SW8463A
TEST METHODS FOR EVALUATING
SOLID WASTE, PHYSICAL/CHEMICAL
METHODS, SW-846, 3RD EDITION,
PROPOSED UPDATE II
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TABLE OF CONTENTS
VOLUME ONE
SECTION A
DISCLAIMER
ABSTRACT
TABLE OF CONTENTS
METHOD INDEX AND CONVERSION TABLE
PREFACE
ACKNOWLEDGEMENTS
CHAPTER ONE -- QUALITY CONTROL f;
• .•»«
1.0 Introduction •
2.0 QA Project Plan
3.0 Field Operations : .
4.0 Laboratory Operations
5.0 Definitions
6.0 References
CHAPTER TWO - CHOOSING THE CORRECT PROCEDURE
- M
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: Add Digestion of Waters for Total Recoverable or
Dissolved Metals for Analysis by Flame Atomic Absorption
(FAA) or Inductively Coupled Plasma {ICP) Spectroscopy
Method 3010A: Acid Digestion of Aqueous Samples and Extracts for Total
Metals for Analysis by Flame Atomic Absorption (FAA) or
Inductively Coupled Plasma (ICP) Spectroscopy
Method 3015: Microwave Assisted Acid Digestion of Aqueous Samples and
Extracts
CONTENTS - 1 Revision 2
November 1992
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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 (GFAA) Spectroscopy
Dissolution Procedure for 011s, Greases, or Waxes
Acid Digestion of Sediments, Sludges, and Soils
Microwave Assisted Add Digestion of Sediments, Sludges,
Soils, and Oils
3.3 Methods for Determination of Metals
Method 6010A: Inductively Coupled Plasma-Atomic Emission Spectres copy
Method 6020: Inductively Coupled Plasma - Mass Spectrometry '
Method 7000A: Atomic Absorption Methods
Method 7020: Aluminum (AA, Direct Aspiration)
Method 7040: Antimony (AA, Direct Aspiration) ,
Method 7041: Antimony (AA, Furnace Technique) '*
Method 7060A: Arsenic (AA, furnace Technique)
Method 7Q61A; ^Arsenic ^AA,.Gaseous Hydride)
Method 7062 r XA^timony'and Ahsenic (AA, Gaseous Borohydride)
Method 7080A: Barium (AA, Direct Aspiration)
Method 7081: Barium (AA, Furnace Technique)
Method 7090: Beryllium (AA, Direct Aspiration)
Method 7091: Beryllium (AA, Furnace Technique) :
Method 7130: Cadmium (AA, Direct Aspiration)
Method 7131A: Cadmium (AA, Furnace Technique)
Method 7140: Calcium (AA, Direct Aspiration)
Method 7190: Chromium (AA, Direct Aspiration) "
Method 7191: Chromium (AA, Furnace Technique)
Method 7195: Chromium, Hexavalent (Coprecipitation)
Method 7196A: Chromium, Hexavalent (Colorimetric)
Method 7197: Chromium, Hexavalent (Chelation/Extractlon)
Method 7198: Chromium, Hexavalent (Differential Pulse Polarography)
Method 7200: Cobalt (AA, Direct Aspiration)
Method 7201: Cobalt (AA, Furnace Technique)
Method 7210: Copper (AA, Direct Aspiration)
Method 7211: Copper (AA, Furnace Technique)
Method 7380: Iron (AA, Direct Aspiration)
Method 7381: Iron (AA, Furnace Technique)
Method 7420: Lead (AA, Direct Aspiration)
Method 7421: Lead (AA, Furnace Technique)
Method 7430: Lithium (AA, Direct Aspiration)
Method 7450: Magnesium (AA, Direct Aspiration)
Method 7460: Manganese (AA, Direct Aspiration)
Method 7461: Ifengail^se fAA, Furnace Technique)
Method 7470A: Mercury in Liquid Waste (Manual Cold-Vapor Technique)
» Method 7471A: Mercury in Solid or Semi solid Waste (Manual Cold-Vapor
•'*'• '•' Technique)
Method 7480: !- Molybdenum (AA, Direct Aspiration)
•». Method 7481: Molybdenum'(AA, Furnace Technique)
Method 7520: 'Nickel fAA, Direct Aspiration)
Method 7550: Osmium (AA, Direct Aspiration)
Method 7610: Potassium (AA, Direct Aspiration)
Method 7740: Selenium (AA, Furnace Technique)
'' 38W •
CONTENTS - 2
Revision 2
November 1992
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Method 7741A: Selenium (AA, Gaseous Hydride)
Method 7742: Selenium (AA, Gaseous Borohydrlde)
Method 7760A: Silver (AA, Direct Aspiration)
Method 7761: Silver (AA, Furnace Technique)
Method 7770: Sodium (AA, Direct Aspiration)
Method 7780: Strontium (AA, Direct Aspiration) r-,*Tft»v:
Method 7840: Thallium (AA, Direct Aspiration) -v-f "-"^
Method 7841: Thallium (AA, Furnace Technique)
Method 7870: Tin (AA, Direct Aspiration) - ' W4 '•'• «H"
Method 7910: Vanadium (AA, Direct Aspiration) J '-*"' ! -^
Method 7911: Vanadium (AA, Furnace Technique) >-,-<,-,
Method 7950: Zinc (AA, Direct Aspiration) ~~~~
Method 7951: Zinc (AA, Furnace Technique) 03'*,' w 3 $5 |-
APPENDIX -- COMPANY REFERENCES .f.
•'•' • i i i inn i • ii i -linn — I"' , t _ ^ ^ \- i
' 1 j . j. '. H''~ S
•:-'.<*''.*-.\' - - o.e
rf ;« . a
J
CONTENTS - 3 Revision 2
November 1992
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SECTION B
pi§c
ABST
TABL
LAMER
lAfr
: o
METHOD
c CONTENTS
NDEX 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 General 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 3520Br 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: Wide-bore Capillary
Column Technique
Method 51001 Determination of the Volatile Organic Concentration of
Waste Samples
Method 5110: "Determination of Organic Phase Vapor Pressure 1n Waste
Samples
4.2.2 Cleanup
Method 3600B: Cleanup
Method 3610A: Alumina Column Cleanup
CONTENTS - 4 Revision 2
November 1992
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Method 3611A:
Method
Method
Method
Method
Method
Method
3620A:
3630B:
3640A:
3650A:
3660A:
3665:
Alumina Column -Cleanup and ^
Petroleum Wastes -=i«it?c#i* •<' -
Flor1s11 Column Cleanup
Silica Gel Cleanup^ '
Gel-Permeation Cleanup
Acid-Base Partition Cleanup
Sulfur Cleanup *
Sulfurlc Add/Permanganate Cleanup
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 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 . • • -*o!«! -j t '>'''*
Halogenated Volatile Organic* by Gas Chromatography
1,2-Dlbromoethane and l,2-Mbromo-3-chloropropane by
Mlcroextraction and Gas Chromatography . ,
Nonhalogenated Volatile Organics^Gas ChroKtography
Aromatic Volatile Organ1cs by Gas Chromatography .
Halogenated Volatlles by Gas ChrttoUgĄaphy< Using
Photo1on1zation and Electrolytic Cond^dtfVnyDetectors
In Series: Capillary Column Technique .
Acroleln and AcrylonltHle by Gas diroiatbgraphy
Acrylonltrlle by Gasrr€nroaatography
Aery 1 amide by Gas Chrofflttograjihy
Phenols by Gas Chromatography
Phthalate Esters 'wfyitsH^
Phthalate Esters by Capillary Gas Chromatography with
Electron Capture Detection {fit/ECD) •-[&* ?-< *
Nltrosamlnes by Gas Chromatography
Organochlorlne Pesticides and^olychlorinated Blphenyls
by Gas Chromatographyv-tt.-n • ^etS f 1 -,s
Organochlorlne Pesticides, Halowaxes and PCBs as
Aroclors by Gas Chro«atography: Capillary Column
Technique ••, ? -; >">%$**n& ;tL .
Nitroaromatics and Cyclic Ketones
Polynuclear Aromatic Hydrocarbons < * '•<• -
Haloethers by Gas Chromatography
Chlorinated Hydrocarbons by Gas Chromatdgraphy
Chlorinated Hydrocarbons 'by Gas Chromatography:
Capillary Column Technique -
Organophosphorus Pesticides
Organophosphorus Compounds by Ga* *thromatograpl>yi
Capillary Column Technique ,
Chlorinated Herbicides ty^sChr()«atWap>y'
Chlorinated Herbicides by flC UsInf^Meth/latlon or
PentafluorobenzylationOer1vatizat1on: ^Capillary Column
Technique 5 '&*-''-'"T
4.3.2 Gas Chromatographic/Mass Spectroscoplc Me
Method 8240B:
Volatile Organlcs
Spectrometry (GC/MS)
by Gas Chromatography/Mass
CONTENTS - 5
Revision 2
November 1992
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Method 8250A: -., Sem1volat1l«^ Organic , Compounds by fits
Chromatography/Mass* Speetrometry (GC/MS)
Method 8260A: Volatile Organic Compounds by Gas CblrWnatograptiy/Hass
Spectrometry (GC/MS): Capillary Column Technique
Method 8270B: Semivolatile Organic Compounds by Gas
Chromatography/Mass Spectrometry (GC/MS): Capillary
Column Technique " , 4f vt. ^
Method 8280: The Analysis of Polychlorinated D1benzo-p-D1ox1ns and
Polychlorinated Dlbenzofurans
Appendix A: Signal -to-Noise Determination Methods +>. \i
Appendix B: Recommended Safety and Handling Procedures for
PCDOs/PCDFs : *.• < . • , ?«.- . • :• u • - i . c . *
Method 8290: Polychlorinated Oibenzodioxins (PCDDs) and
Polychlorinated Dlbenzofurans (PCDFs) by High-Resolution
•i Gas Chromategraphy/H1gh- Resolution Mass Spectrometry
4.3.3 High Performance Liquid Chroma tog raphlc Hethods ; -: -,>^
.:'>->' , . ; -AOSOfi trt*-;.-'^
Method 8310: Polynuclear Aromatic Hydrocarbons rAJSO? ?*oito»*«
Method 831S: Determination of Carbonyl Compounds by High Performance
Liquid Chromatography (HPLC) .
Appendix A: j Recry stall ization of 2,4-D1nitrophenylhydraz1ne
Method 8316: Acrylamide, Acrylonitrile and Acroleln by High
Performance Liquid Chromatography (HPLC): b^':^
Method 8318: N-Methylcarbamates by High Performance Liquid
Chromatography (HPLC) -i •;<#. »«>"*&*.
Method 8321: Solvent Extractable Non-Volatile Compounds by High
Performance Liquid Chromatography/Thermospi*ay/Mass
Spectrometry (HPLC/TSP/MS) or Ultraviolet (UV) Detection
Method 8330: Nitroaromatics and Nitramines by High Performance Liquid
Chromatography (HPLC) , ,'-08 o itsff
Method 8331: Tetrazene by Reverse Phase High Performance Liquid
Chromatography (HPLC)
4.3.4 Fourier Transform infrared Methods ; .»OCP &•«!»«
Method 8410: Gas Chromatography/Fourier Transform Infrared {flC/FT- IR)
Spectrometry> for Semivolatile Organic*:?x Capillary
Column
- •] .^if
4.4 Mi seel 1 aneous Screen 1 ng Methods ; p ~. */1 * 5 • •
Method 3810: - Headspace -. L-.:,;- ., " iV&te hodJsH
Method 3820: Hexadecane Extraction and Screening f of Purge able
Organics - - j -,
Method 8275: Thermal Chromatography/Mass Spectrometry (TC/MS) for
Screening Semivolatile Organic Compounds
APPENDIX - COMPANY REFERENCES
CONTENTS - 6 Revision 2
November 1992
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SECTION C
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 Ml
Method 5050:
Method 9010A:
Method 9012:
Method 9013:
Method 9020B:
Method 9021:
Method 9022:
Method 9030A:
Method 9031:
Method 9035:
Method 9036:
Method 9038:
Method 9056:
Method 9060:
Method 9065:
Method 9066:
Method 9067:
Method 9070:
Method 9071A:
Method 9075:
Method 9076:
• jt>
I'.r.'i
Hq
Bomb Combustion Method for Solid Haste
Total and Amenable Cyanide (Col Diametric, Manual)
Total and Amenable Cyanide (Col or inetric, Automated UV)
Cyanide Extraction Procedure for solids and 011s
Total Orglnlc Hal Jdes itOX) : ,r „.
Purgeable Oraanfc Hflldes (POXJ- "I
Total Organic Hal ides (TOX) by Neutron Activation
Analysis
Sul fides - - -, -
Extractable SulfWes ' l
Sulfate (Colorimetric, Automated, Chloranilate)
Sul fate (Colorimetric, Automated, Methylthymol Blue, AA
H) . r f
Sulfate (Turbid1metr1c) .; , ;
An ion Chromatography Method T._ •
Total Organic Carbon
Phenol 1 cs (Spectrophotooptric, Manual , 4-AAP with
Distillation)^ . , , ,r n
Phenollcs "e1Co1 or 1 metric, Automated 4-AAP with
4Ht ,r^H9> -. - . > >\
Phenol ics (Spectrophotometric, MBTH with Distillation)
Total Recoverable 011 & Grease (Gravimetric, Separatory
Funnel Extraction)
Oil and Grease Extract jon Method for Sludge and Sediment
Samples ' .-i^.j. . -..-...-_ *. ... _ ...... .•
Test Method for Total Chlorine In New and Used Petroleum
Products by X-Ray Fluorescence Spectrometry (XRF)
Test .Method for Total CMorine in New and Used Petroleum
Products by Oxidative Combustion and Microcoulometry
"
CONTENTS •;;?
Revision 2
November 1992
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Method 9077:
Method
Method
Method
Method
Method
Method
Method
Method
9131:
9132:
9200A:
9250:
9251:
9252A:
9253:
9320:
CHAPTER SIX - PROPERTIES
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
Method
1312:
1320:
1330A:
9040A:
9041A:
90458:
9050:
9080:
9081:
9090A:
9095:
9096:
9100:
Method 9310:
Method 9315:
Test Method^ for Total Chlorine in New and Used
Petroled Products (Field Test Kit Methods)
Total CoHform: Multiple Tube Fermentation Technique
Total CoHform: Membrane Filter Technique
Nitrate
Chloride (Color1metric, Automated Ferrlcyanide AAI)
Chloride (ColorlmetHc, Automated Ferrl cyanide AAI I)
Chloride (T1tr1»etr1c, Mercuric Nitrate) ,
Chloride (Tltriaetric, Silver Nitrate)i J
Radlun-228
Synthetic Precipitation Leaching Procedure ,
Multiple Extraction Procedure ;: " I;, (
Extraction Procedure for 01 if Wastes^; *".
pH Electrometric Measuremenf ;.* ^ J } ;
pH Paper Method - -
Soil and Haste pH '
Specific Conductance 9 v9 u.«.
Cation-Exchange Capacity of Soils (Ammonium Acetate)
Cation-Exchange Capacity of Soils (Sodium Acetate)
Conductivity, and intrinsic Permeability
Gross Alpha and Gross Beta x>
Alpha-Emitting Radium Isotopes
CHAPTER SEVEN - INTRODUCTION AND REGULATORY DEFJNiTIONS
7.1 Ignltablllty
7.2 Corrosivity
7.3- Reactivity
Test Method to Determine Hydrogen "Cyanide .Released from Hastes
Test Method to Determine Hydrogen Sulflde Released from Wastes
7.4 Toxicity Characteristic Leaching Procedure
CHAPTER EIGHT - METHODS FOR DETERMINING CHARACTERISTICS *
8.1 Ignltablllty °rd V^°
Method 1010: PensKy-Martens Closed-Cup Method for Determining
Method 1020A: Setaflash Closed-Cup Method for Determining Ign1tab11 Ity
CONTENTS - 8
Revision 2
November 1992
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8.2 Corroslvlty
Method 1110: Corroslvlty Toward Steel
8.3 Reactivity
8.4 Toxlclty
Method 1310A: Extraction Procedure (EP) Toxlclty Test Method and
Structural Integrity Test
Method 1311: Toxlclty Characteristic Leaching Procedure
APPENDIX -- COMPANY REFERENCES
CONTENTS - 9 Revision Z
November 1992
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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 Analyst
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 - 10 Revision
November 19?
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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
Revision 2 methods (methods which have been revised 1n the proposed
Update II package) are designated by the letter "B" In the method
number. Likewise, Revision 1 methods (methods which were previously
revised In the promulgated Update I package or are being revised for
the first time 1n the proposed Update II package) are designated by
the letter "A" In the method number. In order to properly document
the method revision used, the entire method number, Including the
letter designation, must be Identified by the method user. A method
reference found within the text of SU-846 methods and chapters refers
to the latest promulgated revision of the method, even though the
referenced method number 1s without an appropriate letter designation.
CONTENTS - 11 Revision 2
November 1992
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METHOD 8081
ORGANOCHLORINE PESTICIDES. HALOWAXES 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, Halowaxes 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,
Alachlora'b
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
;0-BHCa'b
Y-BHC (Lindane)a>b
6-BHCa'b
Captafolb
Captanb
Chi orobenzi late
a-Chlordaneb
Y~Chlordanea'b
Chloroneb6
Chloropropylateb
Chlorothalonir
DBCPb
DCPAb
4,4'-DDDa/b
15972-60-8
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
2425-06-1
133-06-2
510-15-6
5103-71-9
5103-74-2
2675-77-6
99516-95-7
1897-45-6
96-12-8
1861-32-1
72-54-8
8081 - 1
Revision 0
November 1992
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Compound Name
4,4'-DDEa'b
4,4'-DDTa'b
Dial! ate6
Dichloneb
Dicofolb
Dieldrina'b
Endosulfan Ia'b
Endosulfan Ha'b
Endosulfan sulfatea/b
Endrina'b
Endrin aldehydea/b
Endrin ketoneb
Etridiazoleb
Halowax-1000b
Halowax-1001b
Halowax-1013b
Halowax-1014b
Halowax-1051b
Halowax-1099b
Heptachlora'b
Heptachlor epoxide3'6
Hexachl orobenzeneb
Hexachl orocycl opentadi eneb
Isodrinb
Kepone
Methoxychlor9'6
Mirexb
Nitrofenb
PCNBb
Perthane
Propachlorb
Strobaneb
Toxaphene3'
trans-Nonachlor
tra/7S-Permethrinb
Trifluralinb
a Single-column analysis
b Dual -column analysis
CAS Registry No.
72-55-9
50-29-3
2303-16-4
117-80-6
115-32-2
60-57-1
959-98-8
33213-65-9
1031-07-8
72-20-8
7421-93-4
53494-70-5
2593-15-9
58718-66-4
58718-67-5
12616-35-2
12616-36-3
2234-13-1
39450-05-0
76-44-8
1024-57-3
118-74-1
77-47-4
465-73-6
143-50-0
72-43-5
2385-85-5
1836-75-5
82-68-8
72-56-0
1918-16-17
8001-50-1
8001-35-2
39765-80-5
51877-74-8
1582-09-8
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 new matrix.
8081 - 2
Revision 0
November 1992
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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 listed chemical is a potential method interference when it is not a target
analyte.
1.4 Several multi-component mixtures (i.e., Aroclors, Halowaxes,
Toxaphene and Strobane) 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 (Section 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.
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.
2.0 SUMMARY OF METHOD
2.1 A measured volume or weight of sample (approximately 1 L for liquids,
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.
8081 - 3 Revision 0
November 1992
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After cleanup, the extract is analyzed by injecting a 1-fj.L 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 (Section 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
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 Section 7.
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3.8 The following target analytes coelute using single column analysis:
DB 608 Trifluralin/Diallate 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 Section 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/Dicofol
Perthane/Endrin and Chiorobenzilate/Endosulfan II/Chloropropylate
will also co-elute on DB-5 after moderate deterioration in column
performance.
DB-1701 Chlorothalonil/B-BHC
6-BHC/DCPA/trans-Permethrin
a-Chlordane/trans-Nonachlor
Captan/Dieldrin
Chiorobenzi1 ate/Chioropropylate
Chlorothalonil/B-BHC and a-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).
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3.9.2 DB-5/DB-1701, thick film, fast ramp: See Section 7 and Table
7.
DB-5 Diall ate/a-BHC
Perthane/Endosulfan II
Chiorobenzi1 ate/Chioropropylate
Endrin/Nitrofen
4,4'-DDT/Endosulfan sulfate
Methoxychlor/Di colfol
DB-1701 cc-Chlordane/trans-Nonachlor (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 /zm 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
methylpolysiloxane (DB 608, SPB 608, or equivalent), 25 jum
coating thickness, 1 /itn 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 jum or 0.83 ;um 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
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methylpolysiloxane (DB 1701, or equivalent), 1.0 urn 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
fj,m film thickness, or equivalent.
4.1.2.1.3 30 m x 0.53 mm ID DB-1701 (J&W Scientific),
1.0 /xm 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
Mm film thickness, or equivalent.
4.1.2.2.3 30 m x 0.53 mm ID DB-1701 (J&W Scientific),
1.0 MM film thickness, or equivalent.
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,
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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.
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
standard solutions must be replaced after six months or sooner if
routine QC (Section 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.
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 (e.g., DB-608), two calibration
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mixtures should be prepared for the single component analytes of this
method.
5.5.2 This requirement 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 (Section 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. Prepare the standard to complement the
concentrations found in Section 5.5.
5.6.2 Make a solution of 1 mg/mL of l-bromo-2-nitrobenzene for dual-
column analysis. Dilute it to 5000 ng/VL for spiking, then use a spiking
volume of 10 /A/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 (Section 8.2). Method 3500,
Section 5, indicates the proper procedure for preparing these surrogates.
5.7.2 For the dual column analysis make a solution of 1 mg/mL of 4-
chloro-3-nitrobenzotrifluoride and dilute to 500 ng//iL. Use a spiking
volume of 100 /iL for all 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, Section 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
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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 must be
spiked with the compounds of interest to determine the percent recovery
and the limit of detection for that sample (Section 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
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
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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.
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 multicomponent
organochlorine compounds.
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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 Section
5. Refer to Method 8000 (Section 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 (Section 7.3.3). 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 choice of target analytes chosen for
calibration should be limited to those specified in the project
plan. Sites may require analysis for the organochlorine pesticides
only or the PCBs only. Toxaphene and/or technical Chlordane may
also not be specified at certain sites. In addition, where PCBs are
specified in the project plan, a mixture of 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 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
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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. Section 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
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 Section
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 is 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
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and proceeding with sample analysis. If routine maintenance does not
return the instrument performance to meet the QC requirements (Section
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 Continuation of 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.
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 nl 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.
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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 Section 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.
7.5.8 If compound identification or quantitation are 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 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, PCB, 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
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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 y), 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.
7.6.3.2 Whenever possible, when Chlordane residue does not
resemble technical Chlordane, the analyst should quantitate the
peaks of a-Chlordane,Y-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
PCB involves problems similar to those encountered in the quantisation of
Toxaphene, Strobane, and Chlordane. In each case, the chemical is made up
8081 - 16 Revision 0
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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.
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, use of Aroclor standards is
not appropriate. Several diagnostic peaks useful for identifying
non-Aroclor PCBs are identified in Table 10. Analysts should
examine chromatographs 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
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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, /3, y» and 6) 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.
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
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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 cal ibration
and corrective actions, are found in Method .8000. The following steps are
recommended as additional method QC.
8.2.1 The laboratory control sample (LCS) concentrate (Method 8000)
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 LCS 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.
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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'-DDD, Endrin ketone or Endrin indicates breakdown. If
degradation of either DDT or Endrin exceeds 15%, take corrective action before
proceeding with calibration.
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 (Section 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/ynL in the final extract for each single-component compound.
Ion trap or selected ion monitoring will normally require a minimum
concentration near 1 ng/ L.
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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 ng/ L.
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, 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.
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 method has 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 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 following this method depends
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 ng/kg. 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 spiked
sample was then extracted by Method 3541 (Automated Soxhlet). The data
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represents 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 using the procedure outlined in Method 3541. Multiple extractions
using two different extractors were performed. The extracts were analyzed by
Method 8080. 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 sp,iked 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 8080. These data are listed in Table 16 and were taken from Reference 13.
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.
8081 - 22 Revision 0
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9. Glazer, J.A., et al. "Trace Analyses for Wastewaters"; Environ. Sci. and
Technol. 1981, 15, 1426.
10. Marsden, P.J., "Performance Data for SW-846 Methods 8270, 8081, and 8141,"
EMSL-LV, EPA/600/4-90/015.
11. Marsden, P.J., "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.
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.
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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
Al dnn
a-BHC
B-BHC
5-BHC
Y-BHC (Lindane)
a-Chlordane
y-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 608°
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 1701C
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.
MDLb 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
MDLb 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.
Aqueous MDLs from U.S. EPA Method 8081. Organochlorine Pesticides
and PCBs as Aroclors. Environmental Protection Agency. Office of
Research and Development, Washington, DC 20460.
8081 - 24
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TABLE 1
(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, silica gel 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.
See Table 4 for GC operating conditions.
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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
Aldrin
a-BHC
6-BHC
8-BHC
y-BHC (Lindane)
o-Chlordane
y-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
Liquid = Organic-free
Retention Time (min)
DB 608C DB 5d
14.51
11.43
12.59
13.69
12.46
17.34
21.67
19.09
23.13
19.67
18.27
22.17
24.45
21.37
23.78
13.41
16.62
28.65
MR
MR
MR
MR
MR
MR
MR
MR
reagent water.
14.70
10.94
11.51
12.20
11.71
17.02
20.11
18.30
21.84
18.74
17.62
20.11
21.84
19.73
20.85
13.59
16.05
24.43
MR
MR
MR
MR
MR
MR
MR
MR
MDLb Water
(M9/L)
0.034
0.035
0.023
0.024
0.025
0.037
0.050
0.058
0.081
0.044
0.030
0.040
0.035
0.039
0.050
0.040
0.032
NA
0.086
NA
0.054
NA
NA
NA
NA
0.90
MDLb 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
NA
5.7
NA
57.0
NA
NA
NA
NA
70.0
Solid = Sandy loam soil .
MR = Multiple peak
NA = Data not avai
responses.
Table.
8081 - 26
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TABLE 2
(Continued)
8 Aqueous MDLs from U.S. EPA Method 8081. Organochlorine Pesticides and
PCBs as Aroclors. Environmental Protection Agency. Office of Research
and Development, Washington, DC 20460.
b 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-
1, 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.
c 30 m x 0.25 mm ID DB 608 1 urn film thickness, see Table 4 for GC operating
conditions.
d 30 m x 0.25 mm ID DB 5 1 urn film thickness, see Table 4 for GC operating
conditions.
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TABLE 3
ESTIMATED QUANTITATION LIMITS (EQL) 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
Sample EQLs are highly matrix-dependent. The EQLs listed herein are
provided for guidance and may not always be achievable.
EQL = [Method detection limit for water (Table 1) or (Table 2) wide bore
or narrow bore options] x [Factor (Table 3)]. For nonaqueous samples,
the factor is on a wet-weight basis.
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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
jitm film thickness.
Carrier gas (He)
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
16 psi
2256C
300°C
100°C, hold 2 minutes
100°C to 160°C at 15°C/nrin, followed
by 160°C to 270°C at 5°C/min
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 /xm coating thickness, 1 jiim film thickness
Carrier gas (N2)
Injector temperature
Detector temperature
Initial temperature
Temperature program
Final temperature
20 psi
225bC
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 /xm or 0.83 urn 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 /xm 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
8081 - 29
Revision 0
November 1992
-------
TABLE 4 (Continued)
GC OPERATING CONDITIONS FOR ORGANOCHLORINE COMPOUNDS
SINGLE COLUMN ANALYSIS
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, or equivalent), 1.5 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
6 mL/minute
30 mL/min
205°C
290°C
140°C, hold 2 min
140°C to 240°C at 10°C/min,
hold 5 minutes at 240 C,
240°C to 265°C at 5°C/min
265°C, hold 18 min
8081 - 30
Revision 0
November 1992
-------
TABLE 5
RETENTION TIMES OF THE ORGANOCHLORINE PESTICIDES3
DUAL COLUMN METHOD OF ANALYSIS
Compound
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
14
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
Compound
DBCP
Hexachl orocycl opentadi ene
Etridiazole
Chloroneb
Hexachl orobenzene
Dial late
Propachlor
Trifluralin
a-BHC
PCNB
Y-BHC
Heptachlor
Aldrin
Alachlor
Chlorothalonil
Alachlor
B-BHC
Isodrin
DC PA
6-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' -ODD
Endosulfan II
4, 4' -DDT
Endrin aldehyde
Mirex
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
Revision 0
November 1992
-------
TABLE 5
(Continued)
No.
Compound
CAS No.
DB-5
DB-1701
RT(min) RT(min)
39
40
41
42
43
44
45
IS
SU
Methoxychlor
Captafol
Endrin ketone
trans- Permethrin
Kepone
Dicofol
Dichlone
a, a '-Dibromo-m-xylene
2-Bromobiphenyl
72-43-5
2425-06-1
53494-70-5
51877-74-8
143-50-0
115-32-2
117-80-6
35.33
32.65
33.79
41.50
31.10
35.33
15.17
9.17
8.54
40.31
41.42
42.26
45.81
b
b
b
11.51
12.49
aThe 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-pm 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
November 1992
-------
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 (urn): 1.0
Column 2:
Type: DB-5 (J&W) or equivalent
Dimensions: 30 m x 0.53 mm ID
Film Thickness (urn): 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 \il
Solvent: Hexane
Type of injector: Flash vaporization
Detector type: Dual ECD
Range: 10
Attenuation: 64 (DB-1701)/32 (DB-5)
Type of splitter: Supelco 8 in injection tee
8081 - 33 Revision 0
November 1992
-------
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 (urn): 1.0
Column 2:
Type: DB-5 (J&W) or equivalent
Dimensions: 30 m x 0.53 mm ID
Film Thickness (urn): 1.5
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 \il
Solvent: Hexane
Type of injector: Flash vaporization
Detector type: Dual ECD
Range: 10
Attenuation: 64 (DB-1701)/64 (DB-5)
Type of splitter: J&W Scientific press-fit Y-shaped inlet splitter
8081 - 34 Revision 0
November 1992
-------
TABLE 8 SUMMARY OF RETENTION TIMES (MIN) OF AROCLORS
ON THE DB-5 COLUMN3
DUAL SYSTEM OF ANALYSIS
Peak Aroclor Aroclor Aroclor
No.b 1016 1221 1232
1 5.
2 7.
3 8.41 8.
4 8.77 8.
5 8.98 8.
6 9.71
7 10.49 10.
8 10.58 10.
9 10.90
10 11.23 11.
11 11.88
12 11.99
13 12.27 12.
14 12.66 12.
15 12.98 12.
16 13.18
17 13.61
18 13.80
19 13.96
20 14.48
21 14.63
22 14.99
23 15.35
24 16.01
25
26 16.27
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
85 5.85
63 7 . 64
43 8.43
77 8.78
99 9.00
50 10.50
59 10.59
10.91
24 11.24
11.90
12.00
29 12.29
68 12.69
99 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 eluting at same
retention time
Chlorothalonil (11.18)
Captan (16.21)
gamma -Chi ordane (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.
8081 - 35
(continued)
Revision 0
November 1992
-------
TABLE 8 CONTINUED
Peak
No.
56
57
58
59
60
61
62
63
64
65
66
67
68
69
Aroclor Aroclor Aroclor Aroclor Aroclor Aroclor
1016 1221 1232 1242 1248 1254
23.99
24.27
24.61
24.93
26.22
Aroclor Pesticide eluting 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
November 1992
-------
TABLE 9 SUMMARY OF RETENTION TIMES (MIN) OF AROCLORS
ON THE DB-1701 COLUMN3
DUAL SYSTEM OF ANALYSIS
Peak Aroclor Aroclor Aroclor
No.b 1016 1221 1232
1 4.
2 5.
3 5.
4 5.
5 6.33 6.
6 6.78 6.
7 6.96 6.
8 7.64
9 8.23 8.
10 8.62 8.
11 8.88
12 9.05 9.
13 9.46
14 9.77 9.
15 10.27 10.
16 10.64 10.
17
18 11.01
19 11.09
20 11.98
21 12.39
22
23 12.92
24 12.99
25 13.14
26
27 13.49
28 13.58
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
45 4.45
38
78
86 5.86
34 6.34
78 6.79
96 6.96
23 8.23
63 8.63
8.89
06 9.06
9.47
79 9.78
29 10.29
65 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 eluting at same
1260 retention time
Trifluralin (6.96)
13.52
14.02
14.25
14.56
Chlordane (15.32)
16.61 4,4'-DOE (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
aThe GC operating conditions are given in Table 7.
(continued)
8081 - 37
Revision 0
November 1992
-------
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 eluting 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
November 1992
-------
TABLE 10
PEAKS DIAGNOSTIC OF PCBs OBSERVED IN 0.53 mm ID COLUMN
SINGLE COLUMN ANALYSIS
Peak RT on RT on Retention
No.c DB 608a DB 1701a Aroclor" Window
1 4790 4766 1221 Before 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
Y-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 Y-BHC and heptachlor
epoxide on DB 1701;
heptachlor epoxide and
Y- Chlordane on DB 608
VII 14.81 14.77 1254 Heptachlor epoxide and
Y-Chlordane on DB
1701; a- and Y~
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
November 1992
-------
TABLE 10 (Continued)
PEAKS DIAGNOSTIC OF PCBs OBSERVED IN 0.53 mm ID COLUMN
SINGLE COLUMN ANALYSIS
Peak RT on RT on
No. DB 608a DB 1701a Aroclorb Retention Window
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 Using oven 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.
c These are sequentially numbered from elution order and are not isomer
numbers
8081 - 40 Revision 0
November 1992
-------
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
November 1992
-------
TABLE 12 ANALYTE RECOVERY FROM SEWAGE SLUDGE
Compound Sonication Soxhlet
Hexachloroethane
2-Chloronapthalene
4-Bromodiphenyl ether
a-BHC
g-BHC
Heptachlor
Aldrin
b-BHC
d-BHC
Heptachlor epoxide
Endosulfan I
g-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
November 1992
-------
TABLE 13 ANALYTE RECOVERY FROM DCE STILL BOTTOMS
Compound
Sonication
Soxhlet
Hexachloroethane
2-Chloronapthalene
4-Bromodiphenyl ether
a-BHC
g-BHC
Heptachlor
Aldrin
b-BHC
d-BHC
Heptachlor epoxide
Endosulfan I
g-Chlordane
a-Chlordane
DDE
Dieldrin
Endrin
Endosulfan II
DDT
Endrin aldehyde
ODD
Tetrachl oro-m-xyl ene
Decachlorobiphenyl
Recovery%
70
59
159
55
43
48
48
51
43
47
47
48
45
45
45
50
49
49
40
48
49
17
%RSD
2
3
14
7
6
6
5
7
4
6
4
5
5
4
5
6
5
4
4
5
2
29
Recovery
50
35
128
47
30
55
200
75
119
66
41
47
37
70
58
41
46
40
29
35
176
104
%RSD
30
35
137
25
30
18
258
42
129
34
18
13
21
40
24
23
17
29
20
21
211
93
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 - 43
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November 1992
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TABLE 14
SINGLE LABORATORY ACCURACY DATA FOR THE EXTRACTION OF
ORGANOCHLORINE PESTICIDES FROM SPIKED CLAY SOIL BY METHOD 3541
(AUTOMATED SOXHLET)3
Compound Name Spike Level % Recovery
DB-5 DB-1701
alpha-BHC
delta-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
November 1992
-------
TABLE 15
Single Laboratory Recovery Data for Extraction of
PCBs from Clay and Soil by Method 35413 (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
Recovery
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
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November 1992
-------
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
Recovery
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
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November 1992
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Table 16. Multi-Laboratory Precision and Accuracy Data
for the Extraction of PCBs from Spiked Soil
by Method 3541 (Automated Soxhlet)
PCB Percent Recovery
(%)
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
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
140.9
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
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November 1992
-------
FIGURE 1.
GAS CHROMATOGRAM OF THE MIXED ORGANOCHLORINE PESTICIDE STANDARD
Sta-t ' -ne 0 00 mi-
Sca^e facto" C
End T.me : 33.00 mm
Plot Offset: 20 mv
Low pomt : 2C.OO m
Plot Scale: 400 mv
High Point • •.20.00 mv
Response [mV]
C_H O Lfl O <_J> O (Ji O
oooooooo
Ul
^-3 38
=-4.68
:-7 .99
9.93
-23.18
-23.80
•26.23
-28.64
-0.95
-8 . 60
— 30.19
Column:
Temperature program:
30 m x 0.25 mm ID, DB 5
100°C (hold 2 minutes) to 160°C at l5°C/m1n, then at
5°C/nrin to 270°C; carrier He at 16 psi.
8081 - 48
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November 1992
-------
FIGURE 2.
GAS CHROMATOGRAM OF INDIVIDUAL ORGANOCHLORINE PESTICIDE STANDARD MIX A
S'j-: • -^ ;..; -, ,
;.! - <•>;•-.' • :
E-Xl ':me 55 X »'n
= st Of'se:: :'_ -v
Response
tjl—
H
I
-I
"""^ ro
J 0-
a
a
I ! I I I I
i i i i i i i
17.93
-9.60
12.33
-14.27
-17.08
20.22
0.77
•22.68
-23.73
--28.52
•-9.86
-10.98
-13.58
-17.54
18.47
-19.24
-19.78
-21.13
-2:
-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/nrin to 270°C; carrier He at 16 psi.
8081 - 49
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November 1992
-------
FIGURE 3
GAS CHROMATOGRAM OF INDIVIDUAL ORGANOCHLORINE PESTICIDE STANDARD MIX B
Response [mV]
Q
O
-
O
n
D
9 °
L-i
2.74
;5.13
-6.97
i*.
60
--10.71
14.27
.24
).ll
^20.69
22.00
•11.73
-14.84
-16.23
— 17.08
-17.63
-18.31
19.54
-20.19
--21.03
--22.68
•2.93
— 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/niin, then at
5°C/min to 270°C; carrier He at 16 psi.
8081 - 50
Revision 0
November 1992
-------
FIGURE 4.
GAS CHROMATOGRAM OF THE TOXAPHENE STANDARD
S-.3-" ' -
Sea e -'.1C
End '•« : 33 ""
Plot Of'set: 2C TV
'.o» oo--t • 2C.03 mv
Plot Sca.e: t'j Ml
Poi-t : 8C.3C
Response
•LN! ^ <_?! &r *-J
O O O O O
J
J
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 - 51
Revision 0
November 1992
-------
FIGURE 5.
GAS CHROMATOGRAM OF THE AROCLOR-1016 STANDARD
T n End Time : 33 CC m"
Plot Offset: 20 ^v
Lo* Pom* • 2C.OO m
Plot Scale- 100 mv
2- J1- *"rf
Response [mV]
DO
o
o
-1.81
--, I
^ ,-. ,,-J
I,',
r V
Column:
Temperature program:
-12.95
30 m x 0.25 mm 10 D6 5 fused silica capillary.
100°C (hold 2 minutes) to 160°C at 15°C/irrin, then at
5°C/nrin to 270°C; carrier He at 16 psi.
8081 - 52
Revision 0
November 1992
-------
FIGURE 6.
GAS CHROMATOGRAM OF THE TECHNICAL CHLORDANE STANDARD
o
33 ::
2C nv
Response [mV]
a
o
to
O
O
11
V21.03
^—21.61
-24.33
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
Revision 0
November 1992
-------
OB-1701
L
DB-5
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 ran ID DB-5 (1.5-pm 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.
8081 - 54
Revision 0
November 1992
-------
DB-1701
o
o-
rg
-JJL
DB-5
HI
bi
lit
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-jim 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.
8081 - 55
Revision 0
November 1992
-------
(f
a
<Ł>
rx
to-o
00
!
V
II
s
I
I
c
c
f
r
i
j
1
OB-1701
OB-5
Figure 9. GC/ECO 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-^m film thickness) and
30 m x 0.53 mm 10 DB-1701 (l.0-|>m film thickness) connected to a J&U
Scientific press-fit Y-shaped inlet splitter. Temperature program:
150°C (0.5 min hold) to 190°C (2 min hold) at 12°C/nrin then to 275°C
(10 min hold) at 4°C/min.
8081 - 56
Revision 0
November 1992
-------
r
r.
r~ffl .
f-O V:
DB-1701
(l
f-0
CD
U'ln -V
U-TM 11 «.
vm . h| —
]o> I
I fMi 1 i 0*1 | i
•^*Msi^
o
p"
DB-5
10
— T
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-nm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jim 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 - 57
Revision 0
November 1992
-------
IT
K
•c
— o
ffiD
?JXT.
M
ni
IN
O
1
]
«j
0,'.
I-O
r-
e-i
J
|
4
1
I
,i
(
VI
DB-1701
•5.
DB-5
r
Figure 11. GC/ECD chromatogram of Aroclor 1232 analyzed on a DB-5/DB-1701
fused-slllca open-tubular column pair. The GC operating conditions
were as follows: 30 n x 0.53 mm 10 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 nln hold) at 12°C/min then to 275°C
(10 min hold) at 4°C/min.
8081 - 58
Revision 0
November 1992
-------
r-
0)
3
in
0)
UL
-di in ro
rgr\i ro T
T -O
i
1
1
1
~^l ON
-0
4.
r
1 1
II|J
»
ro
in
DB-1701
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-jim film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-iim film thickness) connected to a J&W
Scientific press-fit Y-shaped Inlet splitter. Temperature program:
150°C (0.5 m1n hold) to 190°C (2 m1n hold) at l2°C/m1n then to 275°C
(10 «1n hold) at 4°C/m1n.
8081 - 59
Revision 0
November 1992
-------
Ill
0
(u
r
in
in
•o i> «<
hi —«n
r«
T OD4U 01
IW O-OkM fc'i
OB-1701
OB H>
— hi
n o»
ry
fM
o-
if-
DB-5
O> IT'
in
T
O ki
l« \f
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-pm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jim 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 m1n hold) at 12°C/nrin then to 275°C
(10 min hold) at 4°C/min.
8081 - 60
Revision 0
November 1992
-------
\r>
K)
at
a
•a
OB-1701
u>
o-
•0
u-
iD • r. OB -
« •« f« * • -
» - «» ®
« a> a
-------
DB-1701
: S
DB-5
mtf * • '
• » ** • i
Figure 15. GC/ECD chroraatogram 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-|im film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-urn 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 m1n hold) at l2°C/m1n then to 275°C
(10 win hold) at 4°C/min.
8081 - 62
Revision 0
November 1992
-------
DB-1701
r-
U1
K)
f-l
rvt
- _
» « r- a- ni
«K -O
CD9
- -^ A
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-|tm film thickness) and
30 m x 0.53 mm 10 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/nrin.
8081 - 63
Revision 0
November 1992
-------
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 (l.C-jim 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/min.
8081 - 64
Revision 0
November 1992
-------
if 9-
-------
DB-1701
» 0-
II «
r- «
•0 Is-
DB-5
•0
•f
Figure 19. GC/ECD chromatogram of Halowax 1013 analyzed on a OB-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-iun film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jim 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 - 66
Revision 0
November 1992
-------
DB-1701
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-jim film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jim film thickness) connected to a J&U
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 - 67
Revision 0
November 1992
-------
OB-1701
= ; * ? 3?5S
.. » . i. , „».
*••»* —» «k •
;:Z3 Si;:
:5«t at ss
DB-5
X
ft
Figure 21. GC/ECD chromatogram of Halowax 1051 analyzed on a 08-5/D6-1701
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 10 OB-1701 (1.0-|im film thickness) connected to a J&U
Scientific press-fit Y-shaped Inlet splitter. Temperature program:
150°C (0.5 mln hold) to 190°C (2 mln hold) at l2°C/m1n then to 275°C
(10 min hold) at 4°C/m1n.
8081 - 68
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November 1992
-------
DB-5
2 4
IS SU
J4 )) 42
'44
It
41
41
40
LU
DB-1701
n
12 1 4 SU 1$ * »
I
Ji ,
•
1
10 II 12 II
t
II
y
^
1
22
_
1
II
»,/» .
I
I
II2
13
J2
ud
M n >« 4
M
I
Ul
0
4)
Zt
Figure 22. GC/ECD chromatogram of the organochlorine pesticides analyzed on a
DB-5/DB-1701 fused-sllica open-tubular column pair. The GC
operating conditions were as follows: 30 m x 0.53 mm ID DB-5 (0.83-
lim 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 mln hold) to 270°C (1 rain hold) at
2.8 C/min.
8081 - 69
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November 1992
-------
MŁTHOD 8081
ORGANOCHLORINE PESTICIDES, HALOWAXES AND PCBs AS AROCLORS BY GAS
CHROMATOGRAPHY: CAPILLARY COLUMN TECHNIQUE
7. 1.1 Choose
appropriate extraction
technique (see Chapter 2)
*
7.1. 2 Add specified
matix spike to sample.
1
7.2 Routine cleanup/
fractfonation.
*
7.3 Set chromatograprfc
conditions.
*
7.4 Refer to Method 8000
for proper calibration
techniques.
7.4.2 Prime or
\
deactivate GC
to cafibralion.
ttf
7.5 Perform GC analysis (see
Method 8000)
Sf-5
/Any*
\ peak
N.feren
No
S1A
/Dotes
N. twt
Noxnpo
No]
.8 >v
impteN^
vart/
.IN.
jduesN^
») /
oenftX^
Yes
Yes
7.5.8 Additional
ctoanup/fractionation
(see Section 7.2)
7.6 Calculation of
toxapnene, chkxdane, PCBs.
DDT, and BHC done here.
8081 - 70
Revision 0
November 1992
-------
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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November 1992
-------
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 50C/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/min 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/nrin, 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/nrin 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/min> 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-jjm film thickness) and 30
m x 0.53 mm ID DB-1701 (1.0-nm 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.
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-jim film thickness) and 30
m x 0.53 mm ID DB-1701 (1.0-nm 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 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-nm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jim film thickness) connected to a J&W
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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-nm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jim 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 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-ym film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jim 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-^m 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/nriri.
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-jim film thickness) and
30 m x 0.53 mm ID DB-1701 (LO-pm 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-pm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-iim 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 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-jim film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-jim 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
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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-pm 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/nn'n then to 275°C
(10 min hold) at 4°C/min.
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-|im film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-pm 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-nm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-pm 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-nm film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-pm 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.
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-|im film thickness) and
30 m x 0.53 mm ID DB-1701 (1.0-nm 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/nnn.
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-pm 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.
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-
[im film thickness) and 30 m x 0.53 mm ID DB-1701 (1.0-fim 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
2.85C/min.
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METHOD 8110
HALOETHERS
1.0 SCOPE AND APPLICATION
1.1 This method covers the determination of certain haloethers. The
following parameters can be determined by this method:
Parameter CAS No.
Bis(2-chloroethoxy) methane 111-91-1
Bis(2-chloroethyl) ether 111-44-4
Bis(2-chloroisopropyl) ether 108-60-1
4-Bromophenyl phenyl ether 101-55-3
4-Chlorophenyl phenyl ether 7005-72-3
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 Step 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 Any modification of this method, beyond those expressly permitted,
shall be considered as major modifications subject to application and approval
of alternate test procedures.
1.5 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 Step 8.2.
1.6 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 be made available to all personnel involved in the chemical
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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 levels 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-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
a halide-specific detector (HSD).
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 Step 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 calibration and 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 Kuderna-Danish (K-D) apparatus
4.1.1 Concentrator tube - 10-mL, graduated (Kontes K-570050-1025 or
equivalent). Calibration must be checked at the volumes employed in the
test. Ground glass stopper is used to prevent evaporation of extracts.
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4.1.2 Evaporative flask - 500 ml (Kontes K-570001-0500 or
equivalent). Attach to concentrator tube with springs.
4.1.3 Snyder column - Three-ball macro (Kontes K-503000-0121 or
equivalent).
4.2 Vials - Amber glass, 10- to 15- ml capacity, with Teflon lined screw-
cap.
4.3 Boiling chips - Approximately 10/40 mesh. Heat to 400eC 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.
4.5 Balance - Analytical, capable of accurately weighing 0.0001 g.
4.6 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.
4.6.1 Column 1 - 1.8 m x 2 mm i.d. 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
Step 7.3.1.
4.6.2 Column 2 - 1.8 m x 2 mm i.d. pyrex glass, packed with Tenax-GC
(60/80 mesh) or equivalent.
4.6.3 Detector - Halide specific: 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 was used to develop the method performance
statements in Section 9.0. Guidelines for the use of alternate detectors
are provided in Step 7.3.1. Although less selective, an electron capture
detector is an acceptable alternative.
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 ASTM Type II Water (ASTM D1193-77 (1983)). All references to water in
the method refer to ASTM Type II unless otherwise specified.
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5.3 Acetone, CHsCOCHs. Pesticide quality or equivalent.
5.4 Hexane, CsHi4. Pesticide quality or equivalent.
5.5 Stock standard solutions (1000 mg/L) - Stock standard solutions can
be prepared from pure standard materials or purchased as certified solutions.
5.5.1 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.
5.5.2 Transfer the stock standard solutions into Teflon lined 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.
5.5.3 Stock standard solutions must be replaced after six months, or
sooner if comparison with check standards indicate a problem.
5.6 Calibration standards - Calibration standards at a minimum of five
concentration levels 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 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.
5.7 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
concentration levels for each analyte of interest as described in Step
5.6.
5.7.2 To each calibration standard, add a known constant amount of
one or more internal standards, and dilute to volume with isooctane.
5.7.3 Analyze each calibration standard according to Section 7.0.
5.8 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.
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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, Step
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 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
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 Step 7.1.2.3. If cleanup is
needed, proceed to Step 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
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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 either Method 3610 or 3620, using the 2-mL hexane
extracts obtained from Step 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
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 Step 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 Step 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.
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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 uL of internal standard to the sample prior to
injection.
7.5.2 Follow Step 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 check
standard after each group of 10 samples in the analysis sequence.
7.5.3 Examples of GC/HSD 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 Step 7.8 of 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,
Step 8.6.
8,2.1 The quality control (QC) reference sample concentrate (Method
8000, Step 8.6) should contain each analyte of interest at 20 ug/mL.
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.
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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, Step 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 This method has been tested for linearity of recovery from spiked
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.
9.3 The U.S. Environmental Protection Agency is in the process of
conducting an interlaboratory method study to fully define the performance of
this method.
10.0 REFERENCES
1. Fed. Reaist. 1984, 49, 43234; October 26.
2. ASTM Annual Book of Standards. Part 31; "Standard Practice for
Preparation of Sample Containers and for Preservation"; ASTM:
Philadelphia, PA, p. 679, 1980; D3694.
3. Carcinogens - Working with Carcinogens; Department of Health, Education,
and Welfare, Public Heath 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." (29CFR1910),
Occupational Safety and Health Administration, OSHA 2206, (Revised
January 1976).
5. Safety in Academic Chemistry Laboratories, 3rd ed.; American Chemical
Society Publication, Committee on Chemical Safety, 1979.
8110 - 8 Revision 0
December 1987
-------
6. 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.
7. 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.
8. ASTM Annual Book of Standards. Part 31; "Standard Practice for Sampling
Water"; ASTM: Philadelphia, PA, p. 76, 1980; D3370.
9. 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.
10. Burke, J.A. "Gas Chromatography for Pesticide Residue Analysis; Some
Practical Aspects"; Journal of the Association of Official Analytical
Chemists 1965, 48, 1037.
11. "EPA Method Validation Study 21 Methods 611 (Halothers)," Report for EPA
Contract 68-03-2633.
12. Rohrbough, W.G.; et al. Reagent Chemicals. American Chemical Society
Specifications, 7th ed.; American Chemical Society: Washington, DC, 1986.
13. 1985 Annual Book of ASTM Standards. Vol. 11.01; "Standard Specification
for Reagent Water"; ASTM: Philadelphia, PA, 1985; D1193-77.
14. "Determination of Haloethers in Industrial and Municipal Wastewaters";
Report for EPA Contract 68-03-2633 (In preparation).
8110 - 9 Revision 0
December 1987
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS
Retention Time
(minutes)
Parameter
Column 1 Column 2
Method
Detection Limit
(ug/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: Supelcoport (100/120 mesh) coated with 3% SP-1000 packed
in a 1.8 m x 2 mm i.d. glass column with helium carrier gas at a flow rate of
40 mL/min. Column temperature: 60°C for 2 minutes after injection then program
at 8°C/min to 230°C and hold for 4 minutes. Under these conditions the
retention time for aldrin is 22.6 minutes.
Column 2 conditions: Tenax-GC (60/80 mesh) packed in a 1.8 m x 2 mm i.d. glass
column with helium carrier gas at 40 mL/min flow rate. Column temperature:
150°C for 4 minutes after injection then program at 16°C/min to 310°C. Under
these conditions the retention time for aldrin is 18.4 minutes.
8110 - 10
Revision 0
December 1987
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TABLE 2.
SINGLE OPERATOR ACCURACY AND PRECISION
Average Standard Spike Number
Percent Deviation Range of Matrix
Parameter Recovery % (ug/L) Analyses Types
Bis(2-chloroethoxy) methane 62 5.3 138 27 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
8110 - 11 Revision 0
December 1987
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FIGURE 1.
GAS CHROMATOGRAM OF HALOTHERS
Column: 3% SP-10OO on Supelcoport
Program. 60°C. -2 minutes 8"'/minute to 230°C.
Detector: He/1 electrolytic conductivity
0 2 4 6 8 10 12 14 16 18 20 22 24
Retention time, minutes
FIGURE 2.
GAS CHROMATOGRAM OF HALOETHERS
Column: Tenax GC
Program: 150°C.-4 minutes 16°/minute to 310°C.
Detector: Hall electrolytic conductivity
*+*
8
12
16
20
24
Retention time, minutes
8110 - 12
Revision 0
December 1987
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METHOD 8110
HALOETHERS
C
Start
7.1.1 Ckooee
appropriate
axtraetioa
procedure
7.1.2 Perform
aolveat exckanfa
aeiaf kexaaa
7 12.4 Perform
micro-K-D procedure
using hexane;
proceed with Method
3610 or 3620
Tee
7.1.2.1 Adjaet
extract volue and
proceed vitk
analyaie or etore
la appropriate
•aaaer
7.S.I lefer to
Table 1 for
recoueadod
operatlag
coaditioae for tae
OC
7.4 lefer to Metkot
(000 for proper
callbratioa
teckaiqaee
7.S.I lefer to
Metkod 8000 for
fildaace oa CC
aaalyeit
7.6.4 lecord itiple
ulnae iajected aad
renltiif peak elze
7.E.S Perfora
appropriate
caleilatioa
(Metkod 1000,
7.8)
Step
Stop
8110 - 13
Revision 0
December 1987
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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 No8 3510 3520 3540 3550 3580
2-Chloronaphthalene
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1,4-Di chl orobenzene
Hexachl orobenzene
Hexachl orobutadi ene
Hexachl orocycl ohexane
Hexachl orocycl opentad i ene
Hexachl oroethane
Pentachlorohexane
Tetrachl orobenzenes
1,2,4-Trichlorobenzene
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 ML 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).
2.2 If interferences are encountered in the analysis, Method 8120 may
also be performed on extracts that have undergone cleanup using Method 3620.
8120A - 1 Revision 1
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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.
4.1.3 Detector - Electron capture (ECO).
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).
8120A - 2 Revision 1
November 1992
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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 nl.
4.7 Syringe - 5 nil.
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 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.
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.
8120A - 3 Revision 1
November 1992
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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 Section 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 Section 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,
Section 4.1.
6.2 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
8120A - 4 Revision 1
November 1992
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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 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
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 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
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
8120A - 5 Revision 1
November 1992
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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 juL 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).
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.
8120A - 6 Revision 1
November 1992
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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 Section 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.
• 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 jug/L. Single operator precision,
8120A - 7 Revision 1
November 1992
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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 (in
preparation).
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 (in preparation).
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 (in preparation).
8120A - 8 Revision 1
November 1992
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TABLE 1.
GAS CHROMATOGRAPHY OF CHLORINATED HYDROCARBONS
Compound
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1 , 4-Di chl orobenzene
Hexachl orobenzene
Hexachl orobutadi ene
Hexachl orocycl ohexane
Hexachl orocycl opentadi ene
Hexachl oroethane
Pentachlorohexane
Tetrachl orobenzenes
1,2,4-Trichlorobenzene
Retention
Col. 1
2.78
6.6
4.5
5.2
5.6"
7.7
ND
4.9
15.5
time (min)
Col. 2
3.6b
9.3
6.8
7.6
10. lb
20.0
16. 5C
8.3
22.3
Method
Detection
limit (/ig/L)
0.94
1.14
1.19
1.34
0.05
0.34
0.40
0.03
0.05
ND = Not determined.
a!50°C column temperature.
b!65°C column temperature.
C100°C column temperature.
8120A - 9
Revision 1
November 1992
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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
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.
8120A - 10 Revision 1
November 1992
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TABLE 3.
QC ACCEPTANCE CRITERIA8
Parameter
2-Chl oronaphthal ene
1 , 2-Di chl orobenzene
1 , 3-Di chl orobenzene
1 , 4-Di chl orobenzene
Hexachl orobenzene
Hexachl orobutadi ene
Hexachl orocycl opentadl ene
Hexachl oroethane
1 , 2 , 4-Tri chl orobenzene
Test
cone.
(M9/L)
100
100
100
100
10
10
10
10
100
s = Standard deviation of four recovery
x = Average recovery
P,PS = Percent recovery
D = Detected; result
for four recovery
measured.
Limit
for s
(M9/L)
37.3
28.3
26.4
20.8
2.4
2.2
2.5
3.3
31.6
Range
for x
(M9/L)
29.5-126.9
23.5-145.1
7.2-138.6
22.7-126.9
2.6-14.8
D-12.7
D-10.4
2.4-12.3
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
measurements, in /ig/L.
measurements, in M9/L.
must be greater than zero.
a Criteria from 40 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.
8120A - 11 Revision 1
November 1992
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TABLE 4.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Parameter
Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1 , 4-Di chl orobenzene
Hexachlorobenzene
Hexachl orobutadi ene
Hexachl orocycl opentadi ene8
Hexachl oroethane
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
fl.21x-l.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' (M§/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'
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 M9/L-
Expected interlaboratory standard deviation of measurements at an
average concentration found of x, in p.g/1.
True value for the concentration, in M9/L.
Average recovery found for measurements of samples containing a
concentration of C, in
Estimates based upon the performance in a single laboratory.
8120A - 12
Revision 1
November 1992
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FIGURE 1
Column: 1.5% 0V-1*1.5% OV-228 en OM Ovom Q
Temperature: 76°C
Dtncter: Electron Capture
4 I 12 16
RETENTION TIME (MINUTES)
20
Gas chromatogram of chlorinatad hydrocarbon* (low molecular wtight compounds).
8120A - 13
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FIGURE 2
Column: 1J% OV-1+1J* OV-22S en QM Chrom Q
Temperature: 160°C
Detector: Electron Capture
i i t t
0 4 • 12 16
RETENTION TIME (MINUTES)
G«s ehromatogram of chlorinated hydrocarbons (high molecular weight compounds).
8120A - 14
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METHOD 8120A
CHLORINATED HYDROCARBONS BY GAS CHROMATOGRAPHY
C
711 Choeoe
appropriate
extraction
procedure (»••
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7 32 I.
cleanup
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7 3.2 Preevee a
teriei of itandard*
through cleanup
procedure: analyse
by CC
7 4 Perfora CC
analyoi.* (aee
Method 8000)
fication
tection prevented
7 S 1 Cleanup uoinc
Method 3420
8120A - 15
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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.8
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
6-Hexachlorocyclohexane (6-BHC) 319-85-7
y-Hexachlorocyclohexane (y-BHC) 58-89-9
&-Hexachlorocyclohexane (6-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-silica 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-silica 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
the nature of interferences in the sample matrix. Table 2 lists the estimated
quantitation limits (EQL) for other matrices.
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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.
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
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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 Section 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
1,2,3,5-tetrachlorobenzene/1,2,4,5-tetrachlorobenzene
1,2,3,4-tetrachlorobenzene/2-chl oronaphthalene
DB 1701 benzyl chloride/1,2-dichlorobenzene/hexachloroethane
benzal chloride/1,2,4-trichl orobenzene/
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 urn or 1.5 jim 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 Section 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 nU 10 jiL (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 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 (Section 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 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.
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/pL. Use a spiking volume
of 10 tiL/mL 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/^L. Use a spiking volume
of 100 |iL for a 1 L aqueous sample. If matrix interferences are a problem, two
alternative surrogates are: alpha, 2,6-trichlorotoluene or
2,3,4,5,6-pentachlorotoluene.
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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.
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 Method 3540
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, Section 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 Section
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.
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.
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7.4.2 Automatic injections of 1 \il are recommended. Hand injections
of no more than 2 pL 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 pL 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 (Section 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
Section 7.5).
7.4.5 Record the volume injected to the nearest 0.05 jiL 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 6C/MS technique (see Section 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
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.
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7.5.2 Suggested chromatograph maintenance: Corrective measures may
require any one or more of the following remedial actions. Also see
Section 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, Section 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, Section 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
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.
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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.
9.3 Single laboratory accuracy data were obtained for chlorinated
hydrocarbons in a clay soil. The spiking concentrations ranged from 500 to 5000
lig/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
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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.
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Table 1
METHOD DETECTION LIMITS FOR CHLORINATED HYDROCARBONS
SINGLE COLUMN METHOD OF ANALYSIS
Compound name
Benzal chloride
Benzotrichloride
Benzyl chloride
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachl orobenzene
Hexachlorobutadiene
a-Hexachlorocyclohexane (a-BHC)
6-Hexachlorocyclohexane (6-BHC)
Y-Hexachlorocyclohexane (y-BHC)
6-Hexachlorocyclohexane (6-BHC)
Hexachl orocycl opentadi ene
Hexachl oroethane
Pentachl orobenzene
1,2, 3, 4-Tetrachl orobenzene
1 ,2,4,5-Tetrachlorobenzene
1,2,3 , 5-Tetrachl orobenzene
1, 2, 4-Trichl orobenzene
1, 2, 3-Trichl orobenzene
1,3, 5 -Trichl orobenzene
CAS Reg. No.
98-87-3
98-07-7
100-44-7
91-58-7
95-50-1
541-73-1
106-46-1
118-74-1
87-68-3
319-84-6
319-85-7
58-89-9
319-86-8
77-47-4
67-72-1
608-93-5
634-66-2
95-94-2
634-90-2
120-82-1
87-61-6
108-70-3
MDLa
(ng/L)
2-5b
6.0
180
1,300
270
250
890
5.6
1.4
11
31
23
20
240
1.6
38
11
9.5
8.1
130
39
12
MDL is the method detection limit for organic-free reagent water. MDL
was determined from the analysis of eight replicate aliquots processed
through the entire analytical method (extraction, Florisil cartridge
cleanup, and GC/ECD analysis).
MDL - Vl.0.99)XSD
where t(n_., 0 99 is the student's t value appropriate for a 99 percent
confidence' interval and a standard deviation with n-1 degrees of
freedom, and SD is the standard deviation of the eight replicate
measurements.
Estimated from the instrument detection limit.
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Table 2
ESTIMATED QUANTITATION LIMIT (EQL) FACTORS FOR VARIOUS MATRICES"
Matrix Factorb
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 miscible with water 100,000
8 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
nonaqueous samples, the factor is on a wet-weight basis.
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Table 3
GAS CHROMATOGRAPHIC RETENTION TIMES FOR CHLORINATED HYDROCARBONS: SINGLE
COLUMN METHOD OF ANALYSIS
Compound
Number
1
2
3
4
5
6
7
8
9
10
12
13
14
15
16
17
18
19
20
21
22
Compound name
Benzal chloride
Benzotrichloride
Benzyl chloride
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachlorobutadiene
a-BHC
Y-BHC
6-BHC
Hexachl orocycl opentadi ene
Hexachloroethane
Pentachl orobenzene
1,2,3 , 4-Tetrachl orobenzene
1,2,4, 5-Tetrachl orobenzene
1,2,3 , 5-Tetrachl orobenzene
1, 2, 4-Trichl orobenzene
1, 2 ,3-Trichl orobenzene
1, 3, 5-Trichl orobenzene
Internal Standards
2,5-Dibromotoluene
1 , 3 , 5-Tri bromobenzene
a , a ' -Di bromo-meta-xyl ene
Surrogates
ot,2,6-Trichlorotoluene
1,4-Dichloronaphthalene
2,3,4,5,6-Pentachlorotoluene
Retention
DB-2103
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
9.55
11.68
18.43
12.96
17.43
18.96
time (min)
DB-WAX0
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
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 jim 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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November 1992
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TABLE 4
RETENTION TIMES OF THE CHLORINATED HYDROCARBONS8
DUAL COLUMN METHOD OF ANALYSIS
Compound
No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
IS
SU
Compound
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Benzyl chloride
1 , 2-Di chl orobenzene
Hexachloroethane
1 , 3 , 5-Tr i chl orobenzene
Benzal chloride
1 , 2 , 4-Tri chl orobenzene
1, 2, 3-Tri chl orobenzene
Hexachl orobutad i ene
Benzotrichloride
1,2,3, 5-Tetrachl orobenzene
1,2,4 , 5-Tetrachl orobenzene
Hexachl orocycl opentad i ene
1,2,3 , 4-Tetrachl orobenzene
2-Chl oronaphthal ene
Pentachl orobenzene
o-BHC
Hexachl orobenzene
6-BHC
Y-BHC
6-BHC
1 , 3 , 5-Tri bromobenzene
1,4-Di chl oronaphthal ene
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
11.83
15.42
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
13.34
17.71
a The GC operating conditions were as follows: 30 m x 0.53 mm ID DB-5
(0.83-jim film thickness) and 30 m x 0.53 mm ID DB-1701 (1.0 \an 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 2eC/min then to
240°C (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
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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 65eC
Temperature program 65°C to 175°C at 4'C/min
Final temperature 175°C, hold 20 minutes.
Injector temperature 220°C
Detector temperature 250°C
Injection volume 1-2 \il
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 60eC
Temperature program 608C to 170°C at 4°C/min
Final temperature 170°C, hold 30 minutes.
Injector temperature 200°C
Detector temperature 230°C
Injection volume 1-2 [iL
8121 - 15 Revision 0
November 1992
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Column 1:
TABLE 6
GC OPERATING CONDITIONS FOR CHLORINATED HYDROCARBONS
DUAL COLUMN METHOD OF ANALYSIS
Type: DB-1701 (J&W Scientific) or equivalent
Dimensions: 30 m x 0.53 mm ID
Film Thickness (urn): 1.0
Column 2:
Type: DB-5 (J&W Scientific) or equivalent
Dimensions: 30 m x 0.53 mm ID
Film Thickness (\an): 0.83
Carrier gas flowrate (mL/min): 6 (Helium)
Makeup gas flowrate (mL/min): 20 (Nitrogen)
Temperature program: 80°C (1.5 min hold) to 125eC (1 min hold) at 2°C/min
then to 240°C (2 min hold) at 5°C/min.
Injector temperature: 250°C
Detector temperature: 320eC
Injection volume: 2 \il
Solvent: Hexane
Type of injector: Flash vaporization
Detector type: Dual ECD
Range: 10
Attenuation: 32 (DB-1701)/32 (DB-5)
Type of splitter: Supelco 8-in injection tee
8121 - 16 Revision 0
November 1992
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Table 7
SUGGESTED CONCENTRATIONS FOR THE CALIBRATION SOLUTIONS8
Concentration (ng/nL)
Benzal chloride
Benzotri chloride
Benzyl chloride
2-Chl oronaphthal ene
l»2-Di chl orobenzene
1 , 3-Di chl orobenzene
1 , 4-Di chl orobenzene
Hexachl orobenzene
Hexachl orobutadi ene
a-BHC
B-BHC
Y-BHC
6-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 orobenzene
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
«c,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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November 1992
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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 chloride6
Benzotrichloride
Benzyl chloride
2-Chloronaphthalene
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
Hexachlorobenzene
Hexachlorobutadiene
a-BHC
6-BHC
y-BHC
6-BHC
Hexachl orocycl opentadi ene
Hexachloroethane
Pentachl orobenzene
1,2,3 , 4-Tetrachl orobenzene
1,2,4, 5-Tetrachl orobenzene6
1,2,3 , 5-Tetrachl orobenzene6
1, 2, 4-Trichl orobenzene
1 , 2 , 3-Tr i chl orobenzene
1 , 3 , 5-Tri chl orobenzene
Amount
(H9)
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 1D
0
0
82
115
102
103
104
116
101
93
100
129
104
102
102
59
96
102
(percent)8
Fraction 2C
0
0
16
95
108
105
71
8 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.
6 This pair cannot be resolved on the DB-210 fused-silica capillary
columns.
8121 - 18
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November 1992
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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)8
Compound Name Spike Level % Recovery
lig/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
November 1992
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1§
-
1 ..1
«
1
1
1 *
aa
I
»
M
0 2
M
21
1
1
1
11
4
»
j
f
4
1
1
1 |
n
i
•
A-
1
J*-
1
I
-------
10
15
13
A
"20 25 30 35 40 4S~
TIM! (mln)
12
A
50
55
Figure 2. GC/ECO chromatogram of Method 8121 composite standard analyzed on a
30 m 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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November 1992
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DB-5
10
OB- 1701
r M u it if is it M it ii n ;o
ILJU
u
uu
JU
Figure 3. 6C/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 \im film
thickness) and 30 m x 0.53 mm ID DB 1701 (1.0 \un 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 240°C (2 min hold) at 5°C/nnn.
8121 - 22
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November 1992
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METHOD 8121
CHLORINATED HYDROCARBONS BY GAS CHROMATOGRAPHY: CAPILLARY COLUMN TECHNIQUE
7.1.1 Choose appropriate
extraction procedure
7.1.2 Add appropriate (piking
compound* to sample prior
to extraction procedure
7.2 Exchange extraction
solvent to hrant during
K-0 proaidun
Following con
on of
meftylene chloride allow K-O
apparatus to drain and cool
7.2.2 Increaae temperature of hot
water bath; add hexane; attach
Snyder column; place apparatus on water
belh; concentrate; remove from
water bath; cool
7.2.3 Remove column; nnse task
and joints with hexane; adjust
extract volume
7.3 Chooae appropriate cleanup
technique, if neceaeary;
fluorosri cleanup is recommended
Refer to Method 3620 or to
Section 7.3.2
723 m further
processing be
performed within
two oW»?
7.2.3 Transfer extract to
Teflon sealed screw-cap
v«ls; refrigerate
7.3.4 Elemental
sulfur removal
required?
7.3.4 Refer to
Method 3660,
Section 7.3
7.3.3 Refer to
Method 3640
8121 - 23
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November 1992
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METHOD 8121
(continued)
7.2.3 Stopper concentrator
and refrigerate
7.4.1 Set column 1 conditions
7.4.2 Set column 2 conditions
7.5.1 Refer to Method 8000 for
calibration 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
8121 - 24
Revision 0
November 1992
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METHOD 8121
(concluded)
0
7.6.3 Record sample volume
injected and resulting peak
sizes
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
Revision 0
November 1992
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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
CAS Registry No.
OP Pesticides
Aspon,
Azinphos-methyl
Azinphos-ethyl8
Bolstar (Sulprofos)
Carbophenothion8
Chlorfenvinphos8
Chlorpyrifos
Chlorpyrifos methyl8
Coumaphos
Crotoxyphos8
Demeton-0°
Demeton-Sc
Diazinon
Dichlorofenthion8
Dichlorvos (DDVP)
Dicrotophos
Dimethoate
Dioxathion8'0
Disulfoton
EPN
Ethion*
Ethoprop
Famphur
Fenithrothion8
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
8141A - 1
Revision 1
November 1992
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Compound Name
CAS Registry No.
Fonophos8
Fenthion
Leptophosa'd
Malathion
Merphosc
Mevinphos6
Monocrotophos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Phosmet8
Phosphamidon8
Ronnel
Stirophos (Tetrachlorovinphos)
Sulfgtepp
TEPPd
Terbufos8
Thionazin8'6 (Zinophos)
Tokuthion (Protothiofos)
Trichlorfon
Trichloronate
Industrial Chemicals
Hexamethylphosphoramide8 (HMPA)
Tri-o-cresylphosphate8' (TOCP)
Triazine Herbicides (NPD only)
Atrazine8
Simazine8
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 can 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 on the use of a dual
column configuration when their instrument is subject to mechanical stress, when
many samples are analyzed over a short time, or when extracts of contaminated
samples are analyzed.
8141A - 2
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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. Section 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 ultrasonic extraction (Method 3550) 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.
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.
8141A - 3 Revision 1
November 1992
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3.0 INTERFERENCES
3.1 Refer to Methods 3500, 3600, and 8000, as well as to Section 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.
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
Tri chloronate/Aspon
Bolstar/Stirophos/Carbophenothion
Phosphamidon/Crotoxyphos
Fensulfothion/EPN
8141A - 4 Revision 1
November 1992
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DB-210 Terbufos/tri-o-cresyl phosphate
Di chlorofenthi on/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
quantitation.
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 amount of
oxidation of the sample and the standard is probably different,
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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 (Section 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 (Section 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 Section 3.0 and Figures 1 through 4 for guidance on selecting
the proper length and diameter for the column(s).
4.1.1.1 Column 1 - 15- or 30-m x 0.53-mm wide-bore
capillary column, 1.0-/zm 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-/zm 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 /im film thickness, chemically bonded with 5%
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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-jim or 1.5-iim 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 (Section 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.
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 (Section 3.0).
4.1.4.4 Electron-capture detectors may be used for a
limited number of analytes (Section 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.
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4.1.5.2 Use of a data system that allows storage of rav
chromatographic data is strongly recommended.
5.0 REAGENTS
5.1 Solvents
5.1.1 Isooctane, (CH3)3CCH2CH(CH3)2 - Pesticide quality 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
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 (Section 8.0) indicates a problem. Standards for easily
hydrolyzed chemicals including TEPP, Methyl Parathion, and Merphos should
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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 [POJ) 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.
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 iiL/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
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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 Section 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-ng/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,"
Section 4.0.
6.2 Extracts are to be refrigerated at 4"C and analyzed within 40 days
of extraction. See Section 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.
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
3550 with methylene chloride/acetone (1:1) as the extraction solvent.
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, Section 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)
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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 6PC 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.
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
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 jiL are recommended. Hand injections
of no more than 2 /nL 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 /il_ of internal
standard to each ml of sample prior to injection. Chromatograms of the
target organophosphorus compounds are shown in Figures 1 through 4.
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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 /uL 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 Section 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
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 Section 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
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columns including the DB-210 and DB-608 are more prone to oxidation.
Oxidized columns will exhibit baselines that rise rapidly during
temperature programming.
7.7 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.
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
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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%.)
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
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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.
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 Table 7.
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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.
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 & T, 1992, vol 26, 921-925.
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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
Azinphos-methyl
Bolstar (Sulprofos)
Chlorpyrlfos
Coumaphos
Demeton, -0, -S
Diazinon
Dichlorvos (DDVP)
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfothion
Fenthion
Malathion
Merphos
Mevinphos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Ronnel
Sulfotepp
TEPPC
Tetrachl orovi nphos
Tokuthion (Protothiofos)0
Trichloronate0
Reagent
Water (3510)8
(M9A)
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
Soil (3540)b
(Mg/kg)
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 - 17
Revision 1
November 1992
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TABLE 2
DETERMINATION OF ESTIMATED QUANTITATION LIMITS
(EQL) FOR VARIOUS MATRICES"
Matrix Factor6
Ground water (Methods 3510 or 3520) 10
Low-concentration soil by Soxhlet and no cleanup 10°
Low-concentration soil by ultrasonic extraction with GPC cleanup 6.7C
High-concentration soil and sludges by ultrasonic extraction 500C
Non-water miscible waste (Method 3580) 1000°
8 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.
c Multiply this factor times the soil MDL.
