L.
Allen, and S.B. Friedman, "Rapid, On-Site Screening Test for
Pentachlorophenol in Soil and Water - PENTA-RISc™", Ensys Inc., Research
Triangle Park, NC 27709
2. J.P. Mapes, K.D. McKenzie, L.R. McClelland, S. Movassaghi, R.A. Reddy, R.L.
Allen, and S.B. Friedman, "PENTA-RISc - An On-Site Immunoassay for
Pentachlorophenol in Soil", Bull. Environ. Contam. Toxicol., 49:334-341,
1992.
3. PENTA-RISc™ Instructions for Use, Ensys Inc.
4010-3
Revision 0
August 1993
-------
Table 1
Cross Reactivity for PCPa
Compound
2,6-Dichlorophenol
2,4,6-Trichlorophenol
2,4,5-Trichlorophenol
2,3, 4-Tri chl orophenol
2,3,5,6-Tetrachlorophenol
Tetrachl orohydroqui none
Concentration (mg/Kg)
in Soil to Cause a
False Positive for PCP
at 0.5 mg/Kg
700
16
100
400
1.2
500
Concentration (ng/L)
in Water to Cause a
False Positive for PCP
at 5 /xg/L
600
100
500
600
7
>1500
a for PENTA RISc Test Kit (EnSys, Inc.)
4010-4
Revision 0
August 1993
-------
Table 2
Comparison of Immunoassay* with GC/MS
Water Matrix
Sample Type
groundwater
process water
wastewater
run-oft
Screening Results (ppm)
0.005
>
>
>
>
>
>
>
0.05
<
>
>
>
>
>
<
>
>
>
>
>
<
>
0.1
>
<
>
>
>
<
>
<
>
0.5
<
<
<
<
<
<
1
>
>
>
<
>
<
<
<
<
<
>
<
>
>
>
5
>
>
<
<
<
>
<
3
>
50
<
<
<
Concentration measured
by GC/MS
3.5
0.35
<0.1
8.2
2.8
2.9
0.21
0.17
0.12
0.6
1.4
<0.1
0.17
<0.1
0.034
0.098
O.OS4
0.086
2.1
0.073
0.026
0.006
0.169
0.239
0.190
0.114
0.346
1.1
19
4.3
Does screening test agree with
GC/MS determination?
no
yes
yes
yes
yes
yes
no
yes
yes
no
yes
yes
yes
yes
yes
yes
yes
yes
no
yes
no
yes
no
yes
no
no
yes
yes
yes
yes
> - screening test Indicates that the sample concentration Is greater than the test concentration
< > screening test Indicates that the sample concentration Is less than the test concentration
• for PENTA RISc Test Kit (EnSys, Inc.)
4010-5
Revision 0
August 1993
-------
Table 3
Comparison of Immunoassay* with GC/MS
Soil Matrix
Screening Results (ppm) |
0.5
>
>
<
<
>
>
>
>
>
>
>
>
>
>
>
>
>
>
>
>
>
>
>
<
<
<
>
<
>
>
<
>
>
5
>
>
<
<
>
<
>
>
<
>
<
>
<
>
>
>
>
>
>
>
>
<
>
<
<
<
<
<
<
>
<
>
>
50
>
<
<
<
>
<
<
<
<
>
<
<
<
<
<
<
<
>
>
>
>
<
<
<
<
<
<
<
<
>
<
>
<
Concentration measured by GC/MS
1100
88
0.31
0.72
315
1.5
6.4
9
1.9
46
<1
21
3.3
4
11
18
33
54
65
74
63
1.1
14.3
<1
<1
<1
3.9
<1
1.4
46
<1
142
106
Does screening test agree with
GC/MS determination?
yes
no
yes
no
yes
yes
yes
yes
yes
no
yes
yes
yes
no
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
yes
no
yes
yes
no
4010-6
Revision 0
August 1993
-------
Table 3
Continued
Screening Results (ppm)
0.5
>
>
>
>
>
<
<
>
5
>
>
<
>
>
<
<
<
50
>
>
<
<
>
<
<
<
Concentration measured by GC/MS
117
56
2.5
3.5
143
nd
0.02
5
Does screening test agree with
GC/MS determination?
yes
yes
yes
no
yes
yes
yes
yes
> - screening test Indicates that the sample concentration Is greater than the test concentration
< - screening test Indicates that the sample concentration Is less than the test concentration
• tor PENTA RISc Test Kit (EnSys, Inc.)