8141A - 18 Revision 1
November 1992
-------
TABLE 3.
RETENTION TIMES FOR METHOD 8141A ANALYTES
EMPLOYING 15-m COLUMNS
Capillary Column
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
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
validated
130°C
3 minutes
5eC/min
180°C
10 minutes
2°C/min
250eC
15 minutes
DB-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
for Famphur.
50°C
1 minute
5°C/min
140'C
10 minutes
10eC/min
2408C
10 minutes
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
50°C
1 minute
5°C/min
140°C
10 minutes
10cC/min
240°C
10 minutes
DB-210
10.66
19.35
36.74
8141A - 19
Revision 1
November 1992
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TABLE 4.
RETENTION TIMES FOR METHOD 8141A ANALYTES
EMPLOYING 30-m COLUMNS3
No. Compound
DB-5
RT (min)
DB-210 DB-608
DB-1
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
Trimethyl phosphate
Dichlorvos (DDVP)
Hexamethyl phosphorami de
Trichlorfon
TEPP
Thionazin
Mevinphos
Ethoprop
Diazlnon
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
Monocrotophosc
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 - 20
Revision 1
November 1992
-------
TABLE 4. (Continued)
RT (min)
No. Compound DB-5 DB-210 DB-608 DB-1
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
IS
SU
SU
SU
Phosphamidon
Crotoxyphos
Leptophos
Fensulfothion
EPN
Phosmet
Azinphos-methyl
Azinphos-ethyl
Famphur
Coumaphos
Atrazine
Simazine
Carbophenothion
Dioxathion
Trithion methyl
Dicrotophos
l-Bromo-2-nitrobenzene
Tri butyl phosphate
Triphenyl phosphate
4-C1 -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
e
e
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
d
e
9.07
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
5.40
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
58C/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.
D 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.
e Not available.
8141A - 21 Revision 1
November 1992
-------
TABLE 5.
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
50eC to 140°C at 5°C/min, hold
for 10 minutes, followed by
140eC to 240eC 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 - 22
Revision 1
November 1992
-------
TABLE 6
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 (pm): 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 |iL
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 - 23 Revision 1
November 1992
-------
TABLE 7
RECOVERY OF METHOD 8141A ANALYTES FROM WATER AND SOIL
Compound
Method 3510
Water Spike
% Recovery
Method 3550
Soil Spike
% Recovery (jug/kg)
Azinphos -methyl
Bolstar (Sulprofos)
Chlorpyrifos
Coumaphos
Demeton, -0, -S
Diazinon
Dichlorvos (DDVP)
Dimethoate
Disulfoton
EPN
Ethoprop
Fensulfothion
Fenthion
Malathion
Merphos
Mevinphos
Monocrotophos
Naled
Parathion, ethyl
Parathion, methyl
Phorate
Ronnel
Sulfotepp
TEPP
Tetrachlorovinphos
Tokuthion
(Protothiofos)
Trichloronate
143 + 8
141 + 8
89 + 6
90 + 6
67 + 11
121 + 10
79 + 11
47 + 3
92 + 7
125 + 9
90 + 6
82 + 12
48 + 10
92 + 6
79
55
18 + 4
ND
94 + 5
46 + 4
77 + 6
97 + 5
85 + 4
55 + 72
90 + 7
45 ± 3
35
1.50
1.63
1.56
1.57
2.05
1.74
14.3
1.60
2.16
1.86
1.83
2.07
2.23
1.80
1.71
21
12.7
2.29
2.02
1.92
2.0
1.57
1.99
19.5
1.54
2.08
1.87
27 + 10
103 + 15
79 + 7
60
16
90 + 14
13 + 9
67
44 + 22
86 + 38
34 + 26
37
35
67
71
23
ND
40
74 + 13
17
51 + 9
84 + 8
68 + 10
7
47 + 24
82
31
50
63
52
52
690
58
475
53
72
62
61
69
74
60
57
700
425
76
67
640
69
52
66
651
51
690
620
ND - Not detected
8141A - 24
Revision 1
November 1992
-------
TABLE 8
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 - 25 Revision 1
November 1992
-------
300.00
250.00
200.00
150.00
100.00
50.00
0.00
I
o
K „.
R
S
«
l
1 3 57 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 4!
Figure 1.Chromatogram of target organophosphorus compounds from a 15-m DB-210
column with FPO detector. More compounds are shown in Figure 2. See Table 3 for
retention times and Table 5 for GC operating conditions.
8141A - 26
Revision 1
November 1992
-------
300.00
250.00
200.00
150.00
100.00
50.00
0.00
\.
1 .
a.
UJ
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 and Table 5 for GC operating conditions.
8141A - 27
Revision 1
November 1992
-------
300.00
250.00
200.00-
150.00-
100.00-
50.00-
0.00^
Ill fi
f» i 11 i n | i ifft
1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31 33 35 37 39 41 43 45
Figure S.Chromatogram of target organophosphorus compounds from a 15-m DB-210
column with NPO detector. More compounds are shown in Figure 4. See Table 3 for
retention times and Table 5 for GC operating conditions.
8141A - 28
Revision 1
November 1992
-------
300.00-
250.00-
200.00-
150.00-
100.00 H
50.00 -H
0.00
i 357911 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 and Table 5 for GC operating conditions.
8141A - 29
Revision 1
November 1992
-------
JU
DB-210
11
41
DB-5
i
M
u
Figure B.Chromatogram of target organophosphorus compounds on a 30-m DB-5/OB-210
column pair with NPD detector, without Simazine, Atrazine and Carbophenothion.
See Table 4 for retention times and Table 6 for GC operating conditions.
8141A - 30
Revision 1
November 1992
-------
-r
IS
DB-210
»
J
u
DB-5
Figure S.Chromatogram of target organophosphorus compounds on a 30-m OB-5/DB-210
column pair with NPD detector, with Simazine, Atrazine and Carbophenothion. See
Table 4 for retention times and Table 6 for GC operating conditions.
8141A - 31
Revision 1
November 1992
-------
METHOD 8141A
ORGANOPHOSPHORUS COMPOUNDS BY GAS CHROMATOGRAPHY:
CAPILLARY COLUMN TECHNIQUE
St»rt
7 1 1 Refer to
Chap ter Two for
guidance on
choosing the
appropr la te
ex t racti on
procedure
? 1 2 Perform
solvent exchange
during K - D
procedures in all
extraction methods
7 2 Select GC
cond i t i ons
1
7 3 Refer to Method
8000 for
ca 1 ibrat i on
techniques
731 Internal or
ex terna 1
calibration may be
used
•*
741 Add internal
standard to sample
7 4 2 Refer to
Method 8000 , Step
7 6- for
ins t ructi ons on
analysis sequence,
di 1 uti ons .
r etenti on times ,
and identification
cr i ter la
7 4 3 Inject sample
1
7 4 5 Record sample
volume injected and
resul ting peak
sizes
,
746 Determine
identi ty and
quantity of each
component peak
refer to Method -
8000, Step 7 8 for
ca Icula t ion
equa ti ons
Yes
751 Perform
appropriate cleanup
752 Reanalyze by
CC
Stc
8141A - 32
Revision 1
November 1992
-------
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
Dichlorprop 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. Section 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
hydroxide, and extraneous organic material is removed by a solvent wash. After
8150B - 1 Revision 2
November 1992
-------
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 (Goerlitz 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 Organic acids, especially chlorinated acids, cause the most direct
interference with the determination. Phenols, including chlorophenols, may also
interfere with this procedure.
3.3 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.
3.4 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.
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 (ECD).
8150B - 2 Revision 2
November 1992
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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 Section 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 Section 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 Section 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.
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).
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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 IJ.L.
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 the nearest
0.0001 g.
4.15 Syringe - 5 ml.
4.16 Glass rod.
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 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.
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5.7.3 Isooctane, (CH3)3CCH2CH(CH3)2 - Pesticide quality or equivalent.
5.7.4 Hexane, C6H14 - Pesticide quality or equivalent.
5.7.5 Diethyl Ether, C2H5OC2Hr. 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, 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.
5.9 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.
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 must be replaced after 1 year,
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.
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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 Section
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 according to Section
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,
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,
Section 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 Section 7.2.2
hydrolysis.
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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) of the well mixed, moist solid sample. Adjust the
pH to 2 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
250 mL of 5% acidified sodium sulfate. 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.
7.2.2 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 for a total of 90 minutes.
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.
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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.
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
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.
7.2.3.3 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
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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 Section 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 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
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 for a total of 60 minutes,
beginning from the time the flask is placed in the water bath.
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
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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
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
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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
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:
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 ampule with several hundred
p.1 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).
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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.
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.
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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:
Analvte Column Analvte 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
7.7 Gas chromatographic analysis
7.7.1 Refer to Method 8000. If the internal standard calibration
technique is used, add 10 juL 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 chloro-phenoxy
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.
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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 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.
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.
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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.
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. 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.
5. "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.
6. Eichelberger, J.W., L.E. Harris, and W.L. Budde, "Reference Compound to
Calibrate Ion Abundance Measurement in Gas Chromatography-Mass
Spectrometry," Analytical Chemistry, 47, 995, 1975.
7. Glaser, J.A. et.al., "Trace Analysis for Wastewaters," Environmental
Science & Technology, 15, 1426, 1981.
8. 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 - 15 Revision 2
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND DETECTION LIMITS
FOR CHLORINATED HERBICIDES
Compound
2,4-D
2,4-DB
2,4,5-T
2,4,5-TP (Silvex)
Dalapon
Dicamba
Dichloroprop
Dinoseb
MCPA
MCPP
Retention
Col. la Col.lb
2.0
4.1
3.4
2.7
-
1.2
-
-
-
-
-
-
-
-
-
4.8
11.2
4.1
3.4
time (min)a
Col. 2 Col. 3
1.6
-
2.4
2.0
5.0
1.0
-
-
-
- -
Method
detection
limit (MQ/L)
1.2
0.91
0.20
0.17
5.8
0.27
0.65
0.07
249
192
aColumn conditions are given in Sections 4.1 and 7.5.
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION
LIMITS (EQL) FOR VARIOUS MATRICES3
Matrix
Factor0
Ground water (based on one liter sample size)
Soil/secliment and other solids
Waste samples
10
200
100,000
aSample EQLs are highly matrix dependent. The EQLs listed herein are provided for
guidance and may not always be achievable.
bEQL = [Method detection limit (Table 1)] X [Factor (Table 2)]. For non-aqueous
samples, the factor is on a wet weight basis.
8150B - 16
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TABLE 3.
SINGLE OPERATOR ACCURACY AND PRECISION8
Compound
2,4-D
Dalapon
2,4-DB
Dicamba
Dichlorprop
Dlnoseb
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
(M9/L)
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 9.
DW = ASTM Type II
MW = Municipal water
8150B - 17
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FIGURE 1.
DIAZOMETHANE GENERATOR
nitrogen
rubber (topper
gloss tubing
tube 1
tube 2
8150B - 18
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FIGURE 2.
GAS CHROMATOGRAM OF CHLORINATED HERBICIDES
Column: 1.5% SP-2250/1.9S* SP-2401 on Suptleoooa (100/120 M«h)
Temperature: Isothermal at 18S°C
Dtttetor: Electron Capture
0 12346
RETENTION TIME (MINUTES)
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FIGURE 3.
GAS CHROMATOGRAM OF CHLORINATED HERBICIDES
Column: 1.5% SP-2250/1.95% SP-2401 on Suptlcoport (100/120 Mt*)
Program: 140°C for 6 Min, 10°C/Minut» to 200°C
Dttcctor: Electron Capturt
468
RETENTION TIME (MINUTES)
10
12
8150B - 20
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FIGURE 4.
GAS CHROMATOGRAM OF DALAPON, COLUMN 3
Column: 0.1% SP-1000 on 80/100 M«sh Ccrfaoprt C
*o«ram: 100°C. 10°C/Min to 180°C
Dractor: Electron Capture
0246
RETENTION TIME (MINUTES)
8150B - 21
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METHOD 81SOB
CHLORINATED HERBICIDES BY GAS CHROMATOGRAPHY
7211
Adjust sampli
pH with HC1
Liquid
sample
7212 Extract
sample three
times with
acetone and
diethyl ether
7214
Combine
extracts
7215 Check
pH of extract,
adjust if
necessary,
Separate layers
7215
Re-extract
and discard
aqueous
phase
7 2 2 Proceed
with
hydrolysis
723 Proceed
with solvent
cleanup
7111 Follow
Method 3S80 for
extraction, using
diethyl ether,
acidified anhydrous
sodium sulfate and
acidified gl.
wool
7 1 1 2 Usi
1 0 mL of
sample for
hydrolysis
V • A
731 Extract
three times
with diethyl
ether
7313
Combine
extracts
732 Proceed
with
hydrolysis
733 Proceed
with solvent
cleanup
8150B - 22
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METHOD 8150B
(Continued)
743 Assembe
diazomethane
bubbler;
generate
diazomethane
7 4
Choose
method for
eaterification
742 Prepare
diazomethane
according to
kit
instructions
7 5 Set
chromatographic
conditions
7 6 Claibrate
according to
Method 8000
762 Choose
appropriate
GC column
7 7 Analyze
by CC (refer
to MEthod
8000)
7 7 7 Do
interference*
prevent peak
detection''
777 Process
series of
standards
through system
cleanup
8150B - 23
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METHOD 8151
CHLORINATED HERBICIDES BY 6C USING METHYLATION OR PENTAFLUOROBENZYLATION
DERIVATIZATION: CAPILLARY COLUMN TECHNIQUE
.0 SCOPE AND APPLICATION
1.1 Method 8151 is a capillary gas chromatographic (GC) method for
etermining certain chlorinated acid herbicides in aqueous, soil and waste
atrices. Specifically, Method 8151 may be used to determine the following
:ompounds:
Compound Name CAS No.1
Acifluorfen
Bentazon
Chloramben
2,4-D
Dalapon
2,4-DB
DCPA diacidb
Dicamba
3,5-Dichlorobenzoic acid
Dichlorprop
Dinoseb
5-Hydroxydicamba
MCPA
MCPP
4-Nitrophenol
Pentachlorophenol
Picloram
2,4,5-TP (Silvex)
2,4,5-T
50594-66-6
25057-89-0
133-90-4
94-75-7
75-99-0
94-82-6
2136-79-0
1918-00-9
51-36-5
120-36-5
88-85-7
7600-50-2
94-74-6
93-65-2
100-02-1
87-86-5
1918-02-1
93-72-1
93-76-5
a 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.
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
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technique. Section 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 aqueou<
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.
1.4 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.
1.5 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.
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
limits of detection 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
8151 - 2 Revision 0
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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.
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.
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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 /xm 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 urn 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 urn 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 p.m film thickness.
4.1.2.2 Megabore Columns
4.1.2.2.1 Primary Column - 30 m x 0.53 mm DB-608 (J&W
Scientific or equivalent) with 0.83 /xm 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 /xm 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.
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 Section 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).
8151 - 4 Revision 0
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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
Section 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.
4.9 Balance - Analytical, capable of accurately weighing to the nearest
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.
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4.12 Sonabox - Recommended with above disrupters for decreasing cavitation
sound (Heat Systems - Ultrasonics, Inc., Model 432B or equivalent).
4.13 Filter paper - Whatman #1, or equivalent.
4.14 pH paper.
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 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
(NaHpPOJ 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.
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.
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5.11 Solvents
5.11.1
equivalent.
5.11.2
5.11.3
5.11.4
5.11.5
Methylene chloride, CH?C1.
Pesticide quality or
Acetone, CH3COCH3. Pesticide quality or equivalent.
Methanol, CH3OH. Pesticide quality or equivalent.
Toluene, C6H5CH3. Pesticide quality or equivalent.
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.
5.11.6
equivalent.
Isooctane, (CH3)3CH2CH(CH3)2. Pesticide quality or
5.11.7 Hexane, C6H14. Pesticide quality or equivalent.
5.11.8
5.11.9
Ethanol, absolute.
C2H5OH
Carbitol
C2H5OCH2CH2OCH2CH20 - optional
(diethylene glycol
for producing alcohol
monoethyl ether),
free diazomethane.
5.12 Stock standard solutions (1000 mg/L) - Can be prepared from pure
standard materials or can be purchased as certified solutions. Commercially
prepared stock standards can be used if they are verified against EPA standards.
If EPA standards are not available for verification, then standards certified by
the manufacturer and verified against a standard made from pure material is
acceptable.
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
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.
8151 - 7
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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 /xL of the
internal standard spiking solution to 10 ml of sample extract results in
a final internal standard concentration of 0.25 M9/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. 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 Section 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
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 juL 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.
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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,
Section 4.1.
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 sulfate, and acidified glass
wool,
• spike the sample with surrogate compound(s) according to
Section 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 Section 7.2.1.8. If the
analysis is for acid herbicides only, proceed to Section 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 Section
7.4.2 and then dilute to 4 mL with acetone).
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) 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 Section 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
8151 - 9 Revision 0
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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 texture, 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 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
Section 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 Section 7.4.4 - Nitrogen
Blow Down.
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 for 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.
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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
Section 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 Section 7.4, Extract
concentration. If additional cleanup is required, proceed to
Section 7.2.1.9.
7.2.1.9 Use this step if additional clean up of the non-
hvdrolyzed herbicides is required:
7.2.1.9.1 Partition the herbicides by extracting the
methylene chloride 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 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.
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
Section 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.
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7.3 Preparation of Aqueous Samples
7.3.1 Separatory Funnel
7.3.1.1 Using a graduated cylinder, measure out a 1 liter
of sample and transfer it into a 2 L separatory funnel. Spike the
sample with surrogate compound(s) according to Section 5.16.1.
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 for 1 hour, shaking the separatory funnel and
contents periodically.
7.3.1.3.2 Add 60 ml of methylene chloride to the
sample bottle to rinse the bottle. 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.
7.3.1.3.3 Add a second 60 ml volume of methylene
chloride to the sample bottle 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
8151 - 12 Revision 0
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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 round bottom flask and funnel with
20 to 30 ml of diethyl ether to complete the quantitative transfer.
Proceed to section 7.4 for extract concentration.
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 (Section 7.4.3) or nitrogen blowdown technique (Section
7.4.4).
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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 di ethyl ether and add to the concentrator tube.
Proceed to Section 7.4.5.
7.4.4 Nitrogen Slowdown 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
operating conditions, the extract should not be allowed to become
dry. Proceed to Section 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 Section
7.5.1. For PFB derivatization proceed with Section 7.5.2.
7.5.1 Diazomethane Derivatization - Two methods may be used for the
generation of diazomethane: the bubbler method (see Figure 1), Section
7.5.1.1, and the Diazald kit method, Section 7.5.1.2.
CAUTION; Diazomethane is a carcinogen and can explode under
certain conditions.
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
8151 - 14 Revision 0
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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 Bubbler method - Assemble the diazomethane bubbler
(see Figure 1).
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.
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.
7.5.1.1.2 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 diazomethane 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. 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.
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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. Analyze by gas chromatography.
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
ML 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.
7.5.2 PFB Method
7.5.2.1 Add 30 ML of 10% K2C03 and 200 jtiL 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.
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 the silica column 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.
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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/min
Helium carrier flow: 30 cm/sec
Injection volume: 2 /zL, 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 /xL, 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 p,L, splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
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 pi, splitless, 45 sec delay
Injector temperature: 250°C
Detector temperature: 320°C
7.6.2 Megabore
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 juL
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 /zL
8151 - 17 Revision 0
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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 pi 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
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.
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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.
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.
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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.
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.
8151 - 20 Revision 0
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Figure 1
DIAZOMETHANE GENERATOR
nitrogen
rubber stopper
tub* 1
tube 2
8151 - 21
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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 diacid6
Dicamba
3,5-Dichlorobenzoic acid
Dichlorprop
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 Samoles
GC/ECD
Estimated
Detection
Limit"
(Mg/kg)
4.0
0.11
0.12
0.38
66
43
0.34
0.16
0.28
GC/MS
Estimated
Identification
Limit6
(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 /iL injected.
Chromatography using narrow bore capillary column, 0.25 pm 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.
40 CFR Part 136, Appendix B (49 FR 43234).
capillary column.
Chromatography using megabore
DCPA monoacid and diacid metabolites included in method scope; DCPA diacid
metabolite used for validation studies. DCPA is a dimethyl ester.
8151 - 22
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TABLE 2
RETENTION TIMES (MINUTES) OF METHYL DERIVATIVES OF CHLORINATED HERBICIDES
Narrow Bore Columns
Megabore Columns
Analyte
Primary8 Confirmation8 Primary6 Confirmation1*
Column Column Column Column
Dalapon
3,5-Dichlorobenzoic acid
4-Nitrophenol
DCAA (surrogate)
Dicamba
Dichlorprop
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
8151 - 23
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TABLE 2 (continued)
Primary Column:
Confirmation Column:
Temperature program:
Helium carrier flow:
Injection volume:
Injector temperature:
Detector temperature:
b 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/"rin
7 mL/min
1 ML
c DCPA monoacid and diacid metabolites included in method scope; DCPA diacid
metabolite used for validation studies. DCPA is a dimethyl ester.
8151 - 24
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TABLE 3
RETENTION TIMES (MINUTES) OF PFB DERIVATIVES OF CHLORINATED HERBICIDES
Gas chromatographic column
Herbicide
Thin-film DB-5a
SP-2250"
Thick-film DB-5C
Dalapon
MCPP
Dicamba
MCPA
Dichlorprop
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
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.
SP-2550 capillary column, 0.25 ^m 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.
DB-5 capillary column, 1.0 ^m 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 - 25
Revision 0
November 1992
-------
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 diacid6
Dlcamba
3,5-Dichlorobenzoic Acid
Dichlorprop
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 - 26
Revision 0
November 1992
-------
TABLE 5
ACCURACY AND PRECISION FOR METHOD 8151
DIAZOMETHANE DERIVATIZATION, CLAY MATRIX
Analyte
Mean8
Percent Recovery
Linear
Concentration
Range
(ng/g)
Percent
Relative0
Standard Deviation
(n-20)
Dicamba
MCPP
MCPA
Dichlorprop
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 on standard solutions and
corrected to 50 g solid samples.
Percent relative standard deviation was calculated on standard solutions,
10 samples high in the linear concentration range, and 10 samples low in
the range.
8151 - 27
Revision 0
November 1992
-------
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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 derrvatizatian
is by diazomelhane
orPFB
7.2.1.1 Weigh sample
and add to beaker;
add add and spike;
mix we*.
LYES
7.2.1.8.1 AddKOHand
water. Reflux tor 2 hrs.
Allow 10 cool.
-------
METHOD 8151
(continued)
Extraction/Hydrolysis of Aqueous Samples and Extract Concentration
7.3.1.1 Measure 1L of
sample and transfer to
a 2Lsep. funnel.
7.3.1.2 Add250gNaCI
to sample and shake
to dissolve
7.3.1.4 Add12Nsulfuric
acid and shake. Add
until pH < 2
7.3.1.3
Does analysis
include heoictde esters?
i
7.3.1.5 Adddiethyl
ether to sample and
extract Save both
phases
7.3.1.3.1 Add6NNaOHto
sample and shake. Add
until pH> 12. Let stand
1hr.
Employ mechanical techniques
to complete phase separation
(e.g. stirring, filtration through
glass wool, centrifugation, or
other physical methods).
Save both phases.
I
7.3.1.3.2 AddMedand
extract by shaking tor
2min. Discard MeCI.
7.3.1.6 Return aqueous phase
to separately funnel and repeat
extraction 2 more times, combine
extracts, and allow extract to
remain in contact with sodium
sulfate for 2 hrs.
7.3.1.7 Pour extract
through glass wool and
proceed to step 7.4.1
Does
difficult
emulsion form
> 1/3 solvent
volume?
Employ mechanical techniques
to complete phase separation
(e.g. stirring, filtration through
glass wool, centrifugafion, or
other physical methods).
Discard Mod.
7.4.1 Place K-D apparatus
in water bath, concentrate
and cool
I
7.4.2 - 7.4.4 Complete
concentration with micro-
Snyder column or nitrogen
blow down.
7.3.1.3.3 Repeat
extraction twice more.
Discard MeCI.
7.4.5 Dilute extract
with 1 ml isooctane and
0.5 ml methanol
8151 - 30
Revision 0
November 1992
-------
METHOD 8151
(continued)
Extract Derivatization
7.4.5 Dilute extract
to 4 ml with acetone
7.5.2.1 Add potassium
carbonate and PFBBr.
Close tube, mix & heat
7.4.5 Dilute extract
to 4 ml with diethyl
ether
7.5.1.1 Assemble the
diazomethane bubbler
(Figure 1)
Oiazald
Kit
7.5.2.2 Evaporate with
nitrogen to 0.5 ml. Add
2 ml hexane and repeat
7.5.2.3 Redissotve the
residue in 2 ml toluene:
hexane (1 :6)
7.5.1.1.1 Add 5 ml to 1st test
tube. Add 1 ml diethyl ether,
1 ml carbitol, 1.5 ml of 37% KOH
and 0.1 - 0.2 g Diazald to the
2nd tube. Bubble with nitrogen
for 10 min or until yellow persists
7.5.2.4 Load sodium
sulfate / silica cleanup
column with residue.
1
.5.1
WHIthe
Bubbler or the
DiazaldKit
metodbe
used?
7.5.1.2 Read kit
instructions
7.5.1.1.2 Remove con-
centrate* lube and seal
it. Store at room temp.
7.5.2.5 Bute column
with enough toluene:
hexane to collect 8ml
eluant
I
7.5.1.2.1 Add 2 ml
diazomethane solution.
Let stand for 10 min
and swirl
7.5.1.1.3 Add silicic acid to
concentrator tube and let stand
until nitrogen evolution has
stopped. Adjust sample volume
to 10 ml with hexane. Stopper.
Immediate analysis is recommended
7.5.1.2.2 Rinse ampule with
diethyl ether and evaporate
to 2 ml to remove diazomethane
Alternatively, silicic acid
may be added.
7.5.2.6 Discard 1st fraction
and continue etutton with
enough toluene: hexane (1 :9)
to collect 8 ml more eluant
Transfer to a 10 ml volumetric
flask and dilute to the mark
with hexane
7.5.1.1.5 If necessary
store at 4 C in the dark
for a max of 28 days.
I
I
7.5.1.2.3 Dilute sample
to 10ml with hexane
7.6.1 & 7.6.2 Set GC
conditions
8151 - 31
Revision 0
November 1992
-------
METHOD 8151
(continued)
Analysis by Gas Chromatography
0
7.7 Internal or external
calibration may be used
(See method 8000).
7.8.1 Add 10 ul internal
standard to the sample
prior to Injection.
NO
7.8.2 See method 8000 for
analysis sequence, appropriate
dilutions, establishing daHy
retention time windows, and
identification criteria. Check
stds every 10 samples.
7.8.4 Record volume
injected and the resulting
peak sizes.
7.8.5 Determine the
identity and quantify
component peaks.
Calculate the correction
for molecular weight of
methyl ester vs herbicide
7.8.6
Have stds
and samples
been prepared and
analyzed the
way?
7.8.6 Calculate con-
centration using procedure
In Method 8000.
7.8.7 Perform further
cleanup if necessary
1
'
8151 - 32
Revision 0
November 1992
-------
METHOD 8240B
VOLATILE QRGANICS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/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:
Analyte
CAS No.1
Appropriate Technique
Direct
Purge-and-Trap Injection
Acetone
Acetonitrile
Acrolein
Acrylonitrile
Allyl alcohol
Ally! chloride
Benzene
Benzyl chloride
Bromoacetone
Bromochloromethane (I.S.)
Bromodi chl oromethane
4-Bromofluorobenzene (surr.)
Bromoform
Bromomethane
2-Butanone
Carbon disulfide
Carbon tetrachloride
Chloral hydrate
Chlorobenzene
Chlorobenzene-dc (I.S.)
Chl orodi bromomethane
Chloroethane
2-Chloroethanol
bis-(2-Chloroethyl) sulfide
2-Chloroethyl vinyl ether
Chloroform
Chl oromethane
Chloroprene
3-Chloropropionitrile
l,2-Dibromo-3-chloropropane
1,2-Dibromoethane
Di bromomethane
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
74-95-3
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
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
pc
a
a
a
8240B - 1
Revision 2
November 1992
-------
Appropriate Technique
Analyte
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
cis-l,3-Dichloropropene
trans -1,3-Di chl oropropene
1,2,3,4-Diepoxybutane
1,4-Difluorobenzene (I.S.)
1,4-Dioxane
Epichlorohydrin
Ethanol
Ethyl benzene
Ethyl ene 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-Picoline
Propargyl alcohol
6-Propiolactone
Propionitrile
n-Propylamine
Pyridine
Styrene
1,1,1 , 2-Tetrachl oroethane
1,1,2, 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
Toluene-d8 (surr.)
1,1,1-Trichloroethane
1,1,2-Trichloroethane
Trichl oroethene
Tri chl orof 1 uoromethane
1,2,3-Trichloropropane
CAS No.b
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
96-18-4
Purge-and-Trap
PP
a
a
a
a
a
a
a
PP
a
a
a
a
PP
i
i
a
PP
a
PP
ND
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
a
Direct
Injection
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
a
8240B - 2
Revision 2
November 1992
-------
Appropriate Technique
Direct
Analyte CAS No. Purge-and-Trap Injection
Vinyl acetate
Vinyl chloride
Xylene (Total)
108-05-4
75-01-4
1330-20-7
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.
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 /xg/kg (wet weight) for soil/sediment
samples, 0.5 mg/kg (wet weight) for wastes, and 5 /zg/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 Method 8240 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 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
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.
8240B - 3 Revision 2
November 1992
-------
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.
4.0 APPARATUS AND MATERIALS
4.1 Microsyringes - 10 /xL, 25 /LtL, 100 juL, 250 jzL, 500 )uL, and 1,000 pi.
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
8240B - 4 Revision 2
November 1992
-------
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 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 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
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
8240B - 5 Revision 2
November 1992
-------
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
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
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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 /xL 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
Bottle Septum (#86600). 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
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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 standards every two months for gases. 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 25% difference.
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 !,2-dich1oroethane-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 Section 5.3, and a
surrogate standard spiking solution should be prepared from the stock at a
concentration of 250 Mg/10 ml in methanol. Each water sample undergoing GC/MS
analysis must be spiked with 10 /ul_ 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
Sections 5.3 and 5.4. 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 /xL of this standard to 5.0 ml of sample or calibration
standard would be the equivalent of 50
5.7 4-Bromofluorobenzene (BFB) standard - A standard solution containing
25 ng/juL 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 Sections 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 (e.g. some or all of the target analytes may be
included). Calibration standards must be prepared daily.
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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 M9/10.0 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
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,
Section 4.1.
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7.0 PROCEDURE
7.1 Direct injection - In very limited applications (e.g. aqueous
process wastes), direct injection of the sample into the GC/MS system with a 10
ML 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 MgA);
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).
7.2 Initial calibration for purge-and-trap procedure
7.2.1 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 7 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.
7.2.2 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 \i\.
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 Section 4.11. Condition the trap overnight at 180°C in the purge mode
with an inert gas flow of at least 20 mL/tnin. 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 jiL or 25 jxL microsyringe equipped with
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a long needle (Section 4.1), take a volume of the secondary dilution
solution containing appropriate concentrations of the calibration
standards (Section 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 ^L of
the internal standard solution (Section 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 Section 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
has a retention time closest to the compound being measured (Section
7.5.2). The RF is calculated as follows:
RF « (AxC(.)/(AuCx)
where:
Ax = Area of the characteristic ion for the compound being
measured.
Ajs = Area of the characteristic ion for the specific internal
standard.
Cu = Concentration of the specific internal standard.
Cx = Concentration of the compound being measured.
7.2.8 The average RRF must be calculated for each compound. 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 RRF
for these compounds should be 0.300 (0.250 for bromoform). These
compounds typically have RRFs 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.
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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 RRFs from the initial calibration, calculate and I
record the percent relative standard deviation (%RSD) for all compounds.
The percent RSD is calculated as follows:
SD
%RSD = ^r~ x 100
RF
where:
RSD = relative standard deviation.
RF = mean of 5 initial RRFs for a compound.
SD = standard deviation of average RRFs for a compound.
SD =
N (x, - x)2
I
i-1 N - 1
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-Di chloropropane,
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 (Section 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 second order regression fit of the five
calibration points. The analyst should select the regression order
which introduces the least calibration error into the quantitation
(Section 7.5.3). If the %RSD is <15%, 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.
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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-brotnofluorobenzene 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.
7.3.2 The initial calibration curve (Section 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 by checking the SPCC (Section 7.3.3) and CCC (Section 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
(0.250 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 Section 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
C,
where:
C, = Calibration Check Compound standard concentration.
Cc = Measured concentration using selected quantisation 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.
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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 check calibration (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 standard check, 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 are 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 BCD (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 Section
7.2.1.
7.4.1.4 BFB tuning criteria and daily GC/MS calibration
criteria must be met (Section 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
Section 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
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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.
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 Section 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 Section 7.4.1.6.
7.4.1.8 Add 10.0 juL of surrogate spiking solution (Section
5.3) and 10 pi of internal standard spiking solution (Section 5.4)
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 ^l of the surrogate spiking
solution to 5 ml of sample is equivalent to a concentration of
50 jLtg/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.
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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 mi flushes of organic-free reagent water (or
methanol followed by organic-free reagent water) to avoid carryover
of pollutant compounds into subsequent analyses.
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 /*L of the matrix
spike solution (Section 5.7) to the 5 ml of sample to be purged.
Disregarding any dilutions, this is equivalent to a concentration
of 50 M9/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 Sections 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 jixL, but not more than 100 /LtL of liquid sample. The sample is
ready for addition of internal and surrogate standards.
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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
extensive cleanup and instrument downtime. Use the screening data to
determine whether to use the low-concentration method (0.005-1 rng/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
Sections 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 water. Replace the plunger and compress the
water to vent trapped air. Adjust the volume to 5.0 ml. Add
10 ML each of surrogate spiking solution (Section 5.3) and
internal standard solution (Section 5.4) to the syringe
through the valve. (Surrogate spiking solution and internal
standard solution may be mixed together.) The addition of 10
/it of the surrogate spiking solution to 5 g of sediment/soil
is equivalent to 50 Mg/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 Section 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
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amenable to percent dry weight determination. Other wastes
should be reported on a wet-weight basis.
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 = q of drv 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 Sections 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
Sections 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 (Section 5.7) to the 5 ml
of organic-free reagent water (Section 7.4.3.1.3). The
concentration for a 5 g sample would be equivalent to 50
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.
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This is purged at ambient temperature. All samples with an expected
concentration of > 1.0 mg/kg should be analyzed by this method.
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 Section 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. Pipet 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: Sections 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 juL aliquot of each of these extracts in Section
7.4.3.2.6 will give a concentration equivalent to 6,200 MQ/kg
of each surrogate standard.
7.4.3.2.4 The GC/MS system should be set up as in
Sections 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
8240B - 19 Revision 2
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ML. All dilutions must keep the response of the major
constituents (previously saturated peaks) in the upper half
of the linear range of the curve.
7.4.3.2.6 Remove the plunger from a 5.0 ml Luerlock
type syringe equipped with a syringe valve and fill until
overflowing with 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 juL
of internal standard solution. Also add the volume of
solvent extract determined in Section 7.4.3.2.5 and a volume
of extraction or dissolution solvent to total 100 i*L
(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
Section 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 /xL 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 (Section 5.3), and 1.0 mL of matrix
spike solution (Section 5.7) as in Section 7.4.3.2.2. This
results in a 6,200 M9/kg concentration of each matrix spike
standard when added to a 4 g sample. Add a 100 /xL aliquot of
this extract to 5 mL of organic-free reagent water for
purging (as per Section 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
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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.
Sufficient GC resolution is achieved if the height of the
valley between two isomer peaks is less fyian 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%).
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(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.
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 Section 7.2.10,
calculate the concentration of each identified analyte in the sample
as follows:
Water
(AJ(IJ
concentration (M9/L) =
(A,.)(RRF)(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.
RRF = Relative Response factor for compound being
measured (Section 7.3.3).
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)
(Ax)ds)(Vt)
concentration (jug/kg) =
where:
Ax, Is, Ajs, RRF, = Same as for water.
Vt = Volume of total extract (/iL) (use 10,000 /xL 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 A and Ajs 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.
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 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 calibration 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 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.
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
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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 Step 7.2.2.
8.4.2 There must be an initial calibration of the GC/MS system as
specified in Step 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 Step 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 /zg/L of each
analyte by adding 200 /iL 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 m
analyzed according to the method beginning in Step 7.4.1. ^
8.5.4 Calculate the average recovery (x) in Mg/L, and the standard
deviation of the recovery (s) in Mg/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
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 Step
8.5.6.1 or 8.5.6.2.
8240B - 24 Revision 2
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8.5.6.1 Locate and correct the source of the problem and
repeat the test for all analytes beginning with Section 8.5.2.
8.5.6.2 Beginning with Section 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 Section
8.5.2.
8.6 The laboratory must, on an ongoing basis, analyze a reagent 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 Step 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 yg/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 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 (Step 8.5.1) appropriate for the background
concentration in the sample. Spike a second 5-mL sample aliquot with 10
\ii 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 20 |ig/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
8240B - 25 Revision 2
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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 Step 8.7.
8.7 If any analyte in a water sample fails the acceptance criteria for
recovery in Step 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 Step 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 \il of the QC
reference sample concentrate (Step 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 Step 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 m
corresponding QC acceptance criteria found in Table 6. Only analytes that
failed the test in Step 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 Section 8.6, 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 + 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.
8240B - 26 Revision 2
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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 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 a different ionization mode using 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 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 M9/L. Single operator precision, overall
precision, and method accuracy were found to be directly related to the
8240B - 27 Revision 2
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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.
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 - 28 Revision 2
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TABLE 1.
RETENTION TIMES AND CHARACTERISTIC IONS FOR VOLATILE COMPOUNDS
Compound
Retention
Time (minutes)
Primary Ion Secondary Ion(s)
Ethyl ene oxide
Chloromethane
Di chl orodi f 1 uoromethane
Bromomethane
Vinyl chloride
Acetonitrile
Chloroethane
Methyl iodide
Methylene chloride
Carbon disulfide
Tr i chl orof 1 uoromethane
Propionitrile
Ally! chloride
1,1-Dichloroethene
Bromochloromethane (I.S.)
Allyl alcohol
trans- 1 , 2-Di chl oroethene
1,2-Dichloroethane
Propargyl alcohol
Chloroform
1 ,2-Dichloroethane-d4(surr)
2-Butanone
Methacrylonitrile
Dibromomethane
2-Chloroethanol
b-Propiolactone
Epichlorohydrin
1 , 1 , 1-Tri chloroethane
Carbon tetrachloride
1,4-Dioxane
Isobutyl alcohol
Bromodi chl oromethane
Chloroprene
l,2:3,4-Diepoxybutane
1 ,2-Dichloropropane
Chloral hydrate (b)
cis-l,3-Dichloropropene
Bromoacetone
Trichloroethene
Benzene
trans -1,3-Di chl oropropene
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 - 29
Revision 2
November 1992
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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 methacrylate
Bromoform
1,1,1 , 2-Tetrachloroethane
l,3-Dichloro-2-propanol
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
1,2,3-Trichloropropane
l,4-Dichloro-2-butene
n-Propylamine
2-Picoline
Toluene
Ethyl methacrylate
Chlorobenzene
Pentachl oroethane3
Ethyl benzene
l,2-Dibromo-3-chloropropane
4-Bromofluorobenzene (surr.)
Benzyl chloride
Styrene
bis-(2-Chloroethyl) sulfide(b)
Acetone
Acrolein
Acrylonitrile
Chlorobenzene-d- (I.S.)
Chi orodi bromomethane
1,1-Dichloroethane
Ethanol
2-Hexanone
lodomethane
4-Methyl -2-pentanone
Toluene-d? (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.
8240B - 30
Revision 2
November 1992
-------
TABLE 2.
ESTIMATED QUANTITATION LIMITS (EQL) FOR VOLATILE ORGANICS8
Estimated
Quantitation
Limits"
Ground water
Volatiles M9/L
Acetone
Acetonitrile
Allyl chloride
Benzene
Benzyl chloride
Bromodi chl oromethane
Bromoform
Bromomethane
2-Butanone
Carbon disulfide
Carbon tetrachloride
Chlorobenzene
Chl orodi bromomethane
Chloroethane
2-Chloroethyl vinyl ether
Chloroform
Chl oromethane
Chloroprene
1 ,2-Dibromo-3-chloropropane
1,2-Dibromoethane
Di bromomethane
l,4-Dichloro-2-butene
Di chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1 Dichloroethene
trans-l,2-Dichloroethene
1,2-Dichloropropane
cis-l,3-Dichloropropene
trans-l,3-Dichloropropene
Ethyl benzene
Ethyl methacrylate
2-Hexanone
Isobutyl alcohol
Methacryl oni tr i 1 e
Methylene 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 - 31 Revision 2
November 1992
-------
TABLE 2.
(Continued)
Estimated
Quantitation
Limits6
Ground water Low Soil/Sediment
Volatiles /ug/L M9/kg
Propionitrile
Styrene
1,1,1, 2-Tetrachl oroethane
1,1,2, 2-Tetrachl oroethane
Tetrachloroethene
Toluene
1 , 1 , 1-Trichloroethane
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. See the following information j
for further guidance on matrix dependent EQLs. I
b EQLs listed for soil/sediment are based on wet weight. Normally data is
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
°EQL = [EQL for low soil sediment (Table 2)] X [Factor]. For non-aqueous
samples, the factor is on a wet weight basis.
8240B - 32 Revision 2
November 1992
-------
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 Extract8
500- 10,000 Mg/kg 100 /xL
1,000- 20,000 M9/kg 50 pi
5,000-100,000 M9/kg 10 /iL
25,000-500,000 M9/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 p.1 added to the syringe.
b Dilute and aliquot of the methanol extract and then take 100 nl for
analysis.
8240B - 33 Revision 2
November 1992
-------
TABLE 5.
VOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES ASSIGNED
FOR QUANTITATION
Bromochloromethane
Acetone
Acrolein
Acrylonitrile
Bromomethane
Carbon disulfide
Chloroethane
Chloroform
Chioromethane
Dichlorodifluoromethane
1,1-Dichloroethane
1,2-Dichloroethane
l,2-Dichloroethane-d4 (surrogate)
1,1-Dichloroethene
trans-1,2-Dichloroethene
lodomethane
Methylene chloride
Tri chl orof1uoromethane
Vinyl chloride
1.4-Difluorobenzene
Benzene
Bromodichloromethane
Bromoform
2-Butanone
Carbon tetrachloride
Chlorodi bromomethane
2-Chloroethyl vinyl ether
Dibromomethane
l,4-Dichloro-2-butene
1,2-Dichloropropane
cis-l,3-Dichloropropene
trans-1,3-Di chloropropene
1,1,1-Tri chloroethane
1,1,2-Trichloroethane
Trichloroethene
Vinyl acetate
Chlorobenzene-de
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-Tri chloropropane
Xylene
8240B - 34
Revision 2
November 1992
-------
TABLE 6.
CALIBRATION AND QC ACCEPTANCE CRITERIA8
Parameter
Benzene
Bromodi chl oromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
2-Chloroethylvinyl ether
Chloroform
Chl oromethane
Di bromochl oromethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
1,1-Dichloroethane
1,2-Dichl oroethane
1,1-Dichloroethene
trans- 1 , 2-Dichl oroethene
1 ,2-Dichloropropane
cis-l,3-Dichloropropene
trans- 1 , 3-Di chl oropropene
Ethyl benzene
Methyl ene chloride
1 , 1,2,2-Tetrachloroethane
Tetrachl oroethene
Toluene
1,1,1 -Tri chl oroethane
1 , 1 , 2-Tr i chl 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
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
Range
for x
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
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
x =
p, ps =
D =
Concentration measured in QC check sample, in M9/L.
Standard deviation of four recovery measurements, in ng/L.
Average recovery for four recovery measurements, in /xg/L.
Percent recovery measured.
Detected; result 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 Mg/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 - 35
Revision 2
November 1992
-------
TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Parameter
Accuracy, as Single analyst Overall
recovery, x' precision, s ' precision,
(Mg/L) ()ug/L) S' (M9/L)
Benzene
Bromodichloromethane
Bromoform
Bromomethane
Carbon tetrachloride
Chlorobenzene
Chl oroethane
2-Chloroethyl vinyl ether8
Chloroform
Chloromethane
Di bromochl oromethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
1,1-Dichloroethane
1,2-Dichloroethane
1,1 -Di chl oroethene
trans - 1,2, -Di chl oroethene
1 , 2-Di chl oropropane8
cis-l,3-Dichloropropenea
trans-l,3-Dichloropropene8
Ethyl benzene
Methyl ene chloride
1,1,2 , 2-Tetrachl oroethane
Tetrachl oroethene
Toluene
1,1,1 -Tri chl oroethane
1,1,2-Trichloroethane
Trichloroethene
Trichlorofluoromethane
Vinyl chloride
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
0.26x-1.74
O.lBx+0.59
0.12X+0.34
0.43x
0.12X+0.25
0.16X-0.09
0.14x+2.78
0.62x
0.16x+0.22
0.37x+2.14
0.17X-0.18
0.22X-1.45
0.14X-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
O.lSx+1.07
0.16X+0.69
0.13x-0.18
O.lSx-0.71
0.12x-0.15
0.14x+0.02
0.13X+0.36
0.33X-1.48
0.48x
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
0.18X+0.16
0.58X+0.43
0.17x+0.49
0.30x-1.20
O.lSx-0.82
0.30X-1.20
O.lSx+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 M9/L.
sr' = Expected single analyst standard deviation of measurements at an
average concentration of x, in Mg/L.
S' = Expected interlaboratory standard deviation of measurements at an
average concentration found of x, in M9/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 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 - 36
Revision 2
November 1992
-------
TABLE 8.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Low/High Low/High
Surrogate Compound Water So11/Sediment
4-Bromofluorobenzene 86-115 74-121
1,2-Dichloroethane-d, 76-114 70-121
Toluene-d8 88-110 81-117
8240B - 37 Revision 2
November 1992
-------
FIGURE 1.
PURGING CHAMBER
GOT m m OO
00
S STAMUS8 »THl
8240B - 38
Revision 2
November 1992
-------
FIGURE 2.
TRAP PACKINGS AND CONSTRUCTION TO INCLUDE
DESORB CAPABILITY FOR METHOD 8240B
PACKING OCTAJL
CONSTRUCTION OCTAJL
8240B - 39
Revision 2
November 1992
-------
FIGURE 3.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - PURGE MODE FOR METHOD 8240B
OPTIONAL **O*T COLUMN
SCLCCTION VALVf
r UOLfO IMJfCnOM POUTS
|— COLUMN QVtN
roocTtcroft
• ANALTT1CAL COLUMN
PUMQf OAt
njOHl CONTROL
taXMOLCCUU
SlCVf FILTW
NOTE:
ALL UNO ITTWf IN
AND QC SHOULD tf HCATB
TO«TC
8240B - 40
Revision 2
November 1992
-------
FIGURE 4.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - OESORB MODE FOR METHOD 8240B
CAfWOGAt
PlOWCOMTHOl
KOUCATOA
StklCTlON VALVt
- COLUMN OVf*
CONFMMATOffy COLUMN
TOOCHCTO*
»
"— ANALYTICAL OOUUMM
JU1P--
TftAPMtfT
NOTl.
ALL UNCS KTWUM
AND OC 1HOULO M HIA1V
TOVC
8240B - 41
Revision 2
November 1992
-------
FIGURE 5.
LOW SOILS IMPINGER
PURGE INLET FITTING
SAMPLE OUTLET FITTING
j • 6mm o D CLASS TUBING
SEPTUM
CAP
40ml VIAL
8240B - 42
Revision 2
November 1992
-------
METHOD 8240B
VOLATILE ORGANICS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS)
7 1
Select
procedure for
introducing
sample into
CC/MS
7 2 1 Set
GC/MS
operating
conditions
Purge-and-trap
7 2.4 Connect
purge-and-trap
device to CC
726 Perform
purge-and-trap
analysis
7 2.8
Calculate RFs
for 5 SPCCs
7 3 Perform
daily
calibration
using SPCCs
and CCCs
/
8240B - 43
Revision 2
November 1992
-------
METHOD 8240B
(continued)
7
7421
3.1-le sample
a', least SO
:old «uth
« a '. e r
Soil/sediment
and waste
samples
7411
Screen sample
us ing Method
3810 or 3820
7417
Per form
secondary
dilutions
7 4 1 8 Add
interna1 standard
and surrogate
spiking solutions
7 4 1 10
Perform
purge • and -trap
procedure
74311
Choose sample
size based on
estimated
concent ra tion
7 4 3 1 3 Add
internal standard
and surrogate
spiking solutions
74315
Determine
percent dry
weight of
sample
74317
Perform
purge-and-trap
procedure
7432 Choose
solvent for
ext faction o r
dilution Weigh
samp 1e
74322 Add
sol vent,
shake
74327
Perform
purge-and- trap
procedure
7 4 1 11
A t tach Iran
to CC ana
perform
ana lysis
7511 Identify
analyt.es ay
comoa r ing *. r.e
sample re t en*, is r.
time ana sarro.e
mas s s pec t ra
7522 Calculate
the concentration
of each identifies
ana 1y te
7524
Report ail
results
J
Stop
8240B - 44
Revision 2
November 1992
-------
METHOD 8250A
SEMIVQLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS1
1.0 SCOPE AND APPLICATION
1.1 Method 8250 is used to determine the concentration of semi volatile
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:
ADorooriate Preparation Technioues
Compounds
Acenaphthene
Acenaphthene-d10 (I.S.)
Acenaphthylene
Acetophenone
Aldrin
4-Aminobiphenyl
Aniline
Anthracene
Aroclor - 1016
Aroclor - 1221
Aroclor - 1232
Aroclor - 1242
Aroclor - 1248
Aroclor - 1254
Aroclor - 1260
Benzidine
Benzoic acid
Benz (a) anthracene
Benzo(b)fluoranthene
Benzo ( k) f 1 uoranthene
Benzo(g,h,i)perylene
Benzo(a)pyrene
Benzyl alcohol
a-BHC
0-BHC
6-BHC
Y-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
CAS No8
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
3510
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
3520
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
3540
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
3550
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
3580
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 - 1
Revision 1
November 1992
-------
Compounds
Appropriate Preparation Techniques
CAS Noa 3510 3520 3540 3550 3580
Chlordane
4-Chloroaniline
1 -Chi oronaphthal ene
2-Chl oronaphthal ene
4-Chloro-3-methyl phenol
2-Chlorophenol
4-Chlorophenyl phenyl ether
Chrysene
Chrysene-d12 (I.S.)
4,4'-DDD
4,4'-DDT
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' -Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorophenol
Dieldrin
Diethyl phthalate
Dimethyl ami noazobenzene
7,12-Dimethylbenz(a)-
anthracene
a, a-Dimethyl phenethyl amine
2,4-Dimethylphenol
Dimethyl phthalate
4, 6-Dinitro-2-methyl phenol
2,4-Dinitrophenol
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di phenyl amine
1 , 2-Di phenyl hydrazi ne
Di-n-octyl phthalate
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Ethyl methanesulfonate
Fluoranthene
Fluorene
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
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
206-44-0
86-73-7
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
X
X
ND
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
ND
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
X
X
ND
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
X
CP
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
8250A - 2
Revision 1
November 1992
-------
Appropriate Preparation Techniaues
Compounds
2-Fluorobiphenyl (surr.)
2-Fluorophenol (surr.)
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachloroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Methoxychlor
3-Methylcholanthrene
Methyl methanesulfonate
2-Methyl naphtha! ene
2-Methylphenol
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-NHrophenol
N-Nitrosodi butyl ami ne
N-Nitrosodimethylamine
N-Nitrosodiphenylamine
N-Nitrosodi-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-Tetrachlorophenol
CAS Noa
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
129-00-0
95-94-3
58-90-2
3510
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
3520
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
ND
ND
3540
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
ND
ND
ND
3550
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
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
X
X
X
X
X
X
X
X
X
X
X
X
ND
X
X
X
X
X
8250A - 3
Revision 1
November 1992
-------
Appropriate Preparation Techniques
Compounds CAS Noa 3510 3520 3540 3550 3580
Toxaphene
2,4,6-Tribromophenol (surr. )
1 , 2 , 4-Tri chl orobenzene
2,4,5-Trichlorophenol
2,4,6-Trichlorophenol
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
X
X
ND
X
X
X
X
X
X
X
X
X
X
X
a 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.
Percent Stability = Average Recovery (Day 7) x 100/Average Recovery (Day 0).
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, y-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, 3-
nitroaniline, 4-chloroaniline, and benzyl alcohol are subject to erratic
chromatographic behavior, especially if the GC system is contaminated with high
boiling material.
8250A - 4 Revision 1
November 1992
-------
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.
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.
8250A - 5 Revision 1
November 1992
-------
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 ptL 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
dichlorodimethylsi 1ane.
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 ni.
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
concentration if they are certified by the manufacturer or by an
independent source.
8250A - 6 Revision 1
November 1992
-------
5.3.2 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.
5.3.3 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
1,4-dichlorobenzene-d,, naphthalene-da, acenaphthene-d10, phenanthrene-d1Q,
chrysene-d12, and perylene-d12. Other compounds may be used as internal standards
as long as the requirements given in Section 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
4,1)00 ng/jul_. Each 1 ml sample extract undergoing analysis should be spiked with
10 /nL of the internal standard solution, resulting in a concentration of 40
ng/juL of each internal standard. Store at 4°C or less when not being used.
5.5 GC/MS tuning standard - A methylene chloride solution containing
50 ng/juL of decafluorotriphenylphosphine (DFTPP) should be prepared. The
standard should also contain 50 ng//uL 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-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.
8250A - 7 Revision 1
November 1992
-------
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,
Section 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, 3550
Waste 3540, 3550, 3580
7.1.1 Direct injection - In very limited applications direct
injection of the sample into the GC/MS 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 where concentrations in
excess of 10,000 /ug/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
"Method 8040 includes a derivatization technique followed by GC/ECD analysis, if
interferences are encountered on GC/FID.
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
8250A - 8 Revision 1
November 1992
-------
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%. 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 Section 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).
7.4.3 Analyze 1 p.1 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 as follows:
RF - (AxCfl)/(Af.Cx)
where:
Ax = Area of the characteristic ion for the compound being
measured.
Ajs = Area of the characteristic ion for the specific internal
standard.
Cx = Concentration of the compound being measured (ng/^L).
Cis = Concentration of the specific internal standard (ng/juL).
8250A - 9 Revision 1
November 1992
-------
7.4.4 A system performance check must be performed to ensure that
minimum average 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 (%RSD =
100[SD/RF]) 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.
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 to injector liner and/or capillary column, then repeat
the calibration procedure beginning with Section 7.5.
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 (Section 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 second order regression fit of the five
calibration points. The analyst should select the regression order
which introduces the least calibration error into the quantitation
(Sections 7.7.2.2 and 7.7.2.3). If the %RSD is <15%, 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 (Section 7.4.3) and
CCC (Section 7.4.4) criteria.
7.5.3 System Performance Check Compounds (SPCCs) - A system
performance check must be made during every 12 hour shift. If the SPCC
8250A - 10 Revision 1
November 1992
-------
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,
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:
ci - cc
% Drift = x 100
where:
Cj = 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 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 (Section 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 standard check, 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 just prior to analysis.
8250A - 11 Revision 1
November 1992
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7.6.3 Analyze the 1 ml extract by GC/MS using the appropriate column
(as specified in Section 4.1.2). The recommended GC/MS operating
conditions to be used are specified in Section 7.3.
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//xL 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 Section 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.
8250A - 12 Revision 1
November 1992
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7.7.1.1.5 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.
7.7.1.2 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 deli sting 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.
8250A - 13 Revision 1
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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. I
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 (7.4.5.2) and the following equation:.
(Ax x Cis)
Cex (mg/L) •
(A,, x RF)
where C is the concentration of the compound in the extract,
and the other terms are as defined in Section 7.4.3.
7.7.2.3 Alternatively, the regression line fitted to the
initial calibration (Section 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 Sections 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 (ng/i) = (Cav x VrJ
V0
where:
V., = extract volume, in mL
.ex
V = 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 (jug/kg) = (C6K x VCM)
s
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 formulas
given above should be used with the following modifications: The
8250A - 14 Revision 1
November 1992
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areas A 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.
7.7.2.6 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 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 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 Section 7.3.1 and 7.4.1.
8.4.2 There must be an initial calibration of the GC/MS system as
specified in Section 7.3.
8.4.3 The GC/MS system must meet the SPCC criteria specified in
Section 7.4.3 and the CCC criteria in Section 7.4.4, each 12 hr.
8250A - 15 Revision 1
November 1992
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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 (QC) check sample concentrate is required containing
each analyte at a concentration of 100 mg/L in acetone. The QC check m
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 M9/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 Section 7.1 with extraction of the samples.
8.5.4 Calculate the average recovery (x) in M9/L, and the standard
deviation of the recovery (s) in M9/U 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 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 A
probability that one or more will fail at least one of the I
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 Step
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 Section
8.5.2.
8.5.6.2 Beginning with Section 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 Section
8.5.2.
8.6 The laboratory must, on an ongoing basis, analyze a reagent 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.
8250A - 16 Revision 1 4
November 1992 ^
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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 Section 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 ng/L or 1 to 5 times higher than
the background concentration determined in Section 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
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 (Section 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 jig/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 (100x'/T) ±
2. 44(1005 '/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 Section 8.7.
8.7 If any analyte fails the acceptance criteria for recovery in Section
8.6, a QC check standard containing each analyte that failed must be prepared and
analyzed.
8250A - 17 Revision 1
November 1992
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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 Step 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 (Step 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 Section 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 (Ps) for each analyte with the
corresponding QC acceptance criteria found in Table 6. Only analytes that
failed the test in Section 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 Section 8.6, calculate A
the average percent recovery (p) and the standard deviation of the percent I
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).
8.9 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
8250A - 18 Revision 1
November 1992
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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.
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.
8250A - 19 Revision 1
November 1992
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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, 4
and J.W. Eichelberger, Unpublished report, October 1980. ™
5. "Interlaboratory 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 - 20 Revision 1
November 1992
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TABLE 1.
CHROMATOGRAPHIC CONDITIONS, METHOD DETECTION LIMITS, AND
CHARACTERISTIC IONS FOR SEMIVOLATILE COMPOUNDS
Retention
Compound Time (min)
Acenaphthene 17.8
Acenaphthene-d10 (I.S.)
Acenaphthylene 17.4
Acetophenone
Aldrin 24.0
4-Aminobi phenyl
Aniline
Anthracene 22.8
Aroclor-10166 18-30
Aroc1or-1221b 15-30
Aroclor-1232b 15-32
Aroclor-1242b 15-32
Aroclor-1248b 12-34
Aroclor-1254b 22-34
Aroclor-1260b 23-32
Benzidine8 28.8
Benzoic acid
Benzo(a)anthracene 31.5
Benzo(b)f!uoranthene 34.9
Benzo(k)fluoranthene 34.9
Benzo(g,h,i)perylene 45.1
Benzo(a)pyrene 36.4
Benzyl alcohol
o-BHC8 21.1
6-BHC 23.4
8-BHC 23.7
Y-BHC (Lindane)8 22.4
Bis(2-chloroethoxy)methane 12.2
Bis(2-chloroethyl) ether 8.4
Bis(2-chloroisopropyl) ether 9.3
Bis(2-ethylhexyl) phthalate 30.6
4-Bromophenyl phenyl ether 21.2
Butyl benzyl phthalate 29.9
Chlordane6 19-30
4-Chloroaniline
1 -Chi oronaphthal ene
2-Chloronaphthalene 15.9
4-Chloro-3-methylphenol 13.2
2-Chlorophenol 5.9
4-Chlorophenyl phenyl ether 19.5
Chrysene 31.5
Chrysene-d12 (I.S.)
4,4'-DDD 28.6
4,4'-DDT 29.3
Method
detection
limit (MQ/L;
1.9
--
3.5
--
1.9
--
--
1.9
30
. -
- -
36
44
--
7.8
4.8
2.5
4.1
2.5
--
--
4.2
3.1
--
5.3
5.7
5.7
2.5
1.9
2.5
--
--
--
1.9
3.0
3.3
4.2
2.5
--
2.8
4.7
Primary
) Ion
154
164
152
105
66
169
93
178
222
190
190
222
292
292
360
184
122
228
252
252
276
252
108
183
181
183
183
93
93
45
149
248
149
373
127
162
162
107
128
204
228
240
235
235
Secondary
Ion(s)
153, 152
162, 160
151, 153
77, 51
263, 220
168, 170
66, 65
176, 179
260, 292
224, 260
224, 260
256, 292
362, 326
362, 326
362, 394
92, 185
105, 77
229, 226
253, 125
253, 125
138, 277
253, 125
79, 77
181, 109
183, 109
181, 109
181, 109
95, 123
63, 95
77, 121
167, 279
250, 141
91, 206
375, 377
129
127, 164
127, 164
144, 142
64, 130
206, 141
226, 229
120, 236
237, 165
237, 165
8250A - 21
Revision 1
November 1992
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TABLE 1.
(Continued)
Retention
Compound Time (min)
Dibenz(a,j)acridine
Dibenz(a,h)anthracene 43.2
Dibenzofuran
Di-n-butyl phthalate 24.7
1,2-Dichlorobenzene 8.4
1,3-Dichlorobenzene 7.4
1,4-Dichlorobenzene 7.8
l,4-Dichlorobenzene-d4 (I.S.)--
3,3'-Dichlorobenzidine 32.2
2,4-Dichlorophenol 9.8
2,6-Dichlorophenol
Dieldrin 27.2
Diethyl phthalate 20.1
p-Dimethylaminoazobenzene
7, 12-Dimethylbenz (a) anthracene- -
o-,o-Dimethylphenethylamine --
2,4-Dimethylphenol 9.4
Dimethyl phthalate 18.3
4,6-Dinitro-2-methylphenol 16.2
2,4-Dinitrophenol 15.9
2,4-Dinitrotoluene 19.8
2,6-Dinitrotoluene 18.7
Diphenylamine
1,2-Diphenylhydrazine
Di-n-octyl phthalate 32.5
Endosulfan Ia 26.4
Endosulfan Ha 28.6
Endosulfan sulfate 29.8
Endrin3 27.9
Endrin aldehyde
Endrin ketone
Ethyl methanesulfonate
Fluoranthene 26.5
Fluorene 19.5
2-Fluorobiphenyl (surr.)
2-Fluorophenol (surr.)
Heptachlor 23.4
Heptachlor epoxide 25.6
Hexachlorobenzene 21.0
Hexachlorobutadiene 11.4
Hexachlorocyclopentadiene3 13.9
Hexachloroethane 8.4
Indeno(l,2,3-cd)pyrene 42.7
Isophorone 11.9
Methoxychlor
Method
detection
limit (M9/L)
--
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
3.7
2.2
Primary
Ion
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
276
82
227
Secondary
Ion(s)
280, 277
139, 279
139
150, 104
148, 111
148, 111
148, 111
150, 115
254, 126
164, 98
164, 98
263, 279
177, 150
225, 77
241, 257
91, 42
107, 121
194, 164
51, 105
63, 154
63, 89
63, 89
168, 167
105, 182
167, 43
339, 341
339, 341
387, 422
82, 81
345, 250
67, 319
109, 97
101, 203
165, 167
171
64
272, 274
355, 351
142, 249
223, 227
235, 272
201, 199
138, 227
95, 138
228
8250A - 22
Revision 1
November 1992
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TABLE 1.
(Continued)
Retention
Compound Time
3-Methyl chol anthrene
Methyl methanesulfonate
2-Methyl naphthalene
2-Methyl phenol
4-Methyl phenol
Naphthalene 12
Naphthalene-d8 (I.S.)
1-Naphthylamine
2-Naphthylamine
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Nitrobenzene 11
Ni trobenzene-d5 ( surr . )
2-Nitrophenol 6
4-Nitrophenol 20
N-Ni troso-di -n-butyl ami ne
N-Ni trosodimethyl ami nea
N-Ni trosodiphenyl ami ne8 20
N-Ni troso-di -N-propyl ami ne
N-Ni trosopi peri dine
Pentachl orobenzene
Pentachl oronitrobenzene
Pentachl orophenol 17
Perylene-d12 (I.S.)
Phenacetin
Phenanthrene 22
Phenanthrene-d10 (I.S.)
Phenol 8
Phenol -d6 (surr.)
2-Picoline
Pronamide
Pyrene 27
Terphenyl-du (surr.)
1 , 2 , 4, 5-Tetrachl orobenzene
2,3,4, 6 -Tetrachl orophenol
Toxaphene 25-34
2,4,6-Tribromophenol (surr.)
1,2,4-Trichlorobenzene 11
2, 4, 5-Trichl orophenol
2,4,6-Trichlorophenol 11
(min)
--
--
--
--
--
.1
--
--
--
--
--
--
.1
--
.5
.3
--
--
.5
--
--
--
--
.5
--
--
.8
--
.0
--
--
--
.3
--
--
--
--
.6
--
.8
Method
detection
Primary
limit (/xg/L) Ion
--
--
--
--
--
1.6
--
--
--
--
--
--
1.9
--
3.6
2.4
--
--
1.9
--
--
--
--
3.6
--
--
5.4
--
1.5
--
--
--
1.9
--
--
--
--
__
1.9
--
2.7
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
159
330
180
196
196
Secondary
Ion(s)
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,
231,
332,
182,
198,
198,
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
233
141
145
200
200
aSee Section 1.3
nhese compounds are mixtures of various isomers.
8250A - 23
Revision 1
November 1992
-------
TABLE 2.
DETERMINATION OF ESTIMATED QUANTITATION LIMITS (EQL)
FOR VARIOUS MATRICES8
Matrix Factor6
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 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.
8250A - 24 Revision 1
November 1992
-------
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 4.
8250A - 25 Revision 1
November 1992
-------
TABLE 4.
CALIBRATION CHECK COMPOUNDS
Base/Neutral Fraction Acid Fraction
Acenaphthene 4-Chloro-3-methylphenol
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-Trichlorophenol
8250A - 26 Revision 1
November 1992
-------
TABLE 5.
SEMIVOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
Phenanthrene-d10 Chrysene-d12 Perylene-d12
4-Aminobiphenyl Benzidine Benzo(b)fluoranthene
Anthracene Benzo(a)anthracene Benzo(k)fluoranthene
4-Bromophenyl phenyl ether Bis(2-ethylhexyl) phthalate Benzo(g,h,i)perylene
Di-n-butyl phthalate Butyl benzyl phthalate Benzo(a)pyrene
4,6-Dinitro-2-methylphenol Chrysene Dibenz(a,j)acridine
Diphenylamine 3,3'-Dichlorobenzidine Dibenz(a,h)anthracene
1,2-Diphenylhydrazine p-Dimethylaminoazobenzene 7,12-Dimethylbenz-
Fluoranthene Pyrene (a)anthracene
Hexachlorobenzene Terphenyl-d14 (surr.) Di-n-octyl phthalate
N-Nitrosodiphenylamine Indeno(l,2,3-cd)pyrene
Pentachlorophenol 3-Methylcholanthrene
Pentachloronitrobenzene
Phenacetin
Phenanthrene
Pronamide
(surr.) = surrogate
8250A - 27 Revision 1
November 1992
-------
TABLE 5.
SEMIVOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
(Continued)
l,4-Dichlorobenzene-D4
Naphthalene-d8
Acenaphthene-d10
Aniline
Benzyl alcohol
Bis(2-chloroethyl) ether
Bi s(2-chloroi sopropylJether
2-Chlorophenol
1,3-Di chlorobenzene
1,4-Di chlorobenzene
1,2-Di chlorobenzene
Ethyl methanesulfonate
2-Fluorophenol (surr.)
Hexachloroethane
Methyl methanesulfonate
2-Methylphenol
4-Methylphenol
N-Nitrosodimethylamine
N-Nitroso-di-n-propylamine
Phenol
Phenol-d6 (surr.)
2-Picoline
Acetophenone
Benzoic acid
Bis(2-chloroethoxy)methane
4-Chloroaniline
4-Chloro-3-methylphenol
2,4-Dichlorophenol
2,6-Dichlorophenol
a, o-Dimethylphenethylami ne
2,4-Dimethylphenol
Hexachlorobutadi ene
Isophorone
2-Methylnaphtha!ene
Naphthalene
Nitrobenzene
Nitrobenzene-d8 (surr.)
2-Nitrophenol
N-Nitroso-di-n-butylamine
N-Nitrosopiperidine
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
(surr.) = surrogate
8250A - 28
Revision 1
November 1992
-------
TABLE 6.
QC ACCEPTANCE CRITERIA8
Compound
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
Test
cone.
(M9/L)
100
100
100
100
100
100
100
100
100
100
100
100
100
100
Bis(2-chloroisopropyl) etherlOO
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-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
Hexachloroethane
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
40
39
32
27
38
32
39
58
23
31
21
55
34
46
41
23
13
33
48
31
32
61
70
16
30
41
32
71
30
26
23
21
29
31
16
32
32
20
37
54
24
26
24
.6
.2
.0
.0
.6
.8
.3
.0
.9
.4
.5
.6
.0
.5
.3
.1
.0
.0
.4
.3
.0
.0
.6
.0
.7
.9
.7
.1
.4
.7
.5
.2
.8
.6
.4
.7
.5
.8
.7
.2
.7
.9
.3
.5
Range
for x
(M9/L)
60
53
7
43
41
42
25
31
41
42
49
62
28
64
64
38
44
19
8
48
16
37
8
44
47
68
18
42
71
70
7
37
55
.1-132.
.5-126.
.2-152.
.4-118.
.8-133.
.0-140.
.2-145.
.7-148.
D-195.
D-139.
.5-130.
D-100.
.9-126.
.2-164.
.8-138.
.9-136.
.9-114.
.5-113.
.4-144.
.1-139.
D-134.
.2-119.
D-170.
D-199.
.4-111.
.6-112.
.7-153.
.3-105.
.2-212.
.3-119.
D-100.
D-100.
.5-126.
.1-136.
.6-131.
D-103.
D-188.
.9-121.
.6-108.
D-172.
.9-109.
.8-141.
.8-102.
.2-100.
3
0
2
0
0
4
7
0
0
9
6
0
0
7
6
8
4
5
7
9
5
7
6
7
0
0
9
7
5
3
0
0
9
7
8
5
8
3
4
2
4
5
2
0
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
40-113
8250A - 29
Revision 1
November 1992
-------
TABLE 6.
QC ACCEPTANCE CRITERIA3
(Continued)
Compound
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-Chlorophenol
2,4-Dimethylphenol
2,4-Dinitrophenol
2-Methyl-4,6-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
100
100
100
100
100
100
100
100
100
Limit
for s
(M9/L)
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)
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>oPs
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
Standard deviation of four recovery measurements, in
Average recovery for four recovery measurements, in
Percent recovery measured.
Detected; result must be greater than zero.
Criteria from 40 CFR Part 136 for Method 625. These criteria are
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 - 30
Revision 1
November 1992
-------
TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION3
Parameter
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benzo(a)anthracene
Chloroethane
Benzo(b)fluoranthene
Benzo(k)fl uoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Butyl benzyl phthalate
6-BHC
6-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'-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
Hexachl orobenzene
Hexachlorobutadiene
Hexachl oroethane
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
0.18x+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
0.12X+0.26
0.24X-0.56
0.33X-0.46
O.lSx-0.10
0.19X+0.92
0.17X+0.67
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
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
O.lGx+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
0.13X+0.61
O.SOx-0.23
0.28x+0.64
0.43x-0.52
0.26X+0.49
0.17x+0.80
8250A - 31
Revision 1
November 1992
-------
TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
(Continued)
Parameter
Indeno(l,2,3-cd)pyrene
Isophorone
Naphthalene
Nitrobenzene
N-Nitroso-di -n-propyl amine
PCB-1260
Phenanthrene
Pyrene
1,2,4-Trichlorobenzene
4-Chl oro-3-methyl phenol
2-Chlorophenol
2,4-Dichlorophenol
2, 4-Dimethyl phenol
2,4-Dinitrophenol
2-Methyl-4,6-dinitrophenol
2-Nitrophenol
4-Nitrophenol
Pentachlorophenol
Phenol
2,4,6-Trichlorophenol
Accuracy, as
recovery, x'
(M9/L)
0.78C-3.10
1.12C+1.41
0.76C+1.58
1.09C-3.05
1.12C-6.2Z
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.lBx+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
O.lGx+2.22
Overall
precision,
S' (M9/L)
0.50X-0.44
0.33x+0.26
0.30X-0.68
0.27X+0.21
0.44X+0.47
0.43X+1.82
O.lBx+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-I-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 ^g/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
True value for the concentration, in M9/L.
Average recovery found for measurements of samples containing a
concentration of C, in M9/L.
8250A - 32
Revision 1
November 1992
-------
TABLE 8.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Low/Med i urn Low/Ned1 urn
Surrogate Compound Water Soil/Sediment
Nitrobenzene-d5 35-114 23-120
2-Fluorobiphenyl 43-116 30-115
p-Terphenyl-du 33-141 18-137
Phenol-d, 10-94 24-113
2-Fluorophenol 21-100 25-121
2,4,6-Tribromophenol 10-123 19-122
8250A - 33 Revision 1
November 1992
-------
METHOD 8250A
SEMIVOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS)
T * & »*«p**«
• amp1• us ±n«
M« tllOd 3 5 4 O
• amp 1• u•i ng
M*tHod 3310
or 3 S 2 O .
•7.1 p r »»ar •
• ample \im Ing
M*thod 3 5 4 O ,
35^0. or
•7 . 3
ac / MS
op • r a t. i n 9
condition*.
7.5 Daily
c a 1 1 b r a t 1 o TI -
Tune CC/MS with
TFTPP and ch«ck
S PCG fc CCC .
8250A - 34
Revision 1
November 1992
-------
METHOD 8250A
continued
1 6 L Screen
en tract.
.n CC/FID
sr CC/PID to
ei imina te
7 6 2
sampl
too • nign
Spilce
B With
internal
standard
7 6 3 Analyze
extract ay CC/MS
j* irg recommended
co i umr
and
ooera t ing
condi 1 i ons
771 Identify
coffloound* by
comparing sample
retention time and
sampi e mas* spectra
to s tandardi
8250A - 35
Revision 1
November 1992
-------
METHOD 8260A
VOLATILE ORGANIC COMPOUNDS BY GAS CHROMAT06RAPHY/MASS SPECTROMETRY fGC/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, mousses, tars,
fibrous wastes, polymeric emulsions, filter cakes, spent carbons, spent
catalysts, soils, and sediments. The following compounds can be determined by
this method:
ADorooriate Techniaue
Analyte
Acetone
Acetonitrile
Acrolein
Acrylonitrile
Allyl 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-Chloro-l,3-butadiene
Chi orodi bromomethane
Chloroethane
2-Chloroethanol
bis-(2-Chloroethyl) sulfide
2-Chloroethyl vinyl ether
Chloroform
Chloromethane
Chloroprene
3-Chloropropene
3-Chloropropionitrile
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
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
107-03-3
505-60-2
110-75-8
67-66-3
74-87-3
126-99-8
107-05-1
542-76-7
Purge-and-Trap
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
i
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
a
pc
a
pc
8260A - 1
Revision 1
November 1992
-------
Appropriate Technique
Analyte
1 , 2-Di bromo-3-chl oropropane
1,2-Dibromoethane
Dibromomethane
1,2-Dichlorobenzene
1 , 3-Di chl orobenzene
1,4-01 chl orobenzene
cis-l,4-D1chloro-2-butene
trans- 1,4-Di chl oro-2-butene
Dichl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
trans- 1, 2-Di chl oroethene
1, 2-Di chl oropropane
l,3-Dichloro-2-propanol
cis-l,3-Dichloropropene
trans-l,3-Dichloropropene
1 , 2 , 3 , 4-Di epoxybutane
Di ethyl ether
1,4-Difluorobenzene (I.S.)