4010-7
Revision 0
August 1993
-------
METHOD 8275
THERMAL CHROMATOGRAPHY/MASS SPECTROMETRY (TC/MS) FOR
SCREENING SEMIVOLATILE ORGANIC COMPOUNDS
1.0 SCOPE AND APPLICATION
1.1 Method 8275 is a screening technique that may be used for the
qualitative identification of semivolatile organic compounds in extracts prepared
from nonaqueous solid wastes and soils. It is not intended for use as a rigorous
quantitative method. Direct injection of a sample may be used in limited
applications. The following analytes can be qualitatively determined by this
method:
Compound Name CAS No.a
2-Chlorophenol 95-57-8
4-Methylphenol 106-44-5
2,4-Dichlorophenol 120-83-2
Naphthalene 91-20-3
4-Chloro-3-methylphenol 59-50-7
1-Chloronaphthalene 90-13-1
2,4-Dinitrotoluene 121-14-2
Fluorene 86-73-7
Diphenylamine 122-39-4
Hexachlorobenzene 118-74-1
Dibenzothiophene 132-65-0
Phenanthrene 85-01-8
Carbazole 86-74-8
Aldrin 309-00-2
Pyrene 129-00-0
Benzo(k)fluoranthene 207-08-9
Benzo(a)pyrene 50-32-8
8 Chemical Abstract Services Registry Number.
1.2 Method 8275 can be used to qualitatively identify most neutral,
acidic, and basic organic compounds that can be thermally desorbed from a sample,
and are capable of being eluted without derivatization as sharp peaks from a gas
chromatographic fused-silica capillary column coated with a slightly polar
silicone.
1.3 This method is restricted to use by or under the supervision of
analysts experienced in the use of gas chromatograph/mass spectrometers and
skilled in the interpretation of mass spectra. Each analyst must demonstrate the
ability to generate acceptable results with this method.
8275 - 1 Revision 0
September 1994
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2.0 SUMMARY OF METHOD
2.1 A portion of the sample (0.010-0.100 g) is weighed into a sample
crucible. The crucible is placed in a pyrocell and heated. The compounds
desorbed from the sample are detected using a flame ionization detector (FID).
The FID response is used to calculate the optimal amount of sample needed for
mass spectrometry. A second sample is desorbed and the compounds are condensed
on the head of a fused silica capillary column. The column is heated using a
temperature program, and the effluent from the column is introduced into the mass
spectrometer.
3.0 INTERFERENCES
3.1 Contamination by carryover can occur whenever low-level samples are
analyzed after high-level samples. Whenever an unusually concentrated sample is
encountered, it should be followed by the analysis of an empty (clean) crucible
to check for cross contamination.
4.0 APPARATUS AND MATERIALS
4.1 Thermal Chromatograph (TC) System
4.1.1 Thermal chromatograph™, Ruska Laboratories, or equivalent.
4.1.2 Column - 30 m x 0.25 mm ID (or 0.32 mm ID), 1 urn film
thickness, si 1icone-coated, fused-silica capillary column (J&W Scientific
DB-5 or equivalent).
4.1.3 Flame Ionization detector (FID).
4.2 Mass Spectrometer (MS) system
4.2.1 Mass Spectrometer - Capable of scanning from 35 to 500 amu
every one second or less, using 70 volts (nominal) electron energy in the
electron impact ionization mode.
4.2.2 TC/MS interface - Any GC-to-MS interface producing acceptable
calibration data in the concentration range of interest may be used.
4.2.3 Data System - A computer must be interfaced to the mass
spectrometer. The data system must allow the continuous acquisition and
storage on machine-readable media of all mass spectra obtained throughout
the duration of the chromatographic program. The computer must have
software that can search any GC/MS data file for ions of a specific mass
(or group of masses) and that can plot such ion abundances versus time or
scan number. This type of plot is defined as an extracted ion
chromatogram (EIC). Software must also be available that allows for
integration of the abundances in, and EIC between, specified time or scan-
number 1imits.
8275 - 2 Revision 0
September 1994
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4.3 Tools and equipment
4.3.1 Fused quartz spatula.
4.3.2 Fused quartz incinerator ladle.
4.3.3 Metal forceps for sample crucible.
4.3.4 Sample crucible storage dishes.
4.3.5 Porous fused quartz sample crucibles with lids.
4.3.6 Sample crucible cleaning incinerator.
4.3.7 Cooling rack.
4.3.8 Microbalance, 1 g capacity, 0.000001 g sensitivity, Mettler
Model M-3 or equivalent.
4.4 Vials - 10 mL, glass with Teflon lined screw-caps or crimp tops.
4.5 Volumetric flasks, Class A - 10 ml to 1000 ml.
5.0 REAGENTS
5.1 Reagent grade inorganic chemicals shall be used in all tests. Unless
otherwise indicated, it is intended that all reagents shall conform to the
specifications of the Committee on Analytical Reagents of the American Chemical
Society, where such specifications are available.
5.2 Solvents
5.2.1 Methanol, CH3OH - Pesticide grade or equivalent.
5.2.2 Acetone, CH3COCH3 - Pesticide grade or equivalent.
5.2.3 Toluene, C6H5CH3 - Pesticide grade or equivalent.
5.2.4 Methylene chloride, CH2C12 - Pesticide grade or equivalent.
5.2.5 Carbon disulfide, CS2 - Pesticide grade or equivalent.
5.2.6 Hexane, C6H14 - Pesticide grade or equivalent.
5.2.7 Other suitable solvents - Pesticide grade or equivalent.
5.3 Stock Standard solutions - Standard solutions may be prepared from
pure standard materials or purchased as certified solutions.
5.3.1 Prepare stock standard solutions by weighing about 0.01 g of
pure material. Dissolve the material in pesticide quality acetone, or
8275 - 3 Revision 0
September 1994
-------
other suitable solvent, and dilute to 10 ml in a volumetric flask. Larger
volumes may be used at the convenience of the analyst.
5.3.2 Transfer the stock standard solutions into glass vials with
Teflon lined screw-caps or crimp tops. Store at -10°C to -20°C or less
and protect from light. Stock standard solutions should be checked
frequently for signs of degradation or evaporation, especially prior to
use in preparation of calibration standards.
5.3.3 Stock standard solutions must be replaced after 1 year, or
sooner if comparison with quality control check samples indicates a
problem.
5.4 Internal Standard solutions - The internal standards recommended are
l,4-dichlorobenzene-d4, naphthalene-d8, acenaphthene-d10, phenanthrene-d10,
chrysene-d12> and perylene-d12. Other compounds may be used as internal standards
as long as the requirements given in Sec. 7 are met. Dissolve about 0.200 g of
each compound with a small volume of carbon disulfide. Transfer to a 50 ml
volumetric flask and dilute to volume with methylene chloride, so that the final
solvent is approximately 20/80 (V/V) carbon disulfide/methylene chloride. Most
of the compounds are also soluble in small volumes of methanol, acetone, or
toluene, except for perylene-d12. Prior to each analysis, deposit about 10 juL
of the internal standard onto the sample in the crucible. Store internal
standard solutions at 4°C or less before, and between, use.
5.5 Calibration standards - Prepare calibration standards within the
working range of the TC/MS system. Each standard should contain each analyte or
interest (e.g. some or all of the compounds listed in Sec. 1.1 may be included).
Each aliquot of calibration standard should be spiked with internal standards
prior to analysis. Stock solutions should be stored at -10°C to -20°C and should
be freshly prepared once a year, or sooner if check standards indicate a problem.
The daily calibration standard should be prepared weekly, and stored at 4°C.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 See the introductory material to this Chapter, Organic Analytes,
Sec. 4.1.
7.0 PROCEDURE
7.1 Crucible Preparation
7.1.1 Turn on the incinerator and let it heat for at least 10
minutes. The bore of the incinerator should be glowing red.
7.1.2 Load the sample crucible and lid into the incinerator ladle
and insert into the incinerator bore. Leave in the incinerator for 5
minutes, then remove and place on the cooling rack.
7.1.3 Allow the crucibles and lids to cool for five minutes before
placing them in the storage dishes.
8275 - 4 Revision 0
September 1994
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CAUTION: Do not touch the crucibles with your fingers. This can
result in a serious burn, as well as contamination of
the crucible. Always handle the sample crucibles and
lids with forceps and tools specified.
7.1.4 All sample crucibles and lids required for the number of
analyses planned should be cleaned and placed in the storage dishes ready
for use.
7.2 Sample Preparation and Loading
7.2.1 The analyst should take care in selecting a sample for
analysis, since the sample size is generally limited to 0.100 g or less.
This implies that the sample should be mixed as thoroughly as possible
before taking an aliquot. Because the sample size is limited, the analyst
may wish to analyze several aliquots for determination.