1,4-Dioxane
Epichlorohydrin
Ethanol
Ethyl acetate
Ethyl benzene
Ethyl ene oxide
Ethyl methacrylate
Hexachlorobutadiene
Hexachloroethane
2-Hexanone
2-Hydroxypropionitrile
lodomethane
Isobutyl alcohol
I sopropyl benzene
Malononitrile
Methacrylonitrile
Methanol
Methyl ene chloride
Methyl iodide
Methyl methacrylate
4-Methyl-2-pentanone (MIBK)
Naphthalene
Nitrobenzene
2-Nitropropane
Pentachloroethane
2-Picoline
Propargyl alcohol
CAS No.b
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
74-88-4
80-62-6
108-10-1
91-20-3
98-95-3
79-46-9
76-01-7
109-06-8
107-19-7
Purge-and-Trap
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
a
PP
a
a
a
i
PP
PP
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
a
a
a
pc
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
8260A - 2
Revision 1
November 1992
-------
Appropriate Technique
Analyte CAS No.b
G-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-Tetrachloroethane 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
Purge-and-Trap
PP
ht
a
i
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
a
.
EQLs.
Direct
Injection
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-
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
n-Butylbenzene
sec-Butyl benzene
tert-Butylbenzene
Chloroacetonitrile
1-Chlorobutane
1,3-Di chloropropane
2,2-Dichloropropane
1,1-Dichloropropene
Fluorobenzene
p-Isopropyltoluene
Methyl acrylate
8260A - 3
Revision 1
November 1992
-------
1-Chlorohexane Methyl-t-butyl ether
2-Chlorotoluene Pentaf1uorobenzene
4-Chlorotoluene n-Propylbenzene
Crotonaldehyde 1,2,3-Tri chlorobenzene
Di bromof1uoromethane 1,2,4-Trimethylbenzene
ci s-1,2-Di chloroethene 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 M9/kg (wet weight) for
soil/sediment samples, 0.5 mg/kg (wet weight) for wastes, and 5 fj.g/1 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.
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
8260A - 4 Revision 1
November 1992
-------
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-
poly tetrafluoroethylene (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
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
8260A - 5 Revision 1
November 1992
-------
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 1 cm into the liner from the oven side of the
injection port, according to
manufacturer's specifications.
Sept.-u.rn BO — GO Oven
mm
Figure 1 Modified Injector
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
8260A - 6 Revision 1
November 1992
-------
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 urn 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 jura 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 urn film thickness.
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.
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. 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.
8260A - 7 Revision 1
November 1992
-------
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 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 Section 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 /iL.
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.
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
8260A - 8 Revision 1
November 1992
-------
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 juL 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.
5.7.2.2 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 methanol. 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
8260A - 9 Revision 1
November 1992
-------
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 two 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 25% 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.
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 /LtL or 500 pi 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.
8260A - 10 Revision 1
November 1992
-------
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 ~30psi.
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
5.7.6.1.10 The following are the recommended gas volumes
spiked in to 5 mLs of water to produce a typical 5-point
calibration:
Gas Calibration
Volume Concentration
40 jiL 20 iig/L
100 ML 50 |ig/L
200 jiL 100 [ig/L
300 |iL 150 jig/L
400 nL 200 jig/L
5.7.6.1.11 The f ol 1 owi ng are the recommended gas vol umes
spiked in to 25-mls of water to produce a typical 5-point
calibration:
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Gas Calibration
Vol time Concentration
10 |iL 1 jig/L
20 jiL 2 jig/L
50 jiL 5 |ig/L
100 jiL 10 [ig/L
250 pL 25 jig/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, 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
M9/10 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-ds, 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 Sections 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.
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/ML 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 Sections 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
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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 (e.g. some or all of the compounds listed in Table
1 may be included). 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 jug/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,
Section 4.1.
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 TCLP
regulatory limits, at concentrations in excess of 10,000 /Lig/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.
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7.2 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
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.
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7.2.6 Direct injection - Column 4
Carrier gas (He) flow rate: 4 mL/min
Column: J&W DB-24, 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
7.3 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
Ion trap only: Axial modulation 4.0 volts
Manifold set 220°C
Emission current 30 amps
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
ML 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 /iL 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.
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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 (Section
7.6.2). The RF is calculated as follows:
RF = (A*Cis)/(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.
Cx = Concentration of the compound being measured.
7.3.4 The average RRF must be calculated and recorded for each
compound. 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 RRFs from the initial calibration, calculate and
record the percent relative standard deviation (%RSD) for all compounds.
The percent RSD is calculated as follows:
% RSD = -2z x 100%
RFX
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where:
RSD = Relative standard deviation.
RFX = mean of 5 initial RRFs for a compound.
SO = standard deviation of average RRFs for a compound.
SD
N (X| - x)2
j
i=l N - 1
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/AU) 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.
If the %RSD is <15%, 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.
7.3.7 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 (Section 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 A
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 0.1
1,1-Dichloroethane 0.1
Bromoform 0.25
Chlorobenzene 0.3
1,1,2,2-Tetrachloroethane 0.3
7.4.4 Calibration Check Compounds (CCCs) - After the system
performance check is met, CCCs listed in Section 7.3.8 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 check calibration (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 standard check, 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 are necessary.
7.4.6 For compounds which exhibit linearity of response, the RRF of
the daily check standard may be used for quantitation, provided the
criteria for SPCCs and CCCs are satisfied.
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.
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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.
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 /xL 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 p.1 of the surrogate spiking solution to 5 ml of sample
is equivalent to a concentration of 50 /xg/L of each surrogate standard.
The addition of 10 /iL 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
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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 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 Sections 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 (Section 5.13) to the 5 ml of sample to be purged. Disregarding
any dilutions, this is equivalent to a concentration of 50 M9/L 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.
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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.
7.6.1.1.5 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
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of sample with the nearest library searches will the mass spectral
Interpretation specialist assign a tentative identification.
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
(AX)(IS)
concentration (M9/L)
(Ais)(RRF)(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.
RRF = 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)ds)(Vt)
concentration (/KjAg) = —
(A1s)(RRF)(V,.)(W8)(D)
where:
AX' ls» A,-s» RRF, = Same as for water.
Vt = Volume of total extract (/xL) (use 10,000 /xL or a
factor of this when dilutions are made).
V,. - Volume of extract added (jtiL) 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
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areas Ax and Ajs should be from the total Ion chromatograms, and the
RRF 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.
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 Sections 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 /zg/L or less of
each analyte by adding 200 juL 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 Section 7.5.1.
8.3.4 Calculate the average recovery (x) in Mg/L, and the standard
deviation of the recovery (s) in fj.g/1, 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 (Section 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 - 24 Revision 1
November 1992
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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 Section
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 Section 8.3.2.
8.3.6.2 Beginning with Section 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 Section
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 ng/L. Single laboratory accuracy and precision data are
8260A - 25 Revision 1
November 1992
-------
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 /xg/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. J. 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 SAIC contract 68-W9-001, 6/92.
8260A - 26 Revision 1
November 1992
-------
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 - 27 Revision 1
November 1992
-------
TABLE 1.
CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS (MDL)
FOR VOLATILE ORGANIC COMPOUNDS ON WIDE BORE CAPILLARY COLUMNS
ANALYTE
Di chl orodi f 1 uoromethane
Chloromethane
Vinyl Chloride
Bromomethane
Chloroethane
Tri chl orof 1 uoromethane
Acrolein
lodomethane
Acetonitrile
Carbon disulfide
Allyl chloride
Methyl ene chloride
1,1-Dichloroethene
Acetone
trans- 1,2-Di chl oroethene
Acrylonitrile
1,1-Dichloroethane
Vinyl acetate
2,2-Dichloropropane
2-Butanone
cis -1,2-Di chl oroethene
Propionitrile
Chloroform
Bromochl oromethane
Methacrylonitrile
1,1,1 -Tri chloroethane
Carbon tetrachloride
1,1-Dichloropropene
Benzene
1,2-Dichloroethane
Tri chl oroethene
1,2-Di chl oropropane
Bromodi chl oromethane
Dibromomethane
Methyl methacrylate
1,4-Dioxane
2-Chloroethyl vinyl ether
4-Methyl -2-pentanone
trans- 1 , 3-Dichl oropropene
Toluene
cis-l,3-Dichloropropene
1 , 1 ,2 -Tri chloroethane
Ethyl methacrylate
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
20.01
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 - 28
Revision 1
November 1992
-------
TABLE 1.
(Continued)
ANALYTE
RETENTION TIME
(minutes)
MDLd
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
I sopropyl benzene (Cumene)
cis-l,4-Dichloro-2-butene
1,1,2 , 2-Tetrachl oroethane
Bromobenzene
1 , 2 , 3-Tri chl oropropane
n-Propyl benzene
2-Chlorotoluene
trans-l,4-Dichloro-2-butene
1,3, 5-Trimethyl benzene
4-Chlorotoluene
Pentachl oroethane
1 , 2 , 4-Trimethyl benzene
sec-Butyl benzene
tert-Butyl benzene
p- I sopropyl toluene
1,3-Dichl orobenzene
1 , 4-Di chl orobenzene
Benzyl chloride
n-Butyl benzene
1 , 2 -Di chl orobenzene
l,2-Dibromo-3-chl oropropane
1 , 2 , 4 -Tr i chl orobenzene
Hexachlorobutadiene
Naphthalene
1, 2, 3-Trichl orobenzene
Column 1"
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 2°
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'e
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 - 29
Revision 1
November 1992
-------
TABLE 1.
(Continued)
ANALYTE
RETENTION TIME
(minutes)
MDL
Column 1"Column 2DColumn 2'
INTERNAL STANDARDS/SURROGATES
1,4-Difluorobenzene 13.26
Chlorobenzene-d5 23.10
l,4-Dichlorobenzene-d4 31.16
4-Bromofluorobenzene 27.83
l,2-Dichlorobenzene-d4 32.30
Dichloroethane-d, 12.08
Dibromofluoromethane
Toluene-d8 18.27
Pentafluorobenzene
Fluorobenzene 13.00
15.71
19.08
23.63
27.25
6.27
14.06
8 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°/nrin.
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°/nrin.
0 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°/nnn, then program to 180°C at 8°/min.
d MDL based on a 25 ml sample volume.
8260A - 30
Revision 1
November 1992
-------
TABLE 2.
CHROMATOGRAPHIC RETENTION TIMES AND METHOD DETECTION LIMITS (MDL)
FOR VOLATILE ORGANIC COMPOUNDS ON NARROW BORE CAPILLARY COLUMNS
ANALYTE
Di chl orodi f 1 uoromethane
Chloromethane
Vinyl chloride
Bromomethane
Chloroethane
Tr i chl orof 1 uoromethane
1,1-Dichloroethene
Methyl ene chloride
trans-l,2-Dichloroethene
1,1 -Di chloroethane
cis-1, 2-Di chl oroethene
2 , 2-Di chl oropropane
Chloroform
Bromochloromethane
1,1,1-Tri chloroethane
1, 2-Di chloroethane
1 , 1 -Di chl oropropene
Carbon tetrachloride
Benzene
1, 2-Di chl oropropane
Trichloroethene
Di bromomethane
Bromodi chloromethane
Toluene
1,1,2-Trichloroethane
1,3-Dichloropropane
Di bromochl oromethane
Tetrachloroethene
1,2-Dibromoethane
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-Trichl oropropane
I sopropyl benzene
RETENTION TIME
(minutes)
Column 3
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
MDLb
(M9/L)
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 - 31
Revision 1
November 1992
-------
TABLE 2.
(Continued)
ANALYTE
RETENTION TIME
MDL
(minutes) (M9/L)
Column 3
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- I sopropyl toluene
1,4-Dichlorobenzene
1 , 2 -Di chl orobenzene
n-Butyl benzene
1 , 2-Di bromo-3-chl oropropane
1 , 2 , 4-Tr i chl orobenzene
Naphthalene
Hexachl orobutadi ene
1, 2, 3-Tri chl orobenzene
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
8 Column 3 - 30 meter x 0.32 mm ID DB-5 capillary with 1 urn film thickness.
b MDL based on a 25 ml sample volume.
8260A - 32
Revision 1
November 1992
-------
TABLE 3.
ESTIMATED QUANTITATION LIMITS FOR VOLATILE ANALYTES"
Estimated
Quantitation
Limits
Ground water Low Soil /Sediment
M9/L
Volume of water purged
All analytes in Table 1
5 ml
5
25 mL
1
5
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 analyte
concentration is selected for 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.
See the following information for further guidance on matrix-dependent
EQLs.
EQLs listed for soil/sediment are based on wet weight. Normally data is
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
CEQL = [EQL for low soil sediment (Table 3)] X [Factor]. For non-aqueous
samples, the factor is on a wet-weight basis.
8260A - 33 Revision 1
November 1992
-------
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
8260A - 34 Revision 1
November 1992
-------
TABLE 5.
CHARACTERISTIC MASSES (M/Z) FOR PURGEABLE ORGANIC COMPOUNDS
Analyte
Primary
Characteristic
Ion
Secondary
Characteristic
Ion(s)
Acetone
Acetonitrile
Acrolein
Acrylonitrile
Allyl alcohol
Allyl 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
l,2-Dibromo-3-chloropropane
Di bromochl oromethane
1,2-Dibromoethane
Di bromomethane
1,2-Dichlorobenzene
1 , 2-Di chl orobenzene-d4
1,3-Dichlorobenzene
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
109
63
83
50
53
54
91
91
75
129
107
93
146
152
146
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
49, 44, 43, 51, 80
111, 158, 160
65, 106
85
52
53, 88, 90, 51
54, 49, 89, 91
126
126
155, 157
127
109, 188
95, 174
111, 148
115, 150
111, 148
8260A - 35
Revision 1
November 1992
-------
TABLE 5.
(continued)
Analyte
Primary
Characteristic
Ion
Secondary
Characteristic
Ion(s)
1,4-Dichlorobenzene
cis-l,4-Dichloro-2-butene
trans- l,4-Dichloro-2-butene
Oi chl orodi f 1 uoromethane
1,1-Dichloroethane
1,2-Dichloroethane
1,1-Dichloroethene
cis-l,2-Dichloroethene
trans-l,2-Dichloroethene
1 , 2 -Di chl oropropane
1,3-Dichloropropane
2 , 2-Di chl oropropane
l,3-Dichloro-2-propanol
1,1-Dichloropropene
cis-l,3-Dichloropropene
trans- 1,3-Di chl oropropene
1 , 2 , 3 , 4-Di epoxybutane
Di ethyl ether
1,4-Dioxane
Epichlorohydrin
Ethanol
Ethyl acetate
Ethyl benzene
Ethyl ene oxide
Ethyl methacryl ate
Hexachl orobutadi ene
Hexachloroethane
2-Hexanone
2 -Hydroxypropi oni tr i 1 e
lodomethane
Isobutyl alcohol
I sopropyl benzene
p- I sopropyl toluene
Malonitrile
Methacryl oni trile
Methyl acrylate
Methyl -t-butyl ether
Methyl ene chloride
Methyl ethyl ketone
Methyl iodide
Methyl methacryl ate
4-Methyl -2-pentanone
Naphthalene
Nitrobenzene
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
111, 148
75, 53, 77, 124,
88, 75
87
65, 83
98
61, 63
61, 98
61, 98
112
78
97
79, 43, 81, 49
110, 77
77, 39
77, 39
55, 57, 56
45, 59
88, 58, 43, 57
57, 49, 62, 51
45, 27, 46
43, 45, 61
106
44, 43, 42
69, 41, 99, 86,
223, 227
166, 199, 203
58, 57, 100
44, 43, 42, 53
127, 141
43, 41, 42, 74
120
134, 91
66, 39, 65, 38
41, 67, 39, 52,
85
57
86, 49
43
142, 127, 141
69, 41, 100, 39
43, 58, 85
-
51, 77
89
114
66
8260A - 36
Revision 1
November 1992
-------
TABLE 5.
(continued)
Analyte
Primary
Characteristic
Ion
Secondary
Characteristic
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-Tri chl orobenzene
1,1,1, 2-Tetrachloroethane
1,1,2 , 2-Tetrachl oroethane
Tetrachloroethene
Toluene
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-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
Pentaf 1 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
8260A - 37
Revision 1
November 1992
-------
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
Bromod i 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-Dlbromoethane
Dibromomethane
1,2-Dichlorobenzene
1,3-Dichlorobenzene
1 , 4-Di chl orobenzene
Di chl orodi f 1 uoromethane
1 , 1 -Di chl orobenzene
1 , 2-Di chl orobenzene
1,1-Dichloroethene
ci s- 1 , 2-Di chl oroethene
trans- 1, 2-Di chl oroethene
1 , 2 -Di chl oropropane
1,3-Dichloropropane
2,2-Dichloropropane
1 , 1 -Di chl oropropene
Ethyl benzene
Hexachl orobutadi ene
I sopropyl benzene
p- I sopropyl tol uene
Methyl ene chloride
Naphthalene
n-Propyl benzene
Styrene
1,1,1 , 2-Tetrachl oroethane
Cone. Number
Range, of Recovery8
M9/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
0.5
- 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
- 10
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
24
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
90
Standard Percent
Deviation Rel Std
of Recovery Dev.
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
6.1
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
6.8
8260A - 38
Revision 1
November 1992
-------
TABLE 6.
(Continued)
Analyte
Cone.
Range,
M9/L
Number
of Recovery8
Samples %
Standard Percent
Deviation Rel Std
of Recovery Dev.
1,1,2,2-Tetrachloroethane
Tetrachloroethene
Toluene
1,2,3-Tri chlorobenzene
1,2,4-Trichlorobenzene
1,1,1-Tri chloroethane
1,1,2-Trichloroethane
Trichloroethene
Tri chlorof1uoromethane
1,2,3-Tri chloropropane
1,2,4-Trimethylbenzene
1,3,5-Trimethylbenzene
Vinyl chloride
o-Xylene
m-Xylene
p-Xylene
0.1
0.5
0.5
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
,5
.5
.5
.5
.5
.5
.5
,5
.5
.5
.1
0.1 -
0.5 -
10
10
10
10
10
10
10
10
10
10
10
10
10
31
10
10
30
24
18
18
18
18
18
24
24
16
18
23
18
18
31
18
91
89
102
109
108
98
104
90
89
108
99
92
98
103
97
104
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
6.3
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 form three concentrations.
8260A - 39
Revision 1
November 1992
-------
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
1 , 2-Di bromo-3-chl oropropane
Di bromochl oromethane
1,2-Dibromoethane
Dibromomethane
1,2-Dichlorobenzene
1 , 3 -Di chl orobenzene
1,4-Di chlorobenzene
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
1,2-Dichloropropane
1,3-Dichl oropropane
2 , 2-Di chl oropropane
1,1-Dichloropropene
Ethyl benzene
Hexachl orobutadi ene
I sopropyl benzene
p- I sopropyl toluene
Methyl ene 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
Rel Std
Dev.
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 - 40
Revision 1
November 1992
-------
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
Trichl orof 1 uoromethane
1,2, 3-Tri chl oropropane
1 , 2 , 4-Trimethyl 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
Recovery8
%
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
Rel Std
Dev
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 - 41
Revision 1
November 1992
-------
TABLE 8.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Surrogate Compound
4-Bromof 1 uorobenzene8
Di bromof 1 uoromethane8
Toluene-d.8
Dichloroethane-d48
Low/High
Water
86-115
86-118
88-110
80-120
Low/High
So 11 /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 Extract8
500 - 10,000 M9/kg 100 ML
1,000 - 20,000 M9A9 50 juL
5,000 - 100,000 jigAg 10 ML
25,000 - 500,000 M9/kg 100 pi of 1/50 dilution"
Calculate appropriate dilution factor for concentrations exceeding this table.
8 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 pi added to the syringe.
b Dilute an aliquot of the solvent extract and then take 100 n\. for
analysis.
8260A - 42 Revision 1
November 1992
-------
TABLE 10
DIRECT INJECTION ANALYSIS OF NEW OIL AT 5
Compound
Acetone
Benzene
n-Butanol*,**
iso-Butanol*,**
Carbon tetrachloride
Carbon disulfide**
Chlorobenzene
Chloroform
1,4-Di chlorobenzene
1,2-Dichloroethane
1,1-Dichloroethene
Diethyl ether
Ethyl acetate
Ethyl benzene
Hexachloroethane
Methylene chloride
Methyl ethyl ketone
MIBK
Nitrobenzene
Pyridine
Tetrachloroethene
Tri chlorof1uoromethane
l,l,2-C!3F3ethane
Toluene
Trichloroethene
Vinyl chloride
o-Xylene
m/p-Xylene
Recovery (%)
91
86
107
95
86
53
81
84
98
101
97
76
113
83
71
98
79
93
89
31
82
76
69
73
66
63
83
84
Alternate mass employed
IS quantitation
Data is taken from Reference 9.
*
**
%RSD
14.8
21.3
27.8
19,
44,
22,
29,
29,
24,
23,
45,
24,
27,
30,
45,
24,
31,
30,
29.
21.
35.
29.
30.1
35.9
27.1
27.6
28.0
29.5
PPM
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
.5
.7
Spike
fppml
5.0
0.5
5.0
5.0
0.5
5.0
5.0
6.0
7.5
0.
0.
5.0
5.0
5.0
3.0
5.0
.0
.0
2.0
5.0
0.7
5.0
5.0
5.0
0.
0.
5.
5.
.5
,2
5.0
10.0
8260A - 43
Revision 1
November 1992
-------
FIGURE 1.
PURGING DEVICE
ixrr IM M 04).
FOAM TRAP
EXIT IM M. 0.0.
10 MM GLASS FWT
MEDIUM FOHOSITV
SAMPUINCET
2-WAY SYMNGE VALVE
17 CM » GAUGE SVMNQE NEEDLE
6 MM O.O HUMCN SCPTUM
MLŁT IM IN 0.0.
IDC IN 00
/^ STAINLESS STIEL
MOLECULAM SIEVE
PURGE GAS FH.TW
PURGE GAS
FLOWCOMTHOL
8260A - 44
Revision 1
November 1992
-------
FIGURE 2.
TRAP PACKING AND CONSTRUCTION TO INCLUDE DESORB CAPABILITY
PACKING DETAIL
5- 5 MM GLASS WOOL
77CMSIUCAQEL
CONSTRUCTION DCTA*.
ANOI
1«FT
13 CM TENAX QC
•- 1 CM 3H OV-1
S MM OLASS WOOL
TMEMMOCOUPL&
OONTHOU0
SEN8OM
ELŁCT10MC
TEMPBMTUHE
OONTMXANO
TUBW«QaCM
a 106 IN. to
0.18 M. OO.
ST,
8260A - 45
Revision 1
November 1992
-------
FIGURE 3.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - PURGE MODE
CARMERGAS
FLOW CONTROL
PRESSURE
REGULATOR
UOUK> INJECTIOM PORTS
COLUMN OVEN
JUUV
CONFIRMATORY COLUMN
TOI
ANALYTICAL COLUMN
PURGE GAil
FLOW i/OnlhOi
1W MOLECULAR
SIEVE FILTER
OPTIONAL *PORT COLUMN
SELECTION VALVE
TRAP INLET
TRAP
22*C
PURGING
DEVICE
NOTE
ALL LINES BETWEEN TRAP
AND OC SHOULD K HEATED
TOIOX.
8260A - 46
Revision 1
November 1992
-------
FIGURE 4.
SCHEMATIC OF PURGE-AND-TRAP DEVICE - DESORB MODE
CAflRERQAS
FLOWCONT
PRESSURE
REGULATOR
UOUIO INJECTION PORTS
COLUMN OVEN
OPTIONAL 4PORT COLUMN
SELECTION VALVE
CONFIRMATORY COLUMN
TO DETECTOR
ANALYTICAL COLUMN
PURGE GAS
FLOW CONTROL
13X MOLECULAR
SIEVE FILTER
/• TRAP INLET
awe
PURGING
OEVCE
NOTE
ALL UNES BETWEEN TRAP
AND OC SHOULD BE HEATH)
TO WC.
8260A - 47
Revision 1
November 1992
-------
FIGURE 5.
GAS CHROMATOGRAM OF VOLATILE ORGANICS
e'
o.
3N3ZN3iO«OTM3 d. -C'3't
©.
s:
3N3ZN380MOX3IM1 -»
3N32N38CWOTHOia -CM
3NWM130M01H3 1 Q - 1 *
3N31AM13W
3N3Hi30«OTM3ia -t 'I
u a
•
I
8260A - 48
Revision 1
November 1992
-------
FIGURE 6.
GAS CHROMATOGRAM OF VOLATILE ORGANICS
3N3ZN38C«0"»OI»U -C
3N3ZN380U01M3U<1 -»
,J
3N3ZN3eOUOX3ia
«
u
- -g
I! i;
Ss „-
• « _ *
~
I
fM
u «
02
i
3N3UA1S * 3N3TAX -
3N«Hl3UOMOlH30MO«fI0
3N3KL30U01H30WJ.3I
3N3ZN3f
uua^ovova
3N3Hl30WOlM3ia -J4T-
3N3HJ.30MOX3IQ -
3GIM01H3 1ANIA
i
1
i
I
8260A - 49
Revision 1
November 1992
-------
FIGURE 7.
GAS CHROMATOGRAM OF VOLATILE ORGANICS
o8
i— "S» ••
r- cc-
•<Ł,
^ •?'
-------
FIGURE 8.
GAS CHROMATOGRAM OF TEST MIXTURE
HI
M M P
i
i
II
I
f
e
J
I
\
if
xr
•i!
aa
*Ł
8260A - 51
Revision 1
November 1992
-------
FIGURE 9.
LOW SOILS IMPINGER
jft
PURGE INLET FITTING
SAMPLE OUTLET FITTING
3- « 6mm 0 0 GLASS TUBING
SEPTUM
CAP
40ml VIAL
8260A - 52
Revision I
November 1992
-------
METHOD 8260A
VOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS)
CAPILLARY COLUMN TECHNIQUE
7.1
Select procedure
for introducing
sample into G
Direct Injection
Purge-and-trap
7.3.4 Calculate RFs
for 5 SPCCs
7.2 Set GC/MS operating
conditions
7.3.1 Tune GC/MS
system with BFB
7.3.2 Assemble purge-and-trap
device and prepare
calibration standards
I
7.3.2.1 Perform purge-
and-trap analysis
7.3.5 Calculate %RSD
ofRFforCCCs
7.4 Perform calibration
verification
7.5 Perform GC/MS analysis
utilizing Methods 5030/8260
7.6.1 Identify analytes by
comparing the sample and
standard mass spectra
I
7.6.2 Calculate the
concentration of each
identified analyte
7.6.2.3 Report all results
Stop
8260A - 53
Revision 1
November 1992
-------
METHOD 8270B
SEMIVOLATILE ORGANIC COMPOUNDS BY
GAS CHROMATOGRAPHY/MASSSPECTROMETRY (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 3520 3540 3550 3580
Acenaphthene
Acenaphthene-d10 (I.S.)
Acenaphthylene
Acetophenone
2 - Acetyl ami nof 1 uorene
l-Acetyl-2-thiourea
Aldrin
2-Aminoanthraquinone
Aminoazobenzene
4-Aminobiphenyl
3-Ami no-9-ethyl carbazol e
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)fluoranthene
Benzo ( k) f 1 uoranthene
Benzo (g , h , i ) peryl ene
Benzo(a)pyrene
p-Benzoquinone
Benzyl alcohol
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
106-51-4
100-51-6
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
OE
X
X
X
X
ND
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
ND
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
ND
ND
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
ND
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
X
X
8270B - 1
Revision 2
November 1992
-------
ADDrooriate Preoaration Technioues
Compounds
a-BHC
0-BHC
5-BHC
y-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
Chi orobenzi late
5-Chloro-2-methylaniline
4-Chl oro-3-methyl phenol
3- (Chloromethyl )pyridine
hydrochloride
1-Chloronaphthalene
2-Chloronaphthalene
2-Chlorophenol
4-Chl oro- 1 , 2-phenyl enedi amine
4-Chloro-l,3-phenylenediamine
4-Chl orophenyl phenyl ether
Chrysene
Chrysene-d12 (I.S.)
Coumaphos
p-Cresidine
Crotoxyphos
2-Cyclohexyl-4,6-dinitro-phenol
4,4'-DDD
4,4'-DDE
4,4'-DDT
Demeton-0
Demeton-S
Diallate (cis or trans)
2,4-Diaminotoluene
Dibenz(a,j)acridine
Dibenz( a, h) anthracene
CAS No8
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
224-42-0
53-70-3
3510 3520 3540 3550
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)
X
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
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
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
ND
X
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
ND
ND
X
X
X
X
X
X
LR
X
X
X
X
X
X
X
X
X
8270B - 2
Revision 2
November 1992
-------
ADorooriate Preparation Techniaues
Compounds
Dibenzofuran
Dibenzo(a,e)pyrene
l,2-Dibromo-3-chloropropane
Di-n-butyl phthalate
Dichlone
1 , 2 -Di chl orobenzene
1 , 3-Di chl orobenzene
1,4-Di chl orobenzene
l,4-Dichlorobenzene-d4 (I.S)
3,3'-Dichlorobenzidine
2,4-Dichlorophenol
2,6-Dichlorophenol
Dichlorovos
Dicrotophos
Dieldrin
Di ethyl phthalate
Diethylstilbestrol
Di ethyl sulfate
Dihydrosaffrole
Dimethoate
3,3'-Dimethoxybenzidine
Dimethyl ami noazobenzene
7,12-Dimethylbenz(a)-
anthracene
3,3'-Dimethylbenzidine
a,a-Dimethylphenethylamine
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-Di phenyl hydrazi ne
Di-n-octyl phthalate
Disulfoton
Endosulfan I
Endosulfan II
Endosulfan sulfate
CAS No8
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
959-98-8
33213-65-9
1031-07-8
3510
X
ND
X
X
OE
X
X
X
X
X
X
X
X
X
X
X
AW,OS(67)
LR
ND
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
X
X
X
3520
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
X
X
X
3540
ND
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
3550
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
3580
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
X
X
X
8270B - 3
Revision 2
November 1992
-------
Appropriate Preparation Techniques
Compounds
CAS No8
3510
3520 3540 3550 3580
Endrin
Endrin aldehyde
Endrin ketone
EPN
Ethion
Ethyl carbamate
Ethyl methanesulfonate
Ethyl parathion
Famphur
Fensulfothion
Fenthion
Fluchloralin
Fluoranthene
Fluorene
2-Fluorobiphenyl (surr.)
2-Fluorophenol (surr.)
Heptachlor
Heptachlor epoxide
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
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
4,4'-Methylenebis(2-chloraniline)101-14-4
4,4'-Methylenebis
(N,N-dimethylaniline)
Methyl methanesulfonate
2 -Methyl naphthal ene
2-Methyl-5-nitroaniline
Methyl parathion
2-Methyl phenol
3-Methyl phenol
101-61-1
66-27-3
91-57-6
99-55-8
298-00-0
95-48-7
108-39-4
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
ND
X
X
X
DC(46)
X
X
HS(5)
HE
X
X
X
X
OE,OS(0)
X
X
X
X
X
X
X
X
X
X
ND
ND
ND
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
ND
X
X
ND
ND
ND
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
ND
ND
ND
ND
ND
ND
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
ND
X
ND
ND
ND
ND
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
X
X
ND
X
X
X
8270B - 4
Revision 2
November 1992
-------
AooroDriate Preoaration Techniaues
Compounds
4 -Methyl phenol
2 -Methyl pyri dine
Mevinphos
Mexacarbate
Mi rex
Monocrotophos
Naled
Naphthalene
Naphtha! ene-dg (I.S.)
1,4-Naphthoquinone
1-Naphthylamine
2-Naphthylamine
Nicotine
5-Nitroacenaphthene
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
5-Nitro-o-anisidine
Nitrobenzene
Nitrobenzene-dc (surr.)
4-Nitrobiphenyl
Nitrofen
2-Nitrophenol
4-Nitrophenol
5-Nitro-o-toluidine
Nitroquinol ine-1-oxide
N-Nitrosodi butyl ami ne
N-Nitrosodiethylamine
N-Nitrosodimethylamine
N-Ni trosomethyl ethyl ami ne
N-Ni trosod i phenyl ami ne
N-Nitrosodi -n-propylamine
N-Nitrosomorpholine
N-Nitrosopiperidine
N-Nitrosopyrrol idine
Octamethyl pyrophosphoramide
4,4'-Oxydianiline
Parathion
Pentachl orobenzene
Pentachloronitrobenzene
Pentachl orophenol
Perylene-d12 (I.S.)
Phenacetin
Phenanthrene
Phenanthrene-d10 (I.S.)
Phenobarbital
Phenol
CAS No8
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
82-68-8
87-86-5
62-44-2
85-01-8
50-06-6
108-95-2
3510
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
X
X
X
X
X
X
X
DC (28)
3520
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
X
X
ND
X
X
ND
X
3540
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
X
ND
X
X
ND
X
3550
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
ND
X
X
ND
X
X
ND
X
3580
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
X
X
X
X
X
X
X
X
8270B - 5
Revision 2
November 1992
-------
Approoriate Preoaration Techniaues
Compounds CAS Noa
Phenol -d6 (surr.)
1,4-Phenylenediamine 106-50-3
Phorate 298-02-2
Phosalone 2310-17-0
Phosmet 732-11-6
Phosphamidon 13171-21-6
Phthalic anhydride 85-44-9
2-Picoline 109-06-8
Piperonyl sulfoxide 120-62-7
Pronamide 23950-58-5
Propylthiouracll 51-52-5
Pyrene 129-00-0
Pyridine 110-86-1
Resorcinol 108-46-3
Safrole 94-59-7
Strychnine , 60-41-3
Sul fall ate 95-06-7
Terbufos 13071-79-9
Terphenyl-du(surr.)
1,2,4, 5-Tetrachl orobenzene 95-94-3
2,3,4,6-Tetrachlorophenol 58-90-2
Tetrachlorvinphos 961-11-5
Tetraethyl dlthiopyrophosphate 3689-24-5
Tetraethyl pyrophosphate 107-49-3
Thionazine 297-97-2
Thiophenol (Benzenethiol) 108-98-5
Toluene diisocyanate 584-84-9
o-Toluidine 95-53-4
Toxaphene 8001-35-2
2,4,6-Tribromophenol (surr.)
1, 2, 4-Trichl orobenzene 120-82-1
2,4,5-Trichlorophenol 95-95-4
2,4,6-Trichlorophenol 88-06-2
Trifluralin 1582-09-8
2,4,5-Trlmethylaniline 137-17-7
Trimethyl phosphate 512-56-1
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
3510 3520 3540 3550 3580
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)
X
X
X
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
ND
ND
ND
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
ND
ND
ND
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
ND
ND
ND
ND
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
X
LR
X
X
a Chemical Abstract Service Registry Number.
AW = Adsorption to walls of glassware during
CP = Nonreproducible chromatographic perform
extraction
ance.
and
storage.
8270B - 6
Revision 2
November 1992
-------
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.
Percent Stability = Average Recovery (Day 7) x 100/Average Recovery (Day 0).
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, y-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,
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 /jg/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.
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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.
4.1.2 Column - 30 m x 0.25 mm ID (or 0.32 mm ID) 1 urn 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 /iL 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.
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
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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.
4.2 Syringe - 10 jiL.
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 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
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 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.
5.3.3 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-d., acenaphthene-d10, phenanthrene-d10,
chrysene-d.2, and perylene-d12 (see Table 5). Other compounds may be used as
internal standards as long as the requirements given in Section 7.3.2 are met.
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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-d.,. 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 /xL of the internal standard solution, resulting in a
concentration of 40 ng//iL of each internal standard. Store at 4°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 4°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
be spiked with 10 ML 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-d6, 2-fluorophenol, 2,4,6-tribromophenol, nitrobenzene-ds,
2-fluorobiphenyl, and p-terphenyl-d,4. 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,
Section 4.1.
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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 nl 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 jug/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, 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
a 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 /xL
Carrier gas: Hydrogen at 50 cm/sec or helium at 30 cm/sec
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
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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 DDD should not exceed 20%. 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.
7.3.2 The internal standards selected in Section 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).
7.3.3 Analyze 1 /il_ 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 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.
Cjs = Concentration of the specific internal standard (ng/jiL).
Cx = Concentration of the compound being measured (ng//iL).
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 =
100[SD/RF]) 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.
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7.3.4.2 If the %RSD of any CCC is 30% or greater, then the
chromatographlc system is too reactive for analysis to begin. Clean
or replace to injector liner and/or capillary column, then repeat
the calibration procedure beginning with section 7.4.
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 (Section 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 second order regression fit of the five
calibration points. The analyst should select the regression order
which introduces the least calibration error into the quantitation
(Section 7.6.2.2 and 7.6.2.3). If the %RSD is <15%, 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
semi volatile 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 (Section 7.4.3) and
CCC (Section 7.4.4) criteria.
7.4.3 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,
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:
ci - cc
% Drift = - x 100
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where:
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 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 check calibration (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 standard check, the mass spectrometer must be inspected for
malfunctions and corrections must be made, as appropriate.
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 nl 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
surrogates (for a 1 fj,i injection). The recommended GC/MS operating
conditions to be used are specified in Section 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//iL 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 Section 7.6. Store the extracts at 4°C, protected from
light in screw-cap vials equipped with unpierced Teflon lined septa.
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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.
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
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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 deli sting 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.
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 cis)
Cex (mg/L) =
(Au x RF)
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where Cex is the concentration of the compound in the extract, and
the other terms are as defined in Section 7.4.3.
7.6.2.3 Alternatively, the regression line fitted to the
initial calibration (Section 7.4.6.1) 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 Sections 7.7.2.4.1 and 7.7.2.4.2.
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) • (CCK x VC)t)
where:
VB, = extract volume, in ml
,ex
V = 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 (Mg/kg) =
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 formulas
given above should be used with the following modifications: The
areas A 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 8080.
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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 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 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 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 Steps 7.3.1 and 7.4.1.
8.4.2 There must be an initial calibration of the GC/MS system as
specified in Step 7.3.
8.4.3 The GC/MS system must meet the SPCC criteria specified in Step
7.4.3 and the CCC criteria in Section 7.4.4, 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 100 mg/L in
acetone. 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 pipet, 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.
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8.5.3 Analyze the well-mixed QC reference samples according to the
method beginning In Step 7.1. with extraction of the samples.
8.5.4 Calculate the average recovery (x) In M9/U 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 Step
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 Step
8.5.2.
8.5.6.2 Beginning with Step 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 Step
8.5.2.
8.6 The laboratory must, on an ongoing basis, analyze a reagent blank,
a matrix spike, and a 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 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 Step 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 |ig/L or 1 to 5 times higher than
8270B - 19 Revision 2
November 1992
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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
lig/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 (Step 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 jig/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
(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 Step 8.7.
8.7 If any analyte in a water sample fails the acceptance criteria for
recovery in Step 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 Step 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.
8270B - 20 Revision 2
November 1992
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8.7.1 Prepare the QC reference sample by adding 1.0 mL of the QC
reference sample concentrate (Step 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 Step 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 Step 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 Jof the same matrix) as in Step 8.6, 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).
8.9 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 Step 8.9.3 must
fall within those given in Table 8 for these matrices.
8270B - 21 Revision 2
November 1992
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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 Mg/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.
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 Soxtec extraction) is
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.
8270B - 22 Revision 2
November 1992
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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. "Interlaboratory 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.
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 - 23 Revision 2
November 1992
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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 -Di chl orobenzene
1,4-Di chl orobenzene-d4 (I.S.)
1,4-Di chl orobenzene
Benzyl alcohol
1,2-Di chl orobenzene
N-Ni trosomethyl ethyl ami ne
Bis(2-chloroisopropyl) ether
Ethyl carbamate
Thiophenol (Benzenethiol)
Methyl methanesulfonate
N-Ni trosodi-n-propyl ami ne
Hexachloroethane
Maleic anhydride
Nitrobenzene
Isophorone
N-Ni trosodi ethyl ami ne
2-Nitrophenol
2, 4-Dimethyl phenol
p-Benzoquinone
Bis(2-chloroethoxy)methane
Benzoic acid
2,4-Dichlorophenol
Trimethyl phosphate
Ethyl methanesulfonate
1,2, 4-Tri chl orobenzene
Naphthalene-d8 (I.S.)
Naphthalene
Hexachlorobutadiene
Tetraethyl pyrophosphate
Di ethyl sulfate
4-Chl oro-3-methyl phenol
2-Methylnaphthalene
2 -Methyl phenol
Hexachloropropene
Hexachl orocycl opentadi ene
N-Ni trosopyrrol idi ne
Acetophenone
4-Methyl phenol
2,4,6-Trichlorophenol
o-Toluidine
3-Methyl phenol
2-Chloronaphthalene
3.75a
5.68
5.77
5.82
5.97
6.27
6.35
6.40
6.78
6.85
6.97
7.22
7.27
7.42
7.48
7.55
7.65
7.65
7.87
8.53
8.70
8.75
9.03
9.13
9.23
9.38
9.48
9.53
9.62
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
13.30
93
93
94
93
128
146
152
146
108
146
88
45
62
110
80
70
117
54
77
82
102
139
122
108
93
122
162
110
79
180
136
128
225
99
139
107
142
107
213
237
100
105
107
196
106
107
162
66,92
66,65
65,66
63,95
64,130
148,111
150,115
148,111
79,77
148,111
42,88,43,56
77,121
62,44,45,74
110,66,109,84
80,79,65,95
42,101,130
201,199
54,98,53,44
123,65
95,138
102,42,57,44,56
109,65
107,121
54,108,82,80
95,123
105,77
164,98
110,79,95,109,140
79,109,97,45,65
182,145
68
129,127
223,227
99,155,127,81,109
139,45,59,99,111,125
144,142
141
107,108,77,79,90
213,211,215,117,106,141
235,272
100,41,42,68,69
71,105,51,120
107,108,77,79,90
198,200
106,107,77,51,79
107,108,77,79,90
127,164
8270B - 24
Revision 2
November 1992
-------
TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary
Ion
Secondary
Ion(s)
N-Nitrosopiperidine 13
1,4-Phenylenediamine 13
1-Chloronaphthalene 13
2-Nitroaniline 13
5-Ch1oro-2-methylaniline 14
Dimethyl phthalate 14
Acenaphthylene 14
2,6-Dinitrotoluene 14
Phthalic anhydride 14
o-Anisidine 15
3-Nitroaniline 15
Acenaphthene-d10 (I.S.) 15
Acenaphthene 15
2,4-Dinitrophenol 15
2,6-Dinitrophenol 15
4-Chloroaniline 15
Isosafrole 15
Dibenzofuran 15
2,4-Diaminotoluene 15
2,4-Dinitrotoluene 15
4-Nitrophenol 15
2-Naphthylamine 16
1,4-Naphthoquinone 16
p-Cresidine 16
Dichlorovos 16
Diethyl phthalate 16
Fluorene 16
2,4,5-Trimethylaniline 16
N-Nitrosodibutylamine 16
4-Chlorophenyl phenyl ether 16
Hydroquinone 16
4,6-Dinitro-2-methylphenol 17
Resorcinol 17
N-Nitrosodiphenylamine 17
Safrole 17
Hexamethyl phosphoramide 17
3-(Chioromethyl)pyridine hydrochloride!7
Diphenylamine 17
1,2,4,5-Tetrachlorobenzene 17
1-Naphthylamine 18
l-Acetyl-2-thiourea 18
4-Bromophenyl phenyl ether 18
Toluene diisocyanate 18
2,4,5-Trichlorophenol 18
Hexachlorobenzene 18
Nicotine 18
Pentachlorophenol 19
.55 114 42,114,55,56,41
.62 108 108,80,53,54,52
.65a 162 127,164
.75 65 92,138
.28 106 106,141,140,77,89
.48 163 194,164
.57 152 151,153
.62 165 63,89
.62 104 104,76,50,148
.00 108 80,108,123,52
.02 138 108,92
.05 164 162,160
.13 154 153,152
.35 184 63,154
.47 162 162,164,126,98,63
.50 127 127,129,65,92
.60 162 162,131,104,77,51
.63 168 139
.78 121 121,122,94,77,104
.80 165 63,89
.80 139 109,65
.00a 143 115,116
.23 158 158,104,102,76,50,130
.45 122 122,94,137,77,93
.48 109 109,185,79,145
.70 149 177,150
.70 166 165,167
.70 120 120,135,134,91,77
.73 84 84,57,41,116,158
.78 204 206,141
.93 110 110,81,53,55
.05 198 51,105
.13 110 110,81,82,53,69
.17 169 168,167
.23 162 162,162,104,77,103,135
.33 135 135,44,179,92,42
.50 92 92,127,129,65,39
.54a 169 168,167
.97 216 216,214,179,108,143,218
.20 143 143,115,89,63
.22 118 43,118,42,76
.27 248 250,141
.42 174 174,145,173,146,132,91
.47 196 196,198,97,132,99
.65 284 142,249
.70 84 84,133,161,162
.25 266 264,268
8270B - 25
Revision 2
November 1992
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TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary Secondary
Ion Ion(s)
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
Dial!ate (trans or cis)
Pentachlorobenzene
5-Nitro-o-anisidine
Pentachloroni trobenzene
4-Nitroquinoline-l-oxide
Di-n-butyl phthalate
2,3,4,6-Tetrachlorophenol
Dihydrosaffrole
Demeton-0
Fluoranthene
1,3,5-Tri n i trobenzene
Dicrotophos
Benzidine
Trifluralin
Bromoxynil
Pyrene
Monocrotophos
Phorate
Sulfall ate
Demeton-S
Phenacetin
Dimethoate
Phenobarbital
Carbofuran
Octamethyl pyrophosphoramide
4-Aminobiphenyl
Dioxathion
Terbufos
o,a-Dimethylphenylamine
Pronamide
Aminoazobenzene
Dichlone
Dinoseb
Disulfoton
Fluchloralin
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
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
8270B - 26 Revision 2
November 1992
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TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary Secondary
Ion Ion(s)
Mexacarbate
4,4'-Oxydianiline
Butyl benzyl phthalate
4-Nitrobiphenyl
Phosphamidon
2-Cyclohexyl-4,6-Dinitrophenol
Methyl parathion
Carbaryl
Dimethyl ami noazobenzene
Propylthiouracil
Benz(a)anthracene
Chrysene-d12 (I.S.)
3,3'-Dichlorobenzidine
Chrysene
Malathion
Kepone
Fenthion
Parathion
Anilazine
Bis(Z-ethylhexyl) phthalate
3,3'-Dimethylbenzidine
Carbophenothion
5-Nitroacenaphthene
Methapyrilene
Isodrin
Captan
Chlorfenvinphos
Crotoxyphos
Phosmet
EPN
Tetrachlorvinphos
Di-n-octyl phthalate
2-Ami noanthraqui none
Barban
Aramite
Benzo(b)fluoranthene
Nitrofen
Benzo(k)fluoranthene
Chiorobenzi late
Fensulfothion
Ethion
Diethylstilbestrol
Famphur
Tri-p-tolyl phosphate
Benzo(a)pyrene
Perylene-d,2 (I.S.)
7,12-Dimetnylbenz(a)anthracene
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
32.75 368 368,367,107,165,198
32.80 252 253,125
33.05 264 260,265
33.25 256 256,241,239,120
8270B - 27
Revision 2
November 1992
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TABLE 1.
(Continued)
Compound
Retention
Time (min.)
Primary Secondary
Ion Ion(s)
5,5-Diphenylhydantoi n
Captafol
Dinocap
Methoxychlor
2-Acetylami nof1uorene
4,4'-Methylenebis(2-chloroaniline)
3,3'-Dimethoxybenzidine
3-Methylcholanthrene
Phosalone
Azinphos-methyl
Leptophos
Mi rex
Tris(2,3-dibromopropyl) phosphate
Dibenz(a>J)acridine
Mestranol
Coumaphos
Indeno(l,2,3-cd)pyrene
Dibenz(a,h)anthracene
Benzo(g,h,i)perylene
1,2:4,5-Di benzopyrene
Strychnine
Piperonyl sulfoxide
Hexachlorophene
Aldrin
Aroclor-1016
Aroclor-1221
Aroclor-1232
Aroclor-1242
Aroclor-1248
Aroclor-1254
Aroclor-1260
a-BHC
0-BHC
8-BHC
y-BHC (Lindane)
4,4'-DDD
4,4'-DDE
4,4'-DDT
Dieldrin
1,2-Di phenylhydrazi ne
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
33.40 180 180,104,252,223,209
33.47 79 79,77,80,107
33.47 69 69,41,39
33.55 227 227,228,152,114,274,212
33.58 181 181,180,223,152
34.38 231 231,266,268,140,195
34.47 244 244,201,229
35.07 268 268,252,253,126,134,113
35.23 182 182,184,367,121,379
35.25 160 160,132,93,104,105
35.28 171 171,377,375,77,155,379
35.43 272 272,237,274,270,239,235
35.68 201 137,201,119,217,219,199
36.40 279 279,280,277,250
36.48 277 277,310,174,147,242
37.08 362 362,226,210,364,97,109
39.52 276 138,227
39.82 278 139,279
41.43 276 138,277
41.60 302 302,151,150,300
45.15 334 334,335,333
46.43 162 162,135,105,77
47.98 196 196,198,209,211,406,408
66 263,220
222 260,292
190 224,260
190 224,260
222 256,292
292 362,326
292 362,326
360 362,394
183 181,109
181 183,109
183 181,109
183 181,109
235 237,165
246 248,176
235 237,165
79 263,279
77 105,182
195 339,341
337 339,341
272 387,422
263 82,81
67 345,250
317 67,319
8270B - 28
Revision 2
November 1992
-------
TABLE 1.
(Continued)
Retention Primary Secondary
Compound Time (min.) Ion Ion(s)
2-Fluorobiphenyl (surr.) -- 172 171
2-Fluorophenol (surr.) -- 112 64
Heptachlor -- 100 272,274
Heptachlor epoxide -- 353 355,351
Nitrobenzene-dc (surr.) -- 82 128,54
N-Nitrosodimetnylamine -- 42 74,44
Phenol-d, (surr.) -- 99 42,71
Terphenyl-d14 (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.
Substitute for the non-specific mixture, tricresyl phosphate.
8270B - 29 Revision 2
November 1992
-------
TABLE 2.
ESTIMATED QUANTITATION LIMITS (EQLs) FOR SEMIVOLATILE ORGANICS8
Estimated
Quantitation
Limitsb
Ground water
Semivolatlles M9/L
Acenaphthene
Acenaphthylene
Acetophenone
2 - Acety 1 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)fluoranthene
Benzo(k)f1uoranthene
Benzole acid
Benzo (g , h , i ) peryl ene
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-Chl oro-2-methyl ani 1 ine
4-Chl oro-3-methyl phenol
3-(Chloromethyl)pyridine hydrochloride
2 -Chi oronaphthal ene
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/Sediment1
M9/kg
660
660
ND
ND
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 - 30 Revision 2
November 1992
-------
TABLE 2.
(Continued)
Estimated
Quantitation
1 J _ • J. **D
Limits
Ground water Low Soil/Sediment1
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-Dimethylbenz(a) anthracene
3, 3' -Dimethyl benzidine
a,a-Dimethylphenethylamine
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-Di phenyl hydantoi n
Di-n-octyl phthalate
M9/L
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
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 - 31
Revision 2
November 1992
-------
Semi vol at lies
Dlsulfoton
EPN
Ethion
Ethyl carbamate
Bis(2-ethy]hexyl) phthalate
Ethyl methanesul f onate
Famphur
Fensulfothion
Fenthion
Fluchloralin
Fluoranthene
Fluorene
Hexachlorobenzene
Hexachlorobutadiene
Hexachl orocycl opentadi ene
Hexachloroethane
Hexachl orophene
Hexachl oropropene
Hexamethylphosphoramide
Hydroquinone
Indeno(l,2,3-cd)pyrene
Isodrin
Isophorone
Isosafrole
Kepone
Leptophos
Malathion
Maleic anhydride
Mestranol
Methapyrilene
Methoxychlor
3-Methyl chol anthrene
4,4'-Methyleneb1s(2-chloroani
Methyl methanesul f onate
2-Methyl naphthalene
Methyl parathion
2-Methyl phenol
3 -Methyl phenol
4-Methyl phenol
Mevlnphos
Mexacarbate
Mi rex
Monocrotophos
Naled
TABLE 2.
(Continued)
Estimated
Quantisation
Limitsb
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
line) NA
10
10
10
10
10
10
10
20
10
40
20
1 /Sediment1
M9/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 - 32
Revision 2
November 1992
-------
Semivolatiles
Naphthalene
1,4-Naphthoquinone
1-Naphthylamine
2-Naphthylamine
Nicotine
5-Ni troacenaphthene
2-Nitr*oaniline
3-Nitroaniline
4-Nitroaniline
5-Nitro-o-anisidine
Nitrobenzene
4-Nitrobiphenyl
Nitrofen
2-Nitrophenol
4-Nitrophenol
5-Nitro-o-toluidine
4-Nitroquinoline-l -oxide
N-Ni trosodi butyl ami ne
N-Nitrosodi ethyl ami ne
N-Nitrosodiphenylamine
N-Ni troso-di-n-propyl ami ne
N-Nitrosopiperidine
N-Nitrosopyrrol idine
Octamethyl pyrophosphoramide
4,4'-Oxydianiline
Parathion
Pentachl orobenzene
Pentachl oron i trobenzene
Pentachl orophenol
Phenacetin
Phenanthrene
Phenobarbital
Phenol
1,4-Phenylenediamine
Phorate
Phosalone
Phosmet
Phosphamidon
Phthalic anhydride
2-Picoline
Piperonyl sulfoxide
Pronamide
Propylthiouracil
Pyrene
TABLE 2.
(Continued)
Estimated
Quantitation
Limits6
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
NO
100
10
100
10
I/Sediment1
/*g/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 - 33
Revision 2
November 1992
-------
TABLE 2.
(Continued)
Estimated
Quantitation
Limits
Semivolatiles
Ground water
M9/L
Low Soil/Sediment1
M9/kg
Pyridine
Resorcinol
Safrole
Strychnine
Sul f al 1 ate
Terbufos
1,2,4, 5 -Tetrachl orobenzene
2,3,4,6 -Tetrachl orophenol
Tetrachl orvinphos
Tetraethyl pyrophosphate
Thionazine
Thiophenol (Benzenethiol)
Toluene diisocyanate
o-Toluidine
1,2, 4 -Tr i chl orobenzene
2, 4, 5-Trichl orophenol
2, 4, 6-Tri chl orophenol
Trifluralin
2,4,5-Trimethylaniline
Trimethyl phosphate
1,3, 5-Tri ni trobenzene
Tri s (2 , 3-di bromopropyl ) phosphate
Tri-p-tolyl phosphate(h)
0,0,0-Tri ethyl phosphorothioate
NO
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 EQLs listed for soil/sediment are based on wet weight. Normally data is
reported on a dry weight basis, therefore, EQLs will be higher based on the
% dry weight of each sample. This is based on a 30 g sample and gel
permeation chromatography cleanup.
b Sample EQLs are highly matrix-dependent. The EQLs listed herein are provided
for guidance and may not always be achievable.
ND = Not determined.
NA = Not applicable.
NT = Not tested.
Other Matrices Factor
High-concentration soil and sludges by sonicator 7.5
Non-water miscible waste 75
1EQL = [EQL for Low Soil/Sediment (Table 2)] X [Factor].
1
8270B - 34
Revision 2
November 1992
-------
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
aSee Reference 4.
TABLE 4.
CALIBRATION CHECK COMPOUNDS
Base/Neutral Fraction
Acid Fraction
Acenaphthene
1,4-Di chlorobenzene
Hexachlorobutadiene
N-Ni trosodi phenylami ne
Di-n-octyl phthalate
Fluoranthene
Benzo(a)pyrene
4-Chloro-3-methylphenol
2,4-Dichlorophenol
2-Nitrophenol
Phenol
Pentachlorophenol
2,4,6-Trichlorophenol
8270B - 35
Revision 2
November 1992
-------
TABLE 5.
SEMIVOLATILE INTERNAL STANDARDS WITH CORRESPONDING ANALYTES
ASSIGNED FOR QUANTITATION
1,4-Dichlorobenzene-d4 Naphtha!ene-da
Acenaphthene-d
10
Aniline
Benzyl alcohol
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl)
ether
2-Chlorophenol
1,3-Dichlorobenzene
1,4-Dichlorobenzene
1,2-Di chlorobenzene
Ethyl methanesulfonate
2-Fluorophenol (surr.)
Hexachloroethan
Methyl methanesulfonate
2-Methyl phenol
4-Methylphenol
N-Nitrosodimethylamine
N-Nitroso-di-n-propyl-
amine
Phenol
Phenol-d6 (surr.)
2-Picoline
Acetophenone
Benzole acid
Bi s(2-chloroethoxy)methane
4-Chloroaniline
4-Chloro-3-rnethyl phenol
2,4-Dichlorophenol
2,6-Dichlorophenol
a,a-Dimethyl-
phenethylamine
2,4-Dimethyl phenol
Hexachlorobutadiene
Isophorone
2-Methylnaphthalene
Naphthalene
Nitrobenzene
Nitrobenzene-d8 (surr.)
2-Nitrophenol
N-Ni trosodi butyl ami ne
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 - 36
Revision 2
November 1992
-------
TABLE 5.
(Continued)
Phenanthrene-d
10
Chrysene-d12
Perylene-d
12
4-Aminobiphenyl
Anthracene
4-Bromophenyl phenyl
ether
Di-n-butyl phthalate
4,6-Dinitro-2-methyl-
phenol
Diphenylamine
Fluoranthene
Hexachlorobenzene
N-N1trosodi phenylami ne
Pentachlorophenol
Pentachloroni trobenzene
Phenacetin
Phenanthrene
Pronamide
Benzidine
Benzo(a)anthracene
Bis(2-ethylhexyl)
phthalate
Butyl benzyl phthalate
Chrysene
3,3'-Dichlorobenzidine
p-Dimethyl ami noazobenzene
Pyrene
Terphenyl-du (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 - 37
Revision 2
November 1992
-------
TABLE 6.
QC ACCEPTANCE CRITERIA8
Compound
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benz(a)anthracene
Benzo (b) f 1 uoranthene
Benzo ( k) f 1 uoranthene
Benzo(a)pyrene
Benzo(ghi)perylene
Benzyl butyl phthalate
/3-BHC
6-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' -DDE
4,4'-DDT
Di benzo ( a , h ) anthracene
Di-n-butyl phthalate
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1,4-Dichlorobenzene
3,3' -Dichlorobenzidine
Dieldrin
Di ethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Endosulfan sulfate
Endrin aldehyde
Fl uoranthene
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»oPs
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 - 38
Revision 2
November 1992
-------
TABLE 6.
(Continued)
Compound
Test
cone.
(M9/L)
Limit
for s
(M9/L)
Range
for x
(M9/L)
Range
P» Ps
(%)
Hexachloroethane 100 24.5
Indeno(l,2,3-cd)pyrene 100 44.6
Isophorone 100 63.3
Naphthalene 100 30.1
Nitrobenzene 100 39.3
N-Nitrosodi-n-propylamine 100 55.4
PCB-1260 100 54.2
Phenanthrene 100 20.6
Pyrene 100 25.2
1,2,4-Trichlorobenzene 100 28.1
4-Chloro-3-methylphenol 100 37.2
2-Chlorophenol ' 100 28.7
2,4-Chlorophenol 100 26.4
2,4-Dimethylphenol 100 26.1
2,4-Dinitrophenol 100 49.8
2-Methyl-4,6-dinitrophenol 100 93.2
2-Nitrophenol 100 35.2
4-Nitrophenol 100 47.2
Pentachlorophenol 100 48.9
Phenol 100 22.6
2,4,6-Trichlorophenol 100 31.7
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
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 = Standard deviation of four recovery measurements, in /ug/L.
x = Average recovery for four recovery measurements, in M9/L.
p, ps = Percent recovery measured.
D = Detected; result must be greater than zero.
a 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 - 39
Revision 2
November 1992
-------
TABLE 7.
METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION8
Compound
Acenaphthene
Acenaphthylene
Aldrin
Anthracene
Benz (a) anthracene
Chloroethane
Benzo(b)fluoranthene
Benzo(k)fluoranthene
Benzo(a)pyrene
Benzo (gh i ) peryl ene
Benzyl butyl phthalate
/3-BHC
6-BHC
Bis(Z-chloroethyl) ether
Accuracy, as
recovery, x'
(M9A)
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
Bis(2-chloroethoxy)methanel.l2C-5.04
Bi s (2-chl oroi sopropyl )
ether
Bis(2-ethylhexyl)
phthalate
4-Bromophenyl phenyl
ether
2 -Chi 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-Dlchlorobenzene
3,3'-Dichlorobenzidine
Dieldrin
Di ethyl phthalate
Dimethyl phthalate
2,4-Dinitrotoluene
2,6-Dinitrotoluene
Di-n-octyl phthalate
Endosulfan sulfate
Endrin aldehyde
Fluoranthene
Fluorene
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
8270B
Single analyst
precision, s '
(Mg/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
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
0.21X+1.19
0.12X+2.47
O.lSx+3.91
0.22X-0.73
0.12X+0.26
- 40
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
O.Slx-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
0.13X+0.61
Revision i
November 1992
-------
TABLE
7.
(Continued)
Compound
Heptachlor
Heptachlor epoxide
Hexachlorobenzene
Hexachl orobutad i ene
Hexachloroethane
Indeno(l,2,3-cd)pyrene
Isophorone
Naphthalene
Nitrobenzene
N-Nitrosodi -n-propyl amine
PCB-1260
Phenanthrene
Pyrene
1 , 2 , 4-Tri chl orobenzene
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'
(M9/L)
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 '
UgA)
0.24x-0.56
0.33x-0.46
0.18x-0.10
0.19x+0.92
0.17x+0.67
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.lBx+0.85
0.23X+0.75
O.lSx+1.46
O.lSx+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' (/ig/L)
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
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'
Expected recovery for one or more measurements of a sample
containing a concentration of C, in
S'
C
x"
Expected single analyst standard deviation of measurements at an
average concentration of x, in jug/L.
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 fig/I.
8270B - 41
Revision 2
November 1992
-------
TABLE 8.
SURROGATE SPIKE RECOVERY LIMITS FOR WATER AND SOIL/SEDIMENT SAMPLES
Low/High Low/High (
Surrogate Compound Water So11/Sediment
Nitrobenzene-d5 35-114 23-120
2-Fluorobiphenyl 43-116 30-115
p-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
8270B - 42 Revision 2
November 1992
-------
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 80
4-Chloro-l,2-phenylenediamine 91
4-Chloro-l,3-phenylenediamine 84
l,2-Dibromo-3-chloropropane 97
2-sec-Butyl-4,6-dinitrophenol 99
Ethyl parathion 100
4,4'-Methylenebis(N,N-dimethylaniline) 108
2-Methyl-5-nitroaniline 99
2-Methylpyridine 80
Tetraethyl dithiopyrophosphate 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 - 43
Revision 2
November 1992
-------
TABLE 10.
AVERAGE PERCENT RECOVERIES AND PERCENT RSDs FOR THE TARGET COMPOUNDS
FROM SPIKED CLAY SOIL AND TOPSOIL BY SOXTEC EXTRACTION WITH HEXANE-ACETONE (1:1)'
Clay Soil
Cmpd
f
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
Average
percent
Compound name recovery
1,3-Dichlorobenzene
1,2-Dichlorobenzene
Nitrobenzene
Benzal chloride
Benzotrichloride
4-Chl oro-2-ni trotol uene
Hexachl orocycl opentadi ene
2,4-Dichloronitrobenzene
3,4-Dichloronitrobenzene
Pentachlorobenzene
2,3,4, 5-Tetrachl oroni 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-Trichlorophenyl-
4'-nitrophenyl ether
Mi rex
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
Topsoil
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 - 44
Revision 2
November 1992
-------
TABLE 11.
SINGLE LABORATORY ACCURACY AND PRECISION DATA FOR THE EXTRACTION
OF SEMIVOLATILE ORGANICS FROM SPIKED CLAY BY
METHOD 3541 (AUTOMATED SOXHLET)8
Compound
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
31
32
33
34
35
36
37
38
39
40
41
Compound name
Phenol
Bis(2-chloroethyl Jether
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
Benzole acid
Bis(2-chloroethoxy)methane
2,4-Dichlorophenol
1 , 2 , 4-Tr i chl orobenzene
Naphthalene
4-Chloroaniline
4-Chloro-3-methyl phenol
2-Methyl naphthalene
Hexachl orocycl opentadi ene
2,4,6-Trichlorophenol
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-Dinitro-2-methyl phenol
N-N i trosodi phenyl ami ne
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 - 45
Revision 2
November 1992
-------
Table 11. (Continued)
Compound
number
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
Compound name
Hexachlorobenzene
Pentachlorophenol
Phenanthrene
Anthracene
Di-n-butyl phthalate
Fluoranthene
Pyrene
Butyl benzyl phthalate
3,3'-Dichlorobenzidine
Benzo(a) anthracene
B1s(2-ethylhexyl) phthalate
Chrysene
Di-n-octyl phthalate
Benzo ( b) f 1 uoranthene
Benzo ( k) f 1 uoranthene
Benzo(a)pyrene
Indeno(l,2,3-cd)pyrene
Dibenzo(a,h) anthracene
Benzo (g,h,i)perylene
1 , 2-Di chl orobenzene
1,3-Dichlorobenzene
1 , 4-Di chl orobenzene
Hexachloroethane
Hexachlorobutadiene
Average
percent
recovery
72.6
62.7
83.9
96.3
78.3
87.7
102
66.3
25.2
73.4
77.2
76.2
83.1
82.7
71.7
71.7
72.2
66.7
63.9
0
0
0
0
0
Percent
RSD
3.7
6.1
5.4
3.9
40
6.9
0.8
5.2
11
3.8
4.8
4.4
4.8
5.0
4.1
4.1
4.3
6.3
8.0
--
--
--
--
8 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 - 46 Revision 2
November 1992
-------
FIGURE 1.
GAS CHROMATOGRAM OF BASE/NEUTRAL AND ACID CALIBRATION STANDARD
s
8270B - 47
Revision 2
November 1992
-------
METHOD 8270B
SEMIVOLATILE ORGANIC COMPOUNDS BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY (GC/MS):
CAPILLARY COLUMN TECHNIQUE
7 1 Prepare
sample using
Method 3540
or 3SSO
? 1 Prepare
sample using
Method 3S10
or 3S20
7 1 Prepare
sample using
Method 3540,
3550 or 3580
7 2 Cleanup
extract
7 3 Set CC/MS
operating
condition*
Perform initial
calibration
7 4 Perform daily
calibration with
SPCCa and CCCs
prior to analysis
of sample*
8270B - 48
Revision 2
November 1992
-------
METHOD 8270B
(Continued)
7 5 4 Dilut«
extract
7
751 Screen
extract on CC/FID
or CC/PID to
eliminate samples
that are too
concentrated
7 5 3 Analyze
extract by CC/MS.
using appropriate
fused•s11 tea
capi 1lary column
7 6 1 Identify
analyte by
cocnparing the
sample and standard
mass spectra
762 Calculate
concentration of
each individual
analyte Report
results
Stop
8270B - 49
Revision 2
November 1992
-------
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
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)
1,2,3,7,8-Pentachlorodibenzo-p-dioxin (PeCDD)
1,2,3,6,7,8-Hexachlorodibenzo-p-dioxin (HxCDD)
1,2,3,4,7,8-Hexachlorodibenzo-p-dioxin (HxCDD)
1,2,3,7,8,9-Hexachlorodibenzo-p-dioxin (HxCDD)
1,2,3,4,6,7,8-Heptachlorodibenzo-p-dioxin (HpCDD)
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-Hexachlorodibenzofuran (HxCDF)
1,2,3,7,8,9-Hexachlorodi benzofuran (HxCDF)
1,2,3,4,7,8-Hexachlorodibenzofuran (HxCDF)
2,3,4,6,7,8-Hexachlorodibenzofuran (HxCDF)
1,2,3,4,6,7,8-Heptachlorodibenzofuran (HpCDF)
1,2,3,4,7,8,9-Heptachlorodibenzofuran (HpCDF)
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.
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 1 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
8290 - 1 Revision 0
November 1992
-------
(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 m
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.
Section 11 of this method discusses safety procedures.
2.0 SUMMARY OF METHOD
2.1 This procedure uses matrix specific extraction, analyte specific
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 ( C.2) 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 M
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) TolueneiSoxhlet extraction for soil, sediment, fly ash and paper
pulp samples;
b) Methylene chloride:!iquid-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.
g) As an option, all solid samples (wet or dry) can be extracted with
toluene using a Soxhlet/Dean Stark extraction system.
8290 - 2 Revision 0
November 1992
-------
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 AX-21 activated carbon on Celite 545*
(or equivalent).
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 AX-21 on Celite 545* (or equivalent).
2.4.2 Fish tissue and paper pulp extracts are subjected to an acid
wash treatment only, prior to chromatography on alumina and
AX-21/Celite 545» (or equivalent).
2.5 The preparation of the final extract for HRGC/HRMS analysis is
accomplished by adding, to the concentrated AX-21/Celite 545* (or equivalent)
column eluate, 10 to 50 pi (depending on the matrix type) of a nonane solution
containing 50 pg/iuL of each of the two recovery standards C12-1,2,3,4-TCDD and
C12-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 One to two /it 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 C-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 identified PCDD/PCDF congeners are identified by their
relative retention times falling 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 C12-OCDD and
the simultaneous detection of the two most abundant ions in the molecular ion
region. Confirmation 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)
calibration curve for each homologue, during which each calibration solution is
analyzed once.
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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 Section 8.1.1.3. While
certain 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 (Section 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). When a column becomes available that resolves
all isomers, then a single analysis on this column can be used instead of
analyses on more than one column.
4.0 APPARATUS AND MATERIALS
4.1 High-Resolution Gas Chromatoqraph/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 In.iection Port - The GC injection port must be designed for
capillary columns. The use of splitless injection techniques is
recommended. On column 1 fj.1 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 n\.).
One juL injections are allowed; however, laboratories must remain
consistent throughout the analyses by using the same injection volume at
all times.
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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
(Section 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 (Section 8.1.1). At the beginning of each 12
hour period (after mass resolution and GC resolution is 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 Section 7.6.
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
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congeners, the sample extract must be reanalyzed on another GC column that
resolves the Isomers. When such a column becomes available, and the
isomer specificity can be documented, the performing laboratory will be
required to use it.
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 Section 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.
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
and hexane and store it inverted or capped with solvent rinsed
aluminum foil in a clean environment.
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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 glass rod
until free of lumps. Store in a glass bottle sealed with a Teflon™ 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, H2SO,, concentrated, ACS grade, specific gravity
1.84.
5.3.2 Potassium hydroxide, KOH, ACS grade, 20 percent (w/v) in
organic-free reagent water.
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.
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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.
or highest available purity.
High purity, distilled in glass
5.5.2 Hexane, C6HK. 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. High purity, distilled in glass or highest
available purity.
5.5.7 Acetone, CH3COCH3.
available purity.
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//iL) and the highest values for the
octachlorinated congeners (1000
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 must be prepared in the analyst's laboratory.
Traceability of standards must be verified against EPA-supplied standard
solutions. 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.
5.6.2 Store the concentration calibration solutions in
mini vials at room temperature in the dark.
1 ml
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
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isomers for the purpose of documenting the chromatographic resolution. The
C12-2,3,7,8-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//iL 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 C12-OCDF is not present in the solution.)
5.9 Recovery Standard Solution - This nonane solution contains two
recovery standards, IOC]2-1,2,3,4-TCDD and 13C12-l,2,3,7,8,9-HxCDD, at a nominal
concentration of 50 pg/juL per compound. 10 to 50 p.1 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,
Section 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
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
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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 must be identified as such.
NOTE: The holding times listed in Section 6.4 are recommendations. PCDDs
and PCDFs are very stable in a variety of matrices, and holding
times under the conditions listed in Section 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 (>25 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 pi pet, 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 (Section 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
(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 (Section 7.2.2), in a tared
200 ml round bottom flask, on a rotary evaporator until a constant weight
(W) is achieved.
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100 (W)
Percent lipid =
10
Dispose of the lipid residue as a hazardous waste if the results of m
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 Section 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 Section 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 (Section 5.8) to the sample. All samples
should be spiked with 100 juL of the sample fortification mixture to give
internal standard concentrations as indicated in Table 1. As an example,
for C12-2,3,7,8-TCDD, a 10 g soil sample requires the addition of 1000 pg
of C12-2,3,7,8-TCDD to give the required 100 ppt fortification level.
The fish tissue sample (20 g) must be spiked with 200 pi of the internal
standard solution, because half of the extract will be used to determine
the lipid content (Section 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 (Section 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 (Section 6.3) and mix thoroughly with a stainless
steel spatula. After breaking up any lumps, place the fish/sodium sulfate
mixture in the Soxhlet apparatus on top of a glasswool 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 mi of toluene).
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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 Section 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 (Section
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 Section 7.5.1.1, but omit the procedures
described in Sections 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 Section 4.3. Teflon" lined caps should be used.
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
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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 pi 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 pipet. 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 (Section
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
methylene 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 Section 7.3.2.6 to the vial. Reduce the volume of the
extract on a water bath (50-60°C) by a gentle stream of purified
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 Section 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.
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7.3.3.4 Calculate the percent lipid content of the
original sample to the nearest 0.1 percent as shown below:
u Y v
lr ext
Lipid content, LC (%) = x 100
U v V
where:
Wlr • weight of the lipid residue to the nearest 0.0001
g calculated from Section 7.3.3.3,
^ext " total volume (100 mL) of the extract in mL from
Section 7.3.2.6,
Wat » weight of the original adipose tissue sample to
the nearest 0.01 g from Section 7.3.2.1, and
Val = volume of the aliquot of the final extract in mL
used for the quantitative measure of the lipid
residue (1.0 mil).
7.3.3.5 Record the lipid residue measured in Section
7.3.3.3 and the percent lipid content from Section 7.3.3.4.
7.3.4 Adipose Tissue Extract Concentration
7.3.4.1 Quantitatively transfer the remaining extract
(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.
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 Section 7.3.5.3. Combine the hexane extracts from
Section 7.3.5.3 with the rinses.
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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 Section 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 (Sections 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 /uL.
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 Section 7.3.5.1.
7.3.5.9 The extract is ready for the column cleanups
described in Sections 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.
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 Section 7.4.4.
NOTE: If the sludge or fuel oil sample dissolves in toluene,
treat it according to the instructions in Section 7.4.2
below. If the labeled sludge sample originates from
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pulp (paper mills), treat It according to the
instructions starting in Section 7.2, but without the
addition of sodium sulfate.
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 Section 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 (j,l sample fortification
solution (Section 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
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 Section 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
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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 Section 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
(Section 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 0.45 urn glass
fiber filter that has been rinsed with toluene. If the suspended
solids content is too great to filter through the 0.45 /xm filter,
centrifuge the sample, decant, and then filter the aqueous phase.
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 Sections 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.
7.4.5.5 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 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
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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
(Section 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
Sections 7.4.5.6 and 7.4.5.8 through 7.4.5.10. Proceed
with Section 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 (Section 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.
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 Section 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.
OTE; If the dried sample is not of free flowing consistency,
more sodium sulfate must be added.
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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
Section 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
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
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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 (Section 5.2.2) can be used
in place of neutral alumina.