7.2.2 The sample should be mixed or ground such that a 0.010 to
0.100 g aliquot can be removed. Remove one sample crucible from the
storage dish and place it on the microbalance. Establish the tare weight.
Remove the sample crucible from the balance with the forceps and place it
on a clean surface.
7.2.3 Load an amount of sample into the sample crucible using the
fused quartz spatula. Place the assembly on the microbalance and
determine the weight of the sample. For severely contaminated samples,
less than 0.010 g will suffice, while 0.050-0.100 g is needed for low
concentrations of contaminants. Place the crucible lid on the crucible;
the sample is now ready for analysis.
7.3 FID Analysis
7.3.1 Load the sample into the TC. Hold the sample at 30°C for 2
minutes followed by linear temperature programmed heating to 260°C at
30°C/minute. Follow the temperature program with an isothermal heating
period of 10 minutes at 260°C, followed by cooling back to 30°C. The total
analysis cycle time is 24.2 minutes
7.3.2 Monitor the FID response in real time during analysis, and
note the highest response in millivolts (mV). Use this information to
determine the proper weight of sample needed for combined thermal
extraction/gas chromatography/mass spectrometry.
7.4 Thermal Extraction/GC/MS
7.4.1 Prepare a calibration curve using a clean crucible and lid by
spiking the compounds of interest at five concentrations into the crucible
and applying the internal standards to the crucible lid. Analyze these
standards and establish response factors at different concentrations.
7.4.2 Weigh out the amount of fresh sample that will provide
approximately 1000 to 3000 mv response. For example, if 0.010 g of sample
gives an FID response of 500 mv, then 0.020 to 0.060 g (0.040 g ± 50 %)
8275 - 5 Revision 0
September 1994
-------
should be used. If 0.100 g gives 8000 mv, then 0.025 g ± 50 % should be
used.
7.4.3 After weighing out the sample into the crucible, deposit the
internal standards (10 juL) onto the sample. Load the crucible into the
pyrocell, using the same temperature program in Sec. 7.3.1. Hold the
capillary at 5°C during this time to focus the released semivolatiles (the
intermediate trap is held at 330°C to pass all compounds onto the column).
Maintain the splitter zone at 310°C, and the GC/MS transfer line at 285°C.
After the isothermal heating period is complete, temperature program the
column from 5°C to 285°C at 10°C/minute and hold at 285°C for 5 minutes.
Acquire data during the entire run time.
7.4.4 If the response for any quantitation ion exceeds the initial
calibration curve range of the TC/MS system, a smaller sample should be
analyzed.
7.5 Data Interpretation
7.5.1 Qualitative Analysis
7.5.1.1 The qualitative identification of compounds
determined by this method is based on retention time, and on
comparison of the sample mass spectrum, after background correction,
with characteristic ions in a reference mass spectrum. The
reference mass spectrum must be generated by the laboratory using
the conditions of this method. The characteristic ions from the
reference mass spectrum are defined to be the three ions of greatest
relative intensity, or any ions over 30% relative intensity if less
than three such ions occur in the reference spectrum. Compounds
should be identified as present when the criteria below are met.
7.5.1.1.1 The intensities of the characteristic ions
of a compound maximize in the same scan or within one scan of
each other. Selection of a peak by a data system target
compound search routine where the search is based on the
presence of a target chromatographic peak containing ions
specific for the target compound at a compound-specific
retention time will be accepted as meeting this criterion.
7.5.1.1.2 The RRT of the sample component is within
± 0.06 RRT units of the RRT of the standard component.
7.5.1.1.3 The relative intensities of the
characteristic ions agree within 30% of the relative
intensities of these ions in the reference spectrum.
(Example: For an ion with an abundance of 50% in the
reference spectrum, the corresponding abundance in a sample
spectrum can range between 20% and 80%.)
7.5.1.1.4 Structural isomers that produce very similar
mass spectra should be identified as individual isomers if
they have sufficiently different GC retention times.
8275 - 6 Revision 0
September 1994
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Sufficient GC resolution is achieved if the height of the
valley between two isomer peaks is less than 25% of the sum of
the two peak heights. Otherwise, structural isomers are
identified as isomeric pairs.