7.5.2.3 Dissolve the residue from Section 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.
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 (Section 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
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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 /zL of the continuing calibration
solution to 950 /uL hexane. Take this solution through
the carbon column cleanup step, concentrate to 50 /ul_
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 (Section 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,
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 fj,m) 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 ML using a stream
of nitrogen and a sand bath. Rinse the rotary evaporator flask
three times with 300 pi of a solution of 1 percent toluene in
methylene chloride, and add the rinses to the concentrate. Add
10 fj.1 of the nonane recovery standard solution for soil, sediment,
water, fish, paper pulp and adipose tissue samples, or 50 nl of the
recovery standard solution for sludge, still bottom and fly ash
samples. Store the sample at room temperature in the dark.
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7.6 Chromatographic/Mass Spectrometric Conditions and Data Acquisition
Parameters
7.6.1 Gas Chromatograph
Column coating: DB-5
Film thickness: 0.25 pm
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. Init. Temp. Final Final
Temp. Hold Time Ramp Temp. Hold
(°C) (min) (°C/min) (°C) 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 (Section
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
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
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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 (Section 7.7.3) does not meet the
required criteria listed in Section 9.4.
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
Section 7.6.2.2.
7.7.1.3 Inject 2 /xL of the GC column performance check
solution (Section 5.7) and acquire SIM mass spectral data as
described earlier in Section 8.1. The total cycle time must be < 1
second. The laboratory must not perform any further analysis until
it is demonstrated and documented that the criterion listed in
Section 8.1.2 was met.
7.7.1.4 By using the same GC (Section 7.6.1) and MS
(Section 7.6.2) conditions that produced acceptable results with the
column performance check solution, analyze a 2 p.1 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).
7.7.1.4.2 The ratio of integrated ion current for the
ions belonging to the carbon-labeled internal and recovery
standards must be within the control limits stipulated in
Table 8.
NOTE: Sections 7.7.1.4.1 and 7.7.1.4.2 require that 17
ion ratios from Section 7.7.1.4.1 and 11 ion
ratios from Section 7.7.1.4.2 be within the
specified control limits simultaneously in one
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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 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 (RRF) for unlabeled target analytes
[RRF(n); n = 1 to 17] relative to their appropriate internal
standards (Table 5) and the nine RRFs for the labeled C12
internal standards [RRF(m); m = 18 to 26)] relative to the
two recovery standards according to the following formulae:
RRF(n) =
RRF(m)
A, x Qis
is
Ais X
Q
rs
where:
rs
is
rs
sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for
unlabeled PCDDs/PCDFs,
sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for the
labeled internal standards,
sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for the
labeled recovery standards,
quantity of the internal standard injected
(pg)>
quantity of the recovery standard injected
(pg), and
quantity of the unlabeled PCDD/PCDF analyte
injected (pg).
The RRF(n) and RRF(m) are dimensionless quantities; the
units used to express Qjs, Qrs and Qx must be the same.
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7.7.1.4.5 Calculate the RRF and their respective
percent relative standard deviations (%RSD) for the five
calibration solutions:
_ 5
RRF(n) = 1/5 S RRFj(n)
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:
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 TCDF), the mean RRF
used will be the same as the mean RRF determined in
Section 7.7.1.4.5.
NOTE: The calibration solutions do not contain
C12-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 "c^CDF with [M]+ of OCDD).
Therefore, the RRF for OCDF is calculated
relative to C12-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 RRF used
for those homologous series will be the mean of the RRFs
calculated for all individual 2,3,7,8-substituted
congeners using the equation below:
1 t
RRF(k) = - S RRFn
t n=1
where:
= 27 to 30 (Table 9), with 27 = PeCDF; 28 =
HxCDF; 29 = HxCDD; and 30 = HpCDF,
= total number of 2,3,7,8-substituted isomers
present in the calibration solutions (Table
5) for eacFT homologous series (e.g., two
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for PeCDF, four for HxCDF, three for HxCDD,
two for HpCDF).
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.
7.7.1.4.7 Relative response factors [RRF(m)] to be
used for the determination of the percent recoveries for the
nine internal standards are calculated as follows:
RRF(m) =
A ffl
Mis
x Q
rs
RRF(m)
where:
m
Ms
rs
RRF(m) =
RRF(m) =
1 5
- S RRFj(m),
5 J-l
18 to 26 (congener type) and j - 1 to 5
(injection number),
sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for a
given internal standard (m = 18 to 26),
sum of the integrated ion abundances of the
quantitation ions (Tables 6 and 9) for the
appropriate recovery standard (see Table 5,
footnotes),
quantities of, respectively, the recovery
standard (rs) and a particular internal
standard (is = m) injected (pg),
relative response factor of a particular
internal standard (m) relative to an
appropriate recovery standard, as
determined from one injection, and
calculated mean relative response factor of
a particular internal standard (m) relative
to an appropriate recovery standard, as
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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 the analysis is
performed.
7.7.2.1 The percent relative standard deviations for the
mean response factors [RRF(n) and RRF(m)] from the 17 unlabeled
standards must not exceed ± 20 percent, and those for the nine
labeled reference compounds must not exceed + 30 percent.
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 isotopic ratios (Table 8) must be within the
specified control limits.
NOTE; If the criterion for acceptable calibration
listed in Section 7.7.2.1 is met, the analyte
specific RRF can then be considered independent
of the analyte quantity for the calibration
concentration range. The mean RRFs will be used
for all calculations until the routine
calibration criteria (Section 7.7.4) are no
longer met. At such time, new mean RRFs 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 ML of the concentration calibration
solution HRCC-3 standard (Table 5). By using the same HRGC/HRMS
conditions as used in Sections 7.6.1 and 7.6.2, determine and
document an acceptable calibration as provided in Section 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 RRFs [RRF(n) for the unlabeled
standards] obtained during the routine calibration runs must be
within + 20 percent of the mean values established during the
initial calibration (Section 7.7.1.4.5).
7.7.4.2 The measured RRFs [RRF(m) for the labeled
standards] obtained during the routine calibration runs must be
within + 30 percent of the mean values established during the
initial calibration (Section 7.7.1.4.7).
7.7.4.3 The ion-abundance ratios (Table 8) must be within
the allowed control limits.
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7.7.4.4 If either one of the criteria in Sections 7.7.4.1
and 7.7.4.2 is not satisfied, repeat one more time. If these
criteria are still not satisfied, the entire routine calibration
process (Section 7.7.1) must be reviewed. It is realized that it
may not always be possible to achieve all RRF criteria. For
example, it has occurred that the RRF criteria for C12-HpCDD and
C12-OCDD were not met, however, the RRF values for the
corresponding unlabeled compounds were routinely within the criteria
established in the method. In these cases, 24 of the 26 RRF
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. 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 RRF
criteria were not met included both the unlabeled and the
corresponding internal standard compounds. If the ion-abundance
ratio criterion (Section 7.7.4.3) is not satisfied, refer to the
note in Section 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 extract (from Section 7.5.3.6) or blank from
storage. With a stream of dry, purified nitrogen, reduce the extract
volume to 10 p.1 to 50 nl.
NOTE: A final volume of 20 juL or more should be used whenever
possible. A 10 fj,i final volume is difficult to handle, and
injection of 2 nl out of 10 juL leaves little sample for
confirmations and repeat injections, and for archiving.
7.8.2 Inject a 2 p.1 aliquot of the extract into the GC, operated
under the conditions that have been established to produce acceptable
results with the performance check solution (Sections 7.6.1 and 7.6.2).
7.8.3 Acquire SIM data according to Sections 7.6.2 and 7.6.3. Use
the same acquisition and mass spectrometer operating conditions previously
used to determine the relative response factors (Sections 7.7.1.4.4
through 7.7.1.4.7). Ions characteristic for 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 (Section
8.1). 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 Section 8.2.2 for
the proper level of reference compound to be metered into the
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ion chamber.) The analyst may be required to monitor a PFK
ion, not 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 a!., 1984, 1987].
Report any discrepancies in the case narrative.
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
labelled 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 C^-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 (Section 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 C12-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 (Section 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.
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7.8.4.2 Ion Abundance Ratios
7.8.4.2.1 The integrated ion current for the two ions
used for quantitation purposes must have a ratio between the
lower and upper limits established for the homologous series
to which the peak is assigned. See Sections 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 Sections 7.8.4.1.1 through 7.8.4.3.1, calculate the concen-
tration of the PCDD or PCDF compounds using the formula:
Cx
Ais x W x RRF(n)
where:
Cx = concentration of unlabeled PCDD/PCDF congeners (or group
of coeluting isomers within an homologous series) in
pg/g,
Ax = sum of the integrated ion abundances of the quantitation
ions (Table 6) for unlabeled PCDDs/PCDFs,
Ais = sum of the integrated ion abundances of the quantitation
ions (Table 6) for the labeled internal standards,
Qjs = quantity, in pg, of the internal standard added to the
sample before extraction,
W = weight, in g, of the sample (solid or liquid), and
RRF = calculated mean relative response factor for the analyte
[RRF(n) with n = 1 to 17; Section 7.7.1.4.5].
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If thejmalyte is identified as one of the 2,3,7,8-substituted PCDDs
or PCDFs, RRF(n) is the value calculated using the equation in
Section_7.7.1.4.5. However, if it is a non-2,3,7,8-substituted congener,
the RRF(k) value is the one calculated using the equation in
Section 7.7.1.4.6.2. [RRF(k) with 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 Qrs
Internal standard percent recovery = —^_ x 100
Qis x Ars x RRF(m)
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
RRF(m) = 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 Section 7.7.1,4.7 [RRF(m) 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//xL
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 (Sections 11.1 to 11.9.3). For the other congeners (including
OCDD), however, report the measured concentration and indicate that the
value exceeds the MCL.
7.9.4 The total concentration for each homologous series of PCDD and
PCDF is calculated by summing up the concentrations of all positively
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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.
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 C-labeled standard),
multiplying that noise height by 2.5, and relating the
product to an estimated concentration that would produce that
peak height.
Use the formula:
EDL (specific 2,3,7,8-subst. PCDD/PCDF) =
2.5 x Ax x Qis
Ais x W x RRF(n)
where:
EDL = estimated detection limit for homologous
2,3,7,8-substituted PCDDs/PCDFs.
A , Ais, W, RRF(n), and Qis retain the same meanings as
defined in Section 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
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S/N in excess of 2.5 and does not meet any of the other
qualitative identification criteria listed in Section 7.8.4,
calculate the "Estimated Maximum Possible Concentration"
(EMPC) according to the expression shown in Section 7.9.1,
except that Ax in Section 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) is calculated as
fol1ows:
I S, - S2 |
RPD = x 100
(S, + S2 ) / 2
S1 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
(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 Sections 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 Section 7.8, and the calculations are
performed as outlined in Section 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
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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
Section 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
(Section 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 Section 8.1.2 are
met and the requirements described in
Section 17.2.2 are followed.
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 Sections 7.8.4.2 and 7.8.4.3,
respectively. In addition, the retention time identification
criterion described in Section 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
Section 4.2. It must be documented that all applicable system performance
criteria (specified in Sections 8.2.1 and 8.2.2) were met before analysis of any
sample is performed. Section 7.6 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.
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8.2.1 GC Column Performance
8.2.1.1 Inject 2 /xL (Section 4.1.1) of the column
performance check solution (Section 5.7) and acquire selected ion
monitoring (SIM) data as described in Section 7.6.2 within a total
cycle time of < 1 second (Section 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.
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 (Section 7.8.4.1) and
quantitative purposes. All peaks (that includes C12-2,3,7,8-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
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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 (Section 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.
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
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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
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.
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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 Section 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
unlabelled compounds and_35_ percent RPD for the 9 labelled reference
compounds, use the mean RRFs from the two daily routine calibration
runs to compute the analyte concentrations, instead of the RRFs
obtained from the initial calibration. A new initial calibration
(new RRFs) 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 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 Section 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 /uL
of the solution containing the nine internal standards
(Table 2) diluted with 1.0 mL acetone (Section 7.1).
8.3.4.1.2 Extract by using the procedures beginning
in Sections 7.4.5 or 7.4.6, as applicable, add 10 /iL of the
recovery standard solution (Section 7.5.3.6) and analyze a
2 fj.i aliquot of the concentrated extract.
8.3.4.1.3 Calculate the concentration (Section 7.9.1)
of 2,3,7,8-substituted PCDDs/PCDFs and the percent recovery
of the internal standards (Section 7.9.2).
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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 juL of the solution containing the nine
internal standards (Table 2).
8.3.4.2.3 Using a KD apparatus, concentrate to
approximately 5 ml.
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 mini vial,
reducing the volume in the mini vial 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 jiL recovery
standard solution (Table 2) and reduce the volume to its
final volume, as necessary (Section 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 /ug/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
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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 (Sections 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").
8.3.6.2 Add an appropriate volume of the matrix spike
fortification solution (Section 5.10) and of the sample
fortification solution (Section 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
Section 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 (Section 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
Sections 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 (Section 7.9.5)
8.5.2 If the first initial identification criteria (Sections
7.8.4.1.1 through 7.8.4.1.4) are met, but the criteria appearing in
Sections 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.
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8.6 Unused portions of samples and sample extracts must 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).
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).
11.0 SAFETY
11.1 The following safety practices are excerpts from EPA Method 613,
Section 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
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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.
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.
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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 m
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.
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.
8290 - 44 Revision 0 m
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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-
lamination 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,L of the sample fortification
solution to each jar containing used or unused wipers (Section 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 Section
7.5.2.2 and follow the steps outlined in Sections 7.5.2.3 through 7.5.2.5.
A.1.3 Add 10 ML of the recovery standard solution as described in
Section 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 /uL 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 Section 7.8. Perform
calculations according to Section 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
r ^
o
Dibenzodioxin
8
6 ^ 0 4
Oibenzof uran
General structures of dibenzo-p-dioxin and dibenzofuran.
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Figure 2.
M/AM
5,600
5,600
8,550
400 ppm
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
GC Column
Performance
11:00 AM
Samples
Method
Blank
8:00 PM
Mass
Resolution
Routine
Calibration
Typical 12 hour analysis sequence of events.
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Figure 4.
6'8'Z'L
8'L't'Z-
B't'Z'l/L't'Z'l
l
fr'C'Z'l
8'9'C'l
o
•I1"
M
O
n
o
o
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 Section 7.6.
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Figure 5.
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/QM of 10,500 (10 percent valley
definition).
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Figure 6.
100
90
80-
70-
60-
50-
40-
30-
20-
10-
E,
jL
T
E«
N
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)
Soil
Sediment
Fly
Water Paper Pulp Ash
Human
Fish Adipose Sludges, Still-
Tissue6 Tissue Fuel Oil Bottom
Lower MCL(a) 0.01 1.0
Upper MCL(a) 2 200
Weight (g) 1000 10
IS Spiking
Levels (ppt)
1
100
Final Extr,
Vol. (AiL)(d) 10-50 10-50
1.0 1.0 1.0 5.0
200 200 200 1000
10 20 10 2
100 100
100
50 10-50 10-50
500
50
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 Section 6.5.
(c) One half of the extract from the 20 g sample is used for determination of
lipid content (Section 7.2.2).
(d) See Section 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/juL; Solvent:
Nonane)
Recovery Standard
Solution
Concentration
(pg//iL; Solvent:
Nonane)
13C12-2,3,7,8-TCDD
13C12-2,3,7,8-TCDF
13C12-1,2,3,4-TCDD
13C12-l,2,3,7,8-PeCDD
13C12-l,2,3,7,8-PeCDF
13C12-l,2,3,6,7,8-HxCDD
;3C12-l,2,3,4,7,8-HxCDF
13C12-l,2,3,7,8,9-HxCDD
;3C12-l,2,3,4,6,7,8-HpCDD
3C12-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
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Table 5.
High-Resolution Concentration Calibration Solutions
Concentration (pg/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
2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
OCDD
OCDF
Internal Standards
"C,,- 2,3,7,8-TCDD
%-2,3,7,8-TCDF
3C12-l,2,3,7,8-PeCDD
3C12-l,2,3,7,8-PeCDF
13C12-l,2,3,6,7,8-HxCDD
13C12-l,2,3,4,7,8-HxCDF
13C.,-l,2,3,4,6,7,8-HpCDD
;3C12-l,2,3,4,6,7,8-HpCDF
13C52-OCDD
Recovery Standards
13C1?-l,2,3,4-TCDD
13C,2-l,2,3,7,8,9-HxCDD(b>
200
200
500
500
500
500
500
500
500
500
500
500
500
500
500
1,000
1,000
50
50
50
50
125
125
125
125
250
50
125
50
50
125
125
125
125
125
125
125
125
125
125
125
125
125
250
250
50
50
50
50
125
125
125
125
250
50
125
10
10
25
25
25
25
25
25
25
25
25
25
25
25
25
50
50
50
50
50
50
125
125
125
125
250
50
125
2.5
2.5
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
6.25
12.5
12.5
50
50
50
50
125
125
125
125
250
50
125
1
1
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
5
5
50
50
50
50
125
125
125
125
250
50
125
-------
Table 6.
Ions Monitored for HRGC/HRMS Analysis of PCDDs/PCDFs
Descriptor
1
2
3
Accurate(a)
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
^3
C12H335C1437C10
C12H335C1337C120
13C12H335C1437C10
13C12H335C1337C120
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 - 59
Revision 0
November 1992
-------
Table 6.
Continued
Descriptor Accurate 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
[442.9278] LOCK
Elemental
Composition
C12H35C1637C10
C12H35C1537C120
13C12H35C170
13C12H35C1637C10
C12H35C1637C102
C12H35C1537C1202
13C12H35C1637C102
13C12H35C1537C1202
C12H35C1737C120
Vl7
C1235C1737C10
C1235C1637C120
C1235C1737C102
C1235C1637C1202
13C1235C1737C102
13c1235ci637ci2o2
C1235C1837C120
^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
(a) The following nuclidic masses were used:
H = 1.007825 0
C = 12.000000 35C1
13C = 13.003355 37C1
F = 18.9984
15.994915
34.968853
36.965903
S = Internal/recovery standard
8290 - 60
Revision 0
November 1992
-------
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
1,2,3,4,6,7
1,2,3,4,6,7,8
1,2,3,4,6,7,8,9
PCDF Positional
First
Eluter
1,3,6,8
1,3,4,6,8
1,2,3,4,6,8
1,2,3,4,6,7,8
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 ispmers, the 1,2,3,4-, 1,2,3,7-, 1,2,3,8-, 2,3,7,8-,
C12-2,3,7,8-, and 1,2,3,9-TCDD isomers must also be present as a check of column
(a)
resolution.
8290 - 61 Revision 0
November 1992
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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
6(a>
7
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
(a) Used only for 13C-HxCDF (IS).
-------
Table 9.
Relative Response Factor [RRF (number)] Attributions
Number Specific Congener Name
1 2,3,7,8-TCDD (and total TCDDs)
2 2,3,7,8-TCDF (and total TCDFs)
3 1,2,3,7,8-PeCDD (and total PeCDDs)
4 1,2,3,7,8-PeCDF
5 2,3,4,7,8-PeCDF
6 1,2,3,4,7,8-HxCDD
7 1,2,3,6,7,8-HxCDD
8 1,2,3,7,8,9-HxCDD
9 1,2,3,4,7,8-HxCDF
10 1,2,3,6,7,8-HxCDF
11 1,2,3,7,8,9-HxCDF
12 2,3,4,6,7,8-HxCDF
13 1,2,3,4,6,7,8-HpCDD (and total HpCDDs)
14 1,2,3,4,6,7,8-HpCDF
15 1,2,3,4,7,8,9-HpCDF
16 OCDD
17 QCDF
18 ;3C12-2,3,7,8-TCDD
19 3C12-2,3,7,8-TCDF
20 3C12-l,2,3,7,8-PeCDD
21 3C2-l,2,3,7,8-PeCDF
22 3C12-l,2,3,6,7,8-HxCDD
23 13C 2-l,2,3,4,7,8-HxCDF
24 3C12-l,2,3,4,6,7,8-HpCDD
25 3C2-l,2,3,4,6,7,8-HpCDF
26 13C12-OCDD
27 Total PeCDFs
28 Total HxCDFs
29 Total HxCDDs
30 Total HpCDFs
8290 - 63 Revision 0
November 1992
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Table 10.
2,3,7,8-TCDD Toxicity Equivalency Factors (TEFs) for the
Polychlorinated Dibenzodioxins and Oibenzofurans
Number
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
Compound(s)
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
TEF
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
8290 - 64
Revision 0
November 1992
-------
Table 11.
Analyte Relative Retention Time Reference Attributions
Analyte Analyte RRT Reference(a>
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
Ca> The retention time of 2,3,4,7,8-PeCDF on the DB-5 column is measured
relative to C12-l,2,3,7,8-PeCDF and the retention time of
1,2,3,4,7,8,9-HpCDF relative to 13C12-l,2,3,4,6,7,8-HpCDF.
8290 - 65 Revision 0
November 1992
-------
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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.8 Analysis Option6
Acetaldehyde
Acetone
Acrolein
Benzaldehyde
Butanal (butyr aldehyde)
Crotonaldehyde
Cyclohexanone
Decanal
2,5-Dimethylbenzaldehyde
Formaldehyde
Heptanal
Hexanal (hexaldehyde)
Isovaleraldehyde
Nonanal
Octanal
Pentanal (valeraldehyde)
Propanal (propionaldehyde)
m-Tolualdehyde
o-Tolualdehyde
p-Tolualdehyde
* Chemical Abstract Services
" Tl% ••*» 1 4 *•• 4- f*Ł ^ •» w»*«* « 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.
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
compounds that have been evaluated using modifications of the
analysis. Refer to the respective option number when choosing the
appropriate analysis technique for a particular compound.
8315 - 1 Revision 0
November 1992
-------
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 Section 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 p.m 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
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
8315 - 2 Revision 0
November 1992
-------
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
Section 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.
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.
8315 - 3 Revision 0
November 1992
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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, 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 pi loop.
4.1.3 Column - 250 mm x 4.6 mm ID, 5 ^m 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.
4.1.6 Helium Gas - 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 jitm polyester membrane filter. (Option 2)
8315 - 4 Revision 0
November 1992
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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 /xm pore size (Fisher Grade G4
or equivalent).
4.2.7 Solid sorbent cartridges - Packed with 2 g CIS (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).
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).
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tops,
4.2.13 Syringes - 5 ml, 500 ML, 100 ML, (Luer-Lok or
equivalent).
4.2.14 Syringe Filters - 0.45 Mm 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
4.6 Balance - Analytical, capable of accurately weighing to the nearest
0.0001 g.
4.7 Glass Funnels
4.8 Polyethylene Gloves - used to handle silica gel cartridges.
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 - Water in which an interferant is not
observed at the method detection limit for the compounds of interest.
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 Section 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.
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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 mi with organic-free reagent
water. 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.
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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 /xl_) 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 Section 5.7.1.1.
5.7.1.1 Standardization of formaldehyde stock solution -
Transfer a 25 ml aliquot of a 0.1 M Na^SOj solution to a beaker and
record the pH. Add a 25.0 ml aliquot of the formaldehyde stock
solution (Section 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:
(30.03)(N HCl)(mL HC1)
Concentration (mg/L) =
25.0 ml
where:
N HC1 = Normality of HC1 solution used (in milli-
equivalents/mL) (1 mmole of HC1 = 1 mini-
equivalent of HC1)
ml HC1= ml of standardized HC1 solution used
30.03 = 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.
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
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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 /ug/L may be
prepared by placing 100 ni 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 jug/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.
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 refrigerated at 4°C. Aqueous samples must be
derivatized and extracted within 3 days of sample collection. Higher molecular
weight aldehydes (heptanal to decanal) evidenced losses greater than 50% in a
liquid matrix over a 3 day holding time. Accordingly, liquid samples should be
derivatized and extracted within 24 hours of sample collection. Likewise, the
holding times of extracts of solid samples should be kept at a minimum. All
derivatized sample extracts should be analyzed within 3 days after preparation.
6.3 Samples 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.
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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.
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 (Section
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
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.
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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 (Section 4.2).
7.3.2 For solid samples, 1 to 10 ml of extract (Section 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 (Section 7.3.4) or liquid-liquid
(Section 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 6 M HC1 or 6 M
NaOH. Add 6 ml of DNPH reagent, seal the container, and place in a
heated (40°C), orbital shaker (Section 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 (Section 4.2.12) for 1 hour. Adjust
the agitation to produce a gentle swirling of the reaction solution.
7.3.4.3 Assemble the vacuum manifold and connect to a
water aspirator or vacuum pump. Attach 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 Add 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 vacuum conditions described in
Section 7.3.4.4, elute each cartridge train with approximately 9 ml
of acetonitrile directly into a 10 ml volumetric flask. Dilute the
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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 Section 7.3.4.5. The presence of this color is not
indicative of non-recovery 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 6 M HC1 or 6 M
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.
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.
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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
Acrodisk 0.45 urn 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.
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 methyl ene chloride. Add the methyl ene 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 Section 7.3.5.5.
If low concentrations of formaldehyde are present, the sample should
be concentrated during the solvent exchange procedure.
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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.
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.
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Column: CIS, 4.6 mm x 250 mm ID, 5 jum 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 ^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/H,0, 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% CH,CN for 5 minutes.
100%/0% to 60%/40% CH3CN/H20, linearly in 1
min.
60%/40% CH,CN/H20 for 15 minutes.
Detector: Ultraviolet, operated at 360 nm
Flow Rate: 1.0 mL/min
Sample Injection volume:25 jiL (suggested)
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 jum 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 Section 7.5.2. Allow the
system to pump for 20-30 minutes at a flow rate of 1.0 mL/min with
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the initial solvent mixture ratio (60%/40% CH3CN/H,0). 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. Suggested chromatographic conditions are
provided in Section 7.5.
7.6.2 Process each calibration standard solution through
derivatization and extraction, using the same procedure employed for
sample processing (Sections 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 Section 7.5, and tabulate peak area
against calibration solution concentration in M9/L-
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 Section 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
Section 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.
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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 Section 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:
RF,
Mean RF = RF
Concentration of standard injected,
Area of signal
ERF,-
N
2 (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.
RF, = 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
fol1ows:
Concentration of aldehydes in jig/L = (RF)(Area of signal)(100/VS)
where:
RF = Mean response factor for a particular analyte.
V = Volume of sample in ml.
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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 = Volume of extraction fluid aliquot in ml .
7.8.4 Calculate the concentration of formaldehyde in stack gas
samples (Method 0011) as follows: (Option 1)
7.8.4.1 Calculation of Total Formaldehyde: To determine
the total formaldehyde in ing , use the following equation:
[g/mole formaldehyde]
Total mg formaldehyde = Cd x V x DF x - x 10 mg//ng
[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(std)
where:
K = 35.31 ft3/m3, if Vm(std) is expressed in
English milts
1.00 m/nr, if Vm(8td) 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:
(Areaa)(RF)(Vola)(MWa)(1000
Conca = x DF
ml/I)
8315 - 18 Revision 0
November 1992
-------
where:
Areaa = Area of the sample peak for analyte "a".
RF = Mean response factor for analyte "a" from the
calibration in jig/L. (See Section 7.8.1)
Vola = Total volume of the sample cartridge eluate in
ml.
MWa = Molecular weight of analyte "a" in g/mole.
MWd = Molecular weight of the DNPH derivative of
analyte "a" in g/mole.
= Total volume of air sampled converted to standard
conditions in liters (L). (To calculate the
concentration at sampling conditions use
V .)(See Section 9.1.3 of Method 0100)
DF = 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, Conca in
ppbv, using the following equation:
(Conca)(22.4)
Conca in ppbv =
(MW8)
where:
Conca = Concentration of analyte "a" in ng/L.
22.4 = Ideal gas law volume (22.4 nL of gas = 1 nmole at
standard conditions).
MWa = 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
inter!aboratory 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|ig/L .
8315 - 19 Revision 0
November 1992
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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.
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
lig/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. Federal Register, Vol. 51, 40643-40652; November 7, 1986.
2. "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 - 20 Revision 0
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TABLE 1.
OPTION 1 - METHOD DETECTION LIMITS8 USING
LIQUID-SOLID EXTRACTION
Analyte
Formaldehyde
Acetaldehyde
Propanal
Crotonaldehyde
Butanal
Cyclohexanone
Pentanal
Hexanal
Heptanal
Octanal
Nonanal
Decanal
Retention Time
(minutes)
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
6.2
43 >
11.0
5.9
6.3
5.8
15.3
10.7
10.0
6.9
13.6
4.4
8 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 jug/L. The MDL was computed as
follows:
MDL = t(N.1f 0.oi)(Std. Dev.)
where:
t(M-i ODD = Tne uPPer first percentile point of the t-
' ' distribution with n-1 degrees of freedom.
Std. Dev. = Standard deviation.
6 The reported MDL is based upon analyses of 3 replicate, fortified
blanks at 250
8315 - 21 Revision 0
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TABLE 2.
OPTION 1 - HETHOD DETECTION LIMITS" USING
LIQUID-LIQUID EXTRACTION A
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)'
23. 2U
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 M9/L. The MDL was computed as follows: A
- Vi, turned. Dev.)
where:
t(M-i ODD = Tne uPPer first percentile point of the t-
' ' distribution with n-1 degrees of freedom.
Std. Dev. - Standard deviation.
The reported MDL is based upon analyses of 3 replicate, fortified
blanks at 250 M9/L.
8315 - 22 Revision 0
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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 HPLC8
Sample Volume (L)b
Compound
Acetaldehyde
Acetone
Acrolein
Benzaldehyde
Butyraldehyde
Crotonaldehyde
2,5-Dimethyl-
benzaldehyde
Formaldehyde
Hexanal
Isovaleraldehyde
Propionaldehyde
m-Tolualdehyde
o-Tolualdehyde
p-Tolualdehyde
Valeraldehyde
JP_
1.36
1.28
1.29
1.07
1.21
1.22
0.97
1.45
1.09
1.15
1.28
1.02
1.02
1.02
1.15
_2P_
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
_3P_
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
40
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
_5P_
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
100
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
0.
14
13
13
11
12
12
10
15
11
11
13
10
10
10
11
200
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
300
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
400
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
500
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
The ppb values are measured at 1 atm and 25°C. The sample cartridge Is
eluted with 5 mL acetonitrile and 25 juL is injected into the HPLC. The
maximum sampling flow through a DNPH-coated Sep-Pak is about 1.5 L/minute.
A sample volume of 1000 L was also done.
of 0.01 ppb for all the target analytes.
The results show a sensitivity
8315 - 23
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TABLE 4.
PERFORMANCE-BASED QC ACCEPTANCE LIMITS CALCULATED
USING THE COLLABORATIVE STUDY DATA
Analyte Spike
Concentration8
Formaldehyde
Propanal
Crotonaldehyde
Butanal
Cyclohexanone
Hexanal
Octanal
Decanal
160
160
160
160
160
160
160
160
Xb
154
148
160
151
169
151
145
153
SRC
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, i»9/L.
Mean recovery calculated using the reagent water, mean recovery, linear
regression equation, jig/L.
Overall standard deviation calculated using the reagent water, overall
standard deviation linear regression equation, jig/L.
Acceptance limits calculated as (X ± 3sR)100/spike concentration.
8315 - 24 Revision 0
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TABLE 5.
WEIGHTED LINEAR REGRESSION EQUATIONS FOR MEAN RECOVERY AND PRECISION (jig/L)
Analyte
Formaldehyde
Applicable
Cone. Range
39.2-2450
X
SR
*r
Reagent Water
0.909C + 8.79
0.185X + 1.98a
0.093X + 5.79
Ground Water
0.870C +14.84
0.177X + 13.85
0.108X + 6.24
Propanal
31.9-2000
X 0.858C + 10.49
SR 0.140X + 1.63
sr 0.056X + 2.76
0.892C + 22.22
0.180X + 12.37
0.146X + 2.08s
Crotonaldehyde
Butanal
Cyclohexanone
Hexanal
Octanal
Decanal
32.4-2030
35.4-2220
31.6-1970
34.1-2130
32.9-2050
33.2-2080
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
sr 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 + 0.378
X 0.856C + 7.88
SR 0.200X + 11.17
sr 0.092X + 1.718
X 0.883C + 12.00
SR 0.225X + 5.52
sr 0.088X + 2.28a
0.971C + 2.94
0.157X + 6.09
0.119X - 2.27
0.925C + 12.71
0.140X + 6.89
0.108X - 1.638
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
8 CODW < 0.50. Variance is not constant over concentration range.
X Mean recovery, ng/L, exclusive of outliers.
s_ Overall standard deviation, (ig/L, exclusive of outliers.
Single-analyst standard deviation, ng/L, exclusive of outliers.
8315 - 25
Revision 0
November 1992
-------
FIGURE la.
OPTION 2 - LIQUID-SOLID PROCEDURAL STANDARD OF GROUP A ANALYTES AT 625
-0.80-
-1.00-
«*
K
-1.40-
-1.80-1
-1.80-1
-2.1
1.00
w
i
a.oo
s.oo
4.00
x 10* Minutes
Retention Time
(minutes)
5.33
11.68
18.13
27.93
36.60
42.99
Analyte
Derivative
Formaldehyde
Propanal
Butanal
Cyclohexanone
Heptanal
Nonanal
8315 - 26
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November 1992
-------
FIGURE Ib.
OPTION 1 - LIQUID-SOLID PROCEDURAL STANDARD OF GROUP B ANALYTES AT 625
-0.80-
3.00
• 10* •inutts
Retention Time
(minutes)
7.50
16.68
26.88
32.53
40.36
45.49
Analyte
Derivative
Acetaldehyde
Crotonaldehyde
Pentanal
Hexanal
Octanal
Decanal
8315 - 27
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November 1992
-------
FIGURE 2a.
OPTION 1 - LIQUID-LIQUID PROCEDURAL STANDARD OF GROUP A ANALYTES AT 625
-2.00
1.00
a.oo
10*
3.00
•inutM
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 - 28
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November 1992
-------
FIGURE 2b.
OPTION 1 - LIQUID-LIQUID PROCEDURAL STANDARD OF GROUP B ANALYTES AT 625 /ig/L
i.oo
•inutts
Retention Time
(minutes)
7.79
17.38
27.22
32.76
40.51
45.62
Analyte
Derivative
Acetaldehyde
Crotonaldehyde
Pentanal
Hexanal
Octanal
Decanal
8315 - 29
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November 1992
-------
FIGURE 3.
OPTION 2 - CHROMATOGRAPHIC SEPARATION OF THE DNPH DERIVATIVES
OF 15 CARBONYL COMPOUNDS
ONPH
Number
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
\J
u
10
20
30
40
Peak Identification
Compound Concentrat ion(no/ul)
Formaldehyde
Acetaldehyde
Acrolein
Acetone
Propanal
Crotonaldehyde
Butanal
Benzaldehyde
Isovaleraldehyde
Pentanal
o-Tolualdehyde
m-Tolualdehyde
p-Tolualdehyde
Hexanal
2,4-Dimethylbenzaldehyde
8315 - 30
1.140
1.000
1.000
1.000
1.000
1.000
0.905
1.000
0.450
0.485
0.515
0.505
0.510
1.000
0.510
Revision 0
November 1992
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METHOD 8315
DETERMINATION OF CARBONYL COMPOUNDS
BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
MeoTa (Option 1)
7.1.1-7.1.1.1
Homogenize sample
and determine dry
weight
7.1.2 Extract
sample tor18
hours; filter and
store extract
7.3.2 Measure 1-10
mL extract; adjust
volume to 100 mL
with water
7.0 What is
the sample
matrix?
7.0te media
solid or
aqueous?
Is sample
dear or sample
complexity
known?
Stack Gas (Option
7.2.2 Centrifuge sample
at 2500 rpm for 10
minutes: decant
and filter
Aqueous
7.3.1 Measure
aliquot o» sample;
adjust volume to
100 ml with water
7.3.5.5 Exchange
solvent to methanol
0
8315 - 31
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November 1992
-------
METHOD 8315
continued
7.4.1.4 Exchange
solvent with methand
as in 7.3.5.5
7.4.1. 5 Store
sample at 4C
i
I
O
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 methytene
chloride; add methytene
chloride extracts to
volumetric flask
I
7.4.1.3 Dilute to volume
with methytene chloride;
mix wett; remove aliquot
7.4.1.4
sample have
a high concentration
of formaldehyde?
7.4.1.4 Dilute
extract with mobile
phase
7.4.1.4 Concentrate
extract during
solvent exchange
process
8315 - 32
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November 1992
-------
METHOD 8315
continued
1
i
7.4.2.2 • 7.4.2.3
Connect sampto cartridge
to dean syringe and
place in syringe rack
i
7.4.2.4 Backflush
cartridge with
acetonitrite
7.4.2.4
Does equate
flow become
blocked?
7.4.24 Displace
trapped air witti
acetonitrite in
syringe using a long-tip
disposable Pasteur pipet
7.4.25 Dilute to 5
mL with acetonitrite;
label flask; pipet 2
aMquotsimo
sample vials
I
7.4.2.6 Store
sample at 4C
\
i
8315 - 33
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November 1992
-------
METHOD 8315
continued
7.5.2 Set LC conditions
to produce appropriate
retention tomes
7.5.1 Set LC
conditions to produce
appropriate retention
Filter and
degas mobile phase
7.6.2 Process caybration
standards through same
processing steps as samples
7.6.3 - 7.6.4
Analyze solvent blank
and calibration standards:
I
7.6.5 Determine response
factor at each concentration
O
7.6.5 Prepare new
calibration
standards
8315 - 34
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November 1992
-------
METHOD 8315
continued
O
1
7.6.6 -7.6.7 Verify
calibration curve every day;
reanalyze 1 calibration
standard alter 10
sample runs or toss
7.7 Analyze samples
byHPLC
7.7.2 Inject a smaller
volume or dilute sample
7.7.4 Further
cleanup to 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 RSD
1
t
7.8.2-7.8.5
Calculate anafyte
concentrations
i
Stop
8315 - 35
Revision 0
November 1992
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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 600C until 95%
of the solvent has evaporated. Repeat the rinsing process as in Section 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 Section 7.0 for Option 2. An acceptable
impurity level is less than 0.025 ng/^L 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 I
waste, repeat the recrystallization as in Section A.4 but rinse with two 25 ml
portions of acetonitrile. Prep and analyze the second rinse as in Section A.5.
A.7 When the impurity level is 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 - 36 Revision 0
November 1992
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METHOD 8316
ACRYLAMIDE. ACRYLONITRILE AND ACROLEIN BY HIGH PERFORMANCE
LIQUID CHROMATOGRAPHY (HPLC1
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 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 Water samples are analyzed by high pressure liquid chromatography
(HPLC). A 200 /iL 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.
4.0 APPARATUS AND MATERIALS
4.1 HPLC system
4.1.1 One high pressure pump.
8316 - 1 Revision 0
November 1992
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4.1.2 Octadecyl Si lane (ODS, C-18) reverse phase HPLC column,
25 cm x 4.6 mm, 10 p.m, (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 urn filter
membrane, or equivalent disposable filter unit.
4.3 Materials
4.3.1 Syringes - 10, 25, 50 and 250 p.1 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 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+% purity.
5.5 Organic-free reagent water. All references to water in this method
refer to organic-free reagent water, as defined in Chapter One.
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 verified against EPA standards. If EPA
8316 - 2 Revision 0
November 1992
-------
standards are not available for verification, then standards certified by the
manufacturer and verified against a standard made from pure material is
acceptable.
5.6.1 Aery1 amide
5.6.1.1 Weigh 0.0100 g of aery1 amide 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 one year,
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 /xL 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 one year,
or sooner if comparison with the check standards indicates a
problem.
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
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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,
Section 4.1.
7.0 PROCEDURE
7.1 HPLC Conditions
Mobile Phase: Degassed organic-free reagent water
Injection Volume: 200 /LtL
Flow Rate: 2.0 mL/min
Pressure: 38 atm
Temperature: 25°C
Detector UV wavelength: 195 nm
7.2 Calibration:
7.2.1 Prepare standard solutions of acrylamide as described in
Section 5.7.1. Inject 200 p.1 aliquots of each solution, in triplicate,
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 Section 7.1. Table 1 provides the retention times that were
obtained under these conditions during method development.
8.0 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.
9.0 METHOD PERFORMANCE
9.1 Method performance data are not available.
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10.0 REFERENCES
1. Hayes, Samm "Acrylamide, 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)
Aery1 amide 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 CHROMATOGRAPHY (HPLC)
S t a. x: t
•7.1 Set
H E> I- C
Condi t ion.
a. 1 i fc> r: a t
\ /•
•7 . 3
t o s ar e
a. 1 y s i
i e:
Stop
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METHOD 8318
N-METHYLCARBAMATES BY HIGH PERFORMANCE LIQUID
CHROMAT06RAPHY (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
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solutions with organic-free reagent water and reanalyzing the sample. If
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 A
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 /*L aliquots and ,
performing multilinear gradients at a constant flow. The system must also I
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 urn).
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 (Section 4.3.11).
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4.3 Materials
4.3.1 HPLC injection syringe - 50 ML.
4.3.2 Filter paper, (Whatman #113 or #114, or equivalent).
4.3.3 Volumetric pipettes, Class A, glass, assorted sizes.
4.3.4 Reverse phase cartridges, (C-18 Sep-PakR [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 ML
displacement, (Gilson Microman [Rainin #M-25] with tips, [Rainin #CP-25],
or equivalent).
4.3.15 Nylon filter unit, 25 mm diameter, 0.45 M") 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, CH3CN - 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
Omnisolv #MX0488-1, or equivalent).
5.2.3 Methylene chloride, CH2C12 - HPLC grade - minimum UV cutoff at
230 nm (EM Omnisolv #0X0831-1, or equivalent).
5.2.4 Hexane, C6H14 - 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-C6H4(CHO)2 - reagent grade (Fisher
#0-4241, or equivalent).
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 Section 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
Section 7.2. If cleanup is not required, proceed directly to
Section 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 Section 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
Sections 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 Section 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 mL 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
separate, drain the acetonitrile layer into the volumetric flask.
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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 Section 7.3.2.
7.2 Cleanup - Pi pet 20.0 ml of the extract into a 20 ml glass vial
containing 100 pi 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 jitm
filter, filter an aliquot of the clean extract directly into a properly labelled
autosampler vial. The extract is now ready for analysis. Proceed to
Section 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 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. (Dilution factor = 0.1). Using a disposable 0.45 /im filter,
filter this extract directly into a properly labelled autosampler vial.
The extract is now ready for analysis. Proceed to Section 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 pm filter, filter
this extract directly into a properly labelled autosampler vial. The
extract is now ready for analysis. Proceed to Section 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
Sections 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 /iL
Solvent delivery system program:
Time Duration
Function Value (min) File
FR 1.0 0
B% 10% 0
20 0
5 0
5 0
3 0
B% 10% 7 0
ALARM 0.01 0
7.4.1.2 Post-column Hydrolysis Parameters (Recommended)
Solution: 0.05 N aqueous sodium hydroxide
Flow Rate: 0.7 mL/min
Temperature: 95° C
Residence Time: 35 seconds (1 mL reaction coil)
7..4.1.3 Post-column Deri vatization Parameters
(Recommended)
Solution: o-phthalaldehyde/2-mercaptoethanol(Section
5.3.1)
Flow Rate: 0.7 mL/min
Temperature: 40° C
Residence time: 25 seconds (1 mL reaction coil)
7.4.1.4 Fluorometer Parameters (Recommended)
Cell: 10 /iL
Excitation wavelength: 340 nm
Emission wavelength: 418 nm cutoff filter
Sensitivity wavelength: 0.5 /uA
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.
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 (Section 5.4.3),
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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 (Section 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
(E RF,)
i
mean RF = RF =
F, - RF)]
%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)
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.
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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 (Section 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 I
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. I
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.
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.
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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 TIMES8 AND
SINGLE OPERATOR METHOD DETECTION LIMITS
Method Detection Limitsb
Compound
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
a-Naphthold
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
(Mg/L)
1.9C
1.7
2.6
2.2
9.4C
2.4
2.0
1.7
3.1
2.5
Soil
(Mg/kg)
44C
12
10C
>50C
12C
17
22
31
32
17
a
b
See Section 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 DATA8 FOR ORGANIC-FREE REAGENT WATER
Compound Recovered % Recovery SD %RSD
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
Methiocarb (Mesurol)
Promecarb
225
244
210
241
224
232
239
242
231
227
75.0
81.3
70.0
80.3
74.7
77.3
79.6
80.7
77.0
75.7
7.28
8.34
7.85
8.53
13.5
10.6
9.23
8.56
8.09
9.43
3.24
3.42
3.74
3.54
6.03
4.57
3.86
3.54
3.50
4.1
Spike Concentration = 300 /jg/L of each compound, n = 10
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TABLE 3
SINGLE OPERATOR AVERAGE RECOVERY AND
PRECISION DATA3 FOR WASTEWATER
Compound
Recovered
% Recovery
Spike Concentration = 300 /xg/L of each compound, n = 10
No recovery
SD
%RSD
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
Methiocarb (Mesurol)
Promecarb
235
247
251
b
258
263
262
262
254
263
78.3
82.3
83.7
-
86.0
87.7
87.3
87.3
84.7
87.7
17.6
29.9
25.4
-
16.4
16.7
15.7
17.2
19.9
15.1
7.49
12.10
10.11
-
6.36
6.47
5.99
6.56
7.83
5.74
8318 - 14
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TABLE 4
SINGLE OPERATOR AVERAGE RECOVERY AND
PRECISION DATA8 FOR SOIL
Compound
Recovered
% Recovery
SD
Spike Concentration = 2.00 mg/kg of each compound, n = 10
%RSD
Aldicarb Sulfone
Methomyl (Lannate)
3-Hydroxycarbofuran
Dioxacarb
Aldicarb (Temik)
Propoxur (Baygon)
Carbofuran (Furadan)
Carbaryl (Sevin)
Methiocarb (Mesurol)
Promecarb
1.57
1.48
1.60
1.51
1.29
1.33
1.46
1.53
1.45
1.29
78.5
74.0
80.0
75.5
64.5
66.5
73.0
76.5
72.5
64.7
0.069
0.086
0.071
0.073
0.142
0.126
0.092
0.076
0.071
0.124
4.39
5.81
4.44
4.83
11.0
9.47
6.30
4.90
4.90
9.61
8318 - 15
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FIGURE 1
100
R
E
S
P
0
N
S
E
TIME (MIH)
1.00 ug/oL EACH OF:
1. ALDICAR3 SULFONE
2. METHOMYL
3. 3-HYDROXYCARBOFURAW
<*. OIOXACAA2
5. ALDICAAB
6.
7.
3.
9.
10.
PROFOXUR
GARBOFURAN
GARSARYL
HETHZOCA&B
PROMECARB
8318 - 16
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METHOD 8321
SOLVENT EXTRACTABLE NON-VOLATILE COMPOUNDS BY
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY/THERMOSPRAY/MASS SPECTROMETRY
(HPLC/TSP/MS) OR ULTRAVIOLET (UV1 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 is
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, it 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
CAS No.8
Azo Dves
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 Dves
Disperse Blue 3
Disperse Blue 14
Disperse Red 60
Coumarin Dyes
(Fluorescent Brighteners)
Fluorescent Brightener 61
Fluorescent Brightener 236
Alkaloids
Caffeine
Strychnine
2872-
3180-
2832-
6439-
730-
5261-
17464-
6535-
85-
52-8
81-2
40-8
53-8
40-5
31-4
91-4
42-8
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.8
Orqanophosphorus Compounds A
Methomyl 16752-77-5 1
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 Phenoxvacid 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 I
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 semivolatile 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.
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
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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 |iL
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 8150 (see Section 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 (Method 3540) or ultrasonic (Method 3550) extraction using
methylene chloride/acetone (1:1) is used for solid samples. A
micro-extraction technique is included for the extraction of Tris-BP from
aqueous and non-aqueous matrices.
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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.
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.
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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 nL 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 urn frit, or equivalent.
4.1.1.2.2 Analytical Column - C18 reverse phase
column, 100 mm x 2 mm ID, 5 \im particle size of ODS-Hypersil;
or C8 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 pL, 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
of producing a calibrated mass spectrum for PEG 400, 600, or 800 (see
Section 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.
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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 pL injection loop, analytical
columns, purging gases, etc. An automatic injector is optional, but is useful
for multiple samples. The columns specified in Section 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.
4.4.5 Wrist shaker - Burrell Model 75 or equivalent.
4.4.6 pH meter.
4.5 Kuderna-Danish (K-D) apparatus (optional).
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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 - 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 ml 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 nl_, 50 nL, 10 \il (Hamilton 701 N or equivalent),
and 50 ^L (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 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.
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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, CH3CQCH3 - Pesticide quality or equivalent.
5.8.4 Diethyl Ether, C2H5OCpH5 - 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 Section 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
chromatography should be employed. Load the solid onto a 3 x 8 inch
silica gel column (Section 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. Commercially prepared
stock standards can be used if they are verified against EPA standards. If EPA
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standards are not available for verification, then standards certified by the
manufacturer and verified against a standard made from pure material is
acceptable.
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,
Section 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, 3550
Waste 3540, 3550, 3580
Samples for the analysis of Tris-(2,3-dibromopropyl)phosphate wastewater
must be prepared according to Section 7.1.1 prior to HPLC/MS analysis. Samples
for the analysis of chlorinated phenoxyacid compounds and their esters must be
prepared according to Section 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/jiL in the 1 mL
extract.
7.1.1.1.2 Remove the gl ass wool pi ug from a di sposable A
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 Section 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 (Section 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 Section 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 is 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 = g of dry sample x 100
g of sample
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 yL of Tris-BP (approximate concentration
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1000 mg/L) to the sample selected for spiking; the amount
added should result in a final concentration of 200 ng/pL 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 Section 7.5,
Method 3510.
7.1.1.2.4 Collect the extract in a 15 ml graduated
glass tube. Proceed as in Section 7.1.1.1.4.
7.1.2 Extraction for chlorinated phenoxyacid compounds - Preparation
of soil, sediment, and other solid samples must follow Method 8150, with
the exception of no hydrolysis or esterification. Section 7.1.2.1
presents an outline of the procedure with the appropriate changes
necessary for determination by Method 8321. Section 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 H2SO,
(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.
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.
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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 5% acidified sodium
sulfate. 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 (Section 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
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
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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 Section
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 Section 7.1.2.1.6 (drying, K-D
concentration, solvent exchange, and final volume
adjustment).
7.2 Prior to HPLC analysis, the extraction solvent must be exchanged to
methanol or acetonitrile (Section 7.1.2.1.9). The exchange is performed using
the K-D procedures listed in all of the extraction methods.
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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
fol1ows:
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, and
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.
Jet 210°C to 220°C.
Source block 230°C to 265°C. (Some compounds may degrade in
the source block at higher temperatures, operator
should use knowledge of chemical properties to
estimate proper source temperature).
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7.4.4 Sample injection volume: 20 \il is necessary in order to
overfill the 10 nL 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 Section 5.14) which has 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: NH,+ (18 amu), NH,+'H,0 (36), CH,OH'NH/ (50) (methanol), or
CH3CN'NH4+ (59) (acetonitrile), and CH3COOH'NH4 (78) (acetic acid). f
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
Section 5.12.
7.5.2.2 Choose the proper ionization conditions, as
outlined in Section 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
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).
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7.5.2.3 If HPLC-UV detection is also being used,
calibrate the instrument by preparing calibration standards as
outlined in Section 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 Section 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 Sections
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 must 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
7.5.2.8 The working calibration curve, or the CF, must be
verified on each working day by the injection of one or more
calibration standards. If the response varies from the predicted
response by more than + 20 percent, a new calibration curve must be
prepared. The % Difference is calculated as follows:
% Difference = (R, - R2)/R1 x 100.
where:
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R., = CF first analysis.
R2 = CF from succeeding analyses.
7.6 Sample Analysis
7.6.1 Once the LC/MS system has been calibrated as outlined in
Section 7.5, then it is ready for sample analysis. It is recommended that
the samples be initially 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 \il 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 \il aliquot of the sample extract from
Section 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.
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8.2.1.2 Check instrument performance. If an instrument
performance problem is identified, correct the problem and
re-analyze the extract.
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 Section 7.5.2.7 for required HPLC/MS parameters for
standard calibration curve %RSD limits.
8.3.2 See Section 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:
o Leaks,
o Proper pressure delivery,
o A dirty guard column; may need replacing or repacking, and
o 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 Section 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.
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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.
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 Section 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 Section
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 Section 7.6.
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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
lab data for Disperse Red 1, Table 11 with organophoshorus pesticides, Table 12
with 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 (3o) of the lowest standard
amount, along with the calculated slope and intercept, was 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 8150
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.; Hickey, 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, 52, 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.
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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.
12. Cotterill, E. G.; Byast, T. H. "HPLC of Pesticide Residues in
Environmental Samples." in Liquid Chromatoqraphv in Environmental
Analysis; Laurence, J. F., Ed.; Humana Press: Clifton, NJ, 1984.
13. Voyksner, R. D. "Thermospray HPLC/MS for Monitoring the Environment." In
Applications of New Mass Spectrometrv 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 Interlaboratory Comparison of an SW-846 Method for the Analysis of the
Chlorinated Phenoxyacid Herbicides by LC/MS", EPA/600/X-90/133, June 1990.
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TABLE 1.
RECOMMENDED HPLC CHROMATOGRAPHIC CONDITIONS
Initial
Mobile
Phase
W
Analytes:
OraanoohosDhorus
Initial
Time
(min)
Compounds
Gradient
(linear)
(min)
Final
Mobile
Phase
(%)
Final
Time
(min)
50/50 0 10
(water/methanol)
Azo Dves (e.g. Disperse Red 1)
50/50 0 5
(water/CH3CN)
Tris-(2.3-dibromopropyl)phosphate
50/50 0 10
(water/methanol)
100 5
(methanol)
100
(CH3CN)
100
(methanol)
Chlorinated phenoxyacidcompounds
75/25 2
(A/methanol)
15 40/60
(A/methanol)
40/60
(A/methanol)
3 5
Where A » 0.01 M ammonium acetate (1% acetic acid)
75/25 10
(A/methanol)
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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 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)
pg
420
1400
4700
115
200
560
EQL(lOs)
P9
600
2000
6700
150
280
800
EQL = Estimated Quantitation Limit
Data from Reference 16.
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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 Recovery
MS CAD
92.5 ± 3.7 87.6 ± 4.6
90.2 ± 4.7 90.4 ± 9.9
2.5% 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
Sample
Spike 1
Spike 2
RPD
HPLC/UV
93.4 ± 0.
96.2 ± 0.
3.0%
Percent Recovery
MS
3 102.0 ± 31
1 79.7 ± 15
25%
CAD
82.7 ± 13
83.7 ± 5.2
1.2%
Data from Reference 16.
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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
0 mg/L Spiking
Concentration
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
November 1992
-------
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
Abundances3
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
November 1992
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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
November 1992
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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
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November 1992
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TABLE 10.
PRECISION AND LIMITS OF DETECTION 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
(ngM)
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
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November 1992
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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
ng/g
50
50
50
50
100
50
50
50
fiq/L
50
50
50
50
100
50
50
50
mq/kg
2
2
2
2
3
2
2
2
Range of
Recovery
(%)
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 -
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
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
November 1992
-------
TABLE 12.
SINGLE OPERATOR ACCURACY AND PRECISION FOR MUNICIPAL
WATER (A), DRINKING WATER (B), CHEMICAL SLUDGE WASTE
Compound
Tris-BP
Data from
Average
Recovery
(%)
(A) 25
(B) 40
(C) 63
Reference 18.
SINGLE
Concentration Average
(ng/nL) Area
50
100
150
200
2675
5091
7674
8379
LOD
(ng/|iL)
33
Standard
Deviation
8.0
5.0
11
TABLE
OPERATOR EQL
Standard
Deviation
782
558
2090
2030
Spike
Amount
(ng/nL)
2
2
100
13.
TABLE FOR
3*Std
Dev.
2347
Lower
EQL
(ng/nL)
113
Range
of %
Recovery
41 - 9.0
50 - 30
84 - 42
TRIS-BP
7*Std
Dev.
5476
Upper
EQL
(ng/|iL)
172
WASTE
(C)
Number of
Analyses
15
12
8
10*Std
Dev.
7823
Data from Reference 18.
EQL = Estimated Quantitation Limit
8321 - 32
Revision 0
November 1992
-------
TABLE 14
LIMITS OF DETECTION 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+NH,)+
238 (M+NH4)+
218 (M NHJ
252 (HjW4)*
232 (M;NHJ;
272 (M^NHj;
286 (M NHJ
228 (M+NH,-NO)+
266 (M*NH,)*
321 (M+H)
372 (M+NHJ +
328 (M+NHJ +
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'HCl )'
184 (M'HCl)"
199 (M'l)'
235 (M'l)"
213 (M'l)'
218 (M'HCl)'
269 (M'l)'
240 (M)"
247 (M'l)'
185 (M'C^O,)"
195 (M'C8H1503r
195 (M'C6Hn02)'
161 (M'C10H1903r
LOD
(ng)
11
3.0
50
28
25
12
6.5
43
19
110
Data from Reference 19.
8321 - 33
Revision 0
November 1992
-------
TABLE 15
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
LOW LEVEL DRINKING WATER
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
63
26
60
78
43
72
62
29
73
ND
73
HIGH LEVEL DRINKING
54
60
67
66
66
61
74
83
91
43
97
LOW LEVEL SAND
117
147
167
142
ND
134
121
199
76
ND
180
22
13
23
21
18
31
14
24
11
ND
17
WATER
30
35
41
33
33
23
35
25
10
9.6
19
26
23
79
39
ND
27
23
86
74
ND
58
5
5
5
5
5
5
5
5
5
5
5
50
50
50
50
50
50
50
50
50
50
50
M9/L
86 - 33
37 - 0
92 - 37
116 - 54
61 - 0
138 - 43
88 - 46
62 - 0
85 - 49
ND
104 - 48
M9/L
103
119
128
122
116
99
132
120
102
56
130
26
35
32
35
27
44
45
52
76
31
76
M9/9
147 - 82
180 -118
280 - 78
192 - 81
ND
171 - 99
154 - 85
245 - 0
210 - 6
ND
239 - 59
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
(a)
'All recoveries are in negative ionization mode, except for 2,4-D,ester.
ND = Not Detected.
8321 - 34
Revision 0
November 1992
-------
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
LOW LEVEL MUNICIPAL
83
ND
ND
ND
ND
27
68
ND
44
ND
29
HIGH LEVEL MUNICIPAL
66
8.7
3.2
10
ND
2.9
6.0
ND
16
ND
1.9
33
27
30
34
37
29
34
22
28
86
3.6
ASH
22
ND
ND
ND
ND
25
38
ND
13
ND
23
ASH
21
4.8
4.8
4.3
ND
1.2
3.1
ND
6.8
ND
1.7
209 -119
276 -187
205 -111
226 -115
161 - 51
204 -131
225 -141
192 -110
140 - 65
418 -166
25 - 17
M9/9
104 - 48
ND
ND
ND
ND
60 - 0
128 - 22
ND
65 - 26
ND
53 - 0
M9/9
96
21
10
16
41
5
0
4.7
ND
3.6- 0
12 - 2.8
ND
23 - 0
ND
6.7- 0
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
(a)All recoveries are in negative ionization mode, except for 2,4-D,ester.
ND = Not Detected.
8321 - 35
Revision 0
November 1992
-------
TABLE 16
MULTILABORATORY ACCURACY AND PRECISION DATA
FOR THE CHLORINATED PHENOXYACID HERBICIDES
Compounds
Spiking
Concentration
Mean % Relative
(% Recovery)8 Standard Deviation
2,4,5-T
2,4,5-T,butoxy
2,4-D
2,4-DB
Dalapon
Dicamba
Dichlorprop
Dinoseb
MCPA
MCPP
Silvex
2,4,5-T
2,4,5-T,butoxy
2,4-D
2,4-DB
Dalapon
Dicamba
Dichlorprop
Dinoseb
MCPA
MCPP
Silvex
2,4,5-T
2,4,5-T,butoxy
2,4-D
2,4-DB
Dalapon
Dicamba
Dichlorprop
Dinoseb
MCPA
MCPP
Silvex
500 mq/L
90
90
86
95
83
77
84
78
89
86
96
50 mg/L
62
85
64
104
121
90
96
86
96
76
65
5 mg/L
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
Data from Reference 20.
8 Mean of duplicate data from 3 laboratories.
b % RSD of duplicate data from 3 laboratories.
8321 - 36
Revision 0
November 1992
-------
TABLE 17
COMPARISON OF LODs: METHOD 8150 vs. METHOD 8321
lonization
Compound
Method 8150
LOD(ng/L)
Method 8321
LOD (jig/L)
Mode
Dalapon
Dlcamba
2,4-D
MCPA
Dichlorprop
MCPP
2,4,5-T
2,4,5-TP (Silvex)
2,4,-DB
Dinoseb
5.8
0.27
1.2
249
0.65
192
0.2
0.17
0.91
1.9
1.1
0.3
0.29
2.8
0.27
0.50
0.65
4.3
0.34
1.9
8321 - 37
Revision 0
November 1992
-------
FIGURE 1.
SCHEMATIC OF THE THERMOSPRAY PROBE AND ION SOURCE
Flangi
K
I
Ion Sampling
Cone
Source
Mounting . Ions
Plate | t
Electron Vaporizer
Beam ^ Probe
I
— LC
Vapor || Heater
Temperature f
T4 Block
Temperature
T.
Vaporizer
Coupling
8321 - 38
Revision 0
November 1992
-------
FIGURE 2.
THERMOSPRAY SOURCE WITH WIRE-REPELLER
(High sensitivity configuration)
CERAMIC INSULATOR
** WIRE REPELLER
8321 - 39
Revision 0
November 1992
-------
FIGURE 3.
THERMOSPRAY SOURCE WITH WIRE-REPELLER
(CAD configuration)
r_
CERAMIC INSULATOR
WIRE REPELLER
8321 - 40
Revision 0
November 1992
-------
METHOD 8321
SOLVENT EXTRACTABLE NON-VOLATILE COMPOUNDS BY
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY/THERMOSPRAY/MASS SPECTROMETRY
(HPLC/TSP/MS) OR ULTRAVIOLET (UV) DETECTION
8321 - 41
Revision 0
November 1992
-------
METHOD 8330
NITROAROMATICS AND NITRAMINES BY HIGH
PERFORMANCE LIQUID CHROMATOGRAPHY (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
Hexahydro-l,3,5-trinitro-l,3,5-triazine
1, 3, 5-Tri nitrobenzene
1,3-Dinitrobenzene
Methyl-2,4,6-trinitrophenylnitramine
Nitrobenzene
2,4,6-Trinitrotoluene
4-Amino-2,6-dinitrotoluene
2-Amino-4, 6-dinitrotoluene
2,4-Dinitrotoluene
2,6-Dinitrotoluene
2-Nitrotoluene
3-Nitrotoluene
4-Nitrotoluene
HMX
RDX
1,3,5-TNB
1,3-DNB
Tetryl
NB
2,4,6-TNT
4-Am-DNT
2-Am-DNT
2,4-DNT
2,6-DNT
2-NT
3-NT
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 Section 5.3.1 and Section 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 Section
11.0 on SAFETY.) Each analyst must demonstrate the ability to generate
acceptable results with this method.
8330 - 1
Revision 0
November 1992
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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 pipet. 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 Section 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. 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
November 1992
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4.0 APPARATUS AND MATERIALS
4.1 HPLC system
4.1.1 HPLC - equipped with a pump capable of achieving 4000 psi, a
100 nl 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 |im), (Supelco LC-18, or equivalent).
4.1.2.2 Secondary column: CN Reverse phase HPLC column,
25 cm x 4.6 mm (5 pm), (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 nL, (Hamilton liquid
syringe or equivalent).
4.3.2 Disposable cartridge filters - 0.45 jim 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.
8330 - 3 Revision 0
November 1992
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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 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
refrigerator at 4°C in the dark. Calculate the concentration of the stock
8330 - 4 Revision 0
November 1992
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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
Section 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 iig/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 ng/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 (Section
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.
8330 - 5 Revision 0
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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 Chapt. 4.
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 sulfate 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 pi pet 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
to a 10 mL graduated cylinder using a Pasteur pipet. At this
8330 - 6 Revision 0
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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 [VJ in the
calculation of concentration after converting to |iL). The
resulting extract, about 5 - 6 ml, is then diluted 1:1 with
reagent water prior to analysis.
7.1.1.1.6 If the diluted extract is turbid, filter it
through a 0.45 - jim 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 Section
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-pm
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 Section 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 8510 prior to
grinding in a mortar and pestle (See Safety Section
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
Teflon-lined cap, vortex swirl for one minute, and place in a
cooled ultrasonic bath for 18 hours.
8330 - 7 Revision 0
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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 (Section 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-jim Teflon filter. Discard first 3
ml and retain remainder in a Teflon-capped vial for RP-HPLC
analysis as in Section 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 jim, (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-jiL
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. Triplicate injections of each
calibration standard over the concentration range of interest are
sequentially injected 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, in triplicate 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 daily calibration must agree within ±15% of the
response factor of the initial calibration. The same criteria is required
8330 - 8 Revision 0
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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 Section 7.2. All positive measurements observed on the C-18 column
must be confirmed by injection onto the CN column.
7.4.2 Follow Section 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 Section 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, Section 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.
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.
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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 multilaboratory 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.W. Schumacher, P.M. 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.
8. Leggett, D.C., T.F. Jenkins and P.H. Miyares (1990) Salting-out solvent
extraction for preconcentration of neutral polar organic solutes from
water. Analytical Chemistry, 62: 1355-1356.
8330 - 10 Revision 0
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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 8510 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 8510 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 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.
8330 - 11 Revision 0
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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 (pg/L)
Low-Level High-Level
13.0
0.84 14.0
0.26 7.3
0.11 4.0
4.0
6.4
0.11 6.9
0.060
0.035
0.31 9.4
0.020 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
8330 - 12
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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
Tetryl
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 2.00 min on LC-CN
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TABLE 3
SINGLE LABORATORY PRECISION OF METHOD FOR SOIL SAMPLES
Spiked Soils
Mean Cone.
(mg/kg) SD
%rsd
Field-Contaminated Soils
Mean Cone.
(mg/kg) SD %rsd
HMX
RDX
46
60
1.7
1.4
3.7
2.3
14
153
104
1.8
21.6
12
12.8
14.1
11.5
1,3,5-TNB
1,3-DNB
Tetryl
2,4,6-TNT
2,4-DNT
8.6
46
3.5
17
40
5.0
0.4
1.9
0.14
3.1
1.4
0.17
4.6
4.1
4.0
17.9
3.5
3.4
877
2.8
72
1.1
2.3
7.0
669
1.0
29.6
0.2
6.0
0.11
0.41
0.61
55
0.44
3.4
7.1
8.3
9.8
18.0
9.0
8.2
42.3
8330 - 14
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TABLE 4
MULTILABORATORY ERROR OF METHOD FOR SOIL SAMPLES
Spiked Soils
Mean Cone.
(mg/kg) SD
HMX
RDX
1,3,5-TNB
1,3-DNB
Tetryl
2,4,6-TNT
2,4-DNT
46
60
8.6
46
3.5
17
40
5.0
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
Mean Cone.
(mg/kg) SD
14
153
104
877
2.8
72
1.1
2.3
7.0
669
1.0
3.7
37.3
17.4
67.3
0.23
8.8
0.16
0.49
1.27
63.4
0.74
Soils
%rsd
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.
(H9/L)
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
8330 - 15
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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 (jig/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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TABLE 7
COMPARISON OF METHOD ACCURACY FOR SOIL AND AQUEOUS SAMPLES
(HIGH CONCENTRATION METHOD)
Recovery (%)
Analyte 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
(pgA)
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
* 1
. -A
*
1 ra ™
— if.
EXPLOSIVES ON A
CN COLUMN
x
A
to
12 14
FIGURE 1
CHROMATOGRAMS FOR COLUMNS DESCRIBED IN SECTION 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
SatlinoOut
7.1.1.1.1 Add 251. 3 gof salt
and 1 L of water sample to a
1 L vol. flask. Mix the contents.
1 1
7.1.1. 12 Add 164 mL of
acetonitrite (ACN) and stir
toMSmins.
t
7.1 .1 .1 .3 Transfer ACN layer
to 100 mL vol. flask. Add 10 mL
ol fresh ACN to 1 L flask and
stir. Transfer 2nd portion and
combine with 1st in 100 mL flask.
\
r
7.1. 1.1.4 Add 84 mL of salt
water to the ACN extract and stir.
Transfer ACN extract to 10 mL
grad. cylinder.
i
7.1. 1.1. 5 Add 1 mL of ACN to
100 mL vol. flask. Stir and
transfer to the 10 mL grad.
cylinder. Record volume.
Oiute 1 :1 with reagent water.
i '
7. 1.1.1. 6 Rlter if turbid.
Transfer to a vial for
RP-HPLC analysis.
7.1.1.1 Sample Filtration:
Pldco 5 mis. sample in
scintillation viat. Add 5 mis.
methane* shake; filler
through 0.5 urn finer. Discard
first 3 mis. Main remainder
for use.
—o
8330 - 20
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METHOD 8330
(continued)
©
7.1.2.1 Sample Homogenization
Air dry sample at room Temp.
or below. Avoid exposure to
direct sunlight. Grind sample
in an acetonitrile rinsed mortar.
7.1.2.2 Sample Extraction
7.1.2.2.1
Place 2 grs. soil subsample,
10 mis. acetonitrile in 15 ml.
glass vial. Cap, vortex swirl,
place in room Temp, or below
ultrasonic bath for 18 hrs.
7.1.2.2.2
Let sola settle. Add 5 mis.
supernatant to 5 mis. calcium
chloride soln. in 20 ml vial.
Shake, let stand 15 mins.
7.1.2.2.3
Filter supernatant through
0.5 um filter. Discard initial
3 mis., retain remainder
for analysis.
0
7.2 Set Chromatographic Conditions
1
p
7.3 Calibration of HPLC
'
7.3.2
Run working stds. in triplicate.
Plot ng. vs. peak area or ht
Curve should be linear with
zero intercept.
•
7.3.3
Analyze midrange calibration
std. at beginning, middle.
and end of sample analyses.
Redo Section 7.3.1 if peak
areas or Ms. do not agree
to w/taW- 20% of initial
calibration values.
1
7.4 Sample Analysis
i
1
7.4.1
Analyze samples. Confirm
measurement w/injection onto
CN column.
1
'
7.4.3
Refer to Table 2 for typical
analyte retention times.
i
8330 - 21
Stop
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METHOD 8331
TETRAZENE BY REVERSE PHASE
HIGH PERFORMANCE LIQUID CHROMAT06RAPHY (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 Noa
Tetrazene 31330-63-9
8 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 p.g/1 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.
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
8331 - 1 Revision 0
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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 ML 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 jum)
(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 /zL.
4.3.2 Filters - 0.5 jum Millex-SR and 0.5 nm 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.
4.4 Preparation
4.4.1 Prepare all materials as described in Chapter 4 for volatile
organics.
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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.
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.
8331 - 3 Revision 0
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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.
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.
8331 - 4 Revision 0
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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 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 urn 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 p.1
UV Detector: 280 nm
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.
8331 - 5 Revision 0
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7.3.2 Continuing Calibration - On a daily basis, inject 250 /nL 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.
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 - 6 Revision 0
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FIGURE 1
I6r-
TNT
12
I
I
ROX
O.O64
Absorbonca Units
8331 - 7
Revision 0
November 1992
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TABLE 1.
METHOD PERFORMANCE, WATER MATRIX
Spike
Cone.
(M9/L)
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 - 8
Revision 0
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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
Repl icate
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
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 - 9
Revision 0
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METHOD 8331
TETRAZENE BY REVERSE PHASE
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC)
7.1.1 Fi 1tar 1o
ml water
sample; discard
11 r a t S ml /
analyze lane 5
7.1.2.1
Determine %
djey weight
7 .1.2.2-7.1.2.5
EXtx ac c 2 g aoil
with so ml
solvent
7 . 1 . 2 . «
1O ml •xertoc
discard 5 ra
«naly xe 1 aa t
ml
7.2 Analyze
samples using
ctironatogzapliic
conditions in
Sect Ion 7.2.1.1
7.3.1 Initial
Calibrat ion i
naly E• working
atandards
(Section 5.3. 3 )
8331 - 10
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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
8410 - 1
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-Chloroaniline
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
Dimethyl phthalate
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
131-11-3
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November 1992
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Compound Name CAS No.8
Diethyl phthalate 84-66-2 I
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
Hexachlorobenzene 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 J
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, 6-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
November 1992
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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 jug/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 Section 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-
el ut ing 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
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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 limited.
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.1 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 /xm 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 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 Section
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.
4.3 Dry Carrier Gas - The carrier gas should be passed through an
efficient cartridge-type drier.
8410 - 4 Revision 0
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4.4 Syringes - 1-p.L, lO-^L.
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 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
prepared at concentrations that will completely bracket the working range
of the chromatographic system (at least one order of magnitude is
suggested).
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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. m
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 - 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. A
Column temperature program: 40-280°C at 10°C/min. I
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 p.1.
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 Aim 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-
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).
8410 - 6 Revision 0 m
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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
Section 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 Section 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 Section
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 Section 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" 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 Section 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.
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 higher in frequency than
those of the condensed phase.
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7.10.5 Quantitation - Although this protocol can be used to
confirm GC/MS identifications, with subsequent quantitation, FT-IR
quantitation guidelines are also provided.
7.10.6 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.
7.10.7 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 concentration).
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.
7.10.8 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 Section 7.5). Collect 16 scans
over the entire detector spectral range. Plot the test and measure the peak-to-
peak noise between 1800 and 2000 cm . 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
8410 - 8 Revision 0
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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 thermostatted, 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.
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; Section 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.
8410 - 9 Revision 0
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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; 2nded.;
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.R.; 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.
8410 - 10 Revision 0
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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(2-chloroethyl) 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-Hexachlorobutadiene
Isophorone
2-Methyl naphthal ene
Naphthalene
Nitrobenzene
N-Nitrosodimethylamine
N-Nitrosodi-n-propylamine
N-Ni trosodi phenyl ami nee
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Phenanthrene
Pyrene
1,2,4-Trichlorobenzene
Identification
ng injected9
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)d
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
40
50(50)
100(50)
50(25)
Limit
M9/LD
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
20
25(25)
50(25)
25(12.5)
vmax, 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
1362
729
820
750
8410 - 11
Revision 0
November 1992
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TABLE 1.
(Continued)
Determined using on-column injection and the^conditions of Section 7.3. A A
medium band HgCdTe detector [3800-700 cm"1; D value (Apeak 1000 Hz, 1) 4.5 x I
1010 cm HzV2W"T] type with a 0.25 mm2 focal chip was used. The GC/FT-IR system
is a 1976 retrofitted model.
Based on a 2 nl injection of a one liter sample that has been extracted and
concentrated to a volume of 1.0 ml.
Most intense 1R peak and suggested quantitation peak.
Values in parentheses were determined with a new (1986) GC/FT-IR system. A
narrow band HgCdTe detector [3800-750cm~1; D value (Apeak 1000 Hz, 1) 4 x 1010
cm Hz W ] was used. Chromatographic conditions are those of Section 7.3.
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 injected3
vmax, cm
Benzoic acid
2-Chlorophenol
4-Chlorophenol
4-Chloro-3-methyl phenol
2 -Methyl phenol
4-Methyl phenol
2,4-Dichlorophenol
2,4-Dinitrophenol
4, 6-Dinitro-2-methyl phenol
2-Nitrophenol
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 Section 7.3.
Based on a 2 ML 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-el uting compounds.
8410 - 13
Revision 0
November 1992
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TABLE 3.
GAS-PHASE GROUP FREQUENCIES
Number of
Functionality Class Compounds
Ether
Ester
Nitro
Nitrile
Ketone
Amide
Al kyne
Acid
Phenol
Aryl , Al kyl
Benzyl, Alkyl
Diaryl
Dial kyl
Alkyl, Vinyl
Unsubstituted Aliphatic
Aromatic
Monosubstituted Acetate
Aliphatic
Aromatic
Aliphatic
Aromatic
Aliphatic (acyclic)
(0,6 unsaturated)
Aromatic
Substituted Acetamides
Aliphatic
Aliphatic
Dimerized-Al iphatic
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, vcm
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
8410 - 14
Revision 0
November 1992
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TABLE 3.
(Continued)
Functionality
Alcohol
Amine
Alkane
Aldehyde
Benzene
Class
Primary Aliphatic
Secondary Aliphatic
Tertiary Aliphatic
Primary Aromatic
Secondary Aromatic
Aliphatic
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, vcm"
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
Revision 0
November 1992
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TABLE 4. FUSED SILICA CAPILLARY COLUMN GC/FT-IR QUANTITATION RESULTS
Concentration
Range, and
Identification
Compound Limit, nga
Acenaphthene
Acenaphthylene
Anthracene
Benzo(a) anthracene
Benzoic acid
Benzo(a)pyrene
Bi s (2-chl oroethoxy)methane
Bis(2-chloroethyl) ether
Bis(2-chloroisopropyl) ether
4-Bromophenyl phenyl ether
Butyl benzyl phthalate
4-Chloroaniline
4-Chl oro-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-Di chl orobenzene
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 -Hexachl orobutad i ene
Hexachl orocycl opentadi ene
Hexachl oroethane
Isophorone
2-Methyl naphthalene
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
Maximum
Absorbance
Correlation
Coefficient
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
Integrated
Absorbance0
Correlation
Coefficient*1
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
November 1992
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TABLE 4. (Continued)
Compound
Concentration
Range, and
Identification
Limit, nga
Maximum
Absorbanceb
Correlation
Coefficient
Integrated
Absorbance0
Correlation
Coefficient01
2-Methyl phenol
4-Methyl phenol
Naphthalene
2-Nitroaniline
3-Nitroaniline
4-Nitroaniline
Nitrobenzene
2-Nitrophenole
4-Nitrophenol
N-Nitrosodimethylamine
N-Nitrosodiphenylamine
N-Nitrosodi-n-propylamine
Pentachlorophenol
Phenanthrene
Phenol
Pyrene
1 ,2,4-Trichlorobenzene
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 Section 7.3.
e Subject to interference from co-eluting compounds.
8410 - 17
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METHOD 8410
GAS CHROMATOGRAPHY/FOURIER TRANSFORM INFRARED (GC/FT-IR)
SPECTROMETRY FOR SEMIVOLATILE ORGANICS: CAPILLARY COLUMN
Start
1 1
7 1 Sample
p r epa ration
prior to
GC/FT-IR
analysis
7 2 Optional
Cel
Permea tion
Cleanup of
ex tracts
7 3 Initial
Ca 1 ibra 1 1 on ,
recommended
CC/FT-IR
conditions
7 6 Adjust
pressure
/7 7 MCtV
/ D«t«ctor N.
/ c«nt«rburst ^
f mt.n.Uy <75*
^v plot max of .
\^rY
Ho
..
7 7 Replace
Source
7 4 Check
detecto r
center burs t
intensi ty
7 8 Frequency
Calibration
.
7 5 Column
Interface
Sens i t i vi ty
C
J)
7 9 Determine
mm identif i -
able quantities
of ana 1 y tes of
interest
791 Prepare
plot of
Iightpipe T v*
MCT centerburst
intense ty
S-Ye
8410 - 18
Revision 0
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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. 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.8
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
a 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.
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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 jum film
thickness, silicone-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 a reconstructed ion
chromatogram (RIC). Software must also be available that allows for
integration of the abundances in, and RIC between, specified time or scan-
number limits.
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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 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
other suitable solvent, and dilute to 10 ml in a volumetric flask. Larger
volumes may be used at the convenience of the analyst.
8275 - 3 Revision 0
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5.3.2 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 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
1,4-dichlorobenzene-d,, naphthalene-da, acenaphthene-d10, phenanthrene-d1Q,
chrysene-d1?, and perylene-d12. Other compounds may be used as internal standards
as long as the requirements given in Section 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-d.,- Prior to each analysis, evaporate about 10 juL
of the internal standard onto the lid of 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 Section 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,
Section 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.
CAUTION: Do not touch the crucibles with your fingers. This can
result in a serious burn, as well as contamination of
8275 - 4 Revision 0
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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 %)
should be used. If 0.100 g gives 8000 mv, then 0.025 g ± 50 % should be
used.
8275 - 5 Revision 0
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7.4.3 After weighing out the sample into the crucible, deposit the
internal standards (10 juL) onto the lid of the crucible. Load the
crucible into the pyrocell, using the same temperature program in Section
7.3.1. Hold the capillary at 5°C during this time to focus the released
semi-volatiles (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.
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.
8275 - 6 Revision 0
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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.
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
8275 - 7 Revision 0
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comparison of sample with the nearest library searches will the mass
spectral interpretation specialist assign a tentative
identification.
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
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TABLE 1
METHOD PERFORMANCE, SOIL MATRIX
Analyte
2-Chlorophenol
4-Methyl phenol
2,4-Dichlorophenol
Naphthalene
4-Chloro-3-methyl -phenol
1 -Chi oronaphthal ene
2,4-Dinitrotoluene
Fluorene
Diphenylamine
Hexachlorobenzene
Dibenzothiophene
Phenanthrene
Carbazole
Aldrin
Pyrene
Benzo(k)fluoranthene
Benzo(a)pyrene
Average
Clay
30
10
23
77
9
96
7
9
5
68
20
11
4
3
7
4
4
% Recovery3
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
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METHOD 8275
THERMAL CHROMATOGRAPHY/MASS SPECTROMETRY (TC/MS) FOR
SCREENING SEMIVOLATILE ORGANIC COMPOUNDS
C
7.1 Px apax
7 . a Px apa r•
a rvd load
mamp1a
fc»x• we1aht
7.3,3 PI »a a
• amp 1 • In
mm Cab 1 1 «
w* iff tit
7.3.1 IT I
7.9.2 U«lnflT
-------
METHOD 5050
BOMB COMBUSTION METHOD FOR SOLID WASTE
1.0 SCOPE AND APPLICATION
1.1 This method describes the sample preparation steps necessary to
determine total chlorine in solid waste and virgin and used oils, fuels and
related materials, including: crankcase, hydraulic, diesel, lubricating and fuel
oils, and kerosene by bomb oxidation and titration or ion chromatography.
Depending on the analytical finish chosen, other halogens (bromine and fluorine)
and other elements (sulfur and nitrogen) may also be determined.
1.2 The applicable range of this method varies depending on the
analytical finish chosen. In general, levels as low as 500 jug/g chlorine in the
original oil sample can be determined. The upper range can be extended to
percentage levels by dilution of the combustate.
1.3 This standard may involve hazardous materials, operations, and
equipment. This standard does not purport to address all of the safety problems
associated with its use. It is the responsibility of the user of this standard
to establish appropriate safety and health practices and determine the
applicability of regulatory limitations prior to use. Specific safety statements
are given in Section 3.0.
2.0 SUMMARY OF METHOD
2.1 The sample is oxidized by combustion in a bomb containing oxygen
under pressure. The liberated halogen compounds are absorbed in a sodium
carbonate/sodium bicarbonate solution. Approximately 30 to 40 minutes are
required to prepare a sample by this method. Samples with a high water content
(> 25%) may not combust efficiently and may require the addition of a mineral oil
to facilitate combustion. Complete combustion is still not guaranteed for such
samples.
2.2 The bomb combustate solution can then be analyzed for the following
elements as their anion species by one or more of the following methods:
Method Title
9252 Chloride (Titrimetric, Mercuric Nitrate)
9253 Chloride (Titrimetric, Silver Nitrate)
9056 Anion Chromatography Method (Chloride, Sulfate, Nitrate,
Phosphate, Fluoride, Bromide)
5050 - 1 Revision 0
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NOTE: Strict adherence to all of the provisions prescribed hereinafter
ensures against explosive rupture of the bomb, or a blowout,
provided the bomb is of proper design and construction and in good
mechanical condition. It is desirable, however, that the bomb be
enclosed in a shield of steel plate at least 1/2 in. (12.7 mm)
thick, or equivalent protection be provided against unforeseeable
contingencies.
3.0 INTERFERENCES
3.1 Samples with very high water content (> 25%) may not combust
efficiently and may require the addition of a mineral oil to facilitate
combustion.
3.2 To determine total nitrogen in samples, the bombs must first be
purged of ambient air. Otherwise, nitrogen results will be biased high.
4.0 APPARATUS AND MATERIALS
4.1 Bomb, having a capacity of not less than 300 mL, so constructed
that it will not leak during the test, and that quantitative recovery of the
liquids from the bomb may be readily achieved. The inner surface of the bomb may
be made of stainless steel or any'other material that will not be affected by the
combustion process or products. Materials used in the bomb assembly, such as the
head gasket and lead-wire insulation, shall be resistant to heat and chemical
action and shall not undergo any reaction that will affect the chlorine content
of the sample in the bomb.
4.2 Sample cup, platinum or stainless steel, 24 mm in outside diameter
at the bottom, 27 mm in outside diameter at the top, 12 mm in height outside, and
weighing 10 to 11 g.
4.3 Firing wire, platinum or stainless steel, approximately No. 26 B
& S gage.
4.4 Ignition circuit, capable of supplying sufficient current to ignite
the nylon thread or cotton wicking without melting the wire.
NOTE: The switch in the ignition circuit shall be of the type that
remains open, except when held in closed position by the operator.
4.5 Nylon sewing thread, or Cotton Wicking, white.
4.6 Funnel, to fit a 100-mL volumetric flask.
4.7 Class A volumetric flasks, 100-mL, one per sample.
4.8 Syringe, 5- or 10-mL disposable plastic.
4.9 Apparatus for specific analysis methods are given in the methods.
4.10 Analytical balance: capable of weighing to 0.0001 g.
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5.0 REAGENTS
5.1 Purity of reagents. 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 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Oxygen. Free of combustible material and halogen compounds,
available at a pressure of 40 atm.
WARNING: Oxygen vigorously accelerates combustion (see Appendix Al.l)
5.4 Sodium bicarbonate/sodium carbonate solution. Dissolve 2.5200 g
NaHC03 and 2.5440 g Na2C03 in reagent water and dilute to 1 L.
5.5 White oil. Refined.
5.6 Reagents and materials for specific analysis methods are given in
the methods.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine.
6.2 Ensure that the portion of the sample used for the test is repre-
sentative of the sample.
6.3 To minimize losses of volatile halogenated solvents that may be
present in the sample, keep the field and laboratory samples as free of headspace
as possible.
6.4 Because used oils may contain toxic and/or carcinogenic substances
appropriate field and laboratory safety procedures should be followed.
7.0 PROCEDURE
7.1 Sample Preparation
7.1.1 Preparation of bomb and sample. Cut a piece of firing wire
approximately 100 mm in length and attach the free ends to the terminals.
Arrange the wire so that it will be just above and not touching the sample
cup. Loop a cotton thread around the wire so that the ends will extend
into the sampling cup. Pi pet 10 mL of the NaHC03/Na?C03 solution into the
bomb, wetting the sides. Take an aliquot of the oil sample of approxi-
mately 0.5 g using a 5- or 10-mL disposable plastic syringe, and place in
the sample cup. The actual sample weight is determined by the difference
5050 - 3 Revision 0
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between the weight of the empty and filled syringe. Do not use more than
1 g of sample.
NOTE: After repeated use of the bomb for chlorine determination, a film
may be noticed on the inner surface. This dullness should be
removed by periodic polishing of the bomb. A satisfactory method
for doing this is to rotate the bomb in a lathe at about 300 rpm
and polish the inside surface with Grit No. 2/0 or equivalent
paper coated with a light machine oil to prevent cutting, and
then with a paste of grit-free chromic oxide and water. This
procedure will remove all but very deep pits and put a high polish
on the surface. Before using the bomb, it should be washed with
soap and water to remove oil or paste left from the polishing
operation. Bombs with porous or pitted surfaces should never be
used because of the tendency to retain chlorine from sample to
sample.
NOTE: If the sample is not readily combustible, other nonvolatile,
chlorine-free combustible diluents such as white oil may be
employed. However, the combined weight of sample and nonvolatile
diluent shall not exceed 1 g. Some solid additives are relatively
insoluble but may be satisfactorily burned when covered with a
layer of white oil.
NOTE: The practice of alternately running samples high and low in
chlorine content should be avoided whenever possible. It is
difficult to rinse the last traces of chlorine from the walls of
the bomb, and the tendency for residual chlorine to carry over
from sample to sample has been observed in a number of
laboratories. When a sample high in chlorine has preceded one low
in chlorine content, the test on the low-chlorine sample should
be repeated, and one or both of the low values thus obtained
should be considered suspect if they do not agree within the
limits of repeatability of this method.
NOTE: Do not use more than 1 g total of sample and white oil or other
chlorine-free combustible material. Use of excess amounts of
these materials could cause a buildup of dangerously high pressure
and possible rupture of the bomb.
7.1.2 Addition of oxygen. Place the sample cup in position
and arrange the thread so that the end dips into the sample. Assemble the
bomb and tighten the cover securely. Admit oxygen slowly (to avoid
blowing the oil from the cup) until a pressure is reached as indicated in
Table 1.
NOTE: Do not add oxygen or ignite the sample if the bomb has been
jarred, dropped, or tiled.
1Emery Polishing Paper grit No. 2/0 may be purchased from the Behr-Manning
Co., Troy, NY.
2Chromic oxide may be purchased from J.T. Baker & Co., Phillipsburg, NJ.
5050 - 4 Revision 0
November 1992
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7.1.3 Combustion. Immerse the bomb in a cold water bath.
Connect the terminals to the open electrical circuit. Close the circuit
to ignite the sample. Remove the bomb from the bath after immersion for
at least 10 minutes. Release the pressure at a slow, uniform rate such
that the operation requires at least 1 min. Open the bomb and examine the
contents. If traces of unburned oil or sooty deposits are found, discard
the determination, and thoroughly clean the bomb before using it again.
7.1.4 Collection of halogen solution. Using reagent water and
a funnel, thoroughly rinse the interior of the bomb, the sample cup, the
terminals, and the inner surface of the bomb cover into a 100-mL
volumetric flask. Dilute to the mark with reagent water.
7.1.5 Cleaning procedure for bomb and sample cup. Remove any
residual fuse wire from the terminals and the cup. Using hot water, rinse
the interior of the bomb, the sample cup, the terminals, and the inner
surface of the bomb cover. (If any residue remains, first scrub the bomb
with Alconox solution). Copiously rinse the bomb, cover, and cup with
reagent water.
7.2 Sample Analysis. Analyze the combustate for chlorine or other
halogens using the methods listed in Step 2.2. It may be necessary to dilute the
samples so that the concentration will fall within the range of standards.
7.3 Calculations. Calculate the concentrations
detected in the sample according to the following equation:
Ccom x Vcom x DF
of each element
(1)
where:
V
DF
W
concentration of element in
concentration of element in
total volume of combustate,
dilution factor
weight of sample combusted,
the sample,
the combustate,
ml
jLtg/mL
Report the concentration of each element detected in the sample in
micrograms per gram.
Example: A 0.5-g oil sample was combusted, yielding 10 ml of combustate.
The combustate was diluted to 100 ml total volume and analyzed for chloride,
which was measured to be 5 ng/ml. The concentration of chlorine in the original
sample is then calculated as shown below:
5 ug x (10 ml) x (10)
ml
0.5 g
(2)
5050 - 5
Revision 0
November 1992
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C0 = l.OOOiifl (3)
g
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures. ™
8.2 One sample in ten should be bombed twice. The results should agree
to within 10%, expressed as the relative percent difference of the results.
8.3 Analyze matrix spike and matrix spike duplicates - spike samples with
the elements of interest at a level commensurate with the levels being
determined. The spiked compounds should be similar to those expected in the
sample. Any sample suspected of containing > 25% water should also be spiked
with organic chlorine.
8.4 For higher levels (e.g.. percent levels), spiking may be
inappropriate. For these cases, samples of known composition should be
combusted. The results should agree to within 10% of the expected result.
8.5 Quality control for the analytical method(s) of choice should be
followed.
9.0 PERFORMANCE
See analytical methods referenced in Step 2.2.
10.0 REFERENCES
1. ASTM Method D 808-81, Standard Test Method for Chlorine in New and Used ^
Petroleum Products (Bomb Method). 1988 Annual Book of ASTM Standards. Volume m
05.01 Petroleum Products and Lubricants. "
2. Gaskill, A.; Estes, E. D.; Hardison, D. L.; and Myers, L. E. Validation
of Methods for Determining Chlorine in Used Oils and Oil Fuels. Prepared for
U.S. Environmental Protection Agency, Office of Solid Waste. EPA Contract No.
68-01-7075, WA 80. July 1988.
5050 - 6 Revision 0
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TABLE 1.
GAGE PRESSURES
Capacity of bomb, ml
Minimum
gage
pressure , atm
Maximum
gage
pressure , atm
300 to 350
350 to 400
400 to 450
450 to 500
38
35
30
27
40
37
32
29
aThe minimum pressures are specified to provide sufficient oxygen for complete
combustion, and the maximum pressures represent a safety requirement. Refer to
manufacturers' specifications for appropriate gage pressure, which may be lower
than those listed here.
APPENDIX
Al. PRECAUTIONARY STATEMENTS
Al.1 Oxygen
vigorously
Warning — Oxygen
accelerates combustion.
Keep oil and grease away. Do
not use oil or grease on regulators,
gages, or control equipment.
Use only with equipment
conditioned for oxygen service by
careful cleaning to remove oil,
grease, and other combustibles.
Keep combustibles away from
oxygen and eliminate ignition
sources.
Keep surfaces clean to prevent
ignition or explosion, or both, on
contact with oxygen.
Always use a pressure
regulator. Release regulator tension
before opening cylinder valve.
All equipment and containers
used must be suitable and recommended
for oxygen service.
Never attempt
oxygen from cylinder
to transfer
in which it is
received to any other cylinder.
not mix gases in cylinders.
Do
Do not drop cylinder. Make
sure cylinder is secured at all
times.
Keep cylinder valve closed when
not in use.
Stand away from outlet when
opening cylinder valve.
For technical use only. Do not
use for inhalation purposes.
Keep cylinder out of sun and
away from heat.
Keep cylinders from corrosive
environment.
Do not use cylinder without
label.
Do not use dented or damaged
cylinders.
See Compressed Gas Association
booklets G-4 and G4.1 for details of
safe practice in the use of oxygen.
5050 - 7
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METHOD 5050
BOMB COMBUSTION METHOD FOR SOLID WASTE
START
1 1 1 Prepare bomb
and sample
i
712 Slowly add
oxygen to sample
cup
..
713 Immerse bomb
in cold wa ter ,
igni te sampl e ,
remove bomb from
water , release
pressure, open bomb
1
714 Rinse bomb.
sampl e cup ,
te rmina is, and bomb
cover with water
—J
|-»
715 Rinse bomb ,
sample cup,
terminals , and bomb
cover with hot
wa ter
7 2 Analyze
combus ta te
I
7 3 Calculate
concentration of
each element
detected
s ^\
/ \
STOP
5050 - 8
Revision 0
November 1992
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METHOD 9010
TOTAL AND AMENABLE CYANIDE
1.0 SCOPE AND APPLICATION
1.1 Method 9010 is used to determine the concentration of inorganic
cyanide (CAS Registry Number 57-12-5) in wastes or leachate. The method
detects inorganic cyanides that are present as either soluble salts or
complexes. It is used to determine values for both total cyanide and cyanide
amenable to chlorination. The "reactive" cyanide content of a waste, that is,
the cyanide content that could generate toxic fumes when exposed to mild
acidic conditions, is not determined by Method 9010 (refer to Chapter Seven,
Step 7.3.3.2). Method 9010 is not intended to determine if a waste is
hazardous by the characteristic of reactivity.
1.2 The titration procedure using silver nitrate with p-dimethylamino-
benzal-rhodanine indicator is used for measuring concentrations of cyanide
exceeding 0.1 mg/L (0.025 mg/250 ml of absorbing liquid).
1.3 The colorimetric procedure is used for concentrations below 1 mg/L of
cyanide and is sensitive to about 0.02 mg/L.
2.0 SUMMARY OF METHOD
2.1 The cyanide, as hydrocyanic acid (HCN), is released from samples
containing cyanide by means of a reflux-distillation operation under acidic
conditions and absorbed in a scrubber containing sodium hydroxide solution.
The cyanide in the absorbing solution is then determined colorimetrically or
titrametrically.
2.2 In the colorimetric measurement, the cyanide is converted to cyanogen
chloride (CNC1) by reaction of cyanide with chloramine-T at a pH less than 8.
After the reaction is complete, color is formed on the addition of pyridine-
barbituric acid reagent. The absorbance is read at 578 nm for the complex
formed with pyridine-barbituric acid reagent and CNC1. To obtain colors of
comparable intensity, it is essential to have the same salt content in both
the sample and the standards.
2.3 The titration measurement uses a standard solution of silver nitrate
to titrate cyanide in the presence of a silver sensitive indicator.
3.0 INTERFERENCES
3.1 Interferences are eliminated or reduced by using the distillation
procedure. Chlorine and sulfide are interferences in Method 9010.
3.2 Oxidizing agents such as chlorine decompose most cyanides. Chlorine
interferences can be removed by adding an excess of sodium arsenite to the
waste prior to preservation and storage of the sample to reduce the chlorine
to chloride which does not interfere.
9010 - 1 Revision 1
December 1987
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3.3 Sulfide interference can be removed by adding an excess of bismuth
nitrate to the waste (to precipitate the sulfide) before distillation.
Samples that contain hydrogen sulfide, metal sulfides, or other compounds that
may produce hydrogen sulfide during the distillation should be treated by the
addition of bismuth nitrate.
3.4 High results may be obtained for samples that contain nitrate and/or
nitrite. During the distillation, nitrate and nitrite will form nitrous acid,
which will react with some organic compounds to form oximes. These compounds
once formed will decompose under test conditions to generate HCN. The
possibility of interference of nitrate and nitrite is eliminated by
pretreatment with sulfamic acid just before distillation. Nitrate and nitrite
are interferences when present at levels higher than 10 mg/L and in
conjunction with certain organic compounds.
3.5 Thiocyanate is reported to be an interference when present at very
high levels. Levels of 10 mg/L were not found to interfere.
3.6 Fatty acids, detergents, surfactants, and other compounds may cause
foaming during the distillation when they are present in large concentrations
and will make the endpoint of the titration difficult to detect. They may be
extracted at pH 6-7.
4.0 APPARATUS AND MATERIALS
4.1 Reflux distillation apparatus such as shown in Figure 1 or Figure 2.
The boiling flask should be of one liter size with inlet tube and provision
for condenser. The gas absorber may be a 270-mL Fisher-Milligan scrubber
(Fisher, Part No. 07-513 or equivalent). The reflux apparatus may be a Wheaton
377160 distillation unit or equivalent.
4.2 Spectrophotometer - Suitable for measurements at 578 nm with a 1.0 cm
cell or larger.
4.3 Hot plate stirrer/heating mantle.
4.4 pH meter.
4.5 Amber light.
4.6 Vacuum source.
4.7 Refrigerator.
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.
9010 - 2 Revision 1
December 1987
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5.2 ASTM Type II Water (ASTM D1193-77 (1983)). All water used in this
method will be Type II unless otherwise specified.
5.3 Reagents for sample collection, preservation, and handling
5.3.1 Sodium arsenite (0.1N), NaAs02- Dissolve 3.2 g NaAs02 in
250 ml water.
5.3.2 Ascorbic acid,
5.3.3 Sodium hydroxide solution (50%), NaOH. Commercially available.
5.3.4 Acetic acid (1.6M) CHsCOOH. Dilute one part of concentrated
acetic acid with 9 parts of water.
5.3.5 2,2,4-Trimethylpentane,
5.3.6 Hexane,
5.3.7 Chloroform,
5.4 Reagents for cyanides amenable to chlorination
5.4.1 Calcium hypochlorite solution (0.35M), Ca(OCl)2- Combine 5 g
of calcium hypochlorite and 100 ml of water. Shake before using.
5.4.2 Sodium hydroxide solution (1.25N), NaOH. Dissolve 50 g of NaOH
in 1 liter of water.
5.4.3 Sodium arsenite (0.1N). See Step 5.3.1.
5.5 Reagents for distillation
5.5.1 Sodium hydroxide (1.25N). See Step 5.4.2.
5.5.2 Bismuth nitrate (0.062M), Bi (NO)3«5H20. Dissolve 30 g
Bi(NO)3-5H20 in 100 ml of water. While stirring, add 250 ml of glacial
acetic acid, CHsCOOH. Stir until dissolved and dilute to 1 liter with
water.
5.5.3 Sulfamic acid (0.4N), H2NSOsH. Dissolve 40 g H2NS03H in
1 liter of water.
5.5.4 Sulfuric acid (18N), H2S04. Slowly and carefully add 500 ml of
concentrated H2S04 to 500 ml of water.
5.5.5 Magnesium chloride solution (2.5M), MgCl2-6H20. Dissolve 510 g
of MgCl2-6H20 in 1 liter of water.
5.6 Reagents for colorimetric determination
5.6.1 Sodium hydroxide solution (0.25N), NaOH. Dissolve 10 g NaOH in
1 liter of water.
9010 - 3 Revision 1
December 1987
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5.6.2 Sodium phosphate monobasic (1M), NaHgPCV^O. Dissolve 138 g
of NaH2P04-H20 in 1 liter of water. Refrigerate this solution.
5.6.3 Chloramine-T solution (0.44%), Cy^ClNNaC^S. Dissolve 1.0 g
of white, water soluble chloramine-T in 100 ml of water and refrigerate
until ready to use.
5.6.4 Pyridine-Barbituric acid reagent, CsHsN ^4^203. Place 15 g
of barbituric acid in a 250-mL volumetric flask and add just enough water
to wash the sides of the flask and wet the barbituric acid. Add 75 ml of
pyridine and mix. Add 15 ml of concentrated hydrochloric acid (HC1), mix,
and cool to room temperature. Dilute to 250 ml with water. This reagent
is stable for approximately six months if stored in a cool, dark place.
5.6.5 Stock potassium cyanide solution (1 ml = 1000 ug CN), KCN.
Dissolve 2.51 g of KCN and 2 g KOH in 900 mL of water. Standardize with
0.0192N silver nitrate, AgNOs. Dilute to appropriate concentration to
achieve 1 ml = 1000 ug of CN.
NOTE: Detailed procedure for AgNOs standardization is described in
"Standard Methods for the Examination of Water and Wastewater",
16th Edition, (1985), Methods 412C and 407A.
5.6.6 Intermediate standard potassium cyanide solution, (1 mL =
100 ug CN), KCN. Dilute 100 ml of stock potassium cyanide solution (1 ml =
1000 ug CN) to 1000 ml with water.
5.6.7 Working standard potassium cyanide solution (1 ml = 10 ug CN),
KCN. Prepare fresh daily by diluting 100 mL of intermediate standard
potassium cyanide solution and 10 ml of IN NaOH to 1 liter with water.
5.7 Reagents for titration procedure
5.7.1 Rhodanine indicator - Dissolve 20 mg of p-dimethylamino-
benzal-rhodanine, Ci2Hi2N20s2> in 1Q0 mL of acetone.
5.7.2 Standard silver nitrate solution (0.0192N), AgN03. Prepare by
crushing approximately 5 g AgNOs and drying to constant weight at 40"C.
Weigh out 3.2647 g of dried AgNOs. Dissolve in 1 liter of water.
NOTE: Detailed procedure for AgNOs standardization is described in
"Standard Methods for the Examination of Water and Wastewater",
16th Edition, (1985), Methods 412C and 407A.
6.0 SAMPLE COLLECTION, PRESERVATION AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine.
6.2 Samples should be collected in plastic or glass containers. All
containers must be thoroughly cleaned and rinsed.
9010 - 4 Revision 1
December 1987
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6.3 Oxidizing agents such as chlorine decompose most cyanides. To
determine whether oxidizing agents are present, test a drop of the sample with
potassium iodide-starch test paper. A blue color indicates the need for
treatment. Add 0.1N sodium arsenite solution a few ml at a time until a drop
of sample produces no color on the indicator paper. Add an additional 5 ml of
sodium arsenite solution for each liter of sample. Ascorbic acid can be used
as an alternative although it is not as effective as arsenite. Add a few
crystals of ascorbic acid at a time until a drop of sample produces no color
on the indicator paper. Then add an additional 0.06 g of ascorbic acid for
each liter of sample volume.
6.4 Aqueous samples must be preserved by adding 50% sodium hydroxide
until the pH is greater than or equal to 12 at the time of collection.
6.5 Samples should be chilled to 4°C.
6.6 When properly preserved, cyanide samples can be stored for up to 14
days prior to analysis.
6.7 Solid and oily wastes may be extracted prior to analysis by the
method in Appendix A. It uses a dilute NaOH solution (pH = 12) as the
extractant. This yields extractable cyanide.
6.8 If fatty acids, detergents, and surfactants are a problem, they may
be extracted using the following procedure. Acidify the sample with acetic
acid (1.6M) to pH 6.0 to 7.0.
CAUTION: The initial reaction product of alkaline chlorination is the very
toxic gas cyanogen chloride; therefore, it is necessary that this
reaction be performed in a hood.
Extract with isooctane, hexane, or chloroform (preference in order named) with
solvent volume equal to 20% of the sample volume. One extraction is usually
adequate to reduce the compounds below the interference level. Avoid multiple
extractions or a long contact time at low pH in order to keep the loss of HCN
at a minimum. When the extraction is completed, immediately raise the pH of
the sample to above 12 with 50% NaOH solution.
CAUTION: This procedure can produce lethal HCN gas.
7.0 PROCEDURE
7.1 Pretreatment for cyanides amenable to chlorination
7.1.1 This test must be performed under amber light. K3[Fe-(CN)s]
may decompose under UV light and hence will test positive for cyanide
amenable to chlorination if exposed to fluorescent lighting or sunlight.
Two identical sample aliquots are required to determine cyanides amenable
to chlorination.
7.1.2 To one 500 ml sample or to a sample diluted to 500 ml, add
calcium hypochlorite solution dropwise while agitating and maintaining the
pH between 11 and 12 with 1.25N sodium hydroxide until an excess of
9010 - 5 Revision 1
December 1987
-------
chlorine is present as indicated by Kl-starch paper turning blue. The
sample will be subjected to alkaline chlorination by this step.
CAUTION: The initial reaction product of alkaline chlorination is
the very toxic gas cyanogen chloride; therefore, it is
necessary that this reaction be performed in a hood.
7.1.3 Test for excess chlorine with Kl-starch paper and maintain
this excess for one hour with continuous agitation. A distinct blue color
on the test paper indicates a sufficient chlorine level. If necessary, add
additional calcium hypochlorite solution.
7.1.4 After one hour, add 1 ml portions of 0.1N sodium arsenite
until Kl-starch paper shows no residual chlorine. Add 5 ml of excess
sodium arsenite to ensure the presence of excess reducing agent.
7.1.5 Test for total cyanide as described below in both the
chlorinated and the unchlorinated samples. The difference of total cyanide
in the chlorinated and unchlorinated samples is the cyanide amenable to
chlorination.
7.2 Distillation Procedure
7.2.1 Place 500 ml of sample, or sample diluted to 500 ml in the one
liter boiling flask. Pipet 50 ml of 1.25N sodium hydroxide into the gas
absorber. If the apparatus in Figure 1 is used, add water until the spiral
is covered. Connect the boiling flask, condenser, gas absorber and vacuum
trap.
7.2.2 Start a slow stream of air entering the boiling flask by
adjusting the vacuum source. Adjust the vacuum so that approximately two
bubbles of air per second enter the boiling flask through the air inlet
tube.
7.2.3 If samples are known to contain sulfide, add 50 mL of 0.062M
bismuth nitrate solution through the air inlet tube. Mix for three
minutes. Use lead acetate paper to check the sample for the presence of
sulfide. A positive test is indicated by a black color on the paper.
7.2.4 If samples are known or suspected to contain nitrate or
nitrite, add 50 ml of 0.4N sulfamic acid solution through the air inlet
tube. Mix for three minutes.
7.2.5 Slowly add 50 ml of 18N sulfuric acid through the air inlet
tube. Rinse the tube with water and allow the airflow to mix the flask
contents for three minutes. Add 20 ml of 2.5M magnesium chloride through
the air inlet and wash the inlet tube with a stream of water.
7.2.6 Heat the solution to boiling. Reflux for one hour. Turn off
heat and continue the airflow for at least 15 minutes. After cooling the
boiling flask, and closing the vacuum source, disconnect the gas absorber.
9010 - 6 Revision 1
December 1987
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7.2.7 Transfer the solution from the absorber into a 250-mL
volumetric flask. Rinse the absorber and add the rinse water to the
volumetric flask. Dilute to volume with water.
7.2.8 If the manual spectrophotometric determination will be
performed, proceed to Step 7.3.1. If the titration procedure will be
performed, proceed to Step 7.7.
7.3 Manual spectrophotometric determination
7.3.1 Pipet 50 ml of the absorber solution into a 100-mL volumetric
flask. If the sample is later found to be beyond the linear range of the
colorimetric determination and redistillation of a smaller sample is not
feasible, a smaller aliquot may be taken. If less than 50 ml is taken,
dilute to 50 ml with 0.25N sodium hydroxide solution.
NOTE: Temperature of reagents and spiking solution can affect the
response factor of the colorimetric determination. The reagents
stored in the refrigerator should be warmed to ambient temperature
before use. Samples should not be left in a warm instrument to
develop color, but instead they should be aliquoted to a cuvette
immediately prior to reading the absorbance.
7.3.2 Add 15 ml of 1M sodium phosphate solution and mix. Add 2 ml of
chloramine-T and mix. Some distillates may contain compounds that have
chlorine demand. One minute after the addition of chloramine-T, test for
excess chlorine with Kl-starch paper. If the test is negative, add 0.5 ml
chloramine-T. After one minute recheck with Kl-starch paper. Continue to
add chloramine-T in 0.5 ml increments until an excess is maintained. After
1 to 2 minutes, add 5 ml of pyridine-barbituric acid solution and mix.
7.3.3 Dilute to 100 ml with water and mix again. Allow 8 minutes for
color development and then read the absorbance at 578 nm in a 1-cm cell
within 15 minutes. The sodium hydroxide concentration will be 0.125N.
7.4 Standard curve for samples without sulfide
7.4.1 Prepare a series of standards by pipetting suitable volumes of
working standard potassium cyanide solution into 250-mL volumetric flasks.
To each flask, add 50 ml of 1.25N sodium hydroxide and dilute to 250 ml
with water. Prepare using the following table. The sodium hydroxide
concentration will be 0.25N.
ml of Working Standard Solution Concentration
(1 ml = 10 uq CN) (uq CN/U
0 Blank
1.0 40
2.0 80
5.0 200
10.0 400
15.0 600
20.0 800
9010 - 7 Revision 1
December 1987
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7.4.2 After the standard solutions have been prepared according to
the table above, pipet 50 ml of each standard solution into a 100-mL
volumetric flask and proceed to Steps 7.3.2 and 7.3.3 to obtain absorbance
values for the standard curve. The final concentrations for the standard
curve will be one half of the amounts in the above table (ranging from 20
to 400 ug/L).
7.4.3 It is recommended that at least two standards (a high and a
low) be distilled and compared to similar values on the curve to ensure
that the distillation technique is reliable. If distilled standards do
not agree within + 10% of the undistilled standards, the analyst should
find the cause of the apparent error before proceeding.
7.4.4 Prepare a standard curve by plotting absorbance of standard
versus the cyanide concentration.
7.4.5 To check the efficiency of the sample distillation, add
cyanide from the working standard to 500 mL of sample to ensure a level of
40 ug/L. Proceed with the analysis as in Step 7.2.1.
7.5 Standard curve for samples with sulfide
7.5.1 It is imperative that all standards be distilled in the same
manner as the samples using the method of standard additions. Standards
distilled by this method will give a linear curve, at low concentrations,
but as the concentration increases, the recovery decreases. It is
recommended that at least five standards be distilled.
7.5.2 Prepare a series of standards similar in concentration to
those mentioned in Step 7.4.1 and analyze as in Step 7.2.1. Prepare a
standard curve by plotting absorbance of standard versus the cyanide
concentration.
7.6 Calculation - If the colorimetric procedure is used, calculate
the cyanide, in ug/L, in the original sample as follows.
CN (ug/L) = A x B x C
D x E
where:
A = ug/L CN read from standard curve.
B = mL of original sample for distillation (500 recommended).
C = mL of sample after distillation (250).
D = mL used for colorimetric analysis (50 recommended).
E = mL of sample after preparation of colorimetric analysis (100).
7.7 Titration Procedure
7.7.1 Transfer the gas absorber solution or a suitable aliquot from
the 250-mL volumetric flask to a 500-mL Erlenmeyer flask. Add 10-12 drops
of the rhodanine indicator.
9010 - 8 Revision 1
December 1987
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7.7.2 Titrate with standard 0.0192N silver nitrate to the first
change in color from yellow to brownish-pink. Titrate a water blank using
the same amount of sodium hydroxide and indicator as in the sample.
7.7.3 The analyst should be familiar with the endpoint of the
titration and the amount of indicator to be used before actually titrating
the samples. A 5-mL buret may be conveniently used to obtain a precise
titration.
7.7.4 Calculation - If the titrimetric procedure is used, calculate
concentration of CN in mg/L in the original sample as follows:
CN (mg/L) = (A - B) 1000 x C
D E
where:
A = ml of AgNOs for titration of sample.
B = mL of AgNOs f°r titration of blank.
C = ml of sample after distillation (250).
D = ml of original sample for distillation (500 recommended).
E = ml of sample taken for titration (250 recommended).
The above equation assumes that the standard silver nitrate concentration
is exactly 0.0192N which is equivalent to one ml of silver nitrate to one
mg of cyanide.
8.0 QUALITY CONTROL
8.1 All quality control data should be maintained and available for easy
reference or inspection.
8.2 Employ a minimum of one reagent blank per analytical batch or once in
every 20 samples to determine if contamination or any memory effects are
occurring.
8.3 Analyze check standards with every analytical batch of samples. If
the standards are not within 15% of the expected value, then the samples must
be reanalyzed.
8.4 Run one replicate sample for every 10 samples. A replicate sample is
a sample brought through the entire sample preparation process. The CV of the
replicates should be 15% or less. If this criterion is not met, the samples
should be reanalyzed.
8.5 The method of standard additions shall be used for the analysis of
all samples that suffer from matrix interferences such as samples which
contain sulfides.
9010 - 9 Revision 1
December 1987
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9.0 METHOD PERFORMANCE
9.1 The titration procedure using silver nitrate is used for measuring
concentrations of cyanide exceeding 0.1 mg/L. The colorimetric procedure is
used for concentrations below 1 mg/L of cyanide and is sensitive to about
0.02 mg/L.
9.2 EPA Method 335.2 (sample distillation with titration) reports that in
a single laboratory using mixed industrial and domestic waste samples at
concentrations of 0.06 to 0.62 mg/L CN, the standard deviations for precision
were + 0.005 to + 0.094, respectively. In a single laboratory using mixed
industrial and domestic waste samples at concentrations of 0.28 and 0.62 mg/L
CN, recoveries (accuracy) were 85% and 102%, respectively.
9.3 In two additional studies using surface water, ground water, and
landfill leachate samples, the titration procedure was further evaluated. The
concentration range used in these studies was 0.5 to 10 mg/L cyanide. The
detection limit was found to be 0.2 mg/L for both total and amenable cyanide
determinations. The precision (CV) was 6.9 and 2.6 for total cyanide
determinations and 18.6 and 9.1 for amenable cyanide determinations. The mean
recoveries were 94% and 98.9% for total cyanide, and 86.7% and 97.4% for
amenable cyanide.
10.0 REFERENCES
1. 1985 Annual Book of ASTM Standards. Vol. 11.01; "Standard Specification
for Reagent Water"; ATSM: Philadelphia, PA, 1985,; D1193-77.
2. 1982 Annual Book ASTM Standards. Part 19; "Standard Test Methods for
Cyanide in Water"; ASTM: Philadelphia, PA, 1982; 2036-82.
3. Bark, L.S.; Higson, H.G. Talanta 1964, 2, 471-479.
4. Britton, P.; Winter, J.; Kroner, R.C. "EPA Method Study 12, Cyanide in
Water"; final report to the U.S. Environmental Protection Agency. National
Technical Information Service: Springfield, VA, 1984; PB80-196674.
5. Casey, J.P.; Bright, J.W.; Helms, B.D. "Nitrosation Interference in
Distillation Tests for Cyanide"; Gulf Coast Waste Disposal Authority:
Houston, Texas.
6. Egekeze, J.O.; Oehne, F.W. J. Anal. Toxicology 1979, 3, 119.
7. Elly, C.T. vL Water Pollution Control Federation 1968, 40, 848-856.
8. Fuller, W. Cyanide in the Environment; Van Zyl, D., Ed.; Proceedings of
Symposium; December, 1984.
9. Gottfried, G.J. "Precision, Accuracy, and MDL Statements for EPA Methods
9010, 9030, 9060, 7520, 7521,7550, 7551, 7910, and 7911"; final report to
the U.S. Environmental Protection Agency. Environmental Monitoring and
Support Laboratory. Biospheric: Cincinnati, OH, 1984.
9010 - 10 Revision 1
December 1987
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10. 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-
11.
12.
13.
14.
600/4-79-020.
Rohrbough, W.G.; et
Specifications, 7th
1986.
al. Reagent Chemicals, American Chemical Society
ed.; American Chemical Society: Washington, DC,
Standard Methods for the Examination of Water and Wastewater. 16th ed.;
Greenberg, A.E.; Trussell, R.R.; Clesceri, L.S., Eds.; American Water
Works Association, Water Pollution Control Federation, American Public
Health Association: Washington, DC, 1985.
Umana, M.; Beach, J.; Sheldon, L. "Revisions to Method 9010"; final
report to the U.S. Environmental Protection Agency. Office of Solid
Waste. Research Triangle Institute: Research Triangle Park, NC, 1986.
Umana, M.; Sheldon, L. "Interim Report: Literature Review"; interim
report to the U.S. Environmental Protection Agency. Office of Solid
Waste. Research Triangle Institute: Research Triangle Park, NC, 1986.
9010 - 11
Revision 1
December 1987
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FIGURE 1.
APPARATUS FOR CYANIDE DISTILLATION
COOLING WATER
INLET TUBE^
SCREW CLAMP
J
TO LOW VACUUM
SOURCE
* ABSORBER
- DISTILLING FLASK
9010 - 12
Revision 1
December 1987
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FIGURE 2.
APPARATUS FOR CYANIDE DISTILLATION
Connecting Tubmg
Condense
Air Inlet Tube
One Liter
Boii.n Flask
SuCt'On
9010 - 13
Revision 1
December 1987
-------
METHOD 9010
TOTAL AND AMENABLE CYANIDE
C
Start
7 1 1 Pirfori tut
under nb.r llfht.
gie two ii»pl«
aliqnoti
721 Place aaiple
ii b
pipe
into
fit!
I"
illBi flail.
1 2SI laOK
abeorber
COB tct bolliBf
coBdeBitr,
biorbtr, and
trap
7.1.2 Add calcine
hypochlorite
•olatioB to o&t
saaplo aliqnot.
Baintaifi pH at
11-12 with 1 2o»
la OH
7 2.6 Xeat loin and
reflux for 1 hour
722 Allow itreal
of air Into boiling
flaik
7 1 3 Teat t
excels chlor
with Il->tar
paper add
additional cal
hypochlonte
if neceatary
T 3 3 Add 0 062M
bumnth nitrate
•oln* iix for 3
•inotei
71.4 After i tear
add lodiu areealt*
7 2 4 Add 0
iBlfaiic acid
•is for 3 ii
41
loin
7 2.4 Do
taaplee contal
altrate aad/or
aitrite?
7 2 < Cool; cloi*
diiconnect gai
abiorber
727 Tranifer ioln
into flalk dilute
with water to
volne
7 1 6 Teat for
total cyanide la
botk eaipla
allqioti.
7.2 6 Slovly add
1SI lalftrlc acid-
riB.ee tab* vita
»ater «ix for 3
alnatei add 2 SM
•afBieiai chloride
vaii vltk water
7.3 1 Iranfer tola
to aaotler flaek
for aanal
e pec topBotoae trie
detariiaatioB
9010 - 14
Revision 1
December 1987
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METHOD 9010
(Continued)
7 3.3 Add 1M iodiu»
phoapkata toll:
• ix: add
chlora»i»a-I »tll
excaaa ii
iiintained: add
pyridiaa-barbltaric
acid tola: lix
7.4.1 Prapart a
aariaa of itandarda
for ataadard curve
preparation
7.3.3 Dllata to
volua vitk vator:
•ix: allow 1
•inataa (or color
developaaat: raad
abaorbaaca
7.4.1 Pipat
appropriata voluoe
of vorkiac atandard
ICI aola iato
flaaka: add 1.3EI
laOH to each
dllata with vatar
to volaaa
743 Pipat
appropriata voloae
of aach itandard
loin Into flaik
obtain abaorbanca
valaea
7.4.3 Dietill a
ataadarda. coapare
to aiailir valaaa
oa carvt
74.4 Prapara a
ataadard carva
746 Ckack
afficlancT of
•aapla diitillatioa
7.7 1 Tranafar xai
abaorbar aola to
flaik- add
rkodaaina
7 E.I Dlatill all
ataadarda
7.7.3 Titrate vltk
itandard 0.01(31
ailvar aitrata:
tltrata vatar blaak
752 Prepare
atandard curva
7 7 3 The anal/it
•hould be faailiar
with titration
procedure before
titrating laaplei
7.6 For colorlaatic
procadara calcalat*
cjraaide la tg/L
7.7.4 Calcalata
coac of cyaaida
aalaf tltrlaatlc
procadaraa
9010 - 15
Revision 1
December 1987
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APPENDIX 9010A
CYANIDE EXTRACTION PROCEDURE FOR SOLIDS AND OILS
1.0 SCOPE AND APPLICATION
1.1 The extraction procedure described in this appendix is designed for
the extraction of soluble cyanides from solid and oil wastes. The method is
applicable to oil, solid, and multiphasic samples. This method is not
applicable to samples containing insoluble cyanide compounds.
2.0 SUMMARY OF METHOD
2.1 If the waste sample contains so much solid, or solids of such a size
as to interfere with agitation and homogenization of the sample mixture in the
distillation flask, or so much oil or grease as to interfere with the
formation of a homogeneous emulsion, the sample must be extracted with water
at pH 10 or greater, and the extract analyzed by Method 9010. Samples that
contain free water must be filtered and separated into an aqueous component
and a combined oil and solid component. The nonaqueous component may then be
extracted, and an aliquot of the extract combined with an aliquot of the
filtrate in proportion to the composition of the sample. Alternatively, the
components may be analyzed separately, and cyanide levels reported for each
component. However, if the sample solids are known to contain sufficient
levels of cyanide (about 50 ug/g) as to be well above the limit of detection,
the extraction step may be deleted and the solids analyzed directly by Method
9010. This can be accomplished by diluting a small aliquot of the waste solid
(1-5 g) in 500 mL water in the distillation flask and suspending the slurry
during distillation with a magnetic stir-bar.
3.0 INTERFERENCES
3.1 Potential interferences that may be encountered during analysis are
discussed in Method 9010.
4.0 APPARATUS AND MATERIALS
4.1 Extractor - Any suitable device that sufficiently agitates a sealed
container of one liter volume or greater. For the purpose of this analysis,
agitation is sufficient when:
1. All sample surfaces are continuously brought into contact with
extraction fluid, and
2. The agitation prevents stratification of the sample and fluid.
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
9010A - 1 Revision 0
December 1987
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high purity to permit its use without lessening the accuracy of the
determination.
5.2 ASTM Type II Water (ASTM D1193-77 (1983)). All references to water in
the method refer to ASTM Type II unless otherwise specified.
5.3 Sodium hydroxide (50% w/v), NaOH. Commercially available.
5.4 n-Hexane, 05^4.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a plan that addresses the
considerations discussed in Chapter 4 of this manual. See Section 6.0 of
Method 9010 for additional guidance.
7.0 PROCEDURE
7.1 If the waste does not contain any free aqueous phase, go to Step 7.5.
If the sample is a homogeneous fluid or slurry that does not separate or
settle in the distillation flask when using a Teflon coated magnetic stirring
bar but mixes so that the solids are entirely suspended, then the sample may
be analyzed by Method 9010 without an extraction step.
7.2 Assemble Buchner funnel apparatus. Unroll glass filtering fiber and
fold the fiber over itself several times to make a pad about 1 cm thick when
lightly compressed. Cut the pad to fit the Buchner funnel. Weigh the pad, then
place it in the funnel. Turn the aspirator on and wet the pad with a known
amount of water.
7.3 Transfer the sample to the Buchner funnel in small aliquots, first
decanting the fluid. Rinse the sample container with known amounts of water
and add the rinses to the Buchner funnel. When no free water remains in the
funnel, slowly open the stopcock to allow air to enter the vacuum flask. A
small amount of sediment may have passed through the glass fiber pad. This
will not interfere with the analysis.
7.4 Transfer the solid and the glass fiber pad to a tared weighing dish.
Since most greases and oils will not pass through the fiber pad, solids, oils,
and greases will be extracted together. If the filtrate includes an oil phase,
transfer the filtrate to a separatory funnel. Collect and measure the volume
of the aqueous phase. Transfer the oil phase to the weighing dish with the
solid.
7.5 Weigh the dish containing solid, oil (if any), and filter pad.
Subtract the weight of the dry filter pad. Calculate the net volume of water
present in the original sample by subtracting the total volume of rinses used
from the measured volume of the filtrate.
9010A - 2 Revision 0
December 1987
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7.6 Place the following in a 1-liter wide-mouthed bottle:
500 ml water
5 ml 50% w/v NaOH
50 ml n-Hexane (if a heavy grease is present)
If the weight of the solids (Step 7.5) is greater than 25 g, weigh
out a representative aliquot of 25 g and add it to the bottle; otherwise
add all of the solids. Cap the bottle.
7.7 The pH of the extract must be maintained above 10 throughout the
extraction step and subsequent filtration. Since some samples may release
acid, the pH must be monitored as follows. Shake the extraction bottle and
after one minute, check the pH. If the pH is below 12, add 50% NaOH in 5 ml
increments until it is at least 12. Recap the bottle, and repeat the procedure
until the pH does not drop.
7.8 Place the bottle or bottles in the tumbler, making sure there is
enough foam insulation to cushion the bottle. Turn the tumbler on and allow
the extraction to run for about 16 hours.
7.9 Prepare a Buchner funnel apparatus as in Step 7.1 with a glass fiber
pad filter.
7.10 Decant the extract
extract is not necessary.
to the Buchner funnel. Full recovery of the
7.11 If the extract contains an oil phase, separate the aqueous phase
using a separatory funnel. Neither the separation nor the filtration are
critical, but are necessary to be able to measure the volume of the aliquot of
the aqueous extract analyzed. Small amounts of suspended solids and oil
emulsions will not interfere.
7.12 At this point, an aliquot of the filtrate of the original sample may
be combined with an aliquot of the extract in a proportion representative of
the sample. Alternatively, they may be analyzed separately and concentrations
given for each phase. This is described by the following equation:
Liquid Sample Aliquot (ml) = Solid Extracted (g) x Total Sample Filtrate (ml)
Extract Aliquot (mL) Total Solid (g) Total Extraction Fluid (ml)
Where the Total Solids are from Step 7.5, weight of solids and oil phase, dry
weight of filter and tared dish subtracted.
The Total Sample Filtrate includes volume of all rinses added to the filtrate.
The Total Extraction Fluid is 500 ml water plus volume of NaOH solution. Does
not include hexane, which is subsequently removed.
9010A - 3
Revision 0
December 1987
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Alternatively, the aliquots may be analyzed separately, concentrations for
each phase reported separately, and the amounts of each phase present in the
sample reported separately.
8.0 QUALITY CONTROL
8.1 Refer to Method 9010.
9.0 METHOD PERFORMANCE
9.1 In a single laboratory study, recoveries of 60 to 90% are reported
for solids and 88 to 92% for oils. The reported CVs are less than 13.
10.0 REFERENCES
Refer to Method 9010.
9010A - 4 Revision 0
December 1987
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APPENDIX 9010A
CYANIDE EXTRACTION PROCEDURE FOR SOLIDS AND OILS
7 1 Analyzi by
Nitkod 9010
7.1 1*
tk* laipl* a
koiof
alarryT
7 2 Aaaaibl* fllt.r
apparatus
7 S «*ifk tka aolid
and oil portioi of
tk* taapl*
7.4 If ai oily
lay«r i> pr*s*at.
s*parata tk* oil
fro* tk* aqntois
lay*r
Oil
T • Prapar*
•xtractloi
apparatis
7 T Adjust tk* pX
7.8 Parfori
•xtraction
7 • Aisaabl* fllt«r
apparataa
Solid
7.10 rilt.r tk*
taapl*
7 10 Discard tk«
solid and oil
layara
7.11 If ti ollT
l«jr«r It prtunl.
••paratt tk« oil
fro* tk* aqaaova
7 13 Analyn tk*
tqiaoti ItTtr* by
Nttkod B010
Oil
9010A - 5
Revision 0
December 1987
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METHOD 9020B
TOTAL ORGANIC HALIDES (TOX)
1.0 SCOPE AND APPLICATION
1.1 Method 9020 determines Total Organic Hal ides (TOX) as chloride in
drinking water and ground waters. The method uses carbon adsorption with a
microcoulometric-titration detector.
1.2 Method 9020 detects all organic halides containing chlorine,
bromine, and iodine that are adsorbed by granular activated carbon under the
conditions of the method. Fluorine-containing species are not determined by this
method.
1.3 Method 9020 is applicable to samples whose inorganic-halide concen-
tration does not exceed the organic-halide concentration by more than 20,000
times.
1.4 Method 9020 does not measure TOX of compounds adsorbed to
undissolved solids.
1.5 Method 9020 is restricted to use by, or under the supervision of,
analysts experienced in the operation of a pyrolysis/microcoulometer and in the
interpretation of the results.
1.6 This method is provided as a recommended procedure. It may be used
as a reference for comparing the suitability of other methods thought to be
appropriate for measurement of TOX (i.e., by comparison of sensitivity, accuracy,
and precision of data). There are three instruments that can be used to carry
out this method. They are the TOX-10 available from Cosa Instruments, and the
DX-20 and DX-20A available from Xertex-Dohrmann Instruments.
2.0 SUMMARY OF METHOD
2.1 A sample of water that has been protected against the loss of
volatiles by the elimination of headspace in the sampling container, and that is
free of undissolved solids, is passed through a column containing 40 mg of
activated carbon. The column is washed to remove any trapped inorganic halides
and is then combusted to convert the adsorbed organohalides to HX, which is
trapped and titrated electrolytically using a microcoulometric detector.
3.0 INTERFERENCES
3.1 Method interferences may be caused by contaminants, reagents,
glassware, and other sample-processing hardware. All these materials must be
routinely demonstrated to be free from interferences under the conditions of the
analysis by running method blanks.
3.1.1 Glassware must be scrupulously cleaned. Clean all
glassware as soon as possible after use by treating with chromate cleaning
solution. This should be followed by detergent washing in hot water.
Rinse with tap water and distilled water and drain dry; glassware which is
9020B - 1 Revision 2
November 1992
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not volumetric should, in addition, be heated in a muffle furnace at 400eC
for 15 to 30 min. (Volumetric ware should not be heated in a muffle
furnace.) Glassware should be sealed and stored in a clean environment
after drying and cooling to prevent any accumulation of dust or other
contaminants. A
3.1.2 The use of high-purity reagents and gases helps to minimize
interference problems.
3.2 Purity of the activated carbon must be verified before use. Only
carbon samples that register less than 1,000 ng CT/40 mg should be used. The
stock of activated carbon should be stored in its granular form in a glass
container with a Teflon seal. Exposure to the air must be minimized, especially
during and after milling and sieving the activated carbon. No more than a 2-wk
supply should be prepared in advance. Protect carbon at all times from all
sources of halogenated organic vapors. Store prepared carbon and packed columns
in glass containers with Teflon seals.
3.3 Particulate matter will prevent the passage of the sample through
the adsorption column. Particulates must, therefore, be eliminated from the
sample. This must be done as gently as possible, with the least possible sample
manipulation, in order to minimize the loss of volatiles. It should also be
noted that the measured TOX will be biased by the exclusion of TOX from compounds
adsorbed onto the particulates. The following techniques may be used to remove
particulates; however, data users must be informed of the techniques used and
their possible effects on the data. These techniques are listed in order of
preference:
3.3.1 Allow the particulates to settle in the sample container
and decant the supernatant liquid into the adsorption system. A
3.3.2 Centrifuge sample and decant the supernatant liquid into ^
the adsorption system.
3.3.3 Measure Purgeable Organic Hal ides (POX) of sample (see SW-
846 Method 9021) and Non-Purgeable Organic Hal ides (NPOX, that is, TOX of
sample that has been purged of volatiles) separately, where the NPOX
sample is centrifuged or filtered.
4.0 APPARATUS AND MATERIALS
4.1 Adsorption system (a schematic diagram of the adsorption system is
shown in Figure 1):
4.1.1 Adsorption module: Pressurized sample and nitrate-wash
reservoirs. (There are three instruments known to EPA at this time that
can be used to carry out this method. They are the TOX-10, available from
Cosa Instruments, and the DX-20 and DX-20A, available from Xertex-Dohrmann
Instruments.)
4.1.2 Adsorption columns: Pyrex, 5-cm-long x 6-mm-O.D. x 2-mm-
I.D.
9020B - 2 Revision 2
November 1992
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4.1.3 Granular activated carbon (GAC): Filtrasorb-400, Calgon-
APC or equivalent, ground or milled, and screened to a 100/200 mesh range.
Upon combustion of 40 mg of GAC, the apparent halide background should be
1,000 ng Cl" equivalent or less.
4.1.4 Cerafelt (available from Johns-Manville) or equivalent:
Form this material into plugs to fit the adsorption module and to hold 40
mg of GAC in the adsorption columns.
CAUTION: Do not touch this material with your fingers. Oily residue will
contaminate carbon.
4.1.5 Column holders.
4.1.6 Class A volumetric flasks: 100-mL and 50-mL.
4.2 Analytical system:
4.2.1 Microcoulometric-titration system: Containing the
following components (a flowchart of the analytical system is shown in
Figure 2):
4.2.1.1 Boat sampler: Muffled at 800°C for at least 2-4
min and cleaned of any residue by vacuuming after each run.
4.2.1.2 Pyrolysis furnace.
4.2.1.3 Microcoulometer with integrator.
4.2.1.4 Titration cell.
4.2.2 Strip-chart recorder.
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 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Sodium sulfite (0.1 M), Na2S03: Dissolve 12.6 g ACS reagent grade
Na2S03 in reagent water and dilute to 1 L.
5.4 Concentrated nitric acid (HN03).
5.5 Nitrate-wash solution (5,000 mg NO//L), KN03: Prepare a nitrate-
wash solution by transferring approximately 8.2 g of potassium nitrate (KN03)
into a 1-liter Class A volumetric flask and diluting to volume with reagent
water.
9020B - 3 Revision 2
November 1992
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5.6 Carbon dioxide (C02): Gas, 99.9% purity.
5.7 Oxygen (02): 99.9% purity.
5.8 Nitrogen (N2): Prepurified. |
5.9 Acetic acid in water (70%), C2H402: Dilute 7 volumes of glacial
acetic acid with 3 volumes of reagent water.
5.10 Trichlorophenol solution, stock (1 fj.1 = 10 ng Cl~): Prepare a
stock solution by accurately weighing accurately 1.856 g of trichlorophenol into
a 100-mL Class A volumetric flask. Dilute to volume with methanol.
5.11 Trichlorophenol solution, calibration (1 pi = 500 ng Cl"), C6H,C130:
Dilute 5 ml of the trichlorophenol stock solution to 100 ml with methanol.
5.12 Trichlorophenol standard, instrument calibration: First, nitrate-
wash a single column packed with 40 mg of activated carbon, as instructed for
sample analysis, and then inject the column with 10 nl of the calibration
solution.
5.13 Trichlorophenol standard, adsorption efficiency (100 /zg CT/liter):
Prepare an adsorption-efficiency standard by injecting 10 /iL of stock solution
into 1 liter of reagent water.
5.14 Blank standard: The methanol used to prepare the calibration
standard should be used as the blank standard.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses I
the considerations discussed in Chapter Nine.
6.2 All samples should be collected in bottles with Teflon septa (e.g.,
Pierce #12722 or equivalent) and be protected from light. If this is not
possible, use amber glass 250-mL bottles fitted with Teflon-lined caps. Foil may
be substituted for Teflon if the sample is not corrosive. Samples must be
preserved by acidification to pH <2 with sulfuric acid, stored at 4°C, and
protected against loss of volatiles by eliminating headspace in the container.
Samples should be analyzed within 28 days. The container must be washed and
muffled at 400°C before use, to minimize contamination.
6.3 All glassware must be dried prior to use according to the method
discussed in Step 3.1.1.
7.0 PROCEDURE
7.1 Sample preparation:
7.1.1 Special care should be taken in handling the sample in
order to minimize the loss of volatile organohalides. The adsorption
procedure should be performed simultaneously on duplicates.
9020B - 4 Revision 2
November 1992
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7.1.2 Reduce residual chlorine by adding sulfite (5 mg sodium
sulfite crystals per liter of sample). Sulfite should be added at the
time of sampling if the analysis is meant to determine the TOX
concentration at the time of sampling. It should be recognized that TOX
may increase on storage of the sample. Samples should be stored at 4°C
without headspace.
7.2 Calibration:
7.2.1 Check the adsorption efficiency of each newly prepared
batch of carbon by analyzing 100 ml of the adsorption efficiency standard,
in duplicate, along with duplicates of the blank standard. The net
recovery should be within 5% of the standard value.
7.2.2 Nitrate-wash blanks (method blanks): Establish the
repeatability of the method background each day by first analyzing several
nitrate-wash blanks. Monitor this background by spacing nitrate-wash
blanks between each group of eight pyrolysis determinations. The nitrate-
wash blank values are obtained on single columns packed with
40 mg of activated carbon. Wash with the nitrate solution, as instructed
for sample analysis, and then pyrolyze the carbon.
7.2.3 Pyrolyze dupl icate instrument-cal ibration standards and the
blank standard each day before beginning sample analysis. The net
response to the calibration standard should be within 3% of the
calibration-standard value. Repeat analysis of the instrument-calibration
standard after each group of eight pyrolysis determinations and before
resuming sample analysis, and after cleaning or reconditioning the
titration cell or pyrolysis system.
7.3 Adsorption procedure:
7.3.1 Connect two columns in series, each containing 40 mg of
100/200-mesh activated carbon.
7.3.2 Fill the sample reservoir and pass a metered amount of
sample through the activated-carbon columns at a rate of approximately 3
mL/min.
NOTE: 100 ml of sample is the preferred volume for concentrations of TOX
between 5 and 500 M9/L, 50 ml for 501 to 1000 M9/L, and 25 ml for
1001 to 2000 M9/L- If the anticipated TOX is greater than 2000
Mg/L, dilute the sample so that 100 ml will contain between 1 and
50 M9 TOX.
7.3.3 Wash the columns-in-series with 2 ml of the 5,000-mg/L
nitrate solution at a rate of approximately 2 mL/min to displace inorganic
chloride ions.
7.4 Pyrolysis procedure:
7.4.1 The contents of each column are pyrolyzed separately.
After being rinsed with the nitrate solution, the columns should be
9020B - 5 Revision 2
November 1992
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protected from the atmosphere and other sources of contamination until
ready for further analysis.
7.4.2 Pyrolysis of the sample is accomplished in two stages. The
volatile components are pyrolyzed in a C02-rich atmosphere at a low
temperature to ensure the conversion of brominated trihalomethanes to a
titratable species. The less volatile components are then pyrolyzed at a
high temperature in an 02-rich atmosphere.
7.4.3 Transfer the contents of each column to the quartz boat for
individual analysis.
7.4.4 Adjust gas flow according to manufacturer's directions.
7.4.5 Position the sample for 2 min in the 200°C zone of the
pyrolysis tube.
7.4.6 After 2 min, advance the boat into the 800°C zone (center)
of the pyrolysis furnace. This second and final stage of pyrolysis may
require from 6 to 10 min to complete.
7.5 Detection: The effluent gases are directly analyzed in the micro-
coulometric-titration cell. Carefully follow manual instructions for optimizing
cell performance.
7.6 Breakthrough: The unpredictable nature of the background bias
makes it especially difficult to recognize the extent of breakthrough of
organohalides from one column to another. All second-column measurements for a
properly operating system should not exceed 10% of the two-column total
measurement. If the 10% figure is exceeded, one of three events could have
happened: (1) the first column was overloaded and a legitimate measure of
breakthrough was obtained, in which case taking a smaller sample may be
necessary; (2) channeling or some other failure occurred, in which case the
sample may need to be rerun; or (3) a high random bias occurred, and the result
should be rejected and the sample rerun. Because it may not be possible to
determine which event occurred, a sample analysis should be repeated often enough
to gain confidence in results. As a general rule, any analysis that is rejected
should be repeated whenever a sample is available. In the event that repeated
analyses show that the second column consistently exceeds the 10% figure and the
total is too low for the first column to be saturated and the inorganic Cl is
less than 20,000 times the organic chlorine value, then the result should be
reported, but the data user should be informed of the problem. If the second-
column measurement is equal to or less than the nitrate-wash blank value, the
second-column value should be disregarded.
7.7 Calculations: TOX as Cl" is calculated using the following formula:
(C, - C3) + (C2 - C3)
= /ig/L Total Organic Halide
9020B - 6 Revision 2
November 1992
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where:
C,, = jug Cl" on the first column in series;
C2 = /ig CT on the second column in series;
C3 = predetermined, daily, average, method-blank value
(nitrate-wash blank for a 40-mg carbon column); and
V = the sample volume in liters.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control guidelines.
8.2 This method requires all samples be run in duplicate.
8.3 Employ a minimum of two blanks to establish the repeatability of
the method background, and monitor the background by spacing method blanks
between each group of eight analytical determinations.
8.4 After calibration, verify calibration with an independently
prepared check standard.
8.5 A matrix spike is run in between every 10 samples and is brought
through the entire sample preparation and analytical process.
9.0 METHOD PERFORMANCE
9.1 Under conditions of duplicate analysis, the method detection limit
is 10 jug/L.
9.2 Analyses of distilled water, uncontaminated ground water, and
ground water from RCRA waste management facilities spiked with volatile
chlorinated organics generally gave recoveries between 75-100% over the
concentration range 10-500 /xg/L. Relative standard deviations were generally
20% at concentrations greater than 25 M9/L. These data are shown in Tables 1
and 2.
10.0 REFERENCES
1. Gaskill, A., Compilation and Evaluation of RCRA Method Performance Data,
Work Assignment No. 2, EPA Contract No. 68-01-7075, September 1986.
2. Stevens, A.A., R.C. Dressman, R.K. Sorrell, and H.J. Brass, Organic Halogen
Measurements: Current Uses and Future Prospects, Journal of the American Water
Works Association, pp. 146-154, April 1985.
3. Tate, C., B. Chow, et al., EPA Method Study 32, Method 450.1, Total Organic
Hal ides (TOX), EPA/600/S4-85/080, NTIS: PB 86 136538/AS.
9020B - 7 Revision 2
November 1992
-------
TABLE 1. METHOD PERFORMANCE DATA8
Spiked
Compound
Bromobenzene
Bromodi chl oromethane
Bromoform
Bromoform
Bromoform
Bromoform
Bromoform
Chloroform
Chloroform
Chloroform
Chloroform
Chloroform
Di bromodi chl oromethane
Di bromodi chl oromethane
Tetrachl oroethyl ene
Tetrachl oroethyl ene
Tetrachl oroethyl ene
trans -Di chl oroethy] ene
trans -Di chl oroethyl ene
trans -Di chl oroethyl ene
Matrix6
D.W.
D.W
D.W.
D.W.
G.W.
G.W.
G.W.
D.W.
D.W.
G.W.
G.W.
G.W.
D.W.
D.W.
G.W.
G.W.
G.W.
G.W.
G.W.
G.W.
TOX
Concentration
(M9/L)
443
160
160
238
10
31
100
98
112
10
30
100
155
374
10
30
101
10
30
98
Percent
Recovery
95
98
110
100
140
93
120
89
94
79
76
81
86
73
79
75
78
84
63
60
aResults from Reference 2.
bG.W. = Ground Water.
D.W. = Distilled Water.
9020B - 8
Revision 2
November 1992
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TABLE 2. METHOD PERFORMANCE DATA8
Sample Unspiked Spike Percent
Matrix TOX (M9/L) Level Recovery
Ground Water 68, 69 100 98, 99
Ground Water 5, 12 100 110, 110
Ground Water 5, 10 100 95, 105
Ground Water 54, 37 100 111, 106
Ground Water 17, 15 100 98, 89
Ground Water 11, 21 100 97, 89
Results from Reference 3.
9020B - 9 Revision 2
November 1992
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Sample
Reservoir
(1 of 4)
Nitrate Wash
Reservoir
GAC Column 1
GAC Column 2
Figure 1. Schematic Diagram of Adsorption System
9020B - 10
Revision 2
November 1992
-------
Sparging
Device
Titration
Cell
Pyrolysis
Furnace
Boat
Inlet
Microcoulometer
with Integrator
Strip Chart
Recorder
Adsorption
Module
Figure 2. Flowchart of Analytical System
9020B - 11
Revision 2
November 1992
-------
START
METHOD 9020B
TOTAL ORGANIC HAL IDES (TOX)
711 Take specia 1
car* in handl ing
•ample to minimize
vola tile 1 033
7 1 2 Add sulfite
to reduce residual
chlorine, store at
4 C without
headspace
721 Check
abs o r pti on
efficiency for each
batch of carbon
7 2 2 Analyze
nitrate-wash blanks
to es tabl ish
backgr ound
723 Pyrolyze
dupl icate
ins t rument
cal ibration and
blank standards
each day
731 Connect in
series two col umns
containing
ac ti va ted ca r bon
•>
7 3 2 Fill sample
sample through
activated carbon
col umns
733 Wash columns
with nitra te
s o 1 u 1 1 on
1 4 1 Protect
col umns f r om
742 Pyrolyze
volatile components
in C02-nch
atmosphere at low
temperature
i
742 Pyrolyze less
at high temperature
in 02 -rich
atmosphere
7 4 3 Transfer
contents of each
col umn to quartz
boat for ana lysis
•*
744 Adjust gas
flow
1
745 Position
sample for 2
minutes in 200 C
zone of pyrolyais
tube
1
746 Advance boa t
into 800 C zone
1
7 5 Analyze
effluent gases in
microcoul omet nc -
titration cell
Si 6 Is 2nd\.
-/^ column >v
f measurement >10% }—
>v of 2 col umn /
^^ total1? /
Yes
7 6 Reject and
repeat
7 6
2nd column
measuramcnt
< nitrate wash
blank'
90208 - 12
Revision 2
November 1992
-------
METHOD 9021
PURGEABLE ORGANIC MAUDES (POX)
1.0 SCOPE AND APPLICATION
1.1 Method 9021 determines organically bound halides (chloride, bromide,
and iodide) purged from a sample of drinking water or ground water. They are
reported as chloride. This method is a quick screening procedure requiring
about 10 minutes. The method uses a sparging device, a pyrolysis furnace, and
a microcoulometric-titration detector.
1.2 Method 9021 detects purgeable organically bound chlorine, bromine,
and iodine. Fluorine containing species are not determined by this method.
Method 9021 measures POX concentrations ranging from 5 to 1,000 ug/L.
1.3 This method provides a recommended procedure. It may be used as a
reference for comparing the suitability of other methods thought to be
appropriate for measurement of POX (i.e. by comparison of sensitivity,
accuracy, and precision of data).
1.4 Method 9021 is restricted to use by, or under the supervision of,
analysts experienced in the operation of a pyrolysis/microcoulometer and in
the interpretation of the results.
2.0 SUMMARY OF METHOD
2.1 A sample of water, protected against the loss of volatiles by the
elimination of headspace in the sampling container, is transferred to a
purging vessel. The volatile organic halides are purged into a pyrolysis
furnace using a stream of C02 and the hydrogen halide (HX) pyrolysis product
is trapped and titrated electrolytically using a microcoulometric detector.
3.0 INTERFERENCES
3.1 Contaminants, reagents, glassware, and other sample processing
hardware may cause interferences. Method blanks must be routinely run to
demonstrate freedom from interferences under the conditions of the analysis.
3.1.1 Glassware must be scrupulously clean. Clean all glassware as
soon as possible after use by treating with chromate cleaning solution.
This should be followed by detergent washing in hot water. Rinse with tap
water and ASTM Type II water and dry at 105eC for 1 hour or until dry.
Glassware which is not volumetric should, in addition, be heated in a
muffle furnace at 300°C for 15 to 30 minutes (volumetric ware should not
be heated in a muffle furnace). Glassware should be sealed and stored in a
clean environment after drying and cooling to prevent any accumulation of
dust or other contaminants.
3.1.2 Use high purity reagents and gases to minimize interference
problems.
9021 - 1 Revision 0
December 1987
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3.1.3 Avoid using non-PTFE (polytetrafluoroethylene) plastic tubing,
non-TFE thread sealants, or flow controllers with rubber components in the
purge gas stream.
3.2 Samples can be contaminated by diffusion of volatile organics
(methylene chloride) through the septum seal into the sample during shipment
and especially during storage. A trip blank prepared from water and carried
through the sampling and handling protocol serves as a check on such
contamination. A trip blank should be run with each analytical batch.
3.3 Contamination by carryover occurs whenever high level and low level
samples are sequentially analyzed. To reduce carryover, the purging device and
sample syringe must be rinsed with water between sample analyses. Whenever an
unusually concentrated sample is encountered, it should be followed by an
analysis of water to check for cross contamination. For samples containing
large amounts of water-soluble materials, suspended solids, high boiling
compounds or high organohalide levels, wash out the purging device with a
detergent solution, rinse it with water, and then dry it in a 105°C oven
between analyses.
3.4 All operations should be carried out in an area where halogenated
solvents, such as methylene chloride, are not being used.
3.5 Residual free chlorine interferes in the method. Free chlorine must
be destroyed by adding sodium sulfite when the sample is collected.
4.0 APPARATUS AND MATERIALS
4.1 Sampling equipment (for discrete sampling)
4.1.1 Vial - 25-mL capacity or larger, equipped with a screw-cap
with hole in center (Pierce #13075 or equivalent).
4.1.2 Septum - Teflon lined silicone (Pierce #12722 or equivalent).
Detergent wash, rinse with tap and ASTM Type II water, and dry at 105eC
for 1 hour before use.
4.2 Analytical system
4.2.1 Microcoulometric-titration system containing the following
components (a schematic diagram of the microcoulometric-titration system
is shown in Figure 1).
4.2.1.1 Purging device.
4.2.1.2 Pyrolysis furnace.
4.2.1.3 Titration cell.
4.2.2 Strip chart recorder (optional) - The recorder is recommended
to make sure the peak in down to baselines before stopping integration.
9021 - 2 Revision 0
December 1987
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4.2.3 Microsyringes - 10-uL and 25-uL with 0.006 in i.d. needle
(Hamilton 702N or equivalent).
4.2.4 Syringe valve - 2 way, with Luer ends.
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 ASTM Type II water (ASTM D1193-77 (1983)). All references to water in
the method refer to ASTM Type II unless otherwise specified.