7.5.1.1.5 Identification is hampered when sample
components are not resolved chromatographically and produce
mass spectra containing ions contributing by more than one
analyte. When gas chromatographic peaks obviously represent
more than one sample component (i.e., a broadened peak with
shoulder(s) or a valley between two or more maxima),
appropriate selection of analyte spectra and background
spectra is important. Examination of extracted ion current
profiles of appropriate ions can aid in the selection of
spectra, and in qualitative identification of compounds. When
analytes coelute (i.e., only one chromatographic peak is
apparent), the identification criteria can be met, but each
analyte spectrum will contain extraneous ions contributed by
the coeluting compound.
7.5.1.2 For samples containing components not associated
with the calibration standards, a library search may be made for the
purpose of tentative identification. The necessity to perform this
type of identification will be determined by the purpose of the
analyses being conducted. Computer generated library search
routines should not use normalization routines that would
misrepresent the library or unknown spectra when compared to each
other. For example, the RCRA permit or waste delisting requirements
may require the reporting of non-target analytes. Only after visual
comparison of sample spectra with the nearest library searches will
the mass spectral interpretation specialist assign a tentative
identification. Guidelines for making tentative identification are:
(1) Relative intensities of major ions in the reference
spectrum (ions > 10% of the most abundant ion) should be present in
the sample spectrum.
(2) The relative intensities of the major ions should agree
within ± 20%. (Example: For an ion with an abundance of 50% in the
standard spectrum, the corresponding sample ion abundance must be
within 30 and 70%).
(3) Molecular ions present in the reference spectrum should
be present in the sample spectrum.
(4) Ions present in the sample spectrum but not in the
reference spectrum should be reviewed for possible background
contamination or presence of coeluting compounds.
(5) Ions present in the reference spectrum but not in the
sample spectrum should be reviewed for possible subtraction from the
sample spectrum because of background contamination or coeluting.
Data system library reduction programs can sometimes create these
discrepancies.
8275 - 7 Revision 0
September 1994
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8.0 QUALITY CONTROL
8.1 Refer to Chapter One and Method 8000 for specific quality control
procedures.
9.0 METHOD PERFORMANCE
9.1 Table 1 presents method performance data, generated using spiked soil
samples. Method performance data in an aqueous matrix are not available.
10.0 REFERENCES
1. Zumberge, J.E., C. Sutton, R.D. Worden, T. Junk, T.R. Irvin, C.B. Henry,
V. Shirley, and E.B. Overton, "Determination of Semi-Volatile Organic
Pollutants in Soils by Thermal Chromatography-Mass Spectrometry (TC/MS):
an Assessment for Field Analysis," in preparation.
8275 - 8 Revision 0
September 1994
-------
TABLE 1
METHOD PERFORMANCE, SOIL MATRIX
Analyte
2-Chlorophenol
4-Methyl phenol
2,4-Dichlorophenol
Naphthalene
4-Chloro-3-methyl -phenol
1-Chloronaphthalene
2,4-Dinitrotoluene
Fluorene
Diphenylamine
Hexachlorobenzene
Dibenzothiophene
Phenanthrene
Carbazole
Aldrin
Pyrene
Benzo ( k) f 1 uoranthene
Benzo(a)pyrene
Average
Clay
30
10
23
77
9
96
7
9
5
68
20
11
4
3
7
4
4
% Recovery"
Silt
22
77
20
120
12
103
10
25
6
64
35
31
8
19
19
9
8
Subsoil
2
7
26
63
9
70
10
19
6
80
50
40
9
15
20
11
11
Mean
Recovery
18
31
23
87
10
90
9
18
6
71
35
24
7
12
15
8
8
Percent theoretical recovery based upon linearity of injections deposited on
the crucible lid (slope and y-intercept). Average of 9 replicates (-10 mg
soil spiked with 50 ppm of analyte); 3 different instruments at 3 different
laboratories.
8275 - 9 Revision 0
September 1994
-------
TABLE 2
CHARACTERISTIC IONS FOR SEMIVOLATILE COMPOUNDS
Compound
Primary
Ion
Secondary
Ion(s)
2-Chlorophenol
4-Methylphenol
2,4-Dichlorophenol
Naphthalene
4-Chloro-3-methyl-phenol
1-Chloronaphthalene
2,4-Dinitrotoluene
Fluorene
Diphenylamine
Hexachlorobenzene
Phenanthrene
Aldrin
Pyrene
Benzo(k)f1uoranthene
Benzo(a)pyrene
128
107
162
128
107
162
165
166
169
284
178
66
202
252
252
64,130
107,108,77,79,90
164,98
129,127
144,142
127,164
63,89
165,167
168,167
142,249
179,176
263,220
200,203
253,125
253,125
8275 - 10
Revision 0
September 1994
-------
METHOD 8275
THERMAL CHROMATOGRAPHY/MASS SPECTROMETRY (TC/MS) FOR
SCREENING SEMIVOLATILE ORGANIC COMPOUNDS
Start
7.1 Prepare
crucible
^
r
7.2.2
Establish
tare weight
of crucible.