5.3 Sodium sulfide, Na2$. Granular, anhydrous.
5.4 Acetic acid in water (70%), CHsCOOH. Dilute 7 volumes of glacial
acetic acid with 3 volumes of water.
5.5 Sodium chloride calibration standard (1 ug Cl~/uL). Dissolve 1.648 g
NaCl in water and dilute to 1 liter.
5.6 Carbon dioxide.
5.7 Methanol, CHsOH. Store away from other solvents.
5.8 Chloroform, CHCls.
5.9 Chloroform (stock) solution (1 uL = 11.2 ug of CHCla or 10 ug C1-).
Prepare a stock solution by delivering accurately 760 uL (1120 mg) of
chloroform into a 100-mL volumetric flask containing approximately 90 ml of
methanol. Dilute to volume with methanol (10,000 mg of chlorine/L).
5.10 Chloroform (calibration) solution (1 uL = 0.1 ug Cl~). Dilute 1 ml
of the chloroform stock solution to 100 mL with methanol (100 mg of
chlorine/L).
5.11 Chloroform Quality Control (QC) reference sample (100 ug/L). Prepare
an aqueous standard by injecting 100 uL of the chloroform calibration standard
(100 mg of C1-/L) into a volumetric flask containing 100 mL of water. Mix and
store in a bottle with zero headspace. Analyze within two hours after
preparation.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine of this manual.
9021 - 3 Revision 0
December 1987
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6.2 All samples should be collected in bottles with Teflon lined silicone
septa (e.g. Pierce #12722 or equivalent) and be protected from light. If this
is not possible, use amber glass 250-mL bottles fitted with Teflon lined caps.
6.3 All glassware must be cleaned prior to use according to the process
described in Step 3.1.1.
6.4 Special care should be taken in handling the sample in order to
minimize the loss of volatile organohalides. This is accomplished through
elimination of headspace and by minimizing the number of transfers.
6.5 Reduce residual chlorine, if present, by adding sodium sulfite (5 mg
of sodium sulfite crystals per liter of sample). Sodium sulfite should be
added to empty sample bottles at the time of sampling. Shake vigorously for 1
minute after bottle has been filled with sample and properly sealed. Samples
should be stored at 4°C without headspace. POX may increase during storage of
the sample.
6.6 All samples must be analyzed within 14 days of collection.
7.0 PROCEDURE
7.1 Calibration.
7.1.1 Assemble the sparging/pyrolysis/microcoulometric-titration
apparatus shown in Figure 1 in accordance with the manufacturer's
specifications. Typically a C02 flow of 150 mL/min and a sparger
temperature of 45 + 5"C are employed. The pyrolysis furnace should be set
at 800 ± 10°C. Attach the titration cell to the pyrolysis tube outlet and
fill with electrolyte (70% acetic acid). Flow rate and temperature changes
will affect the compounds that are purged and change the percent recovery
of marginal compounds. Therefore, these parameters should not be varied.
Adjust gas flow rate according to manufacturer's directions.
7.1.2 Turn on the instrument and allow the gas flow and temperatures
to stabilize. When the background current of the titration cell has
stabilized the instrument is ready for use.
7.1.3 Calibrate the microcoulometric-titration system for Cl"
equivalents by injecting various amounts (1 to 80 uL) of the sodium
chloride calibration standard directly into the titration cell and
integrating the response using the POX integration mode. If desired, the
analog output of the titration cell can be displayed on a strip chart
recorder. The range of sodium chloride amounts should cover the range of
expected sample concentrations and should always be less than 80 ug of
Cl~. The integrated response should read within 2% or 0.05 ug of the
quantity injected (whichever is larger) over the range 1-80 ug Cl~. If
this calibration requirement is not met, then the instrument sensitivity
parameters should be adjusted according to the manufacturer's
specifications to achieve an accurate response.
7.1.4 Check the performance of the analytical system daily by
analyzing three 5-mL aliquots of a freshly prepared 100 ug/mL chloroform
9021 - 4 Revision 0
December 1987
-------
check standard. The mean of these three analyses should be between
0.4-0.55 ug of Cl~ and the percent relative standard deviation should be
5% or less. If these criteria are not met, the system should be checked as
described in the instrument maintenance manual in order to isolate the
problem. Low chloroform recovery can often be traced to a vitrified inlet
tube. The tube should be replaced weekly.
7.1.5 Determine a reagent blank daily by running an analysis with
the purge vessel empty. The reagent blank should be 0.00 ± 0.05 ug of Cl~.
Analyze a calibration blank sample daily. The calibration blank should be
within 0.02 ug of Cl~ of the reagent blank.
7.2 Sample analysis
7.2.1 Select a chloroform spike concentration representative of the
expected levels in the samples. Using the chloroform stock solution,
prepare a spiking solution in methanol which is 500 times more
concentrated than the selected spike concentration. Add 10 uL of the
spiking solution to 5-mL aliquots of the samples chosen for spiking (refer
to Section 8.0, Quality Control, for guidance in selecting the appropriate
number of samples to be spiked).
7.2.2 Allow sample to come to ambient temperature prior to drawing
it into the syringe. Remove the plunger from a 5-mL or 10-mL syringe and
attach a closed syringe valve. If maximum sensitivity is desired and the
sample does not foam excessively, a 10-mL sample aliquot may be analyzed.
Otherwise 5-mL aliquots should be used. Open the sample bottle (or
standard) 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 mL. Since this process of taking an aliquot destroys
the validity of the sample for future analysis, the analyst should fill a
second syringe at this time to protect against possible loss of data (e.g.
accidental spill), or for duplicate anlaysis.
7.2.3 Attach the syringe valve assembly to the syringe valve on the
purging device. Place the pyrolysis/microcoulometer system in the POX
integration mode to activate the integration system. Immediately open the
syringe valves and inject the sample into the purging chamber.
7.2.4 Close both valves and purge the sample for 10 minutes.
7.2.5 After integration is complete, open the syringe valves and
withdraw the purged sample. Flush the syringe and purging device with
water prior to analyzing other samples.
7.2.6 If the integrated response exceeds the working range of the
instrument, prepare a dilution of the sample from the aliquot in the
second syringe with water and reanalyze. The water must meet the criteria
of Step 7.1.5. It may be necessary to heat and purge dilution waters.
9021 - 5 Revision 0
December 1987
-------
7.3 Pyrolysis procedure
7.3.1 Pyrolysis of the purged organic component of the sample is
accomplished by pyrolyzing in a C02-rich atmosphere at a low temperature
to ensure the conversion of brominated trihalomethanes to a titratable
species.
7.4 Directly analyze the effluent gases in the microcoulometric-titration
cell. Carefully follow instrument manual instructions for optimizing cell
performance.
7.5 Calculations - POX as Cl~ is calculated using the following formula:
-5s. x 1000 = g/L Purgeable Organic Halide
where:
Qs = Quantity of POX as ug of Cl~ in the sample aliquot.
V = Volume of sample aliquot in ml.
8.0 QUALITY CONTROL
8.1 All quality control data should be maintained and available for easy
reference or inspection for 3 years. This method is restricted to use by or
under supervision of experienced analysts. Refer to the appropriate section of
Chapter One for additional quality control requirements.
8.2 Analyze a minimum of one reagent blank every 20 samples or per
analytical batch, whichever is more frequent, to determine if contamination or
any memory effects are occurring.
8.3 In addition to the performance check mentioned in Step 7.1.4, verify
calibration with an independently prepared chloroform QC reference sample
every 15 samples.
8.4 Analyze matrix spiked samples for every 10 samples or analytical
batch, whichever is more frequent. The spiked sample is carried through the
whole sample preparation and analytical process.
8.5 Analyze all samples in replicate.
9.0 METHOD PERFORMANCE
9.1 Under conditions of duplicate analysis, the reliable limit of
detection is 5 ug/L.
9.2 Analyses of distilled water, uncontaminated ground water, and ground
water from RCRA waste management facilities spiked with volatile chlorinated
organics generally give recoveries of 44-128% over the concentration range of
29-4500 ug/L. Relative standard deviations are generally less than 20% at
concentrations greater than 25 ug/L. These data are shown in Tables 1 and 2.
9021 - 6 Revision 0
December 1987
-------
10.0 REFERENCES
1. Takahashi, Y.; Moore, R.T.; Joyce, R.J. "Measurement of Total Organic
Hal ides (TOX) and Purgeable Organic Hal ides (POX) in Water Using Carbon
Adsorption and Microcoulometric Determination"; Proceedings from Division
of Environmental Chemistry, American Chemical Society Meeting, March
23-28, 1980.
2. 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.
3. Fed. Regist. 1979, 45, 69468-69473; December 3.
4. Rohrbough, W.G.; et al. Reagent Chemicals, American Chemical Society
Specifications, 7th ed.; American Chemical Society: Washington, DC, 1986.
5. "Development and Evaluation of Methods for Total Organic Halide and
Purgeable Organic Halide in Wastewater"; U.S. Environmental Protection
Agency. Environmental Monitoring and Support Laboratory. Cincinnati, OH,
1984; EPA-600/4-84-008; NTIS-PB-84-134-337.
6. 1985 Annual Book of ASTM Standards. Vol. 11.01; "Standard Specification
for Reagent Water"; ATSM: Philadelphia, PA, 1985; D1193-77.
7. Dohrmann. Rosemount Analytical Division. Santa Clara, CA 95052-8007.
8. Cosa Instruments. Norwood, NJ 21942.
9021 - 7 Revision 0
December 1987
-------
TABLE 1.
PRECISION AND ACCURACY DATA FOR SELECTED PURGEABLE ORGANIC HALIDES
(Reference 5)
Compound
Chloroform
Trichloroethene
Tetrachloroethene
Chlorobenzene
Dosel
(ug/L as Cl")
11
10
10
8
Average
Percent
Recovery
100
60
50
38
Standard
Deviation
14
11
20
20
MDL2
(ug/L)
4.5
2.2
3.2
2.03
Number of
Replicates
7
7
7
7
1 Ten milliliter aliquot of spiked reagent water analyzed.
2 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.
3 Practical MDL probably greater (approximately 5 to 6 ug/L) due to low
recovery.
9021 - 8
Revision 0
December 1987
-------
TABLE 2.
PRECISION AND ACCURACY DATA FOR VARIOUS WATER SAMPLES
(Reference 5)
Sample^
POTW Sewage
Chlorinated
Hydrocarbon
Plant
Wastewater
Chlorinated
Hydrocarbon
Plant
Wastewater
Chlorinated
Hydrocarbon
Plant
Wastewater
Solid Waste
Leachate
Indus trial^
Wastewater
Spike
Component
Chloroform
Chloroform
Chloroform
Chloroform
Background
Level
(/ig/L as C1-)
88
114
32
171
1,1-Dichloro- 171
ethane
Methylene
chloride
510
Average
Spike Level Percent
(A»g/L as Cl') Recovery
29 128
460 77
1500 50
4500 87
800 44
120 65
Number
Standard of
Deviation Replicates
5 3
7 3
4 3
12 3
2 3
12 3
1 Five milliliter sample aliquots analyzed.
2 Diluted 200:1 prior to analysis. Values for this sample are in mg/L for original
sample.
9021 - 9
Revision 0
December 1987
-------
FIGURE 1.
MICROCOULOMETRIC - TITRATION SYSTEM
T
0
o
o
ir>
CVJ
o
o
o
o
00
0)
X
O
a.
3>'|*
<55Ł
9021 - 10
Revision 0
December 1987
-------
METHOD 9021
PURGEABLE ORGANIC HALIDES (POX)
C Start J
7.1.1 Aaaaabla
apparataa: aat
carbon dloxlda flov
rata: tat ipar|*r
and pyrolyala
furaaca taaparatara
T.I. 2 Tara oa
laatriaaat: allov
gaa flov aad
taaparatiraa to
•tabiliza: allov
background cirraat
of titratloa call
to itabillza
T.I. 3 Calibrata tka
aicrocoaloaatric-
tltratioa ayataa)
for Cl aqaivalaata
/T.l.SN.
/ la tka \Io
/raapoaaa vitalaN.
(n or O.Ot t| of >
\ tka aaaatity /
N. lajactad? /
Ny/Taa
T.I. 4 Aaalyza 3
alioaota of
cklorofora ckack
ataadard
1
T.I. 3 Adjaat
liatraaaat
— » aaaaivity
paraaatara:
racallbrata
/I 1.4 la ^V ••
/ 1 ISO <• 51 \
( aad tk« aaaa ) —
\ 0.4-0.55 if /
\z
T.I. 5 Aaalyza vatar
blaak: Datiraiaa
laatraaaat blaak
7.3.1 Salact
apikla| coac: add
apikini tola to
approprlata aaaplaa
7.3.3 Traaafar
aaapla to ayriata
fill aacoad lyrinfa
733 Attack ayrlnfa
ralia aaaaakly to
parfiif davlca: placa
pyrolyaia/
aicrocoaloaatar
ayatai la POI
iatatratloa aoda
lajact taapla iato
parfiaf ckaabar
734 Pirja for 10
alaataa
T 3.5 fitkdrav
partad aaapla:
fliak ayria|a aad
parfiif dovlca vltk
vatar
1
T.I. 4 Ckack ayatai
aa daacrlbac la
laatraaaat
-» aaiataaaaca
aaatal : raaatlyza
ckack ataadard
t
i
>/T.J.«\
Taa / Ooaa ^v
T.a.t Dilata laapla / latafratad \
froa lacoad ayriaca « ( raiponia axcaad )
vltk vatar \ varklaf /
N. raafa' /
xy'Mo
I
7.3.1 Pyrolyala of
tha aaapla ii
accoapliahad by
pyrolyzinf in a
carbon dioxida rich
ataoaphara at a low
tcaparatnra
T.4 Aaalyza tka
afflaaat faaaa in
tka aicrocoolo-
•atrlc-tltratioa
call
T.5 Calcalata POX
aa Cl-
C •- )
9021 - 11
Revision 0
December 1987
-------
METHOD 9030
ACID-SOLUBLE AND ACID-INSOLUBLE SULFIDES
1.0 SCOPE AND APPLICATION
1.1 The distillation procedure
the determination of sulfides in
effluents.
described in
aqueous and
this method is designed for
solid waste materials and
1.2 This method provides only a semi-quantitative determination of
sulfide compounds considered "acid-insoluble" (e.g. CuS and Sn$2) in solid
samples. Recovery has been shown to be 20 to 40% for CuS, one of the most
stable and insoluble compounds, and 40 to 60% for Sn$2 which is slightly more
soluble.
1.3 This method is not applicable to oil or multiphasic samples or
samples not amenable to the distillation procedure. These samples can be
analyzed by Method 9031.
1.4 Method 9030 is suitable for measuring sulfide concentrations in
samples which contain between 0.2 and 50 mg/kg of sulfide.
1.5 This method is not applicable for reactive sulfide. Refer to Chapter
Seven, Step 7.3.4.1 for the determination of reactive sulfide.
1.6 This method measures total sulfide which is usually defined as the
acid-soluble fraction of a waste. If, however, one has previous knowledge of
the waste and is concerned about both soluble sulfides such as H2S, and metal
sulfides, such as CuS and SnS2, then total sulfide is defined as the
combination of both acid-soluble and acid-insoluble fractions. For wastes
where only metal sulfides are suspected to be present, only the acid-insoluble
fraction needs to be performed.
2.0 SUMMARY OF METHOD
2.1 For acid-soluble sulfide samples, separation of sulfide from the
sample matrix is accomplished by the addition of sulfuric acid to the sample.
The sample is heated to 70°C and the hydrogen sulfide (H2S) which is formed is
distilled under acidic conditions and carried by a nitrogen stream into zinc
acetate gas scrubbing bottles where it is precipitated as zinc sulfide.
2.2 For acid-insoluble sulfide samples, separation of sulfide from the
sample matrix is accomplished by suspending the sample in concentrated
hydrochloric acid by vigorous agitation. Tin(II) chloride is present to
prevent oxidation of sulfide to sulfur by the metal ion (as in copper(II)), by
the matrix, or by dissolved oxygen in the reagents. The prepared sample is
distilled under acidic conditions at 100°C under a stream of nitrogen.
Hydrogen sulfide gas is released from the sample and collected in gas
scrubbing bottles containing zinc(II) and a strong acetate buffer. Zinc
sulfide precipitates.
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2.3 The sulfide in the zinc sulfide precipitate is oxidized to sulfur
with a known excess amount of iodine. Then the excess iodine is determined by
titration with a standard solution of phenyl arsine oxide (PAO) or sodium
thiosulfate until the blue iodine starch complex disappears. As the use of
standard sulfide solutions is not possible because of oxidative degradation,
quantitation is based on the PAO or sodium thiosulfate.
3.0 INTERFERENCES
3.1 Aqueous samples must be taken with a minimum of aeration to avoid
volatilization of sulfide or reaction with oxygen, which oxidizes sulfide to
sulfur compounds that are not detected.
3.2 Reduced sulfur compounds, such as sulfite and hydrosulfite, decompose
in acid, and may form sulfur dioxide. This gas may be carried over to the zinc
acetate gas scrubbing bottles and subsequently react with the iodine solution
yielding false high values. The addition of formaldehyde into the zinc acetate
gas scrubbing bottles removes this interference. Any sulfur dioxide entering
the scrubber will form an addition compound with the formaldehyde which is
unreactive towards the iodine in the acidified mixture. This method shows no
sensitivity to sulfite or hydrosulfite at concentrations up to 10 mg/kg of the
interferent.
3.3 Interferences for acid-insoluble sulfides have not been fully
investigated. However, sodium sulfite and sodium thiosulfate are known to
interfere in the procedure for soluble sulfides. Sulfur also interferes
because it may be reduced to sulfide by tin(II) chloride in this procedure.
3.4 The iodometric method suffers interference from reducing substances
that react with iodine, including thiosulfate, sulfite, and various organic
compounds.
3.5 The insoluble method should not be used for the determination of
soluble sulfides because it can reduce sulfer to sulfide, thus creating a
positive interference.
4.0 APPARATUS AND MATERIALS
4.1 Distillation apparatus as shown in Figure 1
4.1.1 Three neck flask - 500-mL, 24/40 standard taper joints.
4.1.2 Dropping funnel - 100-mL, 24/40 outlet joint.
4.1.3 Purge gas inlet tube - 24/40 joint, with coarse frit.
4.1.4 Purge gas outlet - 24/40 joint reduced to 1/4 in. tube.
4.1.5 Gas scrubbing bottles - 125-mL, with 1/4 in. o.d. inlet and
outlet tubes. Impinger tube must be fritted.
4.1.6 Tubing - 1/4 in. o.d. Teflon or polypropylene. Do not use
rubber.
9030 - 2 Revision 1
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NOTE: When analyzing for acid-insoluble sulfides, the distillation
apparatus is identical to that used in the distillation procedure
for acid-soluble sulfides except that the tubing and unions
downstream of the distillation flask must be all Teflon,
polypropylene or other material resistant to gaseous HC1. The
ground glass joints should be fitted with Teflon sleeves to prevent
seizing and to prevent gas leaks. Pinch clamps should also be used
on the joints to prevent leaks.
4.2 Hot plate stirrer.
4.3 pH meter.
4.4 Nitrogen regulator.
4.5 Flowmeter.
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 ASTM Type II water (ASTM D-1193-77 (1983)). All water used in this
method will be Type II unless otherwise specified.
5.3 Zinc acetate solution for sample preservation (2N), Zn(CH3COO)2-2H20.
Dissolve 220 g of zinc acetate dihydrate in 500 ml of water.
5.4 Sodium hydroxide (IN), NaOH. Dissolve 40 g of NaOH in water and
dilute to 1 liter.
5.5 Formaldehyde (37% solution), CH20. This solution is commercially
available.
5.6 Zinc acetate for the scrubber
5.6.1 For acid-soluble sulfides: Zinc acetate solution
(approximately 0.5M). Dissolve about 110 g zinc acetate dihydrate in
200 ml of water. Add 1 ml hydrochloric acid (concentrated), HC1, to
prevent precipitation of zinc hydroxide. Dilute to 1 liter.
5.6.2 For acid-insoluble sulfides: Zinc acetate/sodium acetate
buffer. Dissolve 100 g sodium acetate, NaC2Hs02, and 11 g zinc acetate
dihydrate in 800 mL of water. Add 1 ml concentrated hydrochloric acid and
dilute to 1 liter. The resulting pH should be 6.8.
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5.7 Acid to acidify the sample
5.7.1 For acid-soluble sulfides: Sulfuric acid (concentrated),
5.7.2 For acid-insoluble sulfides: Hydrochloric acid (9.8N), HC1.
Place 200 mL of water in a 1-liter beaker. Slowly add concentrated HC1 to
bring the total volume to 1 liter.
5.8 Starch solution - Use either an aqueous solution or soluble starch
powder mixtures. Prepare an aqueous solution as follows. Dissolve 2 g soluble
starch and 2 g salicylic acid, CyHsOs, as a preservative, in 100 ml hot water.
5.9 Nitrogen.
5.10 Iodine solution (approximately 0.025N)
5.10.1 Dissolve 25 g potassium iodide, KI, in 700 ml of water in a
1-liter volumetric flask. Add 3.2 g iodine, lŁ. Allow to dissolve. Dilute
to 1 liter and standardize as follows. Dissolve approximately 2 g KI in
150 mL of water. Pipet exactly 20 ml of the iodine solution to be titrated
and dilute to 300 ml with water. Titrate with 0.025N standardized
phenylarsine oxide or 0.025N sodium thiosulfate until the amber color
fades to yellow. Add starch indicator solution. Continue titration drop by
drop until the blue color disappears.
5.10.2 Run in replicate.
5.10.3 Calculate the normality as follows.
Normality (\2) = ml of titrant x normality of titrant
sample size in mL
5.11 Sodium sulfide nonahydrate, Na2$-9H20. For the preparation of
standard solutions to be used for calibration curves. Standards must be
prepared at pH > 9 or < 11. Protect standard from exposure to oxygen by
preparing it without headspace. These standards are unstable and should be
prepared daily.
5.12 Tin(II) chloride, SnCle, granular.
5.13 Titrant.
5.13.1 Standard phenylarsine oxide solution (PAO) (0.025N),
This solution is commercially available.
Caution: PAO is toxic.
5.13.2 Standard sodium thiosulfate solution (0.025N),
Dissolve 6.205 ± 0.005 g Na2S203«5H20 in 500 mL water. Add 9 mL IN NaOH
and dilute to 1 liter.
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5.14 Sodium hydroxide (6N), NaOH. Dissolve 240 g of sodium hydroxide in
1 liter of water.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 All aqueous samples and effluents must be preserved with zinc acetate
and sodium hydroxide. Use four drops of 2N zinc acetate solution per 100 mL of
sample. Adjust the pH to greater than 9 with 6N sodium hydroxide solution.
Fill the sample bottle completely and stopper with a minimum of aeration. The
treated sample is relatively stable and can be held for up to seven days. If
high concentrations of sulfide are expected to be in the sample, continue
adding zinc acetate until all the sulfide has precipitated. For solid samples,
fill the surface of the solid with 2N zinc acetate until moistened. Samples
must be cooled to 4°C and stored headspace free.
6.3 Sample Preparation
6.3.1 For an efficient distillation, the mixture in the distillation
flask must be of such a consistency that the motion of the stirring bar is
sufficient to keep the solids from settling. The mixture must be free of
solid objects that could disrupt the stirring bar. Prepare the sample
using one of the procedures in this section then proceed with the
distillation step (Section 7.0).
6.3.2 If the sample is aqueous, shake the sample container to
suspend any solids, then quickly decant the appropriate volume (up to
250 mL) of the sample to a graduated cylinder, weigh the cylinder,
transfer to the distillation flask and reweigh the cylinder to the nearest
milligram. Be sure that a representative aliquot is used, or use the
entire sample.
6.3.3 If the sample is aqueous but contains soft clumps of solid, it
may be possible to break the clumps and homogenize the sample by placing
the sample container on a jar mill and tumble or roll the sample for a few
hours. The slurry may then be aliquotted and weighed as above to the
nearest milligram then diluted with water up to a total volume of 250 mL
to produce a mixture that is completely suspended by the stirring bar.
6.3.4 If the sample is primarily aqueous, but contains a large
proportion of solid, the sample may be roughly separated by phase and the
amount of each phase measured and weighed to the nearest milligram
into the distillation flask in proportion to their abundance in the
sample. Water may be added up to a total volume of 250 mL. As a guideline,
no more than 25 g dry weight or 50 g of sludge can be adequately suspended
in the apparatus.
6.3.5 If the sample contains solid objects that can not be reduced
in size by tumbling, the solids must be broken manually. Clay-like solids
should be cut with a spatula or scalpel in a crystallizing dish. If the
9030 - 5 Revision 1
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solids can be reduced to a size that they can be suspended by the stirring
bar, the solid and liquid can be proportionately weighed.
6.3.6 Non-porous harder objects, for example stones or pieces of
metal, may be weighed and discarded. The percent weight of non-porous
objects should be reported and should be used in the calculation of
sulfide concentration if it has a significant effect on the reported
result.
7.0 PROCEDURE
For acid-soluble sulfide samples, go to 7.1
For acid-insoluble sulfide samples, go to 7.2
7.1 Acid-Soluble Sulfide
7.1.1 In a preliminary experiment, determine the approximate amount
of sulfuric acid required to adjust a measured amount of the sample to pH
less than or equal to 1. The sample size should be chosen so that it
contains between 0.2 and 50 mg of sulfide. Place a known amount of sample
or sample slurry in a beaker. Add water until the total volume is 200 ml.
Stir the mixture and determine the pH. Slowly add sulfuric acid until the
pH is less than or equal to 1.
CAUTION: Toxic hydrogen sulfide may be generated from the acidified
sample. This operation must be performed in the hood and the
sample left in the hood until the sample has been made alkaline
or the sulfide has been destroyed. From the amount of sulfuric
acid required to acidify the sample and the mass or volume of the
sample acidified, calculate the amount of acid required to
acidify the sample to be placed in the distillation flask.
7.1.2 Prepare the gas evolution apparatus as shown in Figure 1 in a
fume hood.
7.1.2.1 Prepare a hot water bath at 70°C by filling a
crystallizing dish or other suitable container with water and place
it on a hot plate stirrer. Place a thermometer in the bath and
monitor the temperature to maintain the bath at 70'C.
7.1.2.2 Assemble the three neck 500-mL flask, fritted gas inlet
tube, and exhaust tube. Use Teflon sleeves to seal the ground glass
joints. Place a Teflon coated stirring bar into the flask.
7.1.2.3 Place into each gas scrubbing bottle 10 + 0.5 ml of the
0.5M zinc acetate solution, 5.0 +0.1 ml of 37% formaldehyde and 100
± 5.0 ml water.
7.1.2.4 Connect the gas evolution flask and gas scrubbing
bottles as shown in Figure 1. Secure all fittings and joints.
9030 - 6 Revision 1
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7.1.3 Carefully place an accurately weighed sample which contains
0.2 to 50 mg of sulfide into the flask. If necessary, dilute to
approximately 200 ml with water.
7.1.4 Place the dropping funnel onto the flask making sure its
stopcock is closed. Add the volume of sulfuric acid calculated in Step
7.1.1 plus an additional 50 ml into the dropping funnel. The bottom
stopcock must be closed.
7.1.5 Attach the nitrogen inlet to the top of the dropping funnel
gas shut-off valve. Turn on the nitrogen purge gas and adjust the flow
through the sample flask to 25 mL/min. The nitrogen in the gas scrubbing
bottles should bubble at about five bubbles per second. Nitrogen pressure
should be limited to approximately 10 psi to prevent excess stress on the
glass system and fittings. Verify that there are no leaks in the system.
Open the nitrogen shut-off valve leading to the dropping funnel. Observe
that the gas flow into the sample vessel will stop for a short period
while the pressure throughout the system equalizes. If the gas flow
through the sample flask does not return within a minute, check for leaks
around the dropping funnel. Once flow has stabilized, turn on magnetic
stirrer. Purge system for 15 minutes with nitrogen to remove oxygen.
7.1.6 Heat sample to 70°C. Open dropping funnel to a position that
will allow a flow of sulfuric acid of approximately 5 mL/min. Monitor the
system until most of the sulfuric acid within the dropping funnel has
entered the sample flask. Solids which absorb water and swell will
restrict fluid motion and, therefore, lower recovery will be obtained.
Such samples should be limited to 25 g dry weight.
7.1.7 Purge, stir, and maintain a temperature of 70°C for a total of
90 minutes from start to finish. Shut off nitrogen supply. Turn off heat.
7.1.8 Proceed to Step 7.3 for the analysis of the zinc sulfide by
titration.
7.2 Acid-Insoluble Sulfide
7.2.1 As the concentration of HC1 during distillation must be within
a narrow range for successful distillation of H2S, the water content must
be controlled. It is imperative that the final concentration of HC1 in the
distillation flask be about 6.5N and that the sample is mostly suspended
in the fluid by the action of the stirring bar. This is achieved by adding
50 mL of water, including water in the sample, 100 mL of 9.8N HC1, and the
sample to the distillation flask. Solids which absorb water and swell will
restrict fluid motion and, therefore, lower recovery will be obtained.
Such samples should be limited to 25 g dry weight.
7.2.2 If the matrix is a dry solid, weigh a portion of the sample
such that it contains 0.2 to 50 mg of sulfide. The solid should be crushed
to reduce particle size to 1 mm or less. Add 50 ml of water.
9030 - 7 Revision 1
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7.2.3 If the matrix is aqueous, then a maximum of 50 g of the sample
may be used. No additional water may be added. As none of the target
compounds are volatile, drying the sample may be preferable to enhance the
sensitivity by concentrating the sample. If less than 50 g of the sample
is required to achieve the 0.2 to 50 mg of sulfide range for the test,
then add water to a total volume of 50 ml,
7.2.4 If the matrix is a moist solid, the water content of the
sample must be determined (Karl Fischer titration, loss on drying, or
other suitable means) and the water in the sample included in the total
50 ml of water needed for the correct HC1 concentration. For example, if a
20 g sample weight is needed to achieve the desired sulfide level of
0.2 to 50 mg and the sample is 50% water then 40 ml rather than 50 ml of
water is added along with the sample and 100 ml of 9.8N HC1 to the
distillation flask.
7.2.5 Weigh the sample and 5 g SnCl2 into the distillation flask.
Use up to 50 ml of water, as calculated above, to rinse any glassware.
7.2.6 Assemble the distillation apparatus as in Figure 1. Place 100
+ 2.0 ml of zinc acetate/sodium acetate buffer solution and 5.0 + 0.1 ml
of 37% formaldehyde in each gas scrubbing bottle. Tighten the pinch clamps
on the distillation flask joints.
7.2.7 Make sure the stopcock is closed and then add 100 ± 1.0 ml of
9.8N HC1 to the dropping funnel. Connect the nitrogen line to the top of
the funnel and turn the nitrogen on to pressurize the dropping funnel
headspace.
7.2.8 Set the nitrogen flow at 25 mL/min. The nitrogen in the gas
scrubbing bottles should bubble at about five bubbles per second. Purge
the oxygen from the system for about 15 minutes.
7.2.9 Turn on the magnetic stirrer. Set the stirring bar to spin as
fast as possible. The fluid should form a vortex. If not, the distillation
will exhibit poor recovery. Add all of the HCL from the dropping funnel to
the flask.
7.2.10 Heat the water bath to the boiling point (100°C). The sample
may or may not be boiling. Allow the purged distillation to proceed for
90 minutes at 100'C. Shut off nitrogen supply. Turn off heat.
7.2.11 Proceed to Step 7.3 for the analysis of the zinc sulfide by
titration.
7.3 Titration of Distillate
7.3.1 Pipet a known amount of standardized 0.025N iodine solution
(See Step 5.10.3) in a 500-mL flask, adding an amount in excess of that
needed to oxidize the sulfide. Add enough water to bring the volume to 100
ml. The volume of standardized iodine solution should be about 65 ml for
samples with 50 mg of sulfide.
9030 - 8 Revision 1
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7.3.2 If the distillation for acid-soluble sulfide is being used,
add 2 mL of 6N HC1. If the distillation for acid-insoluble sulfides is
performed, 10 ml of 6N HC1 should be added to the iodine.
7.3.3 Pipet both of the gas scrubbing bottle solutions to the flask,
keeping the end of the pipet below the surface of the iodine solution. If
at any point in transferring the zinc acetate solution or rinsing the
bottles, the amber color of the iodine disappears or fades to yellow, more
0.025N iodine must be added. This additional amount must be added to the
amount from Step 7.3.1 for calculations. Record the total volume of
standardized 0.025N iodine solution used.
7.3.4 Prepare a rinse solution of a known amount of standardized
0.025N iodine solution, 1 mL of 6N HC1, and water to rinse the remaining
white precipitate (zinc sulfide) from the gas scrubbing bottles into the
flask. There should be no visible traces of precipitate after rinsing.
7.3.5 Rinse any remaining traces of iodine from the gas scrubbing
bottles with water, and transfer the rinses to the flask.
7.3.6 Titrate the solution in the flask with standard 0.025N
phenylarsine oxide or 0.025N sodium thiosulfate solution until the amber
color fades to yellow. Add enough starch indicator for the solution to
turn dark blue and titrate until the blue disappears. Record the volume of
titrant used.
(ml of I2 x N of I2 )-(mL of titrant x N of titrant)x 16.03
8.0
sample weight (kg)
QUALITY CONTROL
= sulfide(mg/kg)
8.1 All quality control data must be maintained and available for
reference or inspection for a period of three years. This method is restricted
to use by or under supervision of experienced analysts. Refer to the
appropriate section of Chapeter One for additional quality control
requirements.
8.2 A reagent blank should be run once in twenty analyses or per
analytical batch, whichever is more frequent.
8.3 Check standards are prepared from water and a known amount of sodium
sulfide. A check standard should be run with each analytical batch of
samples, or once in twenty samples. Acceptable recovery will depend on the
level and matrix.
8.4 A matrix spiked sample should be run for each analytical batch or
every 20 samples, whichever is more frequent, to determine matrix effects. If
recovery is low, acid-insoluble sulfides are indicated. A matrix spiked
sample is a sample brought through the whole sample preparation and analytical
process.
9030 - 9
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9.0 METHOD PERFORMANCE
9.1 Accuracy - Accuracy for this method was determined by three
independent laboratories by measuring percent recoveries of spikes for both
clean matrices (water) and actual waste samples. The results are summarized
below.
For Acid-Soluble Sulfide
Accuracy of titration step only
Lab A 84-100% recovery
Lab B 110-122% recovery
Accuracy for entire method for clean matrices (H20)
Lab C 94-106% recovery
Accuracy of entire method for actual waste samples
Lab C 77-92% recovery
Spiking levels ranged from 0.4 to 8 mg/L
For Acid-Insoluble Sulfide
The percent recovery was not as thoroughly studied for acid-insoluble sulfide
as it was for acid-soluble sulfide.
Accuracy of entire method for synthetic waste samples
Lab C 21-81% recovery
Spiking levels ranged from 2.2 to 22 mg/kg
9.2 Precision - For Acid-Soluble Sulfide
Precision of titration step only
Lab A CV% 2.0 to 37
Lab B CV% 1.1 to 3.8
Precision of entire method for clean matrices (H20)
Lab C CV% 3.0 to 12
Precision of entire method for actual waste samples
Lab C CV% 0.86 to 45
For Acid-Insoluble Sulfide
Precision of entire method with synthetic wastes
Lab C CV 1.2 to 42
9.3 Detection Limit - The detection limit was determined by analyzing
seven replicates at 0.45 and 4.5 mg/L. The detection limit was calculated as
the standard deviation times the student's t-value for a one-tailed test with
n-1 degrees of freedom at 99% confidence level. The detection limit for a
clean matrix (HŁ0) was found to be between 0.2 and 0.4 mg/L.
9030 - 10 Revision 1
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10.0 REFERENCES
1. Test Methods for Evaluating Solid Waste, Physical/Chemical Methods, 2nd
ed.; U.S. Environmental Protection Agency. Office of Solid Waste and
Emergency Response. U.S. Government Printing Office: Washington, DC,
1982, revised 1984; SW-846.
2. 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.
3. CRC Handbook of Chemistry and Physics, 66th ed.; Weast, R., Ed.; CRC: Boca
Raton, FL, 1985.
4. Standard Methods for the Examination of Water and Wastewater, 16th ed.;
Greenberg, A.E.; Trussell, R.R.; Clesceri, L.S., Eds.; American Water
Works Association, Water Pollution Control Federation, American Public
Health Association: Washington, DC, 1985; Methods 427, 427A, 427B, and
427D.
5. Andreae, M.O.; Banard, W.R. Anal. Chem. 1983, 55, 608-612.
6. Barclay, H. Adv. Instrum. 1980, 35(2). 59-61.
7. Bateson, S.W.; Moody, G.J.; Thomas, J.P.R. Analyst 1986, 111, 3-9.
8. Berthage, P.O. Anal. Chim. Acta 1954, 10 310-311.
9. Craig, P.J.; Moreton, P.A. Environ. Techno!. Lett. 1982, 3, 511-520.
10. Franklin, G.O.; Fitchett, A.W. Pulp & Paper Canada 1982, 83(10), 40-44.
11. Fuller, W. In Cyanide in the Environment; Van Zyl, D., Ed.; Proceedings
of Symposium; December, 1984.
12. Gottfried, G.J. "Precision, Accuracy, and MDL Statements for EPA Methods
9010, 9030, 9060, 7520, 7521, 7550, 7551, 7910, and 7911"; final report
to the U.S. Environmental Protection Agency (EMSL-CI); Biopheric.
13. Kilroy, W.P. Talanta 1983, 30(6). 419-422.
14. Kurtenacher, V.A.; Wallak, R. l± Anorg. \L_ Allq. Chem. 1927, 161 202-209.
15. Landers, D.H.; David, M.B.; Mitchell, M.J. Int. J. Anal. Chem 1983, 14,
245-256.
16. Opekar, F.; Brukenstein, S. Anal. Chem. 1984, 56, 1206-1209.
17. Ricklin, R.D.; Johnson, E.L. Anal. Chem. 1983, 55, 4.
9030 - 11 Revision 1
December 1987
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18. Rohrbough, W.G.; et al. Reagent Chemicals. American Chemical Society
Specifications. 7th ed.; American Chemical Society: Washington, DC,
1986.
19. Snedecor, G.W.; Ghran, W.G. Statistical Methods; Iowa State University:
Ames, IA, 1980.
20. Umana, M.; Beach, J.; Sheldon, L. "Revisions to Method 9010"; final
report to the U.S. Environmental Protection Agency on Contract No.
68-01-7266; Research Triangle Institute: Research Triangle Park, NC,
1986; Work Assignment No. 1.
21. Umana, M.; Sheldon, L. "Interim Report: Literature Review"; interim
report to the U.S. Environmental Protection Agency in Contract No.
68-01-7266; Research Triangle Institute: Research Triangle Park, NC,
1986; Work Assignment No. 3.
22. Wang, W.; Barcelona, M.J. Environ. Inter. 1983, 9, 129-133.
23. Wronski, M. Talanta 1981, 28, 173-176.
24. Application Note 156; Princeton Applied Research Corp.: Princeton, NO.
25. Guidelines for Assessing and Reporting Data Quality for Environmental
Measurements; U.S. Environmental Protection Agency. Office of Research
and Development. U.S. Government Printing Office: Washington, DC,
1983.
26. Fed. Regist. 1980, 45(98). 33122.
27. The Analytical Chemistry of Sulfur and Its Compounds. Part I; Karchmer,
J.H., Ed.; Wiley-Interscience: New York, 1970.
28. Methods for the Examination of Water and Associated Materials;
Department of the Environment: England, 1983.
29. "Development and Evaluation of a Test Procedure for Reactivity Criteria
for Hazardous Waste"; final report to the U.S. Environmental Protection
Agency on Contract 68-03-2961; EAL: Richmond, CA.
30. Test Method to Determine Hydrogen Sulfide Released from Wastes; U.S.
Environmental Protection Agency. Office of Solid Waste. Preliminary
unpublished protocol, 1985.
31. 1985 Annual Book of ASTM Standards. Vol. 11.01; "Standard Specification
for Reagent Water"; ASTM: Philadelphia, PA, 1985; D1193-77.
9030 - 12 Revision 1
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FIGURE 1.
GAS EVOLUTION APPARATUS
H2SO4 (HCI for acid inaoluble sulfldet)
N2 Out
Hot Water Bath
with Magnetic Stlrrer
Zinc Acetate
and
Formaldehyde
Scrubbing
•ettfee
Stirring Be/
9030 - 13
Revision 1
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METHOD 9030
ACID-SOLUBLE AND ACID-INSOLUBLE SULFIDES
T 1.1 Ckooaa aaaplo
•i*o(0 3 to SO •(
•tlfldo) placa kaova
aaotat of laapla li
baakar:add
vati
•volttioa apparatu
T.J.I Saapl. aiz*
•ay bo aE-SO|
T 1.3.1 Prtpart lot
wator batk
7.1.3 lolfk
•aiplo(0 3-SO«i of
aalfldo) eriik if
aacttaary :add SO aL
of wator
T.I.3 3 Aiaoablo 3
nock flaik
T.3 4 Dotoralio
•atar caitaat of
•aaplo.laclad* la
total vatar aoodad
(COiL) for correct
IC1 coac
T.3.S Add vator to
aaiplo for a total
volaao of SO aL
9030 - 14
Revision 1
December 1987
-------
METHOD 9030
(continued)
T 1.2 3 Plac« lite
ac«tata tola,
foruld«iyda, aid
v«t«r i> |aa
•cribkiif bottlaa
T 2.3
E0( of
7.1 2 4 Coaaact
flaak aad acmbbiaf
bottlaaratear*
joiata
7.2 E Pl.c* Hiplt
ii fluk.tdd
itiMOM cklorldi
7 1 3 Plic* w«i|h«d
•«ipl> ia
fl««k:dilnt« vitk
uat«r if i«e«t*ary
7.2.6 Ailtiblt
di«tlll»tio«
tpptrttii
zlic >c«tdd
iglfaric «eid (fro*
St.p 7.1.1} to
droppia| fiaul
7 2.7 Add lOOiL of
t.tl HC1 to
droppiaf faaatl
7.1.E Adjut
iltrofti flo*:ck*ck
(or l«*ki.t*Ti o*
•tlrr>r:p«rf«
•ysttB of oxy|«a
for 16 ilnntta
7.2 * S«t aitrofia
flov par(* ayatoi
of oxyioa for IE
•ia«t«a
9030 - 15
Revision 1
December 1987
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METHOD 9030
(continued)
7 i e Haat to
TOdifC Add ailfirlc
tcid to flaak.cloa*
droppia| faaad
v»«a toil of tka
•eld kaa tatarad
th. flaak
T.a » lira 01
•tlrrtr add KC1 to
dlatlllatioa flaak
7 1 T Ptr|«, itir.
aad kaat for 90
•iaotai'shmt off
nltro|<> ttri off
k««t
733 Plp.t
•cribolif bottlt
loll to flttk
T J 10 H«»t vttir
k«tk to toil, allow
dlitlllitioa to
proctod for 90
• i»t>« at
100d<|C tori off
k»t
7 1 S Analyst by
titratloa (St«p
T 3 1)
7.3 1 Plptt kaova
aionat of 0.0361
iodia* tola ia
flaik:brla( to
voloat with vatir
7 3 11 Aaalyz* by
titratloa (SUp
7 3.1)
7.3 1 licord total
rolaM of 0 02SI
iodiaa aola aatd
7 3 3 Add lOaL of
ai HCI
7 3 2 Add 3>L of «»
HCI
7 3.4 Prtpar* ri»««
aola of 0.02BI
lodiaa tola, «l
XC1. aad vatar
736 liaaf tracoi
of iodiao fro*
•cr«bbla|
bottlaa traaaftr
riaaaa to flaik
9030 - 16
Revision 1
December 1987
-------
METHOD 9030
(continued)
7 3.6 Tltrttt toll
vltk PAO or iodlii
thlonlf«t« toll
latil »ib«r color
f»dt§:«dd itarfk
i«dic»tor:titr«t«
Mtll bl»« color
ditapp«tri.rocord
roliM of titrtst
7 3 7 C«lc»l«t« tk*
cote of iilfi.4* li
tk* iupl*
Stop
9030 - 17
Revision 1
December 1987
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METHOD 9031
EXTRACTABLE SULFIDES
1.0 SCOPE AND APPLICATION
1.1 The extraction procedure described in this method is designed for the
extraction of sulfides from matrices that are not directly amenable to the
distillation procedure Method 9030. This method is also not applicable for
reactive sulfide. Refer to Chapter Seven, Step 7.3.4.1 for the determination
of reactive sulfide. This method is applicable to oil, solid, multiphasic, and
all other matrices not amenable to analysis by Method 9030.
1.2 Method 9031 is suitable for measuring sulfide in solid samples at
concentrations above 1 mg/kg.
2.0 SUMMARY OF METHOD
2.1 If the sample contains solids that will interfere with agitation and
homogenization of the sample mixture, or so much oil or grease as to interfere
with the formation of a homogeneous emulsion in the distillation procedure,
the sample must be filtered and the solids extracted with water at pH > 9 or
< 11. The extract is then combined with the filtrate and analyzed by the
distillation procedure. Separation of sulfide from the sample matrix is
accomplished by the addition of sulfuric acid to the sample. The sample is
heated to 70°C and the hydrogen sulfide (H2S) which is formed is distilled
under acidic conditions and carried by a nitrogen stream into zinc acetate gas
scrubbing bottles where it is precipitated as zinc sulfide.
2.2 The sulfide in the zinc sulfide precipitate is oxidized to sulfur
with a known amount of excess iodine. Then the excess iodine is determined by
titration with a standard solution of phenylarsine oxide (PAO) or sodium
thiosulfate until the blue iodine starch complex disappears. The use of
standard sulfide solutions is not possible because of their instability to
oxidative degradation. Therefore quantitation is based on the PAO or sodium
thiosulfate.
3.0 INTERFERENCES
3.1 Samples with aqueous layers must be taken with a minimum of aeration
to avoid volatilization of sulfide or reaction with oxygen which oxidizes
sulfide to sulfur compounds that are not detected.
3.2 Sulfur compounds such as sulfite and hydrosulfite decompose in acid
and may form sulfur dioxide. This gas may be carried over to the zinc acetate
gas scrubbing bottles and subsequently react with the iodine solution yielding
false high values. The addition of formaldehyde into the zinc acetate gas
scrubbing bottles removes this interference. Any sulfur dioxide entering the
scrubber will form an addition compound with the formaldehyde which is
unreactive towards the iodine in the acidified mixture. This method shows no
sensitivity to sulfite or hydrosulfite at concentrations up to 10 mg/kg of the
interferent.
9031 - 1 Revision 0
December 1987
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3.3 The iodometric method suffers interference from reducing substances
that react with iodine including thiosulfate, sulfite, and various organic
compounds.
3.4 Interferences have been observed when analyzing samples with high
metal content such as electroplating waste and chromium containing tannery
waste.
4.0 APPARATUS AND MATERIALS
4.1 Extractor - Any suitable device that sufficiently agitates a sealed
container of one liter volume or greater. For the purpose of this analysis,
agitation is sufficient when all sample surfaces are continuously brought into
contact with extraction fluid, and the agitation prevents stratification of
the sample and fluid. Examples of suitable extractors are shown in Figures 2
and 3. The tumble-extractors turn the extraction bottles end-over-end at a
rate of about 30 rpm. The apparatus in Figure 2 may be easily fabricated from
plywood. The jar compartments must be padded with polyurethane foam to absorb
shock. The drive apparatus is a Norton jar mill.
4.2 Buchner funnel apparatus
4.2.1 Buchner funnel - 500-mL capacity, with 1-liter vacuum
filtration flask.
4.2.2 Glass wool - Suitable for filtering, 0.8 urn diameter such as
Corning Pyrex 3950.
4.2.3 Vacuum source - Preferably a water driven aspirator. A valve
or stopcock to release vacuum is required.
4.3 Distillation apparatus as shown in Figure 1
4.3.1 Three neck flask - 500-mL, 24/40 standard tapered joints.
4.3.2 Dropping funnel - 100-mL, 24/40 outlet joint.
4.3.3 Purge gas inlet tube - 24/40 joint with course frit.
4.3.4 Purge gas outlet - 24/40 joint reduced to 1/4 inch tube.
4.3.5 Gas scrubbing bottles - 125-mL, with 1/4 in. o.d. inlet and
outlet tubes. Impinger tube must not be fritted.
4.3.6 Tubing - 1/4 in. o.d. Teflon or polypropylene. Do not use
rubber.
4.4 Hot plate stirrer.
4.5 pH meter.
4.6 Nitrogen regulator.
9031 - 2 Revision 0
December 1987
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4.7 Flowmeter.
4.8 Separatory funnels - 500-mL.
4.9 Tumbler - See Figures 2 and 3.
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 ASTM Type II Water (ASTM D1193-77 (1983)). All references to water in
the method refer to ASTM Type II unless otherwise specified.
5.3 Zinc Acetate (for sample preservation) (2N), Zn(CH3COO)2'2H20.
Dissolve 220 g of zinc acetate dihydrate in 500 ml of water.
5.4 Sodium Hydroxide (50% w/v in water), NaOH. Commercially available.
5.5 Tin (II) Chloride, SnCl2-2H20, granular.
5.6 n-Hexane, C6Hi4.
5.7 Nitrogen, N2-
5.8 Sulfuric Acid (concentrated), H2S04.
5.9 Zinc Acetate for the scrubber (approximately 0.5M). Dissolve 110 g
zinc acetate dihydrate in 200 ml of water. Add 1 ml concentrated hydrochloric
acid, HC1, to prevent precipitation of zinc hydroxide. Dilute to 1 liter.
5.10 Formaldehyde (37% solution), CH20. Commercially available.
5.11 Starch solution. Use either an aqueous solution or soluble starch
powder mixtures. Prepare an aqueous solution as follows. Dissolve 2 g soluble
starch and 2 g salicylic acid, C/HsOs, as a preservative, in 100 ml hot water.
5.12 Iodine solution (approximately 0.025N). Dissolve 25 g of potassium
iodide, KI, in 700 mL of water in a 1-liter volumetric flask. Add 3.2 g of
iodine, 12. Allow to dissolve. Dilute to 1 liter and standardize as follows.
Dissolve approximately 2 g KI in 150 ml of water. Pipet exactly 20 ml of the
iodine solution to be titrated and dilute to 300 ml with water. Titrate with
0.025N standard phenylarsine oxide, or 0.025N sodium thiosulfate, N32S203,
until the amber color fades. Add starch indicator solution until the solution
turns deep blue. Continue titration drop by drop until the blue color
disappears. Run in replicate. Calculate the normality as follows:
9031 - 3 Revision 0
December 1987
-------
Normality (12) = ml of titrant x normality of titrant
Volume of sample (ml)
5.13 Sodium sulfide nonahydrate N32S-9H20, for the preparation of
standard solutions to be used for calibration curves. Standards must be
prepared at pH > 9 or < 11.
5.14 Titrant.
5.14.1 Standard phenylarsine oxide (PAO) solution (0.025N), CeHsAsO.
This solution is commercially available.
CAUTION: PAO is toxic.
5.14.2 Standard sodium thiosulfate solution (0.025N), N32S203-5H20.
Dissolve 6.205 ± 0.005 g Na2S20s-5H20 in 500 mL of water. Add 9 ml IN NaOH
and dilute to 1 liter.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 All samples must be preserved with zinc acetate and sodium hydroxide.
Use four drops of 2N zinc acetate solution per 100 mL of aqueous or
multiphasic sample. Adjust the pH to greater than 9.0 with 50% NaOH. Fill the
sample bottle completely and stopper with a minimum of aeration. For solid
samples, fill the surface of solid with 2N zinc acetate until moistened.
Samples must be cooled to 4°C during storage.
7.0 PROCEDURE
7.1 Assemble the Buchner funnel apparatus. Unroll the glass wool and fold
the fiber over itself several times to make a pad about 1 cm thick when
lightly compressed. Cut the pad to fit the Buchner funnel. Dry and weigh the
pad, then place it in the funnel. Turn on the aspirator and wet the pad with a
known amount of water.
7.2 Transfer a sample that contains between 1 and 50 mg of sulfide to the
Buchner funnel. Rinse the sample container with known amounts of water and add
the rinses to the Buchner funnel. When no free water remains in the funnel,
slowly open the stopcock to allow air to enter the vacuum flask. A small
amount of sediment may have passed through the glass fiber pad. This will not
interfere with the analysis.
7.3 Transfer the solid and the glass fiber pad to a dried tared weighing
dish. Since most greases and oils will not pass through the fiber pad, solids,
oils, and greases will be extracted together. If the filtrate includes an oil
phase, transfer the filtrate to a separatory funnel. Collect and measure the
volume of the aqueous phase. Transfer the oil phase to the weighing dish with
the solid and glass fiber pad.
9031 - 4 Revision 0
December 1987
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7.4 Weigh the dish containing solid, oil (if any), and glass fiber pad.
Subtract the weight of the dry glass fiber pad. Calculate the volume of water
present in the original sample by subtracting the total volume of rinses from
the measured volume of the filtrate.
7.5 Place the following in a 1-liter wide-mouth bottle:
500 ml water
5 ml 50% w/v NaOH
1 g SnCl2-2H20
50 ml n-hexane (if an oil or grease is present).
Cap the bottle with a Teflon or polyethylene lined cap and shake vigorously to
saturate the solution with stannous chloride. Direct a stream of nitrogen gas
at about 10 mL/min into the bottle for about 1 minute to purge the headspace
of oxygen. If the weight of the solids (Step 7.4) is greater than 25 g, weigh
out a representative aliquot of 25 g and add it to the bottle while still
purging with nitrogen. Otherwise, add all of the solids. Cap the bottle; avoid
the influx of air.
7.6 The pH of the extract must be maintained at > 9 or < 11 throughout
the extraction step and subsequent filtration. Since some samples may release
acid, the pH must be monitored as follows. Shake the extraction bottle and
wait 1 minute. Open the bottle under a stream of nitrogen and check the pH. If
the pH is below 9, add 50% NaOH in 5 ml increments until it is at least 9.
Recap the bottle, and repeat the procedure until the pH does not drop. The
bottle must be thoroughly purged of oxygen before each recapping. Oxygen will
oxidize sulfide to elemental sulfur or other sulfur containing compounds that
will not be detected.
7.7 Place the bottle in the tumbler, making sure there is enough foam
insulation to cushion the bottle. Turn the tumbler on and allow the extraction
to run for about 18 hours.
7.8 Prepare a Buchner funnel apparatus as in Step 7.1 with a glass fiber
pad filter.
7.9 Decant the extract to the Buchner funnel.
7.10 If the extract contains an oil phase, separate the aqueous phase
using a separatory funnel. Neither the separation nor the filtration are
critical, but are necessary to be able to measure the volume of the aqueous
extract analyzed. Small amounts of suspended solids and oil emulsions will not
interfere with the extraction.
7.11 At this point, an aliquot of the filtrate of the original sample may
be combined with an aliquot of the extract in a proportion representative of
the sample. Calculate the proportions as follows:
9031 - 5 Revision 0
December 1987
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Aliquot of the Filtrate(mL) = Solid Extractedfq)a x Total Sample FiltratefmDC
Aliquot of the Extract(mL) Total Solid(g)b Total Extraction Fluid(ml_)d
aFrom Step 7.5. Weight of solid sample taken from extraction.
DFrom Step 7.4. Weight of solids and oil phase with the dry weight of
filter and tared dish subtracted.
clncludes volume of all rinses added to the filtrate (Steps 7.1 and 7.2).
d500 mL water plus total volume of NaOH solution. Does not include hexane,
which is subsequently removed (Steps 7.5 and 7.6).
Alternatively, the samples may be analyzed separately, concentrations for each
phase reported separately, and the amounts of each phase present in the sample
reported separately.
7.12 Distillation of Sulfide
7.12.1 In a preliminary experiment, determine the approximate amount
of sulfuric acid required to adjust a measured amount of the sample to pH
less than or equal to 1. The sample size should be chosen so that it
contains between 0.2 and 50 mg of sulfide. Place a known amount of sample
or sample slurry in a beaker. Add water until the total volume is 200 ml.
Stir the mixture and determine the pH. Slowly add sulfuric acid until the
pH is less than or equal to 1.
CAUTION: Toxic hydrogen sulfide may be generated from the acidified
sample. This operation must be performed in the hood and the
sample left in the hood until the sample has been made alkaline
or the sulfide has been destroyed.
From the amount of sulfuric acid required to acidify the sample and the
mass or volume of the sample acidified, calculate the amount of acid
required to acidify the sample to be placed in the distillation flask.
7.12.2 Prepare the gas evolution apparatus as shown in Figure 1 in a
fume hood.
7.12.2.1 Prepare a hot water bath at 70°C by filling a
crystallizing dish or other suitable container with water and place
it on a hotplate stirrer. Place a thermometer in the bath and monitor
the temperature to maintain the bath at 70eC.
7.12.2.2 Assemble the three neck 500-mL flask, fritted gas
inlet tube, and exhaust tube. Use Teflon sleeves to seal the ground
glass joints. Place a Teflon coated stirring bar into the flask.
7.12.2.3 Place into each gas scrubbing bottle 10 + 0.5 ml of
the 0.5M zinc acetate solution, 5.0 + 0.1 ml of 37% formaldehyde and
100 + 5.0 ml water.
9031 - 6 Revision 0
December 1987
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7.12.2.4 Connect the gas evolution flask and gas scrubbing
bottles as shown in Figure 1. Secure all fittings and joints.
7.12.3 Carefully place an accurately weighed sample which contains
1.0 to 50 tng of sulfide into the flask. If necessary, dilute to
approximately 200 ml with water.
7.12.4 Place the dropping funnel onto the flask making sure its
stopcock is closed. Add the volume of sulfuric acid calculated in Step
7.1.1 plus an additional 50 ml into the dropping funnel. The bottom
stopcock must be closed.
7.12.5 Attach the nitrogen inlet to the top of the dropping funnel
gas shut-off valve. Turn on the nitrogen purge gas and adjust the flow
through the sample flask to 25 mL/min. The nitrogen in the gas scrubbing
bottles should bubble at a rate of about five bubbles per second. Nitrogen
pressure should be limited to approximately 10 psi to prevent excess
stress on the glass system and fittings. Verify that there are no leaks in
the system. Open the nitrogen shut-off valve leading to the dropping
funnel. Observe that the gas flow into the sample vessel will stop for a
short period while the pressure throughout the system equalizes. If the
gas flow through the sample flask does not return within a minute, check
for leaks around the dropping funnel. Once flow has stabilized, turn on
the magnetic stirrer. Purge the system for 15 minutes with nitrogen to
remove oxygen.
7.12.6 Heat sample to 70°C. Open dropping funnel to a position that
will allow a flow of sulfuric acid of approximately 5 mL/min. Monitor the
system until most of the sulfuric acid contained within the dropping
funnel has entered the sample flask. Close the dropping funnel while a
small amount of acid remains. Immediately close the gas shut-off valve to
the dropping funnel.
7.12.7 Purge, stir, and maintain a temperature of 70eC for a total
of 90 minutes from start to finish. Shut off nitrogen supply. Turn off
heat.
7.13 Titration of Distillate
7.13.1 Pi pet a known amount of standardized 0.025N iodine solution
(see Step 5.12) in a 500-mL flask, adding an amount in excess of that
needed to oxidize the sulfide. Add enough water to bring the volume to 100
ml. The volume of standardized iodine solution should be about 65 mL for
samples with 50 mg of sulfide.
7.13.2 Add 2 ml of 6N HC1 to the iodine.
7.13.3 Pipet both of the gas scrubbing bottle solutions into the
flask, keeping the end of the pipet below the surface of the iodine
solution. If at any point in transferring the zinc acetate solution or
rinsing the bottles, the amber color of the iodine disappears or fades to
yellow, more 0.025N iodine must be added. This additional amount must be
9031 - 7 Revision 0
December 1987
-------
added to the amount from Step 7.13.1 for calculations. Record the total
volume of standardized 0.025N iodine solution used.
7.13.4 Prepare a rinse solution of a known amount of standardized
0.025N iodine solution, 1 ml of 6N HC1, and water to rinse the remaining
white precipitate (zinc sulfide) from the gas scrubbing bottles into the
flask. There should be no visible traces of precipitate after rinsing.
7.13.5 Rinse any remaining traces of iodine from the gas scrubbing
bottles with water, and transfer the rinses to the flask.
7.13.6 Titrate the solution in the flask with standard 0.025N
phenylarsine oxide or 0.025N sodium thiosulfate solution until the amber
color fades to yellow. Add enough starch indicator for the solution to
turn dark blue and titrate until the blue disappears. Record the volume of
titrant used.
7.13.7 Calculate the concentration of sulfide in the sample as
follows:
[(ml of I? x N of l2)-(mL of titrant x N of titrant)](16.03)
sample We1ght (kg) =sulfide(mg/kg)
8.0 QUALITY CONTROL
8.1 All quality control data must be maintained and available for
reference or inspection for a period of three years. This method is restricted
to use by or under supervision of experienced analysts. Refer to the
appropriate section of Chapter One for additional quality control
requirements.
8.2 A reagent blank should be run once every twenty analyses or per
analytical batch, whichever is more frequent.
8.3 Check standards are prepared from water and a known amount of sodium
sulfide. A check standard should be run with each analytical batch of samples
or once every twenty samples. Acceptable recovery will depend on level and
matrix.
8.4 A matrix spiked sample should be run for each analytical batch or
every twenty samples, whichever is more frequent, to determine matrix effects.
If recovery is low, acid-insoluble sulfides are indicated. A matrix spiked
sample is a sample brought through the whole sample preparation and analytical
process.
8.5 Verify the calibration with an independently prepared QC reference
sample every twenty samples or once per analytical batch, whichever is more
frequent.
9031 - 8 Revision 0
December 1987
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9.0 METHOD PERFORMANCE
9.1 Accuracy - Accuracy for this method was determined by three
independent laboratories by measuring percent recoveries of spikes for waste
samples. The results are summarized below.
Accuracy for the entire method for four synthetic waste samples
70-104% recovery
9.2 Precision
Precision of entire method for four synthetic waste samples
Percent coefficient of variation 1.0-34
10.0 REFERENCES
1. Test Methods for Evaluating Solid Waste. Physical/Chemical Methods. 3rd ed.;
Emergency Response. U.S. Government Printing Office: Washington, DC,1987;
SW-846; 955-001-00000-1.
2. Methods for Chemical Analysis of Water and Wastes; U.S. Environmental
Protection Agency. Office of Research and Development. Environmental
Monitoring and Support Laboratory. ORD Publications Office. Center for
Environmental Research Information: Cincinati, OH, 1979; EPA-
600/4-79-020, Method 376.1.
3. CRC Handbook of Chemistry and Phvsics. 66th ed.; Weast, R., Ed.; CRC: Boca
Raton, FL, 1985.
4. Standard Methods for the Examination of Water and Wastewater. 16th ed.;
Greenberg, A.E.; Trussell, R.R.; Clesceri, L.S., Eds.; American Water
Works Association, Water Pollution Control Federation, American Public
Health Association: Washington, DC, 1985; Methods 427, 427A, 427B, and
427D.
5. Andreae, M.O.; Bernard, W.R. Anal. Chem. 1983, 55. 608-612.
6. Barclay, H. Adv. Instrum. 1980, 35(2). 59-61.
7. Bateson, S.W.; Moody, G.J.; Thomas, J.P.R. Analyst 1986, 111. 3-9.
8. Berthage, P.O. Anal. Chim. Acta 1954, 10, 310-311.
9. Craig, P.O.; Moreton, P.A. Environ. Techno!. Lett. 1982, 3, 511-520.
10. Franklin, G.O.; Fitchett, A.W. Pulp & Paper Canada 1982, 83(10). 40-44.
11. Fuller, W. Cyanide in the Environment: Van Zyl, D., Ed.; Proceedings of
Symposium; December 1984.
9031 - 9 Revision 0
December 1987
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12. Gottfried, G.J. "Precision, Accuracy, and MDL Statements for EPA Methods
9010, 9030, 9060, 7520, 7521, 7550, 7551, 7910, and 7911"; final report to
the U.S. Environmental Protection Agency (EMSL-CI); Biopheric.
13. Kilroy, W.P. Talanta 1983, 30(6). 419-422.
14. Kurtenacher, V.A.; Wallak, R. Z. Anorg. U. Chem. 1927, 161. 202-209.
15. Landers, D.H.; David, M.B.; Mitchell, M.J. Int. J. Anal. Chem. 1983, 11,
245-256.
16. Opekar, F.; Brukenstein, S. Anal. Chem. 1984, 56, 1206-1209.
17. Ricklin, R.D.; Johnson, E.L. Anal. Chem. 1983, 55, 4.
18. Rohrbough, W.G.; et al. Reagent Chemicals. American Chemical Society
Specifications. 7th ed.; American Chemical Society: Washington, DC, 1986.
19. Snedecor, G.W.; Ghran, W.G. Statistical Methods; Iowa State University
Press: Ames, IA, 1980.
20. Umana, M.; Beach, J.; Sheldon, L. "Revisions to Method 9010"; final report
to the Environmental Protection Agency on Contract No. 68-01-7266;
Research Triangle Institute: Research Triangle Park, NC, 1986; Work
Assignment No. 1.
21. Umana, M.; Sheldon, L. "Interim Report: Literature Review"; interim
report to the U.S. Environmental Protection Agency in Contract No.
68-01-7266; Research Triangle Institute: Research Triangle Park, NC,
1986; Work Assignment No. 3.
22. Wang, W.; Barcelona, M.J. Environ. Inter. 1983, 9, 129-133.
23. Wronksi, M. Talanta 1981, 28, 173-176.
24. Application Note 156; Princeton Applied Research Corp.: Princeton, NJ.
25. Guidelines for Assessing and Reporting Data Quality for Environmental
Measurements; U.S. Environmental Protection Agency
Office of Research and Development: Washington, DC, 1983.
26. Fed. Regist. 1980, 45(98). 33122.
27. The Analytical Chemistry of Sulfur and Its Compounds. Part I; Karchmer,
J.H., Ed.; Wiley-Interscience: New York, 1970.
28. Methods for the Examination of Water and Associated Materials; Department
of the Environment: England, 1983.
29. "Development and Evaluation of a Test Procedure for Reactivity Criteria
for Hazardous Waste"; final report to the U.S. Environmental Protection
Agency on Contract 68-03-2961; EAL: Richmond, CA.
9031 - 10 Revision 0
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30. 1985 Annual Book of ASTM Standards. Vol. 11.01; "Standard Specification
for Reagent Water"; ATSM: Philadelphia, PA, 1985; D1193-77.
9031 - 11 Revision 0
December 1987
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FIGURE 1.
GAS EVOLUTION APPARATUS
H2SO4 (HCI for acid insoluble sulfidea)
N2 in
Hot Water Bath
with Magnetic Stirrer
Out
Zinc Acetate
and
Formaldehyde
Scrubbing
Bettlea
Stirring Bar
9031 - 12
Revision 0
December 1987
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FIGURE 2.
TUMBLER-EXTRACTOR
Foam Lined
1H_ Bottle
with Cap
Jar Mill Drive
Box Wheels Plywood Construction
9031 - 13
Revision 0
December 1987
-------
0
FIGURE 3.
EXTRACTOR
9031 - 14
Revision 0
December 1987
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METHOD 9031
SULFIDES
c
turt
0
T.I Aaaaakla faaaal
apparatus:fora a pad
lea tklek fro* flats
»ool eat tka pad to
fit faaaal:*ol(k pad
•id placa ia
fiaaal v.t pad vltk
kaova aaoaaf of vatar
T.i Place la a 1-t
wit* laatk
bottle:SOOaL vatar.
SaL 601 laQl, 1(
•taaaeM ekleri4a,
aa< (OmL a-kaxaaa
(if ell or fraaaa
la praaaat)
7.2 Traaafar aaapla
(1.0-60»» aalflda)
to fuaal:riaaa
aaapli coatalaar
wltk kaova aaoaata
at vatar:a<4 rlaaaa
to faaaal:fllt«r
aatil ao fr»« vatar
rtaalta ia Iiaaal
T.( Cap kottla vltk
Tafloa lla»« cap
aa4 akaka:llract
altrofta lato
kottla for 1 Blaata
to parga
T.3 Traaafar aolii
aa< flaar pad to a
arlad Ur«4
valfkiaf diak.
T.i «algk oat
2Sf:add to kottlo
•kilo parflaf
T.) Traaafar
flltrata to
loparatorj faaaal:
callact a^aaoaa
pkata aad Maaara
volaaa:traasfor oil
pkaaa to
dlak
74 Valfk dlak aad
coataata:aabtract
flaaa flaar pad (If
aay>-»aktract total
TO 1sat of riaaaa
froa Tolaao of
flltrata
T I Add all
aalldaicap kottla
7.6 pi of axtract
•aat ba > • aad <
lltakaka kottla 1
•laata:opaa aadar
altro|oa:ckack pi
9031 - 15
Revision 0
December 1987
-------
METHOD 9031
(Continued)
7.8 Add SlL
Iliqioti of 601
I.OH lltil pH > 9:
»rj» oxyf.i:recip
ottlii: repeat if
lece.iary
7.7 Pile, kettl* li
tnibl«r:tir» el aid
rii for II koira
7 8 Pr.pare fnaiol
11 in Step 7.1
7.* Decaat (strict
lite final
7.10 Place extract
It •»p«r»tory
final:collect aad
•••••re Teliae e(
eqieeii pkei*
7.11 Coibile
•qieeie extract aid
ericiul niple
filtrite ii
•llqiete
proportloiil te tk«
•npl<: cilcilite
proportioie
7.12.1 Ckoote niple
•lie (1.0-60»|
••Iflde):plice kievi
••out of eaiple ii
keiker:add
viter:*eteir« pR:«dd
ceic eelfirlc icld te
pH-1
7.12.1 Cllcilltl
•Bout ef nlfiric
icid aeeded to
acidify tuple
7.12.2 frepare |ii
•Tolitioi apparatae
(»e Flf.re 1)
7.12.2.1 Prepare
ket vater k*tk
7.12.2.2 Atiaaele 3
•eck flaik
7.12.2.3 Pile, ziic
acetate,
fer>aldekyde, aid
vat.r la »••
kottl.e
7.12.2.4 Collect
fink ««d ecribeiaf
bottl.i:i.car.
Joist.
7.12.3 Place
veifked iiipl* la
flaek:dllat« vltk
vater if lecee.ary
7.12.4 Place
dreppiif fiaiel
oito fleak:add
•ilfiric acid froa
Step 7.12 to
dreppiai filial
0 0
7.12.6 Adjait
aitrotei flow:ch.ck
for leikaitiri on
•tirr.rrpurj.
•yitei of oxyf.n
for 16 iliatei
7.12.6 Heat to
70dofC:add nlfiric
acid to flaik:clo»
droppiig fnael
vkea aeet of the
acid kae eat.r.d
tke flaik
7.12.7 Pirge, itir,
•ad keat for 90
ainitei:ihit off
aitro(*a:tira a!i
h«at
7.12.1 AaalTZe by
tltratiei (Step
7. IS)
7.13.1 Plpet kiova
•ao»t of 0.0261
ledlie eela la
flaak:krii| to
Yol»e vitk vater
9031 - 16
Revision
December
0
1987
-------
METHOD 9031
(Continued)
7.13.3 Pip«t
icribbiag bottlo
•ola to flaak
7.13.E liaao traeii
of iodiao froa
•cr«bbia|
bottl*e:tranafor
riaaoa to flaak
7.13.t Titrate tola
vita PAD or lodiu
tklottlftt* tola
tatil labor color
(•do*.-add itareh
iadleator:tltrato
aatll blao color
diaappoara:record
rol»o of titraat
7 13.7 Calcilato
tko coac of (ilfid
ia tho aaaplo
7.13.3 locord total
rolaao of 0.0361
iodla* aola ttod
CEO
7.13.4 Pr.par.
riaao aola of
0.03EI lodlao aola,
61 MCI. aad water
9031 - 17
Revision 0
December 1987
-------
METHOD 9056
ANION CHROMATOGRAPHY METHOD
1.0 SCOPE AND APPLICATION
1.1 This method addresses the sequential determination of the anions
chloride, fluoride, bromide, nitrate, nitrite, phosphate, and sulfate in the
collection solutions from the bomb combustion of solid waste samples, as well as
all water samples.
1.2 The minimum detection limit (MDL), the minimum concentration of a
substance that can be measured and reported with 99% confidence that the value
is above zero, varies for anions as a function of sample size and the
conductivity scale used. Generally, minimum detectable concentrations are in the
range of 0.05 mg/L for F- and 0.1 mg/L for Br", Cl", N03", N02", P04 , and SO,2"
with a 100-^L sample loop and a 10-umho full-scale setting on the conductivity
detector. Similar values may be achieved by using a higher scale setting and an
electronic integrator. Idealized detection limits of an order of magnitude lower
have been determined in reagent water by using a 1 umho full-scale setting (Table
1).
The upper limit of the method is dependent on total anion concentration and
may be determined experimentally. These limits may be extended by appropriate
dilution.
2.0 SUMMARY OF METHOD
2.1 A small volume of combustate collection solution or other water
sample, typically 2 to 3 ml, is injected into an ion chromatograph to flush and
fill a constant volume sample loop. The sample is then injected into a stream
of carbonate-bicarbonate eluent of the same strength as the collection solution
or water sample.
The sample is pumped through three different ion exchange columns and into
a conductivity detector. The first two columns, a precolumn or guard column and
a separator column, are packed with low-capacity, strongly basic anion exchanger.
Ions are separated into discrete bands based on their affinity for the exchange
sites of the resin. The last column is a suppressor column that reduces the
background conductivity of the eluent to a low or negligible level and converts
the anions in the sample to their corresponding acids. The separated anions in
their acid form are measured using an electrical-conductivity cell. Anions are
identified based on their retention times compared to known standards.
Quantitation is accomplished by measuring the peak height or area and comparing
it to a calibration curve generated from known standards.
3.0 INTERFERENCES
3.1 Any species with a retention time similar to that of the desired ion
will interfere. Large quantities of ions eluting close to the ion of interest
will also result in an interference. Separation can be improved by adjusting the
eluent concentration and/or flow rate.
9056 - 1 Revision 0
November 1992
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Sample dilution and/or the use of the method of standard additions can also
be used.
For example, high levels of organic acids may be present in industrial
wastes, which may interfere with inorganic anion analysis. Two common species,
formate and acetate, elute between fluoride and chloride.
3.2 Because bromide and nitrate elute very close together, they are
potential interferents for each other. It is advisable not to have Br"/N03"
ratios higher than 1:10 or 10:1 if both anions are to be quantified. If nitrate
is observed to be an interference with bromide, use of an alternate detector
(e.g.. electrochemical detector) is recommended.
3.3 Method interferences may be caused by contaminants in the reagent
water, reagents, glassware, and other sample processing apparatus that lead to
discrete artifacts or elevated baseline in ion chromatograms.
3.4 Samples that contain particles larger than 0.45 urn and reagent
solutions that contain particles larger than 0.20 urn require filtration to
prevent damage to instrument columns and flow systems.
3.5 If a packed bed suppressor column is used, it will be slowly consumed
during analysis and, therefore, will need to be regenerated. Use of either an
anion fiber suppressor or an anion micromembrane suppressor eliminates the time-
consuming regeneration step through the use of a continuous flow of regenerant.
4.0 APPARATUS AND MATERIALS
4.1 Ion chromatograph, capable of delivering 2 to 5 ml of eluent per
minute at a pressure of 200 to 700 psi (1.3 to 4.8 MPa). The chromatograph shall
be equipped with an injection valve, a 100-juL sample loop, and set up with the
following components, as schematically illustrated in Figure 1.
4.1.1 Precolumn, a guard column placed before the separator column
to protect the separator column from being fouled by particulates or
certain organic constituents (4 x 50 mm, Dionex P/N 030825 [normal run],
or P/N 030830 [fast run], or equivalent).
4.1.2 Separator column, a column packed with low-capacity
pellicular anion exchange resin that is styrene diyinylbenzene-based has
been found to be suitable for resolving F", CT, N02", P04 , Br", N03", and
SO/ (see Figure 2) (4 x 250 mm, Dionex P/N 03827 [normal run], or P/N
030831 [fast run], or equivalent).
4.1.3 Suppressor column, a column that is capable of converting
the eluent and separated anions to their respective acid forms (fiber,
Dionex P/N 35350, micromembrane, Dionex P/N 38019 or equivalent).
4.1.4 Detector, a low-volume, flowthrough, temperature-
compensated, electrical conductivity cell (approximately 6 pi volume,
Dionex, or equivalent) equipped with a meter capable of reading from 0 to
1,000 /xseconds/cm on a linear scale.
9056 - 2 Revision 0
November 1992
-------
4.1.5 Pump, capable of delivering a constant flow of approximately
2 to 5 mL/min throughout the test and tolerating a pressure of 200 to 700
psi (1.3 to 4.8 MPa).
4.2 Recorder, compatible with the detector output with a full-scale
response time in 2 seconds or less.
4.3 Syringe, minimum capacity of 2 ml and equipped with a male pressure
fitting.
4.4 Eluent and regenerant reservoirs, suitable containers for storing
eluents and regenerant. For example, 4 L collapsible bags can be used.
4.5 Integrator, to integrate the area under the chromatogram. Different
integrators can perform this task when compatible with the electronics of the
detector meter or recorder. If an integrator is used, the maximum area
measurement must be within the linear range of the integrator.
4.6 Analytical balance, capable of weighing to the nearest 0.0001 g.
4.7 Pipets, Class A volumetric flasks, beakers: assorted sizes.
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 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One. Column life may be extended by passing
reagent water through a 0.22-/im filter prior to use.
5.3 Eluent, 0.003M NaHC03/0.0024M Na2C03. Dissolve 1.0080 g of sodium
bicarbonate (0.003M NaHC03) and 1.0176 g of sodium carbonate (0.0024M Na2C03) in
reagent water and dilute to 4 L with reagent water.
5.4 Suppressor regenerant solution. Add 100 ml of IN H2S04 to 3 L of
reagent water in a collapsible bag and dilute to 4 L with reagent water.
5.5 Stock solutions (1,000 mg/L).
5.5.1 Bromide stock solution (1.00 ml = 1.00 mg Br"). Dry
approximately 2 g of sodium bromide (NaBr) for 6 hours at 150°C, and cool
in a desiccator. Dissolve 1.2877 g of the dried salt in reagent water,
and dilute to 1 L with reagent water.
5.5.2 Chloride stock solution (1.00 ml = 1.00 mg Cl"). Dry sodium
chloride (NaCl) for 1 hour at 600°C, and cool in a desiccator. Dissolve
1.6484 g of the dry salt in reagent water, and dilute to 1 L with reagent
water.
9056 - 3 Revision 0
November 1992
-------
5.5.3 Fluoride stock solution (1.00 ml = 1.00 mg F"). Dissolve
2.2100 g of sodium fluoride (NaF) in reagent water, and dilute to 1 L with
reagent water. Store in chemical-resistant glass or polyethylene.
5.5.4 Nitrate stock solution (1.00 ml = 1.00 mg NO,"). Dry
approximately 2 g of sodium nitrate (NaNO?) at 105°C for 24 hours.
Dissolve exactly 1.3707 g of the dried salt in reagent water, and dilute
to 1 L with reagent water.
5.5.5 Nitrite stock solution (1.00 ml = 1.00 mg N02"). Place
approximately 2 g of sodium nitrate (NaN02) in a 125 ml beaker and dry to
constant weight (about 24 hours) in a desiccator containing concentrated
H2SO,. Dissolve 1.4998 g of the dried salt in reagent water, and dilute
to 1 L with reagent water. Store in a sterilized glass bottle.
Refrigerate and prepare monthly.
NOTE: Nitrite is easily oxidized, especially in the presence of moisture,
and only fresh reagents are to be used.
NOTE: Prepare sterile bottles for storing nitrite solutions by heating for
1 hour at 170°C in an air oven.
5.5.6 Phosphate stock solution (1.00 ml = 1.00 mg P043"). Dissolve
1.4330 g of potassium dihydrogen phosphate (KH2P04) in reagent water, and
dilute to 1 L with reagent water. Dry sodium sulfate (Na2S04) for 1 hour
at 105°C and cool in a desiccator.
5.5.7 Sulfate stock solution (1.00 ml = 1.00 mg S042"). Dissolve
1.4790 g of the dried salt in reagent water, and dilute to 1 L with
reagent water.
5.6 Anion working solutions. Prepare a blank and at least three
different working solutions containing the following combinations of anions. The
combination anion solutions must be prepared in Class A volumetric flasks. See
Table 2.
5.6.1 Prepare a high-range standard solution by diluting the
volumes of each anion specified in Table 2 together to 1 L with reagent
water.
5.6.2 Prepare the intermediate-range standard solution by diluting
10.0 ml of the high-range standard solution (see Table 2) to 100 ml with
reagent water.
5.6.3 Prepare the low-range standard solution by diluting 20.0 ml
of the intermediate-range standard solution (see Table 2) to 100 ml with
reagent water.
5.7 Stability of standards. Stock standards are stable for at least 1
month when stored at 4°C. Dilute working standards should be prepared weekly,
except those that contain nitrite and phosphate, which should be prepared fresh
daily.
9056 - 4 Revision 0
November 1992
-------
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 Analyze the samples as soon as possible after collection. Preserve
by refrigeration at 4°C.
7.0 PROCEDURE
7.1 Calibration
7.1.1 Establish ion chromatographic operating parameters
equivalent to those indicated in Table 1.
7.1.2 For each analyte of interest, prepare calibration standards
at a minimum of three concentration levels and a blank by adding
accurately measured volumes of one or more stock standards to a Class A
volumetric flask and diluting to volume with reagent water. If the
working range exceeds the linear range of the system, a sufficient number
of standards must be analyzed to allow an accurate calibration curve to be
established. One of the standards should be representative of a concen-
tration near, but above, the method detection limit if the system is
operated on an applicable attenuator range. The other standards should
correspond to the range of concentrations expected in the sample or should
define the working range of the detector. Unless the attenuator range
settings are proven to be linear, each setting must be calibrated
individually.
7.1.3 Using injections of 0.1 to 1.0 mL (determined by injection
loop volume) of each calibration standard, tabulate peak height or area
responses against the concentration. The results are used to prepare a
calibration curve for each analyte. During this procedure, retention
times must be recorded. The retention time is inversely proportional to
the concentration.
7.1.4 The working calibration curve must be verified on each
working day, or whenever the anion eluent is changed, and for every batch
of samples. If the response or retention time for any analyte varies from
the expected values by more than + 10%, the test must be repeated, using
fresh calibration standards. If the results are still more than + 10%, an
entirely new calibration curve must be prepared for that analyte.
7.1.5 Nonlinear response can result when the separator column
capacity is exceeded (overloading). Maximum column loading (all anions)
should not exceed about 400 ppm.
7.2 Analyses
7.2.1 Sample preparation. When aqueous samples are injected, the
water passes rapidly through the columns, and a negative "water dip" is
observed that may interfere with the early-eluting fluoride and/or
chloride ions. The water dip should not be observed in the combustate
9056 - 5 Revision 0
November 1992
-------
samples; the collecting solution is a concentrated eluent solution that
will "match" the eluent strength when diluted to 100-mL with reagent water
according to the bomb combustion procedure. Any dilutions required in
analyzing other water samples should be made with the eluent solution.
The water dip, if present, may be removed by adding concentrated eluent to
all samples and standards. When a manual system is used, it is necessary
to micropipet concentrated buffer into each sample. The recommended
procedures follow:
(1) Prepare a 100-mL stock of eluent 100 times normal concentration by
dissolving 2.5202 g NaHCO, and 2.5438 g Na2C03 in 100-mL reagent
water. Protect the volumetric flask from air.
(2) Pipet 5 mL of each sample into a clean polystyrene micro-beaker.
Micropipet 50 /xL of the concentrated buffer into the beaker and stir
well.
Dilute the samples with eluent, if necessary, to concentrations within the
linear range of the calibration.
7.2.2 Sample analysis.
7.2.2.1 Start the flow of regenerant through the
supressor column.
7.2.2.2 Set up the recorder range for maximum sensitivity
and any additional ranges needed.
7.2.2.3 Begin to pump the eluent through the columns.
After a stable baseline is obtained, inject a midrange standard. If
the peak height deviates by more than 10% from that of the previous
run, prepare fresh standards.
7.2.2.4 Begin to inject standards starting with the
highest concentration standard and decreasing in concentration. The
first sample should be a quality control reference sample to check
the calibration.
7.2.2.5 Using the procedures described in Step 7.2.1,
calculate the regression parameters for the initial standard curve.
Compare these values with those obtained in the past. If they
exceed the control limits, stop the analysis and look for the
problem.
7.2.2.6 Inject a quality control reference sample. A
spiked sample or a sample of known content must be analyzed with
each batch of samples. Calculate the concentration from the
calibration curve and compare the known value. If the control
limits are exceeded, stop the analysis until the problem is found.
Recalibration is necessary.
7.2.2.7 When an acceptable value has been obtained for
the quality control sample, begin to inject the samples.
9056 - 6 Revision 0
November 1992
-------
7.2.2.8 Load and inject a fixed amount of well-mixed
sample. Flush injection loop thoroughly, using each new sample.
Use the same size loop for standards and samples. Record the
resulting peak size in area or peak height units. An automated
constant volume injection system may also be used.
7.2.2.9 The width of the retention time window used to
make identifications should be based on measurements of actual
retention time variations of standards over the course of a day.
Three times the standard deviation of a retention time can be used
to calculate a suggested window size for a compound. However, the
experience of the analyst should weigh heavily in the interpretation
of chromatograms.
7.2.2.10 If the response for the peak exceeds the working
range of the system, dilute the sample with an appropriate amount of
reagent water and reanalyze.
7.2.2.11 If the resulting chromatogram fails to produce
adequate resolution, or if identification of specific anions is
questionable, spike the sample with an appropriate amount of
standard and reanalyze.
NOTE: Retention time is inversely proportional to concentration. Nitrate
and sulfate exhibit the greatest amount of change, although all
anions are affected to some degree. In some cases, this peak
migration can produce poor resolution or misidentification.
7.3 Calculation
7.3.1 Prepare separate calibration curves for each anion of
interest by plotting peak size in area, or peak height units of standards
against concentration values. Compute sample concentration by comparing
sample peak response with the standard curve.
7.3.2 Enter the calibration standard concentrations and peak
heights from the integrator or recorder into a calculator with linear
least squares capabilities.
7.3.3 Calculate the following parameters: slope (s), intercept
(I), and correlation coefficient (r). The slope and intercept define a
relationship between the concentration and instrument response of the
form:
yf = s, x, + I (1)
where: y,- = predicted instrument response
s,- = response slope
X,- = concentration of standard i
I = intercept
9056 - 7 Revision 0
November 1992
-------
Rearrangement of the above equation yields the concentration corresponding
to an instrumental measurement:
(2)
where:
Xj = calculated concentration for a sample
yj = actual instrument response for a sample
Sj and I are calculated slope and intercept from calibration above.
7.3.4 Enter the sample peak height into the calculator, and
calculate the sample concentration in milligrams per liter.
8.0 QUALITY CONTROL
8.1 All quality control data should be maintained and available for easy
reference and inspection. Refer to Chapter One for additional quality control
guidelines.
8.2 After every 10 injections, analyze a midrange calibration standard.
If the instrument response has changed by more than 5%, recalibrate.
8.3 Analyze all samples in duplicate. Take the duplicate sample through
the entire sample preparation and analytical process.
8.4 A matrix spiked sample should be run for each analytical batch or
twenty samples, whatever is more frequent, to determine matrix effects.
9.0 METHOD PERFORMANCE
9.1 Single-operator accuracy and precision for reagent, drinking and
surface water, and mixed domestic and industrial wastewater are listed in Table
3.
9.2 Combustate samples. These data are based on 41 data points obtained
by six laboratories who each analyzed four used crankcase oils and three fuel oil
blends with crankcase in duplicate. The oil samples were combusted using Method
5050. A data point represents one duplicate analysis of a sample. One data
point was judged to be an outlier and was not included in the results.
9.2.1 Precision. The precision of the method as determined by the
statistical examination of inter! aboratory test results is as follows:
Repeatability - The difference between successive results obtained
by the sample operator with the same apparatus under constant operating
conditions on identical test material would exceed, in the long run, in
the normal and correct operation of the test method, the following values
only in 1 case in 20 (see Table 4):
9056 - 8 Revision 0
November 1992
-------
Repeatability - 20.9
*where x is the average of two results in M9/9-
Reproducibilitv - The difference between two single and independent
results obtained by different operators working in different laboratories
on identical test material would exceed, in the long run, the following
values only in 1 case in 20:
Reproducibili ty - 42.1 /x*
*where x is the average value of two results in M9/9-
9.2.2 Bias. The bias of this method varies with concentration,
as shown in Table 5:
Bias = Amount found - Amount expected
10.0 REFERENCES
1. Environmental Protection Agency. Test Method for the Determination of
Inorganic Anions in Water by Ion Chromatography. EPA Method 300.0. EPA-600/4-
84-017. 1984.
2. Annual Book of ASTM Standards, Volume 11.01 Water D4327, Standard Test
Method for Anions in Water by Ion Chromatography, pp. 696-703. 1988.
3. Standard Methods for the Examination of Water and Wastewater, Method 429,
"Determination of Anions by Ion Chromatography with Conductivity Measurement,"
16th Edition of Standard Methods.
4. Dionex, 1C 16 Operation and Maintenance Manual, PN 30579, Dionex Corp.,
Sunnyvale, CA 94086.
5. Method detection limit (MDL) as described in "Trace Analyses for
Wastewater," J. Glaser, D. Foerst, G. McKee, S. Quave, W. Budde, Environmental
Science and Technology, Vol. 15, Number 12, p. 1426, December 1981.
6. Gaskill, A.; Estes, E. D.; Hardison, D. L.; and Myers, L. E. Validation
of Methods for Determining Chlorine in Used Oils and Oil Fuels. Prepared for
U.S. Environmental Protection Agency Office of Solid Waste. EPA Contract No. 68-
01-7075, WA 80. July 1988.
9056 - 9 Revision 0
November 1992
-------
TABLE 1.
CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION
LIMITS IN REAGENT WATER
Analyte
Fluoride
Chlorine
Nitrite-N
0-Phosphate-P
Nitrate-N
Sulfate
Retention8
time
min
1.2
3.4
4.5
9.0
11.3
21.4
Relative
retention
time
1.0
2.8
3.8
7.5
9.4
17.8
Method6
detection limit,
mg/L
0.005
0.015
0.004
0.061
0.013
0.206
Standard conditions:
Columns - As specified in 4.1.4
Detector - As specified in 4.1.4
Eluent - As specified in 5.3
"Concentrations of mixed standard (mg/L):
Fluoride 3.0
Chloride 4.0
Nitrite-N 10.0
Sample loop - 100 /zL
Pump volume - 2.30 mL/min
0-Phosphate-P 9.0
Nitrate-N 30.0
Sulfate 50.0
calculated from data obtained using an attentuator setting of 1 umho full
scale. Other settings would produce an MDL proportional to their value.
9056 - 10
Revision 0
November 1992
-------
TABLE 2.
PREPARATION OF STANDARD SOLUTIONS FOR INSTRUMENT CALIBRATION
Hiah-ranqe standard (see 5.6.1)
Milliliters
of each
stock solution
Fluoride (F')
Chloride (Cl~)
Nitrite (N02~)
Phosphate (P043')
Bromide (Br")
Nitrate (N03~)
Sulfate (S042")
(1.00 mL =
1.00 mg)
diluted to
1,000 mL
10
10
20
50
10
30
100
An ion
concentration
mg/L
10
10
20
50
10
30
100
Intermediate-
range standard,
mg/L
(see 5.6.2)
1.0
1.0
2.0
5.0
1.0
3.0
10.0
Low-range
standard,
mg/L (see
5.6.3)
0.2
0.2
0.4
1.0
0.2
0.6
2.0
9056 - 11
Revision 0
November 1992
-------
TABLE 3.
SINGLE-OPERATOR ACCURACY AND PRECISION
Sample
Analyte type
Chloride
Fluoride
Nitrate-N
Nitrite-N
0-Phosphate-P
Sulfate
RW
DM
SW
WW
RW
DM
SW
WW
RW
DW
SW
WW
RW
DW
SW
WW
RW
DE
SW
WW
RW
DW
SW
WW
Spike
mg/L
0.050
10.0
1.0
7.5
0.24
9.3
0.50
1.0
0.10
31.0
0.50
4.0
0.10
19.6
0.51
0.52
0.50
45.7
0.51
4.0
1.02
98.5
10.0
12.5
Number
of
replicates
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
7
Mean
recovery,
%
97.7
98.2
105.0
82.7
103.1
87.7
74.0
92.0
100.9
100.7
100.0
94.3
97.7
103.3
88.2
100.0
100.4
102.5
94.1
97.3
102.1
104.3
111.6
134.9
Standard
deviation,
mg/L
0.0047
0.289
0.139
0.445
0.0009
0.075
0.0038
0.011
0.0041
0.356
0.0058
0.058
0.0014
0.150
0.0053
0.018
0.019
0.386
0.020
0.04
0.066
1.475
0.709
0.466
RW = Reagent water.
DW = Drinking water.
SW = Surface water.
WW = Wastewater.
9056 - 12
Revision 0
November 1992
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TABLE 4.
REPEATABILITY AND REPRODUCIBILITY FOR CHLORINE IN
USED OILS BY BOMB OXIDATION AND ION CHROMATOGRAPHY
Average value, Repeatability, Reproducibility,
M9/g
500
1,000
1,500
2,000
2,500
3,000
467
661
809
935
1,045
1,145
941
1,331
1,631
1,883
2,105
2,306
TABLE 5.
RECOVERY AND BIAS DATA FOR CHLORINE IN USED OILS BY
BOMB OXIDATION AND ION CHROMATOGRAPHY
Amount Amount
Expected found Bias, Percent,
Mg/g Mg/g bias
320 567 247 +77
480 773 293 +61
920 1,050 130 +14
1,498 1,694 196 +13
1,527 1,772 245 +16
3,029 3,026 -3 0
3,045 2,745 -300 -10
9056 - 13 Revision 0
November 1992
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FIGURE 1
SCHEMATIC OF ION CHROMATOGRAPH
WAS IT
(1) Eluent reservoir
(2) Pump
(3) Precolumn
(4) Separator column
(5) Suppressor column
(6) Detector
(7) Recorder or integrator, or both
9056 - 14
Revision 0
November 1992
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FIGURE 2
TYPICAL ANION PROFILE
a 12
MINUTES
18
20
9056 - 15
Revision 0
November 1992
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METHOD 9056
ANION CHROMATOGRAPHY METHOD
START
711 Establish ion
chroma tog r a phi c
operating
712 Prepare
calibration
s tanda rds at a
minimum of three
concentration
levels and a blank
1
713 Prepare
calibration curves
7 1 4 Verify the
each working day or
whenever the anion
eluent is changed ,
and for every batch
of samples
/ 7 2 1 Are \v*
C samples j-
^. aqueous*7 S
No
722 Analyze
standards beginning
with the highes t
concentration and
decreasing in
concentration
7 2 1 If a dilution
— » dilution should be
made with eluent
soluti on
7 2 1 Add
concentrated eluent
s tanda rds to remove
water dip
1
p*
7225 Compare
results to
if results exceed
identify problem
bef o r e proceeding
I
7226 Inject a
spiked sample of
calculate the cone
from the calibration
curve , if resul t
exceeds contro I
limits, find problem
bef o re proceeding
7227 Begin
sample analysis
1
7228 Analyze all
samples in same
manner
/I 2 2 10 N.
f for peak exceed y-
X. range? >^
No
731 Prepare
sample calibration
curves for each
anion of interest
and compute sample
concentration
es 7 2 2 10 Dilute
— * sample with reagent
water
•* instrumental
( STOP j
9056 - 16
Revision 0
November 1992
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METHOD 9071A
OIL AND GREASE EXTRACTION METHOD FOR SLUDGE AND SEDIMENT SAMPLES
1.0 SCOPE AND APPLICATION
1.1 Method 9071 is used to recover low levels of oil and grease (10
mg/L) by chemically drying a wet sludge sample and then extracting via the
Soxhlet apparatus.
1.2 Method 9071 is used when relatively polar, heavy petroleum
fractions are present, or when the levels of nonvolatile greases challenge the
solubility limit of the solvent.
1.3 Specifically, Method 9071 is suitable for biological lipids,
mineral hydrocarbons, and some industrial wastewaters.
1.4 Method 9071 is not recommended for measurement of low-boiling
fractions that volatilize at temperatures below 70°C.
2.0 SUMMARY OF METHOD
2.1 A 20-g sample of wet sludge with a known dry-solids content is
acidified to pH 2.0 with 0.3 mL concentrated HC1.
2.2 Magnesium sulfate monohydrate will combine with 75% of its own
weight in water in forming MgS04 • 7H20 and is used to dry the acidified sludge
sample.
2.3 Anhydrous sodium sulfate is used to dry samples of soil and
sediment.
2.4 After drying, the oil and grease are extracted with
trichlorotrifluoroethane (Fluorocarbon-113) using the Soxhlet apparatus.
3.0 INTERFERENCES
3.1 The method is entirely empirical, and duplicate results can be
obtained only by strict adherence to all details of the processes.
3.2 The rate and time of extraction in the Soxhlet apparatus must be
exactly as directed because of varying solubilities of the different greases.
3.3 The length of time required for drying and cooling extracted
material must be constant.
3.4 A gradual increase in weight may result due to the absorption of
oxygen; a gradual loss of weight may result due to volatilization.
Replacement solvent will be specified in a forthcoming regulation.
9071A - 1 Revision 1
November 1992
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4.0 APPARATUS AND MATERIALS
4.1 Extraction apparatus: Soxhlet.
4.2 Analytical balance.
4.3 Vacuum pump or some other vacuum source.
4.4 Extraction thimble: Filter paper.
4.5 Glass wool or small glass beads to fill thimble.
4.6 Grease-free cotton: Extract nonabsorbent cotton with solvent.
4.7 Beaker: 150-mL.
4.8 pH Indicator to determine acidity.
4.9 Porcelain mortar.
4.10 Extraction flask: 150-mL.
4.11 Distilling apparatus: Waterbath at 70°C.
4.12 Desiccator.
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 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Concentrated hydrochloric acid (HC1).
5.4 Magnesium sulfate monohydrate: Prepare MgS04 • H20 by spreading a
thin layer in a dish and drying in an oven at 150°C overnight.
5.5 Sodium sulfate, granular, anhydrous (Na?S04): 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.
9071A - 2 Revision 1
November 1992
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5.6 Trichlorotrif1uoroethane (1,1,2-trichloro-1,2,2-trif1uoroethane):
Boiling point, 47°C. The solvent should leave no measurable residue on
evaporation; distill if necessary.2
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Transfers of the solvent trichlorotrifluoroethane should not
involve any plastic tubing in the assembly.
6.2 Sample transfer implements: Implements are required to transfer
portions of solid, semisolid, and liquid wastes from sample containers to
laboratory glassware. Liquids may be transferred using a glass hypodermic
syringe. Solids may be transferred using a spatula, spoon, or coring device.
6.3 Any turbidity or suspended solids in the extraction flask should
be removed by filtering through grease-free cotton or glass wool.
7.0 PROCEDURE
7.1 Determination of Sample Dry Weight Fraction
7.1.1 Weigh 5-10 g of the sample into a tared crucible.
Determine the dry weight fraction of the sample by drying overnight at
105°C.
NOTE: The drying oven should be contained in a hood or vented.
Significant laboratory contamination may result from a heavily
contaminated hazardous waste sample.
Allow to cool in a desiccator before weighing:
dry weight fraction = q of dry sample
g of sample
7.2 Sample Handling
7.2.1 Sludge Samples
7.2.1.1 Weigh out 20 ± 0.5 g of wet sludge with a known
dry-weight fraction (Section 7.1). Place in a 150-mL beaker.
7.2.1.2 Acidify to a pH of 2 with approximately 0.3 mL
concentrated HC1.
7.2.1.3 Add 25 g prepared Mg2S04 • H20 and stir to a
smooth paste.
7.2.1.4 Spread paste on sides of beaker to facilitate
evaporation. Let stand about 15-30 min or until substance is
solidified.
Replacement solvent will be specified in a forthcoming regulation.
9071A - 3 Revision 1
November 1992
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7.2.1.5 Remove solids and grind to fine powder in a
mortar.
7.2.1.6 Add the powder to the paper extraction thimble.
7.2.1.7 Wipe beaker and mortar with pieces of filter
paper moistened with solvent and add to thimble.
7.2.1.8 Fill thimble with glass wool (or glass beads).
7.2.2 Sediment/Soil Samples
7.2.2.1 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.
7.2.2.2 Blend 10 g of the solid sample of known dry
weight fraction 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.
7.3 Extraction
7.3.1 Extract in Soxhlet apparatus using trichlorotrifluorocarbon
at a rate of 20 cycles/hr for 4 hr.
7.3.2 Using grease-free cotton, filter the extract into a pre-
weighed 250-mL boiling flask. Use gloves to avoid adding fingerprints to
the flask.
7.3.3 Rinse flask and cotton with solvent.
7.3.4 Connect the boiling flask to the distilling head and
evaporate the solvent by immersing the lower half of the flask in water at
70°C. Collect the solvent for reuse. A solvent blank should accompany
each analytical batch of samples.
7.3.5 When the temperature in the distilling head reaches 50°C
or the flask appears dry, remove the distilling head. To remove solvent
vapor, sweep out the flask for 15 sec with air by inserting a glass tube
that is connected to a vacuum source. Immediately remove the flask from
the heat source and wipe the outside to remove excess moisture and
fingerprints.
7.3.6 Cool the boiling flask in a desiccator for 30 min and
weigh.
7.3.7 Calculate oil and grease as a percentage of the total dry
solids. Generally:
% of oil and grease = gain in weight of flask (q) x 100
wt. of wet solids (g) x dry weight fraction
9071A - 4 Revision 1
November 1992
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8.0 QUALITY CONTROL
8.1 All quality control data should be maintained and available for
easy reference and inspection. Refer to Chapter One for additional quality
control guidelines.
8.2 Employ a minimum of one blank per analytical batch or twenty
samples, whichever is more frequent, to determine if contamination has occurred.
8.3 Run one matrix duplicate and matrix spike sample every twenty
samples or analytical batch, whichever is more frequent. Matrix duplicates and
spikes are brought through the whole sample preparation and analytical process.
8.3 The use of corn oil is recommended as a reference sample solution.
9.0 METHOD PERFORMANCE
9.1 The two oil and grease methods (Methods 9070 and 9071) in this
manual were tested on sewage by a single laboratory. When 1-liter portions of
the sewage were dosed with 14.0 mg of a mixture of No. 2 fuel oil and Wesson oil,
the recovery was 93%, with a standard deviation of + 0.9 mg/L.
10.0 REFERENCES
1. Blum, K.A. and M.J. Taras, "Determination of Emulsifying Oil in Industrial
Wastewater," JWPCF Research Suppl., 40, R404 (1968).
2. Standard Methods for the Examination of Water and Wastewater, 14th ed.,
p. 515, Method 502A (1975).
9071A - 5 Revision 1
November 1992
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METHOD 9071A
OIL AND GREASE EXTRACTION METHOD FOR SLUDGE SAMPLES
Start
7 2 L 1 Weigh
a sample of
wet sludge
and place in
beaker
7212
Acidify to pH
2
7 2 1 3 Add
and stir
magnesium
sulfate
monohydra te
7 2 1 4 Let
sample
mix lure
solidify
7215
Remove and
grind solids
to a f me
powder
7 1 Determine
dry weight of
sample
Sludge
7221 Decant
wa ter, mix
sample; discard
fo reign ob jects
7222 Blend
with sodium
sulfate, add
to extraction
thimble
9071A - 6
Revision 1
November 1992
-------
METHOD 9071A
OIL AND GREASE EXTRACTION METHOD FOR SLUDGE SAMPLES
(Continued)
7217 Wipe
beaker and
mortar add
to thimble
7 2 1
6 Add
powder to
paper
ex t raction
thimble
7218 Fill
thimble with
glass wool
731 Extract
in Soxhlet
4 hours
7 3 2 Filter
ex tract into
boil ing flask
733 Rinse
flask with
solvent
1
734
Evapora te and
collect
solvent vapor
1
735 Remove
(
736 Cool
boi 1 ing f lask
7 3 7
Calculate %
oil and
grease
STOP
^>
9071A - 7
Revision 1
November 1992
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METHOD 9075
TEST METHOD FOR TOTAL CHLORINE IN NEW AND USED
PETROLEUM PRODUCTS BY X-RAY FLUORESCENCE SPECTROMETRY (XRF)
1.0 SCOPE AND APPLICATION
1.1 This test method covers the determination of total chlorine in new
and used oils, fuels, and related materials, including crankcase, hydraulic,
diesel, lubricating and fuel oils, and kerosene. The chlorine content of
petroleum products is often required prior to their use as a fuel.
1.2 The applicable range of this method is from 200 /zg/g to percent
levels.
2.0 SUMMARY OF METHOD
2.1 A well-mixed sample, contained in a disposable plastic sample cup,
is loaded into an X-ray fluorescence (XRF) spectrometer. The intensities of the
chlorine K alpha and sulfur K alpha lines are measured, as are the intensities
of appropriate background lines. After background correction, the net inten-
sities are used with a calibration equation to determine the chlorine content.
The sulfur intensity is used to correct for absorption by sulfur.
3.0 INTERFERENCES
3.1 Possible interferents include metals, water, and sediment in the oil.
Results of spike recovery measurements and measurements on diluted samples can
be used to check for interferences.
Each sample, or one sample from a group of closely related samples, should
be spiked to confirm that matrix effects are not significant. Dilution of
samples that may contain water or sediment can product incorrect results, so
dilution should be undertaken with caution and checked by spiking. Sulfur
interferes with the chlorine determination, but a correction is made.
Spike recovery measurements of used crankcase oil showed that diluting
samples five to one allowed accurate measurements on approximately 80% of the
samples. The other 20% of the samples were not accurately analyzed by XRF.
3.2 Water in samples absorbs X-rays due to chlorine. For this inter-
ference, using as short an X-ray counting time as possible is beneficial. This
appears to be related to stratification of samples into aqueous and nonaqueous
layers while in the analyzer.
Although a correction for water may be possible, none is currently
available. In general, the presence of any free water as a separate phase or a
water content greater than 25% will reduce the chlorine signal by 50 to 90%.
9075 - 1 Revision 0
November 1992
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4.0 APPARATUS AND MATERIALS
4.1 XRF spectrometer, either energy dispersive or wavelength dispersive.
The instrument must be able to accurately resolve and measure the intensity of
the chlorine and sulfur lines with acceptable precision.
4.2 Disposable sample cups with suitable plastic film such as Mylar*.
5.0 REAGENTS
5.1 Purity of reagents. 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 Mineral oil, mineral spirits or paraffin oil, sulfur and chlorine
free, for preparing standards and dilutions.
5.3 1-Chlorodecane (Aldrich Chemical Co.), 20.1% chlorine, or similar
chlorine compound.
5.4 Di-n-butyl sulfide (Aldrich Chemical Co.), 21.9% sulfur by weight.
5.5 Quality control standards such as the standard reference materials
NBS 1620, 1621, 1622, 1623, and 1624, sulfur in oil standards, and NBS 1818,
chlorine in oil standards.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine.
6.2 The collected sample should be kept headspace free prior to prepara-
tion and analysis to minimize volatilization losses of organic halogens. Because
waste oils may contain toxic and/or carcinogenic substances, appropriate field
and laboratory safety procedures should be followed.
6.3 Laboratory sampling of the sample should be performed on a well-mixed
sample of oil. The mixing should be kept to a minimum and carried out as nearly
headspace free as possible to minimize volatilization losses of organic halogens.
6.4 Free water, as a separate phase, should be removed and cannot be
analyzed by this method.
7.0 PROCEDURE
7.1 Calibration and standardization.
7.1.1 Prepare primary calibration standards by diluting the
chlorodecane and n-butyl sulfide with mineral spirits or similar material.
9075 - 2 Revision 0
November 1992
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7.1.2 Prepare working calibration standards that contain sulfur,
chlorine, or both according to the following table:
Cl:
S:
1.
2.
3.
4.
500, 1,000, 2,000, 4,000, and 6,000 jug/9
0.5, 1.0, and 1.5% sulfur
0.5% S, 1,000 M9/9 Cl
0.5% S, 4,000 M9/9 Cl
1.0% S, 500 ng/g Cl
1.0% s, 2,000 jug/g ci
5. 1.0% s, 6,000 Mg/g ci
6. 1.5% s, 1,000 jug/g ci
7. 1.5% S, 4,000 Atg/g Cl
8. 1.5% s, 6,000 jug/g ci
Once the correction factor for sulfur interference with chlorine is
determined, fewer standards may be required.
7.1.3 Measure the intensity of the chlorine K alpha line and the
sulfur K alpha line as well as the intensity of a suitably chosen
background. Based on counting statistics, the relative standard deviation
of each peak measurement should be 1% or less.
7.1.4 Determine the net chlorine and sulfur intensities by
correcting each peak for background. Do this for all of the calibration
standards as well as for a paraffin blank.
7.1.5 Obtain a linear calibration curve for sulfur by performing
a least squares fit of the net sulfur intensity to the standard concentra-
tions, including the blank. The chlorine content of a standard should
have little effect on the net sulfur intensity.
7.1.6 The calibration equation for chlorine must include a
correction term for the sulfur concentration. A suitable equation
follows:
Cl = (ml + b) (1 + k*S) (1)
where:
I = net chlorine intensity
m, b, k* = adjustable parameters.
Using a least squares procedure, the above equation or a suitable
substitute should be fitted to the data. Many XRF instruments are
equipped with suitable computer programs to perform this fit. In any
case, the resulting equation should be shown to be accurate by analysis of
suitable standard materials.
7.2 Analysis.
7.2.1 Prepare a calibration curve as described in Step 7.1. By
periodically measuring a very stable sample containing both sulfur and
chlorine, it may be possible to use the calibration equations for more
than 1 day. During each day, the suitability of the calibration curve
should be checked by analyzing standards.
9075 - 3
Revision 0
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7.2.2 Determine the net chlorine and sulfur intensities for a
sample in the same manner as was done for the standards.
7.2.3 Determine the chlorine and sulfur concentrations of the
samples from the calibration equations. If the sample concentration for
either element is beyond the range of the standards, the sample should be
diluted with mineral oil and reanalyzed.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 One sample in ten should be analyzed in triplicate and the relative
standard deviation reported. For each triplicate, a separate preparation should
be made, starting from the original sample.
8.3 Each sample, or one sample in ten from a group of similar samples,
should be spiked with the elements of interest by adding a known amount of
chlorine or sulfur to the sample. The spiked amount should be between 50% and
200% of the sample concentration, but the minimum addition should be at least
five times the limit of detection. The percent recovery should be reported and
should be between 80% and 120%. Any sample suspected of containing >25% water
should also be spiked with organic chlorine.
8.4 Quality control standard check samples should be analyzed every day
and should agree within 10% of the expected value of the standard.
9.0 METHOD PERFORMANCE
9.1 These data are based on 47 data points obtained by seven laboratories
who each analyzed four used crankcase oils and three fuel oil blends with
crankcase in duplicate. A data point represents one duplicate analysis of a
sample. Two data points were determined to be outliers and are not included in
these results.
9.2 Precision. The precision of the method as determined by the
statistical examination of interlaboratory test results is as follows:
Repeatability - The difference between successive results obtained by the
same operator with the same apparatus under constant operating conditions on
identical test material would exceed, in the long run, in the normal and correct
operation of the test method, the following values only in 1 case in 20 (see
Table 1):
Repeatability - 5.72 /x*
*where x is the average of two results in M9/9-
Reproducibility - The difference between two single and independent
results obtained by different operators working in different laboratories
9075 - 4 Revision 0
November 1992
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on identical test material would exceed, in the long run, the following
values only in 1 case in 20:
Reproducibility - 9.83 /x*
*where x is the average value of two results in M9/9-
9.3 Bias. The bias of this test method varies with concentration, as
shown in Table 2:
Bias = Amount found - Amount expected.
10.0 REFERENCE
1. Gaskill, A.; Estes, E.D.; Hardison, D.L.; and Myers, I.E. Validation of
Methods for Determining Chlorine in Used Oils and Oil Fuels. Prepared for
U.S. Environmental Protection Agency, Office of Solid Waste. EPA Contract
No. 68-01-7075, WA 80. July 1988.
9075 - 5 Revision 0
November 1992
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TABLE 1. REPEATABILITY AND REPRODUCIBILITY
FOR CHLORINE IN USED OILS BY
X-RAY FLUORESCENCE SPECTROMETRY
Average value, Repeatability, Reproducibility,
M9/9 M9/9 M9/9
500 128 220
1,000 181 311
1,500 222 381
2,000 256 440
2,500 286 492
3,000 313 538
TABLE 2. RECOVERY AND BIAS DATA FOR CHLORINE IN
USED OILS BY X-RAY FLUORESCENCE SPECTROMETRY
Amount Amount
expected, found, Bias, Percent
M9/9 M9/9 M9/9 bias
320 278 -42 -13
480 461 -19 -4
920 879 -41 -4
1,498 1,414 -84 -6
1,527 1,299 -228 -15
3,029 2,806 -223 -7
3,045 2,811 -234 -8
9075 - 6 Revision 0
November 1992
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METHOD 9075
TEST METHOD FOR TOTAL CHLORINE IN NEW AND USED
PETROLEUM PRODUCTS BY X-RAY FLUORESCENCE SPECTROMETRY
(XRF)
START
711-712
Prepare calibration
standards
713 Measure
intensi ty of
3tanda rds and
backgr ound
1 4 Determine net
in tens11 y for
3 tandards and a
pa raff in blank
715-716
Cons t ruct
calibration curves
for sulfur and
chi or me
7 2 1 Check
calibration curves
periodica11y
throughout the day
722 Determine net
chlorine and sulfur
intensities for
samp 1e
7 2 3 De termine
chlorine and sulfur
concentrations from
calibration curves
7 2 3 Dilute sample
with minera1 oil
9075 - 7
Revision
November
0
1992
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METHOD 9076
TEST METHOD FOR TOTAL CHLORINE IN NEW AND USED PETROLEUM
PRODUCTS BY OXIDATIVE COMBUSTION AND MICROCOULOMETRY
1.0 SCOPE AND APPLICATION
1.1 This test method covers the determination of total chlorine in new
and used oils, fuels and related materials, including crankcase, hydraulic,
diesel, lubricating and fuel oils, and kerosene by oxidative combustion and
microcoulometry. The chlorine content of petroleum products is often required
prior to their use as a fuel.
1.2 The applicable range of this method is from 10 to 10,000 p,g/g
chlorine.
2.0 SUMMARY OF METHOD
2.1 The sample is placed in a quartz boat at the inlet of a high-
temperature quartz combustion tube. An inert carrier gas such as argon, carbon
dioxide, or nitrogen sweeps across the inlet while oxygen flows into the center
of the combustion tube. The boat and sample are advanced into a vaporization
zone of approximately 300°C to volatilize the light ends. Then the boat is
advanced to the center of the combustion tube, which is at 1,000'C. The oxygen
is diverted to pass directly over the sample to oxidize any remaining refractory
material. All during this complete combustion cycle, the chlorine is converted
to chloride and oxychlorides, which then flow into an attached titration cell
where they quantitatively react with silver ions. The silver ions thus consumed
are coulometrically replaced. The total current required to replace the silver
ions is a measure of the chlorine present in the injected samples.
2.2 The reaction occurring in the titration cell as chloride enters is:
Cl" + Ag+ --> AgCl (1)
The silver ion consumed in the above reaction is generated coulometrically
thus:
Ag° > Ag+ + e" (2)
2.3 These microequivalents of silver are equal to the number of micro-
equivalents of titratable sample ion entering the titration cell.
3.0 INTERFERENCES
3.1 Other titratable halides will also give a positive response. These
titratable halides include HBr and HI (HOBr + HOI do not precipitate silver).
Because these oxyhalides do not react in the titration cell, approximately 50%
microequivalent response is detected from bromine and iodine.
3.2 Fluorine as fluoride does not precipitate silver, so it is not an
interferant nor is it detected.
9076 - 1 Revision 0
November 1992
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3.3 This test method is applicable in the presence of total sulfur
concentrations of up to 10,000 times the chlorine level.
4.0 APPARATUS AND MATERIALS1
4.1 Combustion furnace. The sample should be oxidized in an electric
furnace capable of maintaining a temperature of 1,000'C to oxidize the organic
matrix.
4.2 Combustion tube, fabricated from quartz and constructed so that a
sample, which is vaporized completely in the inlet section, is swept into the
oxidation zone by an inert gas where it mixes with oxygen and is burned. The
inlet end of the tube connects to a boat insertion device where the sample can
be placed on a quartz boat by syringe, micropipet, or by being weighed
externally. Two gas ports are provided, one for an inert gas to flow across the
boat and one for oxygen to enter the combustion tube.
4.3 Microcoulometer, Stroehlein Coulomat 702 CL or equivalent, having
variable gain and bias control, and capable of measuring the potential of the
sensing-reference electrode pair, and comparing this potential with a bias
potential, and applying the amplified difference to the working-auxiliary
electrode pair so as to generate a titrant. The microcoulometer output signal
shall be proportional to the generating current. The microcoulometer may have
a digital meter and circuitry to convert this output signal directly to nanograms
or micrograms of chlorine or micrograms per gram chlorine.
4.4 Titration cell. Two different configurations have been applied to
coulometrically titrate chlorine for this method.
4.4.1 Type I uses a sensor-reference pair of electrodes to detect
changes in silver ion concentration and a generator anode-cathode pair of
electrodes to maintain constant silver ion concentration and an inlet for
a gaseous sample from the pyrolysis tube. The sensor, reference, and
anode electrodes are silver electrodes. The cathode electrode is a
platinum wire. The reference electrode resides in a saturated silver
acetate half-cell. The electrolyte contains 70% acetic acid in water.
4.4.2 Type II uses a sensor-reference pair of electrodes to
detect changes in silver ion concentration and a generator anode-cathode
pair of electrodes to maintain constant silver ion concentration, an inlet
for a gaseous sample that passes through a 95% sulfuric acid dehydrating
tube from the pyrolysis tube, and a sealed two-piece titration cell with
an exhaust tube to vent fumes to an external exhaust. All electrodes can
be removed and replaced independently without reconstructing the cell
assembly. The anode electrode is constructed of silver. The cathode
electrode is constructed of platinum. The anode is separated from the
cathode by a 10% KNO, agar bridge, and continuity is maintained through an
aqueous 10% KN03 salt bridge. The sensor electrode is constructed of
1Three commercial analyzers fulfill the requirements for apparatus Steps
4.1 through 4.4 and have been found satisfactory for this method. They are
the two Dohrmann Models DX-20B and MCTS-20 and Mitsubishi Model TSX-10
available from Cosa Instrument.
9076 - 2 Revision 0
November 1992
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silver. The reference electrode is a silver/silver chloride ground glass
sleeve, double- junction electrode with aqueous 1M KN03 in the outer chamber
and aqueous 1M KC1 in the inner chamber.
4.5 Sampling syringe, a microliter syringe of 10 p.1 capacity capable of
accurately delivering 2 to 5 /iL of a viscous sample into the sample boat.
4.6 Micropipet, a positive displacement micropipet capable of accurately
delivering 2 to 5 pi of a viscous sample into the sample boat.
4.7 Analytical balance. When used to weigh a sample of 2 to 5 mg onto
the boat, the balance shall be accurate to ± 0.01 mg. When used to determine the
density of the sample, typically 8 g per 10 ml, the balance shall be accurate to
± 0.1 g.
4.8 Class A volumetric flasks: 100 ml.
5.0 REAGENTS
5.1 Purity of Reagents. 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 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Acetic acid, CH3C02H. Glacial.
5.4 Isooctane, (CH3)2CHCH2C(CH3)3 (2,2,4-Trimethylpentane) .
5.5 Chlorobenzene, C6H5C1 .
5.6 Chlorine, standard stock solution - 10,000 ng Cl//iL, weigh
accurately 3.174 g of chlorobenzene into 100-mL Class A volumetric flask. Dilute
to the mark with isooctane.
5.7 Chlorine, standard solution. 1,000 ng Cl/ML, pipet 10.0 ml of
chlorine stock solution (Step 5.6) into a 100-mL volumetric flask and dilute to
volume with isooctane.
5.8 Argon, helium, nitrogen, or carbon dioxide, high-purity grade (HP)
used as the carrier gas. High-purity grade gas has a minimum purity of 99.995%.
5.9 Oxygen, high-purity grade (HP), used as the reactant gas.
5.10 Gas regulators. Two-stage regulator must be used on the reactant and
carrier gas.
5.11 Cell Type 1.
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5.11.1 Cell electrolyte solution. 70% acetic acid: combine 300
ml reagent water with 700 ml acetic acid (Step 5.3) and mix well.
5.11.2 Silver acetate, CH3C02Ag. Powder purified for saturated
reference electrode.
5.12 Cell Type 2.
5.12.1 Sodium acetate, CH3C02Na.
5.12.2 Potassium nitrate, KN03.
5.12.3 Potassium chloride, KC1.
5.12.4 Sulfuric acid (concentrated), H2S04.
5.12.5 Agar, (jelly strength 450 to 600 g/cm2).
5.12.6 Cell electrolyte solution - 85% acetic acid: combine 150
ml reagent water with 1.35 g sodium acetate (Step 5.12.1) and mix well;
add 850 ml acetic acid (Step 5.3) and mix well.
5.12.7 Dehydrating solution - Combine 95 ml sulfuric acid (Step
5.12.4) with 5 ml reagent water and mix well.
CAUTION: This is an exothermic reaction and may proceed with bumping unless
controlled by the addition of sulfuric acid. Slowly add sulfuric
acid to reagent water. Do not add water to sulfuric acid.
5.12.8 Potassium nitrate (10%), KNO?. Add 10 g potassium nitrate
(Step 5.12.2) to 100 ml reagent water and mix well.
5.12.9 Potassium nitrate (1M), KN03. Add 10.11 g potassium
nitrate (Step 5.12.2) to 100 ml reagent water and mix well.
5.12.10 Potassium chloride (1M), KC1. Add 7.46 g potassium
chloride (Step 5.12.3) to 100 ml reagent water and mix well.
5.12.11 Agar bridge solution - Mix 0.7 g agar (Step 5.12.5), 2.5g
potassium nitrate (Step 5.12.2), and 25 ml reagent water and heat to
boiling.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine.
6.2 Because the collected sample will be analyzed for total halogens, it
should be kept headspace free and refrigerated prior to preparation and analysis
to minimize volatilization losses of organic halogens. Because waste oils may
contain toxic and/or carcinogenic substances, appropriate field and laboratory
safety procedures should be followed.
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6.3 Laboratory subsampling of the sample should be performed on a well-
mixed sample of oil.
7.0 PROCEDURES
7.1 Preparation of apparatus.
7.1.1 Set up the analyzer as per the equipment manufacturer's
instructions.
7.1.2 Typical operating conditions: Type 1.
Furnace temperature 1,000'C
Carrier gas flow 43 cm /min
Oxygen gas flow 160 cm3/min
Coulometer
Bias 250 mV
Gain 25%
7.1.3 Typical operating conditions: Type 2.
Furnace temperature H-l 850°C
H-2 1,000°C
Carrier gas flow 250 cm3/min
Oxygen gas flow 250 cm /min
Coulometer
End point potential (bias) 300 mV
Gain G-l 1.5 coulombs/A mV
G-2 3.0 coulombs/A mV
G-3 3.0 coulombs/A mV
ES-1 (range 1) 25 mV
ES-2 (range 2) 30 mV
NOTE: Other conditions may be appropriate. Refer to the instrumentation manual.
7.2 Sample introduction.
7.2.1 Carefully fill a 10-^L syringe with 2 to 5 /xL of sample
depending on the expected concentration of total chlorine. Inject the
sample through the septum onto the cool boat, being certain to touch the
boat with the needle tip to displace the last droplet.
7.2.2 For viscous samples that cannot be drawn into the syringe
barrel, a positive displacement micropipet may be used. Here, the 2-5 /xL
of sample is placed on the boat from the micropipet through the opened
hatch port. The same technique as with the syringe is used to displace
the last droplet into the boat. A tuft of quartz wool in the boat can aid
in completely transferring the sample from the micropipet into the boat.
NOTE: Dilution of samples to reduce viscosity is not recommended due to
uncertainty about the solubility of the sample and its chlorinated
constituents. If a positive displacement micropipet is not available,
dilution may be attempted to enable injection of viscous samples.
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7.2.3 Alternatively, the sample boat may be removed from the
instrument and tared on an analytical balance. A sample of 2-5 mg is
accurately weighed directly into the boat and the boat and sample returned
to the inlet of the instrument.
2-5 IJ.L = 2-5 mg
NOTE: Sample dilution may be required to ensure that the titration system is not
overloaded with chlorine. This will be somewhat system dependent and
should be determined before analysis is attempted. For example, the MCTS-
20 can titrate up to 10,000 ng chlorine in a single injection or weighed
sample, while the DX-20B has an upper limit of 50,000 ng chlorine. For 2
to 5 juL sample sizes, these correspond to nominal concentrations in the
sample of 800 to 2,000 ;ug/g and 4,000 to 10,000 jug/g, respectively. If
the system is overloaded, especially with inorganic chloride, residual
chloride may persist in the system and affect results of subsequent
samples. In general, the analyst should ensure that the baseline returns
to normal before running the next sample. To speed baseline recovery, the
electrolyte can be drained from the cell and replaced with fresh
electrolyte.
NOTE: To determine total chlorine, do not extract the sample either with reagent
water or with an organic solvent such as toluene or isooctane. This may
lower the inorganic chlorine content as well as result in losses of
volatile solvents.
7.2.4 Follow the manufacturer's recommended procedure for moving
the sample and boat into the combustion tube.
7.3 Calibration and standardization.
7.3.1 System recovery - The fraction of chlorine in a standard
that is titrated should be verified every 4 hours by analyzing the
standard solution (Step 5.7). System recovery is typically 85% or better.
The pyrolysis tube should be replaced whenever system recovery drops below
75%.
NOTE: The 1,000 ng/g system recovery sample is suitable for all systems except
the MCTS-20 for which a 100 /xg/g sample should be used.
7.3.2 Repeat the measurement of this standard at least three
times.
7.3.3 System blank - The blank should be checked daily with
isooctane. It is typically less than 1 /ug/g chlorine. The system blank
should be subtracted from both samples and standards.
7.4 Calculations.
7.4.1 For systems that read directly in mass units of chloride,
the following equations apply:
Chlorine, M9/g (wt/wt) = ' B &
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or
where:
Display
V
D
RF
B
Chlorine, /xg/g (wt/wt)
Display
- B
(4)
Integrated value in nanograms (when the integrated values are
displayed in micrograms, they must be multiplied by 103)
DisplayB = blank measurement Displays = sample measurement
Volume of sample injected in microliters
VB = blank volume
Vs = sample volume
Density of sample, grams per cubic centimeters
DB = blank density Ds = sample density
Recovery factor = ratio of chlorine
determined in standard minus the system
blank, divided by known standard content
System blank, ng/g chlorine
Found - Blank
Known
Display,,
M
Mass of sample, mg
7.4.2 Other systems internally compensate for recovery factor,
volume, density, or mass and blank, and thus read out directly in parts
per million chlorine units. Refer to instrumentation manual.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
8.2 Each sample should be analyzed twice. If the results do not agree
to within 10%, expressed as the relative percent difference of the results,
repeat the analysis.
8.3 Analyze matrix spike and matrix spike duplicates - spike samples with
a chlorinated organic at a level of total chlorine commensurate with the levels
being determined. The spike recovery should be reported and should be between
80 and 120% of the expected value. Any sample suspected of containing >25% water
should also be spiked with organic chlorine.
9.0 METHOD PERFORMANCE
9.1 These data are based on 66 data points obtained by 10 laboratories
who each analyzed four used crankcase oils and three fuel oil blends with
crankcase in duplicate. A data point represents one duplicate analysis of a
sample. One laboratory and four additional data points were determined to be
outliers and are not included in these results.
9.2 Precision. The precision of the method as determined by the
statistical examination of interlaboratory test results is as follows:
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Repeatability - The difference between successive results obtained by the
same operator with the same apparatus under constant operating conditions on
identical test material would exceed, in the long run, in the normal and correct
operation of the test method the following values only in 1 case in 20 (see Table
1):
Repeatability - 0.137 x*
*where x is the average of two results in Mg/9-
Reproducibilitv - The difference between two single and independent
results obtained by different operators working in different laboratories
on identical test material would exceed, in the long run, the following
values only in 1 case in 20:
Reproducibility - 0.455 x*
*where x is the average value of two results in /ig/g.
9.3 Bias. The bias of this test method varies with concentration, as
shown in Table 2:
Bias = Amount found - Amount expected
10.0 REFERENCE
1. Gaskill, A.; Estes, E.D.; Hardison, D.L.; and Myers, I.E. "Validation of
Methods for Determining Chlorine in Used Oils and Oil Fuels." Prepared
for U.S. Environmental Protection Agency, Office of Solid Waste. EPA
Contract No. 68-01-7075, WA80. July 1988.
2. Rohrobough, W.G.; et al. Reagent Chemicals, American Chemical Society
Specifications, 7th ed.; American Chemical Society: Washington, DC, 1986.
3. Standard Instrumentation, 3322 Pennsylvania Avenue, Charleston, WV 25302.
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TABLE 1.
REPEATABILITY AND REPRODUCIBILITY FOR CHLORINE IN
USED OILS BY MICROCOULOMETRIC TITRATION
Average value Repeatability, Reproducibility,
M9/9 M9/9 M9/9
500
1,000
1,500
2,000
2,500
3,000
69
137
206
274
343
411
228
455
683
910
1,138
1,365
TABLE 2.
RECOVERY AND BIAS DATA FOR CHLORINE IN USED OILS
BY MICROCOULOMETRIC TITRATION
Amount
expected,
M9/9
320
480
920
1,498
1,527
3,029
3,045
Amount
found
M9/9
312
443
841
1,483
1,446
3,016
2,916
Bias,
M9/9
-8
-37
-79
-15
-81
-13
-129
Percent
bias
-3
-8
-9
-1
-5
0
-4
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METHOD 9076
TEST METHOD FOR TOTAL CHLORINE IN NEW AND USED PETROLEUM
PRODUCTS BY OXIDATIVE COMBUSTION AND MICROCOULOMETRY
722 Inject
sample into
cool boat
with
micropipet
724 Move
sample and
boat into
combua to. on
tube
721 Inject
sample into
cool boat
with syringe
7 3 1 Verify
sys tern
recovery
every 4 hours
732 Repeat
s tandard
measurement
at least
three times
733 Check
sys tern blank
daily with
isooctane
7 4 Calculate
chl o r me
concent ration
STOP
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METHOD 9077
TEST METHODS FOR TOTAL CHLORINE IN NEW AND USED PETROLEUM
PRODUCTS (FIELD TEST KIT METHODS)
1.0 SCOPE AND APPLICATION
1.1 The method may be used to determine if a new or used petroleum
product meets or exceeds requirements for total halogen measured as chloride.
An analysis of the chlorine content of petroleum products is often required prior
to their use as a fuel. The method is specifically designed for used oils
permitting onsite testing at remote locations by nontechnical personnel to avoid
the delays for laboratory testing.
1.2 In these field test methods, the entire analytical sequence,
including sampling, sample pretreatment, chemical reactions, extraction, and
quantification, are combined in a single kit using predispensed and encapsulated
reagents. The overall objective is to provide a simple, easy to use procedure,
permitting nontechnical personnel to perform a test with analytical accuracy
outside of a laboratory environment in under 10 minutes. One of the kits is
preset at 1,000 ng/g total chlorine to meet regulatory requirements for used
oils. The other kits provide quantitative results over a range of 750 to 7,000
and 300 to 4,000
2.0 SUMMARY OF METHOD
2.1 The oil sample (around 0.4 g by volume) is dispersed in a solvent
and reacted with a mixture of metallic sodium catalyzed with naphthalene and
diglyme at ambient temperature. This process converts all organic halogens to
their respective sodium halides. All halides in the treated mixture, including
those present prior to the reaction, are then extracted into an aqueous buffer,
which is then titrated with mercuric nitrate using diphenyl carbazone as the
indicator. The end point of the titration is the formation of the blue-violet
mercury diphenyl carbazone complex. Bromide and iodide are titrated and reported
as chloride.
2.2 Reagent quantities are preset in the fixed end point kit (Method
A) so that the color of the solution at the end of the titration indicates
whether the sample is above 1,000 Mg/g chlorine (yellow) or below 1,000 /ug/g
chlorine (blue).
2.3 The first quantitative kit (Method B) involves a reverse titration
of a fixed volume of mercuric nitrate with the extracted sample such that the end
point is denoted by a change from blue to yellow in the titration vessel over the
range of the kit (750 to 7,000 Mg/g)- The final calculation is based on the
assumption that the oil has a specific gravity of 0.9.
2.4 The second quantitative kit (Method C) involves a titration of the
extracted sample with mercuric nitrate by means of a 1-mL microburette such that
the end point is denoted by a change from pale yellow to red-violet over the
range of the kit (300 to 4,000 ng/g). The concentration of chlorine in the
original oil is then read from a scale on the microburette.
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NOTE: Warning—All reagents are encapsulated or contained within
ampoules. Strict adherence to the operational procedures included
with the kits as well as accepted safety procedures (safety glasses
and gloves) should be observed.
NOTE: Warning—When crushing the glass ampoules, press firmly in the
center of the ampoule once. Never attempt to recrush broken glass
because the glass may come through the plastic and cut fingers.
NOTE: Warning—In case of accidental breakage onto skin or clothing, wash
with large amounts of water. All the ampoules are poisonous and
should not be taken internally.
NOTE: Warning—The gray ampoules contain metallic sodium. Metallic
sodium is a flammable water-reactive solid.
NOTE: Warning—Do not ship kits on passenger aircraft. Dispose of used
kits properly.
NOTE: Caution—When the sodium ampoule in either kit is crushed, oils
that contain more than 25% water will cause the sample to turn
clear to light gray. Under these circumstances, the results may
be biased excessively low and should be disregarded.
3.0 INTERFERENCES
3.1 Free water, as a second phase, should be removed. However, this
second phase can be analyzed separately for chloride content if desired.
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METHOD A
FIXED END POINT TEST KIT METHOD
4.0A APPARATUS AND MATERIALS
4.1A The CLOR-D-TECT 10001 is a complete self-contained kit. It
includes: a sampling tube to withdraw a fixed sample volume for analysis; a
polyethylene test tube #1 into which the sample is introduced for dilution and
reaction with metallic sodium; and a polyethylene tube #2 containing a buffered
aqueous extractant, the mercuric nitrate titrant, and diphenyl carbazone
indicator. Included are instructions to conduct the test and a color chart to
aid in determining the end point.
5.0A REAGENTS
5.1A Purity of reagents. 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 determina-
tion.
5.2A All necessary reagents are contained within the kit.
5.3A The kit should be examined upon opening to see that all of the
components are present and that all the ampoules (4) are in place and not
leaking. The liquid in Tube #2 (yellow cap) should be approximately 1/2 in.
above the 5-mL line and the tube should not be leaking. The ampoules are not
supposed to be completely full.
6.0A SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1A All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine.
6.2A Because the collected sample will be analyzed for total halogens,
it should be kept headspace free and refrigerated prior to preparation and
analysis to minimize volatilization losses of organic halogens. Because waste
oils may contain toxic and/or carcinogenic substances, appropriate field and
laboratory safety procedures should be followed.
7.0A PROCEDURE
7.1A Preparation. Open analysis carton, remove contents, mount plastic
test tubes in the provided holder. Remove syringe and glass sampling capillary
from foil pouch.
Available from Dexsil Corporation, One Hamden Park Drive, Hamden, CT 06517.
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NOTE: Perform the test in a warm, dry area with adequate light. In cold
weather, a truck cab is sufficient. If a warm area is not available, Step
7.3A should be performed while warming Tube #1 in palm of hand.
7.2A Sample introduction. Remove white cap from Tube #1. Using the
plastic syringe, slowly draw the oil up the capillary tube until it reaches the
flexible adapter tube. Wipe excess oil from the tube with the provided tissue,
keeping capillary vertical. Position capillary tube into Tube #1, and detach
adapter tubing, allowing capillary to drop to the bottom of the tube. Replace
white cap on tube. Crush the capillary by squeezing the test tube several times,
being careful not to break the glass reagent ampoules.
7.3A Reaction. Break the lower (colorless) capsule containing the clear
diluent solvent by squeezing the sides of the test tube. Mix thoroughly by
shaking the tube vigorously for 30 seconds. Crush the upper grey ampoule
containing metallic sodium, again by squeezing the sides of the test tube. Shake
vigorously for 20 seconds. Allow reaction to proceed for 60 seconds, shaking
intermittently several times while timing with a watch.
NOTE: Caution—Always crush the clear ampoule in each tube first. Otherwise,
stop the test and start over using another complete kit. False (low)
results may occur and allow a contaminated sample to pass without
detection if clear ampoule is not crushed first.
7.4A Extraction. Remove caps from both tubes. Pour the clear buffered
extraction solution from Tube #2 into Tube #1. Replace the white cap on Tube #1,
and shake vigorously for 10 seconds. Vent tube by partially unscrewing the
dispenser cap. Close cap securely, and shake for an additional 10 seconds. Vent
again, tighten cap, and stand tube upside down on white cap. Allow phases to
separate for 2 minutes.
7.5A Analysis. Put filtration funnel into Tube #2. Position Tube #1
over funnel and open nozzle on dispenser cap. Squeeze the sides of Tube #1 to
dispense the clear aqueous lower phase through the filter into Tube #2 to the 5-
mL line on Tube #2. Remove the filter funnel. Replace the yellow cap on Tube
#2 and close the nozzle on the dispenser cap. Break the colorless lower capsule
containing mercuric nitrate solution by squeezing the sides of the tube, and
shake for 10 seconds. Then break the upper colored ampoule containing the
diphenylcarbazone indicator, and shake for 10 seconds. Observe color
immediately.
7.6A Interpretation of results
7.6.1A Because all reagent levels are preset, calculations are not
required. A blue solution in Tube #2 indicates a chlorine content in the
original oil of less than 1,000 M9/9> and a yellow color indicates that
the chlorine concentration is greater than 1,000 M9/9- Refer to the color
chart enclosed with the kit in interpreting the titration end point.
7.6.2A Report the results as < or > 1,000 ^9/9 chlorine in the oil
sample.
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8.0A QUALITY CONTROL
8.1A Refer to Chapter One for specific quality control procedures.
8.2A Each sample should be tested two times. If the results do not
agree, then a third test must be performed. Report the results of the two that
agree.
9.0A METHOD PERFORMANCE
9.1A No formal statement is made about either the precision or bias of
the overall test kit method for determining chlorine in used oil because the
result merely states whether there is conformance to the criteria for success
specified in the procedure, (i.e., a blue or yellow color in the final solution).
In a collaborative study, eight laboratories analyzed four used crankcase oils
and three fuel oil blends with crankcase in duplicate using the test kit. Of the
resulting 56 data points, 3 resulted in incorrect classification of the oil's
chlorine content (Table 1). A data point represents one duplicate analysis of
a sample. There were no disagreements within a laboratory on any duplicate
determinations.
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TABLE 1.
PRECISION AND BIAS INFORMATION FOR METHOD A-
FIXED END POINT TEST KIT METHOD
Expected
concentration,
M9/9
Percent agreement
Expected results, Percent
correct8 Within Between
320
480
920
1,498
1,527
3,029
3,045
< 1,000
< 1,000
< 1,000
> 1,000
> 1,000
> 1,000
> 1,000
100
100
100
87
75
100
100
100
100
100
100
100
100
100
100
100
100
87
75
100
100
aPercent correct—percent correctly identified as above or below
1,000 ng/g.
bPercent agreement--percent agreement within or between laboratories.
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START
METHOD 9077 A
FIXED END POINT TEST KIT METHOD
7 1ft Open test bit
7 2A Draw oil into
capillary tuba,
ramov* excess oil.
drop capillary tuba
into Tuba fl and
cap Tuba f\. crush
capillary tuba
7 3A Braak
colorless capsule,
miM. crush gray
capsule, mm. allow
reaction to proceed
for 60 sec
7 4A Pour Tub. /2
solution into Tub*
/I, mix, vent;
allow phaa«» to
««parat«
7 SA filter aqueous
lower phase in Tube
fl into Tube t2.
remove filter
funnel, break
colorless capsule,
miK, break upper
colored capsule,
mm. observe color
761 Chlorine
content is > 1000
ug/g
761 Chlorine
content n < 1000
ug/g
762 Report
results
STOP
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METHOD B
REVERSE TITRATION QUANTITATIVE END POINT TEST KIT METHOD
4.OB APPARATUS AND MATERIALS
4.IB QuantiClor2 kit components (see Figure 1).
4.1.IB Plastic reaction bottle: 1 oz, with flip-top dropper cap
and a crushable glass ampoule containing sodium.
4.1.2B Plastic buffer bottle: contains 9.5 ml of aqueous buffer
solution.
4.1.3B Titration vial: contains buffer bottle and indicator-
impregnated paper.
4.1.4B Glass vial: contains 2.0 ml of solvents.
4.1.5B Micropipet and plunger, 0.25 ml.
4.1.6B Activated carbon filtering column.
4.1.7B Titret and valve assembly.
4.2B The reagents needed for the test are packaged in disposable
containers.
4.3B The procedure utilizes a Titret. Titrets* are hand-held,
disposable cells for titrimetric analysis. A Titret is an evacuated glass
ampoule (13 mm diameter) that contains an exact amount of a standardized liquid
titrant^ A flexible valve assembly is attached to the tip of the ampoule.
Titrets* employ the principle of reverse titration; that is, small doses of
sample are added to the titrant to the appearance of the end point color. The
color change indicates that the equivalency point has been reached. The flow of
the sample into the Titret may be controlled by using an accessory called a
Titrettor™.
5.OB REAGENTS
5.IB The crushable glass ampoule, which is inside the reaction bottle,
contains 85 mg of metallic sodium in a light oil dispersion.
5.2B The buffer bottle contains 0.44 g of NaH2P04 • 2H20 and 0.32 ml of
HN03 in distilled water.
5.3B The glass vial contains 770 mg Stoddard Solvent (CAS No. 8052-41-
3), 260 mg toluene, 260 mg butyl ether, 260 mg diglyme, 130 mg naphthalene, and
70 mg demulsifier.
2Quanti-Chlor Kit, Titrets*, and Titrettor™ are manufactured by Chemetrics,
Inc., Calverton, VA 22016. U.S. Patent No. 4,332,769.
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5.4B The Titret contains 1.12 mg mercuric nitrate in distilled water.
5.5B The indicator-impregnated paper contains approximately 0.3 mg of
diphenylcarbazone and 0.2 mg of brilliant yellow.
6.OB SAMPLE COLLECTION, PRESERVATION, AND HANDLING
See Section 6.0A of Method A.
7.OB PROCEDURE
7.IB Shake the glass vial and pour its contents into the reaction
bottle.
7.2B Fill the micropipet with a well-shaken oil sample by pulling the
plunger until its top edge is even with the top edge of the micropipet. Wipe off
the excess oil and transfer the sample into the reaction bottle (see Figure 2.1).
7.3B Gently squeeze most of the air out of the reaction bottle (see
Figure 2.2). Cap the bottle securely, and shake vigorously for 30 seconds.
7.4B Crush the sodium ampoule by pressing against the outside wall of
the reaction bottle (see Figure 2.3).
CAUTION: Samples containing a high percentage of water will generate heat
and gas, causing the reaction bottle walls to expand. To release
the gas, briefly loosen the cap.
7.5B Shake the reaction bottle vigorously for 30 seconds.
7.6B Wait 1 minute. Shake the reaction bottle occasionally during this
time.
7.7B Remove the buffer bottle from the titration vial, and slowly pour
its contents into the reaction bottle (see Figure 2.4).
7.8B Cap the reaction bottle and shake gently for a few seconds. As
soon as the foam subsides, release the gas by loosening the cap. Tighten the
cap, and shake vigorously for 30 seconds. As before, release any gas that has
formed, then turn the reaction bottle upside down (see Figure 2.5).
7.9B Wait 1 minute.
7.10B While holding the filtering column in a vertical position, remove
the plug. Gently tap the column to settle the carbon particles.
7.11B Keeping the reaction bottle upside down, insert the flip top into
the end of the filtering column and position the column over the titration vial
(see Figure 2.6). Slowly squeeze the lower aqueous layer out of the reaction
bottle and into the filtering column. Keep squeezing until the first drop of oil
is squeezed out.
9077 - 9 Revision 0
November 1992
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NOTE: Caution--The aqueous layer should flow through the filtering column into
the titration vial in about 1 minute. In rare cases, it may be necessary
to gently tap the column to begin the flow. The indicator paper should
remain in the titration vial.
7.12B Cap the titration vial and shake it vigorously for 10 seconds.
7.13B Slide the flexible end of the valve assembly over the tapered tip
of the Titret so that it fits snugly (see Figure 3.1).
7.14B Lift (see Figure 3.2) the control bar and insert the assembled
Titret into the Titrettor™.
7.15B Hold the Titrettor™ with the sample pipe in the sample, and press
the control bar to snap the pre-scored tip of the Titret (see Figure 3.3).
NOTE: Caution—Because the Titret is sealed under vacuum, the fluid inside may
be agitated when the tip snaps.
7.16B With the tip of the sample pipe in the sample, briefly press the
control bar to pull in a SMALL amount of sample (see Figure 3.3). The contents
of the Titret will turn purple.
CAUTION: During the titration, there will be some undissolved powder inside
the Titret. This does not interfere with the accuracy of the
test.
7.17B Wait 30 seconds.
7.18B Gently press the control bar again to allow another SMALL amount
of the sample to be drawn into the Titret.
CAUTION: Do not press the control bar unless the sample pipe is immersed
in the sample. This prevents air from being drawn into the
Titret.
7.19B After each addition, rock the entire assembly to mix the contents
of the Titret. Watch for a color change from purple to very pale yellow.
7.20B Repeat Steps 7.18B and 7.19B until the color change occurs.
CAUTION: The end point color change (from purple to pale yellow) actually
goes through an intermediate gray color. During this intermediate
stage, extra caution should be taken to bring in SMALL amounts of
sample and to mix the Titret contents well.
7.21B When the color of the liquid in the Titret changes to PALE YELLOW,
remove the Titret from the Titrettor™. Hold the Titret in a vertical position
and carefully read the test result on the scale opposite the liquid level.
7.22B Calculation
7.22.IB To obtain results in micrograms per gram total chlorine,
multiply scale units on the Titret by 1.3 and then subtract 200.
9077 - 10 Revision 0
November 1992
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8.OB QUALITY CONTROL
8.IB Refer to Chapter One for specific quality control procedures.
8.2B Each sample should be tested two times. If the results do not
agree to within 10%, expressed as the relative percent difference of the results,
a third test must be performed. Report the results of the two that agree.
9.OB METHOD PERFORMANCE
9.IB These data are based on 49 data points obtained by seven
laboratories who each analyzed four used crankcase oils and three fuel oil blends
with crankcase in duplicate. A data point represents one duplicate analysis of
a sample. There were no outlier data points or laboratories.
9.2B Precision. The precision of the method as determined by the
statistical examination of interlaboratory test results is as follows:
Repeatability - The difference between successive results obtained
by the same operator with the same apparatus under constant
operating conditions on identical test material would exceed, in
the long run, in the normal and correct operation of the test
method, the following values only in 1 case in 20 (see Table 2):
Repeatability - 0.31 x*
*where x is the average of two results in
Reproducibilitv - The difference between two single and
independent results obtained by different operators working in
different laboratories on identical test material would exceed,
in the long run, the following values only in 1 case in 20:
Repioducibility - 0.60 x*
*where x is the average value of two results in jug/9-
9.3B Bias. The bias of this test method varies with concentration, as
shown in Table 3:
Bias = Amount found - Amount expected
9077 - 11 Revision 0
November 1992
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TABLE 2.
REPEATABILITY AND REPRODUCIBILITY FOR CHLORINE IN USED
OILS BY THE QUANTITATIVE END POINT TEST KIT METHOD
Average value, Repeatability, Reproducibility,
1,000
1,500
2,000
2,500
3,000
310
465
620
775
930
600
900
1,200
1,500
1,800
TABLE 3.
RECOVERY AND BIAS DATA FOR CHLORINE IN USED OILS BY THE
QUANTITATIVE END POINT TEST KIT METHOD
Amount
expected,
M9/9
320 (< 750)a
480 (< 750)a
920
1,498
1,527
3,029
3,045
Amount
found,
M9/9
776
782
1,020
1,129
1,434
1,853
2,380
Bias,
M9/9
+16
+32
+100
-369
-93
-1,176
-665
Percent
bias
+3
+4
+11
-25
-6
-39
-22
The lower limit of the kit is 750
9077 - 12 Revision 0
November 1992
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Reaction bottle
Valyeassembly
p__^
Micro pipet
Figure 1. Components of CHEMetrics Total Chlorine in Waste Oil Test Kit
(Cat. No. K2610).
9077 - 13
Revision 0
November 1992
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Push plunger
down to
transfer
sample
Figure 2.1
Figure 2.2
*• Crush
Figure 2.3
Buffer Bottle
Figure 2.4
Reaction bottle
upsidedown in
component tray
Figure 2.5
Aqueous
Layer
Filtering Column
Figure 2.6
Titration Vial
Figure 2. Reaction-Extraction Procedure.
9077 - 14
Revision 0
November 1992
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Attaching
the Valve
Assembly
Figure 3.1
Valve
Assembly
Snapping
the Tip
Figure 3.2
Titret
Lift control bar
Performing the
Analysis
Figure 3.3
Watch for
color change
here
Press control bar
Sample pipe
Sample - ^
Reading
the Result
Figure 3.4
Read
scale units
when color
changes
permanently
Figure 3. Titration Procedure
9077 - 15
Revision 0
November 1992
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METHOD 9077 B
REVERSE TITRATION QUANTITATIVE END POINT TEST KIT METHOD
START
I
7 IB Shake glass
reaction bottle
1
7 2B Fill
micropipet with
oil, transfer oil
to reaction bottle
1
f r om reac 1 1 on
bottle , cap , miK
1
7 4B Crush sodium
ampoule
1
7 SB - 7 6B Shake
reaction bottle for
30 seconds, wait
one minute
1
7 7B Pour buffer
into reaction
bottle
-
7 8B - 7 9B Shake
gently, release
gas, shake, ^release
upside down, wait
one minute
1
7 10B Prepare
filter ing col umn
• 1
7 11B Filter lower
aqueous layer
through filtering
column into
ti tration vial
1
7 12B Shake vial
1
7 13B Assemble
valve assembly over
Titret
1
7 14B Insert Titret
into Ti trettor
7 15B Snap tip of
Titret
7 16B - 7 20B Pull
smal1 amount of
sample into Titret.
mix, wait 30
seconds, repeat
proce»» unt11 color
changes from purple
to pale yellow
7 21B When color
changes to pal*
yel1ow, remove
Titret, record teit
result from Titr«t
7 22B Calculate
concentration of
chlorine in ug/g
STOP
9077 - 16
Revision 0
November 1992
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METHOD C
DIRECT TITRATION QUANTITAVE END POINT TEST KIT METHOD
4.0C APPARATUS AND MATERIALS
4.1C The CHLOR-D-TECT Q40003 is a complete self-contained kit. It
includes: a sampling syringe to withdraw a fixed sample volume for analysis; a
polyethylene test tube #1 into which the sample is introduced for dilution and
reaction with metallic sodium; a polyethylene tube #2 containing a buffered
aqueous extractant and the diphenylcarbazone indicator; a microburette containing
the mercuric nitrate titrant; and a plastic filtration funnel. Also included are
instructions to conduct the test.