1
f
7.2.3 Place
sample in
crucible; establish
weight.
^
r
7.3.1 FID
Analysis using
linear temp.
programmed
heating.
\r
7.3.2 Using
FID response,
determine
sample weight
for TE/GC/MS.
>fc
•
7.4.1 f
cahbr
cur
>
repare
ve.
f
7.4.2 Prepare
amount of
sample for
appropriate
FID response.
^
r
7.4.3 Weigh
sample into
crucible; use
temp, program
in Sec. 7.3.1 .
/7.4.4\
/ Is N.
/ quantitation N^
f ion > initial
N. calib. curve .
N. range of /
XTC/MS?/
] No
I
7.5.1
Qualitative
Identification.
1
*
.Yes
r
7.4.4 Use
smaller
sample.
Stop
8275 - 11
Revision 0
September 1994
-------
APPENDIX
COMPANY REFERENCES
The following listing of frequently-used addresses is provided for the
convenience of users of this manual. No endorsement is intended or implied.
Ace Glass Company
1342 N.W. Boulevard
P.O. Box 688
Vineland, NJ 08360
(609) 692-3333
Aldrlch Chemical Company
Department T
P.O. Box 355
Milwaukee, WI 53201
Alpha Products
5570 - T W. 70th Place
Chicago, IL 60638
(312) 586-9810
Barneby and Cheney Company
E. 8th Avenue and N. Cassidy Street
P.O. Box 2526
Columbus, OH 43219
(614) 258-9501
Bio - Rad Laboratories
2200 Wright Avenue
Richmond, CA 94804
(415) 234-4130
Burdick & Jackson Lab Inc.
1953 S. Harvey Street
Muskegon, MO 49442
Calgon Corporation
P.O. Box 717
Pittsburgh, PA 15230
(412) 777-8000
Conostan Division
Conoco Speciality Products, Inc.
P.O. Box 1267
Ponca City, OK 74601
(405) 767-3456
COMPANIES - 1
Revision 0
Date September 1986
-------
Corning Glass Works
Houghton Park
Corning, NY 14830
(315) 974-9000
Dohrmann, Division of Xertex Corporation
3240 - T Scott Boulevard
Santa Clara, CA 95050
(408) 727-6000
(800) 538-7708
E. M. Laboratories, Inc.
500 Executive Boulevard
Elmsford, NY 10523
Fisher Scientific Co.
203 Fisher Building
Pittsburgh, PA 15219
(412) 562-8300
General Electric Corporation
3135 Easton Turnpike
Fairfield, CT 06431
(203) 373-2211
Graham Manufactory Co., Inc.
20 Florence Avenue
Batavia, NY 14020
(716) 343-2216
Hamilton Industries
1316 18th Street
Two Rivers, WI 54241
(414) 793-1121
ICN Life Sciences Group
3300 Hyland Avenue
Costa Mesa, CA 92626
Johns - Manville Corporation
P.O. Box 5108
Denver, CO 80217
Kontes Glass Company
8000 Spruce Street
Vineland, NJ 08360
Milllpore Corporation
80 Ashby Road
Bedford, MA 01730
(617) 275-9200
(800) 225-1380
COMPANIES - 2
Revision
Date September 1986
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National Bureau of Standards
U.S. Department of Commerce
Washington, DC 20234
(202) 921-1000
Pierce Chemical Company
Box 117
Rockford, IL 61105
(815) 968-0747
Scientific Glass and Instrument, Inc.
7246 - T Wynnwood
P.O. Box 6
Houston, TX 77001
(713) 868-1481
Scientific Products Company
1430 Waukegon Road
McGaw Park, IL 60085
(312) 689-8410
Spex Industries
3880 - T and Park Avenue
Edison, NJ 08820
Waters Associates
34 - T Maple Street
Mil ford, MA 01757
(617) 478-2000
(800) 252-4752
Whatman Laboratory Products, Inc.
Clifton, NJ 07015
(201) 773-5800
COMPANIES - 3 ' • -
'"'" Revision 0
* •"• Date September 1986
&U.S. GOVERNMENT PRINTING OFFICE : 1987 O - 169-932
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