5.0C REAGENTS
5.1C All necessary reagents are contained within the kit. The diluent
solvent containing the catalyst, the metallic sodium, and the diphenylcarbazone
are separately glass-encapsulated in the precise quantity required for analysis.
A predispensed volume of buffer is contained in the second polyethylene tube.
Mercuric nitrate titrant is also supplied in a sealed titration burette.
5.2C The kit should be examined upon opening to see that all of the
components are present and that all ampoules (3) are in place and not leaking.
The liquid in Tube #2 (clear cap) should be approximately 1/2 in. above the 5-mL
line and the tube should not be leaking. The ampoules are not supposed to be
completely full.
6.0C SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1C See Section 6.0A of Method A.
7.0C PROCEDURE
7.1C Preparation. Open analysis carton, remove contents, mount plastic
test tubes in the provided holder.
NOTE: Perform the test in a warm, dry area with adequate light. In cold
weather, a truck cab is sufficient. If a warm area is not available, Step
7.3C should be performed while warming Tube #1 in palm of hand.
7.2C Sample introduction. Unscrew the white dispenser cap from Tube #1.
Slide the plunger in the empty syringe a few times to make certain that it slides-
easily. Place the top of the syringe in the oil sample to be tested, and pull
back on the plunger until it reaches the stop and cannot be pulled further.
Remove the syringe from the sample container, and wipe any excess oil from the
outside of the syringe with the enclosed tissue. Place the tip of the syringe
in Tube #1, and dispense the oil sample by depressing the plunger. Replace the
white cap on the tube.
'Available from Dexsil Corporation, One Hamden Park Drive, Hamden, CT 06517.
9077 - 17 Revision 0
November 1992
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7.3C Reaction. Break the lower (colorless) capsule containing the clear
diluent solvent by squeezing the sides of the test tube. Mix thoroughly by
shaking the tube vigorously for 30 seconds. Crush the upper grey ampoule
containing metallic sodium, again by squeezing the sides of the test tube. Shake
vigorously for 20 seconds. Allow reaction to proceed for 60 seconds, shaking
intermittently several times while timing with a watch.
CAUTION: Always crush the clear ampoule in each tube first. Otherwise, stop
the test and start over using another complete kit. False (low)
results may occur and allow a contaminated sample to pass without
detection if clear ampoule is not crushed first.
7.4C Extraction. Remove caps from both tubes. Pour the clear buffered
extraction solution from Tube #2 into Tube #1. Replace the white cap on Tube #1,
and shake vigorously for 10 seconds. Vent tube by partially unscrewing the
dispenser cap. Close cap securely, and shake for an additional 10 seconds. Vent
again, tighten cap, and stand tube upside down on white cap. Allow phases to
separate for 2 minutes.
NOTE: Tip Tube #2 to an angle of only about 45*. This will prevent the holder
from sliding out.
7.5C Analysis. Put filtration funnel into Tube #2. Position Tube #1
over funnel and open nozzle on dispenser cap. Squeeze the sides of Tube #1 to
dispense the clear aqueous lower phase through the filter into Tube #2 to the 5-
mL line on Tube #2. Remove the filter funnel, and close the nozzle on the
dispenser cap. Place the plunger rod in the titration burette and press until
it clicks into place. Break off (do not pull off) the tip on the titration
burette. Insert the burette into Tube #2, and tighten the cap. Break the
colored ampoule, and shake gently for 10 seconds. Dispense titrant dropwise by
pushing down on burette rod in small increments. Shake the tube gently to mix
titrant with solution in Tube #2 after each increment. Continue adding titrant
until solution turns from yellow to red-violet. An intermediate pink color may
develop in the solution, but should be disregarded. Continue titrating until a
true red-violet color is realized. The chlorine concentration of the original
oil sample is read directly off the titrating burette at the tip of the black
plunger. Record this result immediatley as the red-violet color will fade with
time.
8.0C QUALITY CONTROL
8.1C Refer to Chapter One for specific quality control procedures.
8.2C Each sample should be tested two times. If the results do not
agree to within 10%, expressed as the relative percent difference of the results,
a third test must be performed. Report the results of the two that agree.
9.0C METHOD PERFORMANCE
9.1C These data are based on 96 data points obtained by 12 laboratories
who each analyzed six used crankcase oils and two fuel oil blends with crankcase
in duplicate. A data point represents one duplicate analysis of a sample.
9077 - 18 Revision 0
November 1992
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9.2C Precision. The precision of the method as determined by the
statistical examination of interlaboratory test results is as follows:
Repeatability - The difference between successive results obtained
by the same operator with the same apparatus under constant
operating conditions on identical test material would exceed, in
the long run, in the normal and correct operation of the test
method, the following values only in 1 case in 20 (see Table 4):
Repeatability - 0.175 x*
*where x is the average of two results in
Reproducibility - The difference between two single and independent
results obtained by different operators working in different
laboratories on identical test material would exceed, in the long
run, the following values only in 1 case in 20:
Reproducibility - 0.331 x*
*where x is the average value of two results in vg/g.
9.3C Bias. The bias of this test method varies with concentration, as
shown in Table 5:
Bias = Amount found - Amount expected
10.0 REFERENCE
1. Gaskill, A.; Estes, E.D.; Hardison, D.L.; and Myers, I.E. Validation of
Methods for Determining Chlorine in Used Oils and Oil Fuels. Prepared for
U.S. Environmental Protection Agency, Office of Solid Waste. EPA Contract
No. 68-01-7075, wA 80. July 1988.
9077 - 19 Revision 0
November 1992
-------
TABLE 4.
REPEATABILITY AND REPRODUCIBILITY FOR CHLORINE IN USED
OILS BY THE QUANTITATIVE END POINT TEST KIT METHOD
Average value, Repeatability, Reproducibility,
M9/g
500
1,000
1,500
2,000
2,500
3,000
4,000
88
175
263
350
438
525
700
166
331
497
662
828
993
1,324
TABLE 5.
RECOVERY AND BIAS DATA FOR CHLORINE IN USED OILS BY THE
QUANTITATIVE END POINT TEST KIT METHOD
Amount
expected,
M9/9
664
964
1,230
1,445
2,014
2,913
3,812
4,190
Amount
found,
M9/9
695
906
1,116
1,255
1,618
2,119
2,776
3,211
Bias,
M9/g
31
-58
-114
-190
-396
-794
-1,036
-979
Percent
bias
+5
-6
-9
-13
-20
-27
-27
-23
9077 - 20 Revision 0
November 1992
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METHOD 9077 C
DIRECT TITRATION QUANTITAVE END POINT TEST KIT METHOD
START
7 1C Open teat kit
7 2C Draw oil into
ayringe, remove
excesa oil,
dispense oil into
Tube /I
7 3C Break
colorless capaule,
mix, crush grey
capaule. mix, allow
reaction to proceed
for 60 seconds
7 4C Pour Tube #2
solution into Tube
#1, mix; vent,
al1DW phases to
separata
7 5C Filter aqueous
lower phase in Tube
#1 into Tube #2.
remove fi1ter
funnel
7 5C Place piunger
in titraton
burette, press,
break off burette
tip. inaert burette
in Tube #2, break
co 1 o red atnpoul e ,
shake
7 SC Dispense
tit rant, shake,
repeat process
unti1 a o1ution
turna from yel1 aw
to red-violet
7 5C Record level
from titrating
bure t te
STOP
9077 - 21
Revision 0
November 1992
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METHOD 9090
COMPATIBILITY TEST FOR WASTES AND MEMBRANE LINERS
1.0 SCOPE AND APPLICATION
1.1 Method 9090 is intended for use in determining the effects of
chemicals in a surface impoundment, waste pile, or landfill on the physical
properties of flexible membrane liner (FML) materials intended to contain
them. Data from these tests will assist in deciding whether a given liner
material is acceptable for the intended application.
2.0 SUMMARY OF METHOD
2.1 In order to estimate waste/liner compatibility, the liner material
is immersed in the chemical environment for minimum periods of 120 days at
room temperature (23 + 2°C) and at 50 ± 2°C. In cases where the FML will be
used in a chemical environment at elevated temperatures, the immersion testing
shall be run at the elevated temperatures if they are expected to be higher
than 50°C. Whenever possible, the use of longer exposure times is
recommended. Comparison of measurements of the membrane's physical
properties, taken periodically before and after contact with the waste fluid,
is used to estimate the compatibility of the liner with the waste over time.
3.0 INTERFERENCES (Not Applicable)
4.0 APPARATUS AND MATERIALS
NOTE: In general, the following definitions will be used in this method:
1. Sample -- a representative piece of the liner material proposed for
use that is of sufficient size to allow for the removal of
all necessary specimens.
2. Specimen -- a piece of material, cut from a sample, appropriately
shaped and prepared so that it is ready to use for a test.
4.1 Exposure tank - Of a size sufficient to contain the samples, with
provisions for supporting the samples so that they do not touch the bottom or
sides of the tank or each other, and for stirring the liquid in the tank. The
tank should be compatible with the waste fluid and impermeable to any of the
constituents they are intended to contain. The tank shall be equipped with a
means for maintaining the solution at room temperature (23 ± 2°C) and 50 + 2"C
and for preventing evaporation of the solution (e.g., use a cover equipped
with a reflux condenser, or seal the tank with a Teflon gasket and use an
airtight cover). Both sides of the liner material shall be exposed to the
chemical environment. The pressure inside the tank must be the same as that
outside the tank. If the liner has a side that (1) is not exposed to the
waste in actual use and (2) is not designed to withstand exposure to the
chemical environment, then such a liner may be treated with only the barrier
surface exposed.
9090 - 1 Revision 1
December 1987
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4.2 Stress-strain machine suitable for measuring elongation, tensile
strength, tear resistance, puncture resistance, modulus of elasticity, and ply
adhesion.
4.3 Jig for testing puncture resistance for use with FTMS 101C, Method
2065.
4.4 Liner sample labels and holders made of materials known to be
resistant to the specific wastes.
4.5 Oven at 105 ± 2°C.
4.6 Dial micrometer.
4.7 Analytical balance.
4.8 Apparatus for determining extractable content of liner materials.
NOTE: A minimum quantity of representative waste fluid necessary to conduct
this test has not been specified in this method because the amount will
vary depending upon the waste compostion and the type of liner
material. For example, certain organic waste constituents, if present
in the representative waste fluid, can be absorbed by the liner
material, thereby changing the concentration of the chemicals in the
waste, this change in waste composition may require the waste fluid to
be replaced at least monthly in order to maintain representative
conditions in the waste fluid. The amount of waste fluid necessary to
maintain representative waste conditions will depend on factors such as
the volume of constituents absorbed by the specific liner material and
the concentration of the chemical constituents in the waste.
5.0 REAGENTS (Not Applicable)
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 For information on what constitutes a representative sample of the
waste fluid, refer to the following guidance document:
Permit Applicants' Guidance Manual for Hazardous Waste Land Treatment,
Storage, and Disposal Facilities; Final Draft; Chap. 5, pp. 15-17;
Chap. 6, pp. 18-21; and Chap. 8, pp. 13-16, May 1984.
7.0 PROCEDURE
7.1 Obtain a representative sample of the waste fluid. If a waste
sample is received in more than one container, blend thoroughly. Note any
signs of stratification. If stratification exists, liner samples must be
placed in each of the phases. In cases where the waste fluid is expected to
stratify and the phases cannot be separated, the number of immersed samples
per exposure period can be increased (e.g. if the waste fluid has two phases,
then 2 samples per exposure period are needed) so that test samples exposed at
each level of the waste can be tested. If the waste to be contained in the
9090 - 2 Revision 1
December 1987
-------
land disposal unit is in solid form, generate a synthetic leachate (see Step
7.9.1).
7.2 Perform the following tests on unexposed samples of the polymeric
membrane liner material at 23 ± 2'C (see Steps 7.9.2 and 7.9.3 below for
additional tests suggested for specific circumstances). Tests for tear
resistance and tensile properties are to be performed according to the
protocols referenced in Table 1. See Figure 1 for cutting patterns for
nonreinforced liners, Figure 2 for cutting patterns for reinforced liners, and
Figure 3 for cutting patterns for semicrystalline liners. (Table 2, at the end
of this method, gives characteristics of various polymeric liner materials.)
1. Tear resistance, machine and transverse directions, three specimens
each direction for nonreinforced liner materials only. See Table 1
for appropriate test method, the recommended test speed, and the
values to be reported.
2. Puncture resistance, two specimens, FTMS 101C, Method 2065. See
Figure 1, 2, or 3, as applicable, for sample cutting patterns.
3. Tensile properties, machine and transverse directions, three tensile
specimens in each direction. See Table 1 for appropriate test
method, the recommended test speed, and the values to be reported.
See Figure 4 for tensile dumbbell cutting pattern dimensions for
nonreinforced liner samples.
4. Hardness, three specimens, Duro A (Duro D if Duro A reading is
greater than 80), ASTM D2240. The hardness specimen thickness for
Duro A is 1/4 in., and for Duro D it is 1/8 in. The specimen
dimensions are 1 in. by 1 in.
5. Elongation at break. This test is to be performed only on membrane
materials that do not have a fabric or other nonelastomeric support
as part of the liner.
6. Modulus of elasticity, machine and transverse directions, two
specimens each direction for semicrystalline liner materials only,
ASTM D882 modified Method A (see Table 1).
7. Volatiles content, SW 870, Appendix III-D.
8. Extractables content, SW 870, Appendix III-E.
9. Specific gravity, three specimens, ASTM D792 Method A.
10. Ply adhesion, machine and transverse directions, two specimens each
direction for fabric reinforced liner materials only, ASTM D413
Machine Method, Type A -- 180 degree peel.
11. Hydrostatic resistance test, ASTM D751 Method A, Procedure 1.
9090 - 3 Revision 1
December 1987
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7.3 For each test condition, cut five pieces of the lining material of a
size to fit the sample holder, or at least 8 in. by 10 in. The fifth sample
is an extra sample. Inspect all samples for flaws and discard unsatisfactory
ones. Liner materials with fabric reinforcement require close inspection to
ensure that threads of the samples are evenly spaced and straight at 90°.
Samples containing a fiber scrim support may be flood-coated along the exposed
edges with a solution recommended by the liner manufacturer, or another
procedure should be used to prevent the scrim from being directly exposed.
The flood-coating solution will typically contain 5-15% solids dissolved in a
solvent. The solids content can be the liner formula or the base polymer.
Measure the following:
1. Gauge thickness, in. -- average of the four corners.
2. Mass, Ib. -- to one-hundredth of a Ib.
3. Length, in. -- average of the lengths of the two sides plus the
length measured through the liner center.
4. Width, in. -- average of the widths of the two ends plus the width
measured through the liner center.
NOTE: Do not cut these liner samples into the test specimen shapes shown in
Figure 1, 2, or 3 at this time. Test specimens will be cut as
specified in Step 7.7, after exposure to the waste fluid.
7.4 Label the liner samples (e.g. notch or use metal staples to identify
the sample) and hang in the waste fluid by a wire hanger or a weight.
Different liner materials should be immersed in separate tanks to avoid
exchange of plasticizers and soluble constituents when plasticized membranes
are being tested. Expose the liner samples to the stirred waste fluid held at
room temperature and at 50 + 2°C.
7.5 At the end of 30, 60, 90, and 120 days of exposure, remove one liner
sample from each test condition to determine the membrane's physical
properties (see Steps 7.6 and 7.7). Allow the liner sample to cool in the
waste fluid until the waste fluid has a stable room temperature. Wipe off as
much waste as possible and rinse briefly with water. Place wet sample in a
labeled polyethylene bag or aluminum foil to prevent the sample from drying
out. The liner sample should be tested as soon as possible after removal from
the waste fluid at room temperature, but in no case later than 24 hours after
removal.
7.6 To test the immersed sample, wipe off any remaining waste and rinse
with deionized water. Blot sample dry and measure the following as in Step
7.3:
1. Gauge thickness, in.
2. Mass, Ib.
9090 - 4 Revision 1
December 1987
-------
3. Length, in.
4. Width, in.
7.7 Perform the following tests on the exposed samples (see Steps 7.9.2
and 7.9.3 below for additional tests suggested for specific circumstances).
Tests for tear resistance and tensile properties are to be performed according
to the protocols referenced in Table 1. Die-cut test specimens following
suggested cutting patterns. See Figure 1 for cutting patterns for
nonreinforced liners, Figure 2 for cutting patterns for reinforced liners, and
Figure 3 for semi crystal line liners.
1. Tear resistance, machine and transverse directions, three specimens
each direction for materials without fabric reinforcement. See Table 1 for
appropriate test method, the recommended test specimen and speed of test, and
the values to be reported.
2. Puncture resistance, two specimens, FTMS 101C, Method 2065. See
Figure 1, 2, or 3, as applicable, for sample cutting patterns.
3. Tensile properties, machine and transverse directions, three
specimens each direction. See Table 1 for appropriate test method, the
recommended test specimen and speed of test, and the values to be reported.
See Figure 4 for tensile dumbbell cutting pattern dimensions for nonreinforced
liner samples.
4. Hardness, three specimens, Duro A (Duro D if Duro A reading is
greater than 80), ASTM 2240. The hardness specimen thickness for Duro A is
1/4 in., and for Duro D is 1/8 in. The specimen dimensions are 1 in. by 1 in.
5. Elongation at break. This test is to be performed only on membrane
materials that do not have a fabric or other nonelastomeric support as part of
the liner.
6. Modulus of elasticity, machine and transverse directions, two
specimens each direction for semi crystal line liner materials only, ASTM D882
modified Method A (see Table 1).
7. Volatiles content, SW 870, Appendix III-D.
8. Extractables content, SW 870, Appendix III-E.
9. Ply adhesion, machine and transverse directions, two specimens each
direction for fabric reinforced liner materials only, ASTM D413 Machine
Method, Type A -- 180 degree peel.
10. Hydrostatic resistance test, ASTM D751 Method A, Procedure 1.
7.8 Results and reporting
9090 - 5 Revision 1
December 1987
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7.8.1 Plot the curve for each property over the time period 0 to
120 days and display the spread in data points.
7.8.2 Report all raw, tabulated, and plotted data. Recommended
methods for collecting and presenting information are described in the
documents listed under Step 6.1 and in related agency guidance manuals.
7.8.3 Summarize the raw test results as follows:
1. Percent change in thickness.
2. Percent change in mass.
3. Percent change in area (provide length and width dimensions).
4. Percent retention of physical properties.
5. Change, in points, of hardness reading.
6. The modulus of elasticity calculated in pounds-force per
square inch.
7. Percent volatiles of unexposed and exposed liner material.
8. Percent extractables of unexposed and exposed liner material.
9. The adhesion value, determined in accordance with ASTM D413,
Step 12.2.
10. The pressure and time elapsed at the first appearance of
water through the flexible membrane liner for the hydrostatic
resistance test.
7.9 The following additional procedures are suggested in specific
situations:
7.9.1 For the generation of a synthetic leachate, the Agency
suggests the use of the Toxicity Characteristic Leaching Procedure (TCLP)
that was proposed in the Federal Register on June 13, 1986, Vol. 51,
No. 114, p. 21685.
7.9.2 For semi crystal line membrane liners, the Agency suggests the
determination of the potential for environmental stress cracking. The
test that can be used to make this determination is either ASTM D1693 or
the National Bureau of Standards Constant Tensile Load. The evaluation
of the results should be provided by an expert in this field.
7.9.3 For field seams, the Agency suggests the determination of
seam strength in shear and peel modes. To determine seam strength in
peel mode, the test ASTM D413 can be used. To determine seam strength in
shear mode for nonreinforced FMLs, the test ASTM D3083 can be used, and
for reinforced FMLs, the test ASTM D751, Grab Method, can be used at a
9090 - 6 Revision 1
December 1987
-------
speed of 12 in. per minute. The evaluation of the results should be
provided by an expert in this field.
8.0 QUALITY CONTROL
8.1 Determine the mechanical properties of identical nonimmersed and
immersed liner samples in accordance with the standard methods for the
specific physical property test. Conduct mechanical property tests on
nonimmersed and immersed liner samples prepared from the same sample or lot of
material in the same manner and run under identical conditions. Test liner
samples immediately after they are removed from the room temperature test
solution.
9.0 METHOD PERFORMANCE
9.1 No data provided.
10.0 REFERENCES
1. None required.
9090 - 7 Revision 1
December 1987
-------
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9090 - 8
Revision 1
December 1987
-------
TABLE 2.
POLYMERS USED IN FLEXIBLE MEMBRANE LINERS
Thermoplastic Materials (TP)
CPE (Chlorinated polyethylene)3
A family of polymers produced by a chemical reaction of chlorine on
polyethylene. The resulting thermoplastic elastomers contain 25 to 45%
chlorine by weight and 0 to 25% crystal 1inity.
CSPE (Chlorosulfonated polyethylene)3
A family of polymers that are produced by the reaction of polyethylene
with chlorine and sulfur dioxide, usually containing 25 to 43% chlorine
and 1.0 to 1.4% sulfur. Chlorosulfonated polyethylene is also known as
hypalon.
EIA (Ethylene interpolymer alloy)3
A blend of EVA and polyvinyl chloride resulting in a thermoplastic
elastomer.
PVC (Polyvinyl chloride)3
A synthetic thermoplastic polymer made by polymerizing vinyl chloride
monomer or vinyl chloride/vinyl acetate monomers. Normally rigid and
containing 50% of plasticizers.
PVC-CPE (Polyvinyl chloride - chlorinated polyethylene alloy)3
A blend of polyvinyl chloride and chlorinated polyethylene.
TN-PVC (Thermoplastic nitrile-polyvinyl choloride)3
An alloy of thermoplastic unvulcanized nitrile rubber and polyvinyl
chloride.
Vulcanized Materials (XL)
Butyl rubber3
A synthetic rubber based on isobutylene and a small amount of isoprene to
provide sites for vulcanization.
3Also supplied reinforced with fabric.
9090 - 9 Revision 1
December 1987
-------
TABLE 2. (Continued)
EPDM (Ethylene propylene diene monomer)a>b
A synthetic elastomer based on ethylene, propylene, and a small amount of
nonconjugated diene to provide sites for vulcanization.
CM (Cross-linked chlorinated polyethylene)
No definition available by EPA.
CO, ECO (Epichlorohydrin polymers)3
Synthetic rubber, including two epichlorohydrin-based elastomers that are
saturated, high-molecular-weight aliphatic polyethers with chloromethyl
side chains. The two types include homopolymer (CO) and a copolymer of
epichlorohydrin and ethylene oxide (ECO).
CR (Polychloroprene)a
Generic name for a synthetic rubber based primarily on chlorobutadiene.
Polychloroprene is also known as neoprene.
Semi crystalline Materials (CX)
HOPE - (High-density polyethylene)
A polymer prepared by the low-pressure polymerization of ethylene as
the principal monomer.
HOPE - A (High-density polyethylene/rubber alloy)
A blend of high-density polyethylene and rubber.
LLDPE (Liner low-density polyethylene)
A low-density polyethylene produced by the copolymerization of ethylene
with various alpha olefins in the presence of suitable catalysts.
PEL (Polyester elastomer)
A segmented thermoplastic copolyester elastomer containing recurring
long-chain ester units derived from dicarboxylic acids and long-chain
glycols and short-chain ester units derived from dicarboxylic acids and
low-molecular-weight diols.
aAlso supplied reinforced with fabric.
^Also supplied as a thermoplastic.
9090 - 10 Revision 1
December 1987
-------
TABLE 2. (Continued)
PE-EP-A (Polyethylene ethylene/propylene alloy)
A blend of polyethylene and ethylene and propylene polymer resulting in a
thermoplastic elastomer.
T-EPDM (Thermoplastic EPDM)
An ethylene-propylene diene monomer blend resulting in a thermoplastic
elastomer.
9090 - 11 Revision 1
December 1987
-------
Figure 1 .
Suggested pattern for cutting test specimens from
nonreinforced cross linked or thermoplastic Immersed
liner samples.
10'
Puncture test specimens
Tear test specimens
Volatlles test specimen
test specimens
9090 - 12
Not to scale
Revision 1
December 1987
-------
Figure 2 .
Suggested pattern for cutting test specimens from
fabric reinforced Immersed Hner samples. Note: To
avoid edge effects, cut specimens 1/8 - 1/4 Inch In
from edge of Immersed sample.
10'
\/
Volatile* Jest specimen
Puncture test specimens
9090 - 13
Not to scale
Revision 1
December 1987
-------
Figure 3 .
Suggested pattern for cutting test specimens from
semi crystal line immersed liner samples. Note: To
avoid edge effects, cut specimens 1/8 - 1/4 Inch
1n from edge of Immersed sample.
10*
Modulus of elasticity
test specimens
Tensile test specimens
Volatlles test specimen
Puncture test specimens
Tear test specimens
9090 - 14
Not to seal*
Revision 1
December 1987
-------
Figure 4.
Die for tensile dumbbell (nonreinforced liners) having the following
t
1
wo
I
i
N
s/
1
J
w
f
1
_ 1
Q
LO
s
V
W - Width of narrow section
L • Length of narrow section
WO - Width overall
LO - Length overall
G - Gage length
D Distance between g<.ps
0.25 inches
1.25 inches
0 625 inches
3.50 inches
1.00 inches
2 00 .nches
9090 - 15
Revision 1
December 1987
-------
METHOD 9090
COMPATIBILITY TEST FOR WASTES AND MEMBRANE LINERS
7. 1
Obtain sample
of waste fluid
O
7.S
Determine
membrane
physical
properties at
30 day
intervals
7.Z
Perform
tests on
unexposed
samples of
liner materiel
7.6
To test
exposed
specimens.
measure gauge
thickness, mass.
length, width
7.3
1 Cut
Pieces of
lining material
for each tect
condition
7. 4
7.7
Perform tests
on exposed
samples
Label
test specimens
and expose
to waste flula
7.8
Report ana
evaluate data
0
f Stop J
9090 - 16
Revision 1
December 1987
-------
METHOD 9200A
NITRATE
1.0 SCOPE AND APPLICATION
1.1 This method is applicable to the analysis of groundwater, drinking,
surface, and saline waters, and domestic and industrial wastes. Modifications
can be made to remove or correct for turbidity, color, salinity, or dissolved
organic compounds in the sample.
1.2 The applicable range of concentration is 0.1 to 2 mg N03-N per liter
of sample.
2.0 SUMMARY OF METHOD
2.1 This method is based upon the reaction of the nitrate ion with
brucine sulfate in a 13 N H2S04 solution at a temperature of 100'C. The color
of the resulting complex is measured at 410 nm. Temperature control of the color
reaction is extremely critical.
3.0 INTERFERENCES
3.1 Dissolved organic matter will cause an off color in 13 N H2S04 and
must be compensated for by additions of all reagents except the brucine-
sulfanilic acid reagent. This also applies to natural color, not due to
dissolved organics, that is present.
3.2 If the sample is colored or if the conditions of the test cause
extraneous coloration, this interference should be corrected by running a
concurrent sample under the same conditions but in the absence of the brucine-
sulfanilic acid reagent.
3.3 Strong oxidizing or reducing agents cause interference. The
presence of oxidizing agents may be determined by a residual chlorine test;
reducing agents may be detected with potassium permanganate.
3.3.1 Oxidizing agents' interference is eliminated by the
addition of sodium arsenite.
3.3.2 Reducing agents may be oxidized by addition of H202.
3.4 Ferrous and ferric ion and quadrivalent manganese give slight
positive interferences, but in concentrations less than 1 mg/L these are
negligible.
3.5 Uneven heating of the samples and standards during the reaction
time will result in erratic values. The necessity for absolute control of
temperature during the critical color development period cannot be too strongly
emphasized.
9200A - 1 Revision 1
November 1992
-------
4.0 APPARATUS AND MATERIALS
4.1 Spectrophotometer or filter photometer suitable for measuring
absorbance at 410 nm.
4.2 Sufficient number of 40- to 50-mL glass sample tubes for reagent
blanks, standards, and samples.
4.3 Neoprene-coated wire racks to hold sample tubes.
4.4 Water bath suitable for use at 100°C. This bath should contain a
stirring mechanism so that all tubes are at the same temperature and should be
of sufficient capacity to accept the required number of tubes without a
significant drop in temperature when the tubes are immersed.
4.5 Water bath suitable for use at 10-15°C.
4.6 Analytical balance: capable of weighing to 0.0001 g.
4.7 Class A volumetric flasks: 1 L.
4.8 pH Indicator paper.
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 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Sodium chloride solution (30%): Dissolve 300 g NaCl in reagent
water and dilute to 1 liter with reagent water.
5.4 Sulfuric acid solution: Carefully add 500 ml concentrated H2S04 to
125 ml reagent water. Cool and keep tightly stoppered to prevent absorption of
atmospheric moisture.
5.5 Brucine-sulfanilic acid reagent: Dissolve 1 g brucine sulfate --
(C23H2fiN204)2 • H2S04 • 7H20 --and 0.1 g sulfanilic acid (NH2C6H4SO,H • H20) in
70 mLTiot reagent water. Add 3 ml concentrated HC1, cool, mix, and dilute to 100
ml with reagent water. Store in a dark bottle at 5°C. This solution is stable
for several months; the pink color that develops slowly does not affect its
usefulness. Mark bottle with warning. "CAUTION: Brucine Sulfate is toxic; do
not ingest."
5.6 Potassium nitrate stock solution (1.0 ml = 0.1 mg N03-N): Dissolve
0.7218 g anhydrous potassium nitrate (KN03) in reagent water and dilute to 1
9200A - 2 Revision 1
November 1992
-------
liter in a Class A volumetric flask. Preserve with 2 ml chloroform per liter.
This solution is stable for at least 6 months.
5.7 Potassium nitrate standard solution (1.0 ml = 0.001 mg N03-N):
Dilute 10.0 mL of the stock solution (Step 5.6) to 1 liter in a Class A
volumetric flask. This standard solution should be prepared fresh weekly.
5.8 Acetic acid (1+3): Dilute 1 volume glacial acetic acid (CH3COOH)
with 3 volumes of reagent water.
5.9 Sodium hydroxide (1 N): Dissolve 40 g of NaOH in reagent water.
Cool and dilute to 1 liter with reagent water.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 Analysis should be performed within 48 hours. If analysis can be
done within 24 hours, the sample should be preserved by refrigeration at 4°C.
When samples must be stored for more than 24 hours, they should be preserved with
sulfuric acid (2 ml/I concentrated H2S04) and refrigerated.
7.0 PROCEDURE
7.1 Adjust the pH of the samples to approximately 7 with acetic acid
(Step 5.8) or sodium hydroxide (Step 5.9). If necessary, filter to remove
turbidity. Sulfuric acid can be used in place of acetic acid, if preferred.
7.2 Set up the required number of sample tubes in the rack to handle
the reagent blank, standards, and samples. Space tubes evenly throughout the
rack to allow for even flow of bath water between the tubes. This should assist
in achieving uniform heating of all tubes.
7.3 If it is necessary to correct for color or dissolved organic matter
which will cause color on heating, run a set of duplicate samples with all of the
reagents, except the brucine-sulfanilic acid.
7.4 Pipet 10.0 mL of standards and samples or an aliquot of the samples
diluted to 10.0 mL into the sample tubes.
7.5 If the samples are saline, add 2 mL of the 30% sodium chloride
solution (Step 5.3) to the reagent blank, standards, and samples. For freshwater'
samples, sodium chloride solution may be omitted. Mix contents of tubes by
swirling; place rack in cold-water bath (0-10°C).
7.6 Pipet 10.0 mL of sulfuric acid solution (Step 5.4) into each tube
and mix by swirling. Allow tubes to come to thermal equilibrium in the cold
bath. Be sure that temperatures have equilibrated in all tubes before
continuing.
7.6.1 Add 0.5 mL brucine-sulfanilic acid reagent (Step 5.5) to
each tube (except the interference control tubes) and carefully mix by swirling;
place the rack of tubes in the 100'C water bath for exactly 25 minutes.
9200A - 3 Revision 1
November 1992
-------
CAUTION: Immersion of the tube rack into the bath should not decrease the
temperature of the bath by more than 1-2°C. In order to keep this
temperature decrease to an absolute minimum, flow of bath water
between the tubes should not be restricted by crowding too many
tubes into the rack. If color development in the standards reveals
discrepancies in the procedure, the operator should repeat the
procedure after reviewing the temperature control steps.
7.7 Remove rack of tubes from the hot-water bath, immerse in the cold-
water bath, and allow to reach thermal equilibrium (20-25'C).
7.8 Read absorbance against the reagent blank at 410 nm using a 1-cm
or longer cell.
7.9 Calculation:
7.9.1 Obtain a standard curve by plotting the absorbance of
standards run by the above procedure against mg/L N03-N. (The color
reaction does not always follow Beer's law.)
7.9.2 Subtract the absorbance of the sample without the brucine-
sulfanilic reagent from the absorbance of the sample containing brucine-
sulfanilic acid and determine'mg/L N03-N. Multiply by an appropriate
dilution factor if less than 10 ml of sample is taken.
8.0 QUALITY CONTROL
8.1 All quality control data should be maintained and available for
easy reference or inspection.
8.2 Linear calibration curves must be composed of a minimum of a blank
and five standards. A set of standards must be included with each batch of
samples.
8.3 Dilute samples if they are more concentrated than the highest
standard or if they fall on the plateau of a calibration curve.
8.4 After calibrating, verify calibration with an independently
prepared check standard.
8.5 Run one matrix spike and matrix spike duplicate sample for every
10 samples. Matrix spikes and matrix spike duplicates are brought through the
whole sample preparation and analytical process.
9.0 METHOD PERFORMANCE
9.1 Twenty-seven analysts in fifteen laboratories analyzed natural-
water samples containing exact increments of inorganic nitrate, with the
following results:
9200A - 4 Revision 1
November 1992
-------
Increment as
Nitrogen, Nitrate
(mg/L N)
Precision as
Standard Deviation
(mg/L N)
Accuracy as
Bias Bias
(%) (mg/L N)
0.16
0.19
1.08
1.24
0.092
0.083
0.245
0.214
-6.79
+8.30
+4.12
+2.82
-0.01
+0.02
+0.04
+0.04
10.0 REFERENCES
1. Annual Book of ASTM Standards, Part 31, "Water," Standard D992-71, p. 363
(1976).
2. Jenkins, D. and L. Medsken, "A Brucine Method for the Determination of
Nitrate in Ocean, Estuarine, and Fresh Water," Anal.Chem., 36. P- 610 (1964).
3. Standard Methods for the Examination of Water and Wastewater, 14th ed., p.
427, Method 419D (1975).
9200A - 5
Revision 1
November 1992
-------
METHOD 9200A
NITRATE
START
7 3 Run
duplicates with
all reagents
except brucine
9ul f am lie acid
? 5 Add 30%
sodium chloride
solution, mix,
place in cold
water bath
7 1 Adjust pH
of samples to
7, filter if
necessary
7 2 Set up
sample tubes
in rack
7 4 Pipette
standards and
samples into
sample tubes
7 6 Pipette
sulfuric acid
solution into
each tube,
mi x
7 D 1 Add
brucine
sulfanilic
acid reagent
to each tube
761 Bathe
rack of tubes
in 1QOC water
for 25 mm
7 7 Immerse
tubes in cold
water, al1ow to
reach therma1
equi1ibrlum
7 8 Read
absorbanco
agains t
reagent blank
at 410 nm
791 Obtain a
std absorbance
curve and
calculate mg/L
nitrate
STOP
9200A - 6
Revision 1
November 1992
-------
METHOD 9252A
CHLORIDE (TITRIMETRIC. MERCURIC NITRATE)
1.0 SCOPE AND APPLICATION
1.1 This method is applicable to ground water, drinking, surface, and
saline waters, and domestic and industrial wastes.
1.2 The method is suitable for all concentration ranges of chloride
content; however, in order to avoid large titration volume, a sample aliquot
containing not more than 10 to 20 mg Cl" per 50 ml is used.
1.3 Automated titration may be used.
2.0 SUMMARY OF METHOD
2.1 An acidified sample is titrated with mercuric nitrate in the
presence of mixed diphenylcarbazone-bromophenol blue indicator. The end point
of the titration is the formation of the blue-violet mercury diphenylcarbazone
complex.
3.0 INTERFERENCES
3.1 Anions and cations at concentrations normally found in surface
waters do not interfere. However, at the higher concentration often found in
certain wastes, problems may occur.
3.2 Sulfite interference can be eliminated by oxidizing the 50 mi of
sample solution with 0.5-1 ml of H202.
4.0 APPARATUS AND MATERIALS
4.1 Standard laboratory titrimetric equipment, including 1 mL or 5 mL
microburet with 0.01 mL gradations.
4.2 Class A volumetric flasks: 1 L and 100 mL.
4.3 pH Indicator paper.
4.4 Analytical balance: capable of weighing to 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 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
9252A - 1 Revision 1
November 1992
-------
5.3 Standard sodium chloride solution, 0.025 N: Dissolve 1.4613 g ±
0.0002 g of sodium chloride (dried at 600°C for 1 hr) in chloride-free water in
a 1 liter Class A volumetric flask and dilute to the mark with reagent water.
5.4 Nitric acid (HNO,) solution: Add 3.0 ml concentrated nitric acid
to 997 ml of reagent water ("3 + 997" solution).
5.5 Sodium hydroxide (NaOH) solution (10 g/L): Dissolve approximately
10 g of NaOH in reagent water and dilute to 1 L with reagent water.
5.6 Hydrogen peroxide (H202): 30%.
5.7 Hydroquinone solution (10 g/L): Dissolve 1 g of purified
hydroquinone in reagent water in a 100 ml Class A volumetric flask and dilute to
the mark.
5.8 Mercuric nitrate titrant (0.141 N): Dissolve 24.2 g Hg(N03)2 • H20
in 900 ml of reagent water acidified with 5.0 ml concentrated HN03 in a 1 liter
volumetric flask and dilute to the mark with reagent water. Filter, if
necessary. Standardize against standard sodium chloride solution (Step 5.3)
using the procedures outlined in Section 7.0. Adjust to exactly 0.141 N and
check. Store in a dark bottle. A 1.00 ml aliquot is equivalent to 5.00 mg of
chloride.
5.9 Mercuric nitrate titrant (0.025 N): Dissolve 4.2830 g Hg(N03)2 •
H20 in 50 mL of reagent water acidified with 0.05 ml of concentrated
HN03 (sp. gr. 1.42) in a 1 liter volumetric flask and dilute to the mark with
reagent water. Filter, if necessary. Standardize against standard sodium
chloride solution (Step 5.3) using the procedures outlined in Section 7.0.
Adjust to exactly 0.025 N and check. Store in a dark bottle.
5.10 Mercuric nitrate titrant (0.0141 N): Dissolve 2.4200 g Hg(N03)2 •
H20 in 25 ml of reagent water acidified with 0.25 ml of concentrated HN03 (sp.
gr. 1.42) in a 1 liter Class A volumetric flask and dilute to the mark with
reagent water. Filter, if necessary. Standardize against standard sodium
chloride solution (Step 5.3) using the procedures outlined in Section 7.0.
Adjust to exactly 0.0141 N and check. Store in a dark bottle. A 1 ml aliquot
is equivalent to 500 M9 of chloride.
5.11 Mixed indicator reagent: Dissolve 0.5 g crystalline diphenylcar-
bazone and 0.05 g bromophenol blue powder in 75 ml 95% ethanol in a 100 ml Class
A volumetric flask and dilute to the mark with 95% ethanol. Store in brown
bottle and discard after 6 mo.
5.12 Alphazurine indicator solution: Dissolve 0.005 g of alphazurine
blue-green dye in 95% ethanol or isopropanol in 100 ml Class A volumetric flask
and dilute to the mark with 95% ethanol or isopropanol.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
9252A - 2 Revision 1
November 1992
-------
6.2 There are no special requirements for preservation.
7.0 PROCEDURE
7.1 Place 50 ml of sample in a vessel for titration. If the concentra-
tion is greater than 20 mg/L chloride, use 0.141 N mercuric nitrate titrant (Step
5.8) in Step 7.6, or dilute sample with reagent water. If the concentration is
less than 2.5 mg/L of chloride, use 0.0141 N mercuric nitrate titrant (Step 5.10)
in Step 7.6. Using a 1 ml or 5 ml microburet, determine an indicator blank on
50 mi chloride-free water using Step 7.6. If the concentration is less than 0.1
mg/L of chloride, concentrate an appropriate volume to 50 mL.
7.2 Add 5 to 10 drops of mixed indicator reagent (Step 5.11); shake or
swirl solution.
7.3 If a blue-violet or red color appears, add HN03 solution (Step 5.4)
dropwise until the color changes to yellow. Proceed to Step 7.5.
7.4 If a yellow or orange color forms immediately on addition of the
mixed indicator, add NaOH solution (Step 5.5) dropwise until the color changes
to blue-violet; then add HN03 solution (Step 5.4) dropwise until the color
changes to yellow.
7.5 Add 1 mL excess HN03 solution (Step 5.4).
7.6 Titrate with 0.025 N mercuric nitrate titrant (Step 5.9) until a
blue-violet color persists throughout the solution. If volume of titrant exceeds
10 mL or is less than 1 mL, use the 0.141 N or 0.0141 N mercuric nitrate
solutions, respectively. If necessary, take a small sample aliquot. Alphazurine
indicator solution (Step 5.12) may be added with the indicator to sharpen the end
point. This will change color shades. Practice runs should be made.
Note: The use of indicator modifications and the presence of heavy metal
ions can change solution colors without affecting the accuracy of
the determination. For example, solutions containing alphazurine
may be bright blue when neutral, grayish purple when basic, blue-
green when acidic, and blue-violet at the chloride end point.
Solutions containing about 100 mg/L nickel ion and normal mixed
indicator are purple when neutral, green when acidic, and gray at
the chloride end point. When applying this method to samples that
contain colored ions or that require modified indicator, it is
recommended that the operator become familiar with the specific
color changes involved by experimenting with solutions prepared as
standards for comparison of color effects.
7.6.1 If chromate is present at <100 mg/L and iron is not
present, add 5-10 drops of alphazurine indicator solution (Step 5.12) and
acidify to a pH of 3 (indicating paper). End point will then be an olive-
purple color.
7.6.2 If chromate is present at >100 mg/L and iron is not
present, add 2 mL of fresh hydroquinone solution (Step 5.7).
9252A - 3 Revision 1
November 1992
-------
7.6.3 If ferric ion is present use a volume containing no more
than 2.5 mg of ferric ion or ferric ion plus chromate ion. Add 2 mL fresh
hydroquinone solution (Step 5.7).
7.6.4 If sulfite ion is present, add 0.5 mL of H202 solution
(Step 5.6) to a 50 ml sample and mix for 1 min.
7.7 Calculation:
(A - B)N x 35,450
mg chloride/liter =
mL of sample
where:
A = mL titrant for sample;
B = mL titrant for blank; and
N = normality of mercuric nitrate titrant.
8.0 QUALITY CONTROL
8.1 All quality control data should be maintained and available for
easy reference or inspection. Refer to Chapter One for specific quality control
guidelines.
8.2 Analyze a standard reference material to ensure that correct
procedures are being followed and that all standard reagents have been prepared
properly.
8.3 Employ a minimum of one blank per analytical batch or twenty
samples, whichever is more frequent, to determine if contamination has occurred.
8.4 Run one matrix spike and matrix duplicate every analytical batch
or twenty samples, whichever is more frequent. Matrix spikes and duplicates are
brought through the whole sample preparation and analytical process.
9.0 METHOD PERFORMANCE
9.1 Water samples—A total of 42 analysts in 18 laboratories analyzed
synthetic water samples containing exact increments of chloride, with the results
shown in Table 1.
In a single laboratory, using surface water samples at an average
concentration of 34 mg CT/L, the standard deviation was +1.0.
A synthetic unknown sample containing 241 mg/L chloride, 108 mg/L Ca, 82
mg/L Mg, 3.1 mg/L K, 19.9 mg/L Na, 1.1 mg/L nitrate N, 0.25 mg/L nitrate N, 259
mg/L sulfate and 42.5 mg/L total alkalinity (contributed by NaHCO,) in reagent
water was analyzed in 10 laboratories by the mercurimetric method, with a
relative standard deviation of 3.3% and a relative error of 2.9%.
9252A - 4 Revision 1
November 1992
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9.2 Oil combustates--These data are based on 34 data points obtained by
five laboratories who each analyzed four used crankcase oils and three fuel oil
blends with crankcase oil in duplicate. The samples were combusted using Method
5050. A data point represents one duplicate analysis of a sample. One data
point was judged to be an outlier and was not included in these results.
9.2.1 Precision and bias.
9.2.1.1 Precision. The precision of the method as determined
by the statistical examination of interlaboratory test results is as
fol1ows:
Repeatability - The difference between successive results
obtained by the same operator with the same apparatus under constant
operating conditions on identical test material would exceed, in the
long run, in the normal and correct operation of the test method, the
following values only in 1 case in 20 (see Table 2):
Repeatability - 7.61 /x*
*where x is the average of two results in M9/9-
Reproducibility - The difference between two single and
independent results obtained by different operators working in
different laboratories on identical test material would exceed, in
the long run, the following values only in 1 case in 20:
Reproducibility - 20.02 /x*
*where x is the average value of two results in /xg/g.
9.2.1.2 Bias. The bias of this method varies with
concentration, as shown in Table 3:
Bias = Amount found - Amount expected
10.0 REFERENCES
1. Annual Book of ASTM Standards, Part 31, "Water," Standard D512-67, Method
A, p. 270 (1976).
2. Standard Methods for the Examination of Water and Wastewater, 15th ed.,
(1980).
3. U.S. Environmental Protection Agency, Methods for Chemical Analysis of
Water and Wastes, EPA 600/4-79-020 (1983), Method 325.3.
9252A - 5 Revision 1
November 1992
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TABLE 1. ANALYSES OF SYNTHETIC WATER SAMPLES
FOR CHLORIDE BY MERCURIC NITRATE METHOD
Increment as
Chloride
(mg/L)
Precision as
Standard Deviation
(mg/L)
Accuracy as
Bias
Bias
(mg/L)
17
18
91
97
382
398
1.54
1.32
2.92
3.16
11.70
11.80
+2.16
+3.50
+0.11
-0.51
-0.61
-1.19
+0.4
+0.6
+0.1
-0.5
-2.3
-4.7
TABLE 2. REPEATABILITY AND REPRODUCIBILITY
FOR CHLORINE IN USED OILS BY BOMB
OXIDATION AND MERCURIC NITRATE TITRATION
Average value,
Repeatability, Reproducibility,
500
1,000
1,500
2,000
2,500
3,000
170
241
295
340
381
417
448
633
775
895
1,001
1,097
9252A - 6
Revision 1
November 1992
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TABLE 3. RECOVERY AND BIAS DATA FOR CHLORINE IN
USED OILS BY BOMB OXIDATION AND
MERCURIC NITRATE TITRATION
Amount Amount
expected, found, Bias, Percent
M9/9 M9/9 M9/9 bias
320 460 140 +44
480 578 98 +20
920 968 48 +5
1,498 1,664 166 +11
1,527 1,515 - 12 - 1
3,029 2,809 -220 - 7
3,045 2,710 -325 -11
9252A - 7 Revision 1
November 1992
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METHOD 9252
CHLORIDE (TITRIMETRIC, MERCURIC NITRATE)
( STURT J
7 1 Place 50 ml
sample in Vit.rjl.ion
vessel. determine
concentration of
mercuric nitrate
titrant to use in
Step 7 6. determine
an indicator blank
7 2
to
Add indicator
sanpL* . »HaU«
7 4 Add sodium
KydroKid* untx1
•ample i*
blu«-viol»t. add
nitric acid unti 1
sampl* la y*lloo
7 3 Add nitric acid
unlj. 1 yampl* i>
ymi lo*
—
7 5 Add 1 ml nitric
acid
7 6 Tilrat* with
mercuric nitrat*
until blu«-viol«t
color p«r»i»ta
? 7 CalculaV*
concentration of
chloride in *anpl«
C
9252A - 8
Revision 1
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METHOD 9253
CHLORIDE (TITRIMETRIC, SILVER NITRATE)
1.0 SCOPE AND APPLICATION
1.1 This method is intended primarily for oxygen bomb combustates or
other waters where the chloride content is 5 mg/L or more and where interferences
such as color or high concentrations of heavy metal ions render Method 9252
impracticable.
2.0 SUMMARY OF METHOD
2.1 Water adjusted to pH 8.3 is titrated with silver nitrate solution
in the presence of potassium chromate indicator. The end point is indicated by
persistence of the orange-silver chromate color.
3.0 INTERFERENCES
3.1 Bromide, iodide, and sulfide are titrated along with the chloride.
Orthophosphate and polyphosphate interfere if present in concentrations greater
than 250 and 25 mg/L, respectively. Sulfite and objectionable color or turbidity
must be eliminated. Compounds that precipitate at pH 8.3 (certain hydroxides)
may cause error by occlusion.
3.2 Residual sodium carbonate from the bomb combustion may react with
silver nitrate to produce the precipitate, silver carbonate. This competitive
reaction may interfere with the visual detection of the end point. To remove
carbonate from the test solution, add small quantities of sulfuric acid followed
by agitation.
4.0 APPARATUS AND MATERIALS
4.1 Standard laboratory titrimetric equipment, including 1 mL or 5 mL
microburet with 0.01 mL gradations, and 25 mL buret.
4.2 Analytical balance: capable of weighing to 0.0001 g.
4.3 Class A volumetric flask: 1 L.
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 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Hydrogen peroxide (30%), H202.
9253 - 1 Revision 0
November 1992
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5.4 Phenolphthalein indicator solution (10 g/L).
5.5 Potassium chromate indicator solution. Dissolve 50 g of potassium
chromate (K2Cr04) in 100 ml of reagent water and add silver nitrate (AgN03) until
a slightly red precipitate is produced. Allow the solution to stand, protected
from light, for at least 24 hours after the addition of AgN03. Then filter the
solution to remove the precipitate and dilute to 1 L with reagent water.
5.6 Silver nitrate solution, standard (0.025N). Crush approximately
5 g of silver nitrate (AgNO,) crystals and dry to constant weight at 40°C.
Dissolve 4.2473 + 0.0002 g of the crushed, dried crystals in reagent water and
dilute to 1 L with reagent water. Standardize against the standard NaCl
solution, using the procedure given in Section 7.0.
5.7 Sodium chloride solution, standard (0.025N). Dissolve 1.4613 g ±
0.0002 g of sodium chloride (dried at 600°C for 1 hr) in chloride-free water in
a 1 liter Class A volumetric flask and dilute to the mark with reagent water.
5.8 Sodium hydroxide solution (0.25N). Dissolve approximately 10 g of
NaOH in reagent water and dilute to 1 L with reagent water.
5.9 Sulfuric acid (1:19), H2S04. Carefully add 1 volume of concentrated
sulfuric acid to 19 volumes of reagent water, while mixing.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must have been collected using a sampling plan that
addresses the considerations discussed in Chapter Nine of this manual.
6.2 There are no special requirements for preservation.
7.0 PROCEDURE
7.1 Pour 50 mL or less of the sample, containing between 0.25 mg and
20 mg of chloride ion, into a white porcelain container. Dilute to approximately
50 mL with reagent water, if necessary. Adjust the pH to the phenolphthalein end
point (pH 8.3) using H2S04 (Step 5.9) or NaOH solution (Step 5.8).
7.2 Add approximately 1.0 mL of K2Cr04 indicator solution and mix. Add
standard AgN03 solution dropwise from a 25 mL buret until the orange color
persists throughout the sample when illuminated with a yellow light or viewed
with yellow goggles.
7.3 Repeat the procedure described in Steps 7.1 and 7.2 using exactly
one-half as much original sample, diluted to 50 mL with halide-free water.
7.4 If sulfite ion is present, add 0.5 mL of H202 to the samples
described in Steps 7.2 and 7.3 and mix for 1 minute. Adjust the pH, then proceed
as described in Steps 7.2 and 7.3.
9253 - 2 Revision 0
November 1992
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7.5 Calculation
7.5.1 Calculate the chloride ion concentration in the original
sample, in milligrams per liter, as follows:
Chloride (mg/L) = [(V, - V2) x N x 71,000] / S
where:
V, = Milliliters of standard AgNO, solution added in titrating
the sample prepared in Step 7.1.
V2 = Milliliters of standard AgNO, solution added in titrating
the sample prepared in Step 7.3.
N = Normality of standard AgN03 solution.
S = Milliliters of original sample in the 50 ml test sample
prepared in Step 7.1.
71,000 = 2 x 35,500 mg CT/equivalent, since V., - 2V2.
8.0 QUALITY CONTROL
8.1 All quality control data should be maintained and available for
easy reference or inspection. Refer to Chapter One for specific quality control
guidelines.
8.2 Analyze a standard reference material to ensure that correct
procedures are being followed and that all standard reagents have been prepared
properly.
8.3 Employ a minimum of one blank per analytical batch or twenty
samples, whichever is more frequent, to determine if contamination has occurred.
8.4 Run one matrix spike and matrix duplicate every analytical batch
or twenty samples, whichever is more frequent. Matrix spikes and duplicates are
brought through the whole sample preparation and analytical process.
9.0 METHOD PERFORMANCE
9.1 These data are based on 32 data points obtained by five
laboratories who each analyzed four used crankcase oils and three fuel oil blends
with crankcase in duplicate. The samples were combusted using Method 5050. A
data point represents one duplicate analysis of a sample. Three data points were
judged to be outliers and were not included in these results.
9.1.1 Precision. The precision of the method as determined by
the statistical examination of interlaboratory test results is as follows:
Repeatability - The difference between successive results obtained
by the same operator with the same apparatus under constant operating
9253 - 3 Revision 0
November 1992
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conditions on identical test material would exceed, in the long run, in
the normal and correct operation of the test method, the following values
only in 1 case in 20 (see Table 1):
Repeatability - 0.36 x*
*where x is the average of two results in M9/g.
Reproducibilitv - The difference between two single and independent
results obtained by different operators working in different laboratories
on identical test material would exceed, in the long run, the following
values only in 1 case in 20:
Reproducibility - 0.71 x*
where x is the average of two results in ;ug/g.
9.1.2 Bias. The bias of this method varies with concentration,
as shown in Table 2:
Bias = Amount found - Amount expected
10.0 REFERENCES
1. Rohrbough, W.G.; et al. Reagent Chemicals, American Chemical Society
Specifications, 7th ed.; American Chemical Society: Washington, DC, 1986.
2. 1985 Annual Book of ASTM Standards. Vol. 11.01; "Standard Specification for
Reagent Water"; ASTM: Philadelphia, PA, 1985; D1193-77.
3. Gaskill, A.; Estes, E. D.; Hardison, D. L.; and Myers, L. E. "Validation
of Methods for Determining Chlorine in Used Oils and Oil Fuels," Prepared for
U.S. Environmental Protection Agency, Office of Solid Waste. EPA Contract No.
68-01-7075, WA 80. July 1988.
9253 - 4 Revision 0
November 1992
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TABLE 1.
REPEATABILITY AND REPRODUCIBILITY FOR CHLORINE IN USED
OILS BY BOMB OXIDATION AND SILVER NITRATE TITRATION
Average value
(M9/9)
Repeatability
(M9/9)
Reproducibility
(M9/9)
500
1,000
1,500
2,000
2,500
3,000
180
360
540
720
900
1,080
355
710
1,065
1,420
1,775
2,130
TABLE 2.
RECOVERY AND BIAS DATA FOR CHLORINE IN USED OILS BY
BOMB OXIDATION AND SILVER NITRATE TITRATION
Amount
expected
(M9/9)
320
480
920
1,498
1,527
3,029
3,045
Amount
found
(M9/9)
645
665
855
1,515
1,369
2,570
2,683
Bias,
(M9/9)
325
185
-65
17
-158
-460
-362
Percent
bias
+102
+39
-7
+1
-10
-15
-12
9253 - 5
Revision 0
November 1992
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METHOD 9253
CHLORIDE (TITRIMETRIC, SILVER NITRATE)
START
7 1 Place 50 ml
>ampIe in porcelain
container
7 4 Add hydrogen
peroxide, mm for 1
minute
7 1 Adjust pH to
8 3
7 2 Add 1 0 mL
potassium chromate
stir, add si1ver
nit rat* unt11
orange color
persis ts
7 3 Repeat step*
7 1 and 7 2 with
1/2 as much sampla
diluted to SO mL
7 5 Calculate
concent ration of
chloride in sample
STOP
9253 - 6
Revision 0
November 1992
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METHOD 1310
EXTRACTION PROCEDURE (EP) TOXICITY TEST METHOD
AND STRUCTURAL INTEGRITY TEST
1.0 SCOPE AND APPLICATION
1.1 This method is employed to determine whether a waste exhibits the
characteristic of Extraction Procedure Toxicity (see Chapter 7, Step 7.4 for
interim guidance).
1.2 The procedure may also be used to simulate the leaching which a
waste will undergo if disposed of in a sanitary landfill. Method 1310 is
applicable to liquid, solid, and multiphase samples.
2.0 SUMMARY OF METHOD
2.1 If a representative sample of the waste contains > 0.5% solids, the
solid phase of the sample is ground to pass a 9.5 mm sieve and extracted with
deionized water which is maintained at a pH of 5 ± 0.2, with acetic acid.
Wastes that contain < 0.5% solids are not subjected to extraction but are
directly analyzed. Monolithic wastes which can be formed into a cylinder
3.3 cm (dia) x 7.1 cm, or from which such a cylinder can be formed which is
representative of the waste, may be evaluated using the Structural Integrity
Procedure instead of being ground to pass a 9.5-mm sieve.
3.0 INTERFERENCES
3.1 Potential interferences that may be encountered during analysis are
discussed in the individual analytical methods.
4.0 APPARATUS AND MATERIALS
4.1 Extractor - For purposes of this test, an acceptable extractor is
one that will impart sufficient agitation to the mixture to (1) prevent
stratification of the sample and extraction fluid and (2) ensure that all
sample surfaces are continuously brought into contact with well-mixed
extraction fluid. Examples of suitable extractors are shown in Figures 1-3 of
this method and are available from: Associated Designs & Manufacturing Co.,
Alexandria, Virginia; Glas-Col Apparatus Co., Terre Haute, Indiana; Millipore,
Bedford, Massachusetts; and Rexnard, Milwaukee, Wisconsin.
4.2 pH meter or pH controller - Accurate to 0.05 pH units with
temperature compensation.
4.3 Filter holder - Capable of supporting a 0.45-um filter membrane and
of withstanding the pressure needed to accomplish separation. Suitable filter
holders range from simple vacuum units to relatively complex systems that can
exert up to 5.3 kg/cm3 (75 psi) of pressure. The type of filter holder used
depends upon the properties of the mixture to be filtered. Filter holders
known to EPA and deemed suitable for use are listed in Table 1.
1310 - 1 Revision 1
December 1987
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4.4 Filter membrane - Filter membrane suitable for conducting the
required filtration shall be fabricated from a material that (1) is not
physically changed by the waste material to be filtered and (2) does not
absorb or leach the chemical species for which a waste's EP extract will be
analyzed. Table 2 lists filter media known to the agency to be suitable for
solid waste testing.
4.4.1 In cases of doubt about physical effects on the filter,
contact the filter manufacturer to determine if the membrane or the
prefilter is adversely affected by the particular waste. If no
information is available, submerge the filter in the waste's liquid
phase. A filter that undergoes visible physical change after 48 hours
(i.e. curls, dissolves, shrinks, or swells) is unsuitable for use.
4.4.2 To test for absorbtion or leaching by the filter:
4.4.2.1 Prepare a standard solution of the chemical species
of interest.
4.4.2.2 Analyze the standard for its concentration of the
chemical species.
4.4.2.3 Filter the standard and reanalyze. If the
concentration of the filtrate differs from that of the original standard,
then the filter membrane leaches or absorbs one or more of the chemical
species and is not usable in this test method.
4.5 Structural integrity tester - A device meeting the specifications
shown in Figure 4 and having a 3.18-cm (1.25-in) diameter hammer weighing 0.33
kg (0.73 Ib) with a free fall of 15.24 cm (6 in) shall be used. This device
is available from Associated Design and Manufacturing Company, Alexandria, VA
22314, as Part No. 125, or it may be fabricated to meet these specifications.
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 ASTM Type II Water (ASTM Dl193-77 (1983)). All references to water
in the method refer to ASTM Type II unless otherwise specified.
5.3 Acetic acid (0.5N), CHsCOOH. This can be made by diluting
concentrated glacial acetic acid (17.5N) by adding 57 ml glacial acetic acid
to 1,000 ml of water and diluting to 2 liters. The glacial acetic acid should
be of high purity and monitored for impurities.
1310 - 2 Revision 1
December 1987
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5.4 Analytical standards should be prepared according to the applicable
analytical methods.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine of this manual.
6.2 Preservatives must not be added to samples.
6.3 Samples can be refrigerated if it is determined that refrigeration
will not affect the integrity of the sample.
7.0 PROCEDURE
7.1 If the waste does not contain any free liquid, go to Step 7.9. If
the sample is liquid or multiphase, continue as follows. Weigh filter
membrane and prefilter to ± 0.01 g. Handle membrane and prefliters with blunt
curved-tip forceps or vacuum tweezers, or by applying suction with a pipet.
7.2 Assemble filter holder, membranes, and prefilters following the
manufacturer's instructions. Place the 0.45-um membrane on the support screen
and add prefilters in ascending order of pore size. Do not prewet filter
membrane.
7.3 Weigh out a representative subsample of the waste (100 g minimum).
7.4 Allow slurries to stand, to permit the solid phase to settle.
Wastes that settle slowly may be centrifuged prior to filtration.
7.5 Wet the filter with a small portion of the liquid phase from the
waste or from the extraction mixture. Transfer the remaining material to the
filter holder and apply vacuum or gentle pressure (10-15 psi) until all liquid
passes through the filter. Stop filtration when air or pressurizing gas moves
through the membrane. If this point is not reached under vacuum or gentle
pressure, slowly increase the pressure in 10-psi increments to 75 psi. Halt
filtration when liquid flow stops. This liquid will constitute part or all of
the extract (refer to Step 7.16). The liquid should be refrigerated until
time of analysis.
NOTE: Oil samples or samples containing oil are treated in exactly the
same way as any other sample. The liquid portion of the sample is
filtered and treated as part of the EP extract. If the liquid
portion of the sample will not pass through the filter (usually
the case with heavy oils or greases), it should be carried through
the EP extraction as a solid.
7.6 Remove the solid phase and filter media and, while not allowing them
to dry, weigh to ± 0.01 g. The wet weight of the residue is determined by
calculating the weight difference between the weight of the filters (Step 7.1)
and the weight of the solid phase and the filter media.
1310 - 3 Revision 1
December 1987
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7.7 The waste will be handled differently from this point on, depending
on whether it contains more or less than 0.5% solids. If the sample appears
to have < 0.5% solids, determine the percent solids exactly (see Note below)
by the following procedure:
7.7.1 Dry the filter and residue at 80°C until two successive
weighings yield the same value.
7.7.2 Calculate the percent solids, using the following equation:
1 sol'ds = wei9nt °f filtered solid and filters - tared weight of filters ,g0
initial weight of waste material
NOTE: This procedure is used only to determine whether the solid must be
extracted or whether it can be discarded unextracted. It is not
used in calculating the amount of water or acid to use in the
extraction step. Do not extract solid material that has been
dried at 80°C. A new sample will have to be used for extraction
if a percent solids determination is performed.
7.8 If the solid constitutes < 0.5% of the waste, discard the solid and
proceed immediately to Step 7.17, treating the liquid phase as the extract.
7.9 The solid material obtained from Step 7.5 and all materials that do
not contain free liquids should be evaluated for particle size. If the solid
material has a surface area per g of material > 3.1 cm2 or passes through a
9.5-mm (0.375-in.) standard sieve, the operator should proceed to Step 7.11.
If the surface area is smaller or the particle size larger than specified
above, the solid material is prepared for extraction by crushing, cutting, or
grinding the material so that it passes through a 9.5-mm (0.375-in.) sieve or,
if the material is in a single piece, by subjecting the material to the
"Structural Integrity Procedure" described in Step 7.10.
7.10 Structural Integrity Procedure (SIP)
7.10.1 Cut a 3.3-cm diameter by 7.1-cm long cylinder from the
waste material. If the waste has been treated using a fixation process,
the waste may be cast in the form of a cylinder and allowed to cure for
30 days prior to testing.
7.10.2 Place waste into sample holder and assemble the tester.
Raise the hammer to its maximum height and drop. Repeat 14 additional
times.
7.10.3 Remove solid material from tester and scrape off any
particles adhering to sample holder. Weigh the waste to the nearest
0.01 g and transfer it to the extractor.
7.11 If the sample contains > 0.5% solids, use the wet weight of the
solid phase (obtained in Step 7.6) to calculate the amount of liquid and acid
to employ for extraction by using the following equation:
1310 - 4 Revision 1
December 1987
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W = Wf - Wt
where :
W = Wet weight in g of solid to be charged to extractor.
Wf = Wet weight in g of filtered solids and filter media.
Wt = Weight in g of tared filters.
If the waste does not contain any free liquids, 100 g of the material will be
subjected to the extraction procedure.
7.12 Place the appropriate amount of material (refer to Step 7.11) into
the extractor and add 16 times its weight with water.
7.13 After the solid material and water are placed in the extractor, the
operator should begin agitation and measure the pH of the solution in the
extractor. If the pH is > 5.0, the pH of the solution should be decreased to
5.0 + 0.2 by adding 0.5N acetic acid. If the pH is < 5.0, no acetic acid
should be added. The pH of the solution should be monitored, as described
below, during the course of the extraction, and, if the pH rises above 5.2,
0.5N acetic acid should be added to bring the pH down to 5.0 + 0.2. However,
in no event shall the aggregate amount of acid added to the solution exceed
4 ml of acid per g of solid. The mixture should be agitated for 24 hours and
maintained at 20-40eC (68-104T) during this time. It is recommended that the
operator monitor and adjust the pH during the course of the extraction with a
device such as the Type 45-A pH Controller, manufactured by Chemtrix, Inc.,
Hillsboro, Oregon 97123, or its equivalent, in conjunction with a metering
pump and reservoir of 0.5N acetic acid. If such a system is not available,
the following manual procedure shall be employed.
7.13.1 A pH meter should be calibrated in accordance with the
manufacturer's specifications.
7.13.2 The pH of the solution should be checked, and, if necessary,
0.5 N acetic acid should be manually added to the extractor until the pH
reaches 5.0 ± 0.2. The pH of the solution should be adjusted at 15-, 30-,
and 60-minute intervals, moving to the next longer interval if the pH
does not have to be adjusted > 0.5 pH units.
7.13.3 The adjustment procedure should be continued for at least 6
hours.
7.13.4 If, at the end of the 24-hour extraction period, the pH of
the solution is not below 5.2 and the maximum amount of acid (4 ml per g
of solids) has not been added, the pH should be adjusted to 5.0 + 0.2 and
the extraction continued for an additional 4 hours, during which the pH
should be adjusted at 1-hour intervals.
7.14 At the end of the extraction period, water should be added to the
extractor in an amount determined by the following equation:
1310 - 5 Revision 1
December 1987
-------
V = (20)(W) - 16(W) - A
where:
V = ml water to be added.
W = Weight in g of solid charged to extractor.
A = ml of 0.5N acetic acid added during extraction.
7.15 The material in the extractor should be separated into its
component liquid and solid phases in the following manner:
7.15.1 Allow slurries to stand to permit the solid phase to settle
(wastes that are slow to settle may be centrifuged prior to filtration)
and set up the filter apparatus (refer to Steps 4.3 and 4.4).
7.15.2 Wet the filter with a small portion of the liquid phase from
the waste or from the extraction mixture. Transfer the remaining
material to the filter holder and apply vacuum or gentle pressure
(10-15 psi) until all liquid passes through the filter. Stop filtration
when air or pressurizing gas moves through the membrane. If this point
is not reached under vacuum or gentle pressure, slowly increase the
pressure in 10-psi increments to 75 psi. Halt filtration when liquid
flow stops.
7.16 The liquids resulting from Steps 7.5 and 7.15 should be combined.
This combined liquid (or waste itself, if it has < 0.5% solids, as noted in
Step 7.8) is the extract and should be analyzed for the presence of any of the
contaminants specified in 40 CFR Part 261.24 using the analytical procedures
as designated in Step 7.17.
7.17 The extract is then prepared and analyzed using the appropriate
analytical methods described in Chapters Three and Four of this manual.
NOTE: If the EP extract includes two phases, concentration of
contaminants is determined by using a simple weighted average.
For example: An EP extract contains 50 ml of oil and 1,000 mL of
an aqueous phase. Contaminant concentrations are determined for
each phase. The final contamination concentration is taken to be:
(50)(contaminant cone, in oil) + (1,000)(contaminant cone, of aqueous phase)
1050
7.18 The extract concentrations are compared with the maximum
contamination limits listed in 40 CFR Part 261.24. If the extract
concentrations are greater than or equal to the respective values, the waste
then is considered to exhibit the characteristic of Extraction Procedure
Toxicity.
1310 - 6 Revision 1
December 1987
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8.0 QUALITY CONTROL
8.1 All quality control data should be maintained and available for easy
reference or inspection.
8.2 Employ a minimum of one blank per sample batch to determine if
contamination or any memory effects are occurring.
8.3 All quality control measures described in Chapter One and in the
referenced analytical methods should be followed.
9.0 METHOD PERFORMANCE
9.1 The data tabulated in Table 3 were obtained from records of state
and contractor laboratories and are intended to show the precision of the
entire method (1310 plus analysis method).
10.0 REFERENCES
1. Rohrbough, W.G.; et al. Reaoent Chemicals. American Chemical Society
Specifications, 7th ed.; American Chemical Society: Washington, DC, 1986.
2. 1985 Annual Book of ASTM Standards. Vol. 11.01; "Standard Specification
for Reagent Water"; ASTM: Philadelphia, PA, 1985; D1193-77.
3. Gaskill, A., Compilation and Evaluation of RCRA Method Performance Data,
Work Assignment No. 2, EPA Contract No. 68-01-7075, September 1986.
1310 - 7 Revision 1
December 1987
-------
TABLE 1. ERA-APPROVED FILTER HOLDERS
Manufacturer
Size
Model No.
Comments
Vacuum Filters
Nalgene
500 mL
Nuclepore
Millipore
Pressure Filters
Nuclepore
Micro Filtration
Systems
Millipore
47 mm
47 mm
142 mm
142 mm
142 mm
44-0045
410400
XX10 047 00
425900
302300
YT30 142 HW
Disposable plastic unit,
including prefilter, filter
pads, and reservoir; can be
used when solution is to
be analyzed for inorganic
constituents.
1310 - 8
Revision 1
December 1987
-------
METHOD 1330
EXTRACTION PROCEDURE FOR OILY WASTES
1.0 SCOPE AND APPLICATION
1.1 Method 1330 is used to determine the mobile metal concentration (MMC)
in oily wastes.
1.2 Method 1330 is applicable to API separator sludges, rag oils, slop
oil emulsions, and other oil wastes derived from petroleum refining.
2.0 SUMMARY OF METHOD
2.1 The sample is separated into solid and liquid components by
filtration.
2.2 The solid phase is placed in a Soxhlet extractor, charged with
tetrahydrofuran, and extracted. The THF is removed, the extractor is then
charged with toluene, and the sample is reextracted.
2.3 The EP method (Method 1310) is run on the dry solid residue.
2.4 The original liquid, combined extracts, and EP leachate are analyzed
for the EP metals.
3.0 INTERFERENCES
3.1 Matrix interferences will be coextracted from the sample. The extent
of these interferences will vary considerably from waste to waste, depending
on the nature and diversity of the particular refinery waste being analyzed.
4.0 APPARATUS AND MATERIALS
4.1 Soxhlet extraction apparatus.
4.2 Vacuum pump or other source of vacuum.
4.3 Buchner funnel 12.
4.4 Electric heating mantle.
4.5 Paper extraction thimble.
4.6 Filter paper.
4.7 Muslin cloth disks.
4.8 Evaporative flask - 250-mL.
4.9 Balance - Analytical, capable of weighing to + 0.5 mg.
1330 - 1 Revision 1
December 1987
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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 ASTM Type II Water (ASTM 01193-77 (1983)). All references to water in
the method refer to ASTM Type II unless otherwise specified.
5.3 Tetrahydrofuran, C^sO.
5.4 Toluene, CeHsC^.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 Samples must be collected in glass containers having a total volume
of at least 150 mL. No solid material should interfere with sealing the
sample container.
6.2 Sampling devices should be wiped clean with paper towels or absorbent
cloth, rinsed with a small amount of hexane followed by acetone rinse, and
dried between samples. Alternatively, samples can be taken with disposable
sampling devices in beakers.
7.0 PROCEDURE
7.1 Separate the sample (minimum 100 g) into its solid and liquid
components. The liquid component is defined as that portion of the sample
which passes through a 0.45 urn filter media under a pressure differential of
75 psi.
7.2 Determine the quantity of liquid (mL) and the concentration of the
toxicants of concern in the liquid phase (mg/L).
7.3 Place the solid phase into a Soxhlet extractor, charge the
concentration flask with 300 mL tetrahydrofuran, and extract for 3 hours.
7.4 Remove the flask containing tetrahydrofuran and replace it with one
containing 300 mL toluene.
7.5 Extract the solid a second time, for 3 hours, with the toluene.
7.6 Combine the tetrahydrofuran and toluene extracts.
7.7 Analyze the combined extracts for the toxicants of concern.
7.8 Determine the quantity of liquid (mL) and the concentration of the
toxicants of concern in the combined extracts (mg/L).
1330 - 2 Revision 1
December 1987
-------
7.9 Take the solid material remaining in the Soxhlet thimble and dry it
at 100"C for 30 minutes.
7.10 Run the EP (Method 1310) on the dried solid.
7.11 Calculate the mobile metal concentration (MMC) in mg/L using the
following formula:
MMC • 1.000 x
Ul + L2 + L3)
where:
Ql = Mass of toxicant in initial liquid phase of sample (amount of
liquid x concentration of toxicant) (mg).
Q2 = Mass of toxicant in combined organic extracts of sample
(amount of liquid x concentration of toxicant) (mg).
Q3 = Mass of toxicant in EP extract of solid (amount of extract x
concentration of toxicant) (mg).
LI = Volume of initial liquid (ml).
[_2 = Volume of liquid in EP (ml) = 20 x [weight of dried solid from
Step 7.8 (g)].
13 = Volume of liquid in THF and toluene extract (ml).
8.0 QUALITY CONTROL
8.1 Any reagent blanks or replicate samples should be subject to exactly
the same analytical procedures as those used on actual samples.
8.2 Refer to Chapter One for specific quality control procedures.
9.0 METHOD PERFORMANCE
9.1 No data provided.
10.0 REFERENCES
1. Rohrbough, W.G.; et al . Reagent Chemicals, American Chemical Society
Specifications, 7th ed.; American Chemical Society: Washington, DC,
1986.
2. 1985 Annual Book of ASTM Standards, Vol. 11.01; "Standard Specification
for Reagent Water"; ASTM: Philadelphia, PA, 1985; D1193-77.
1330 - 3 Revision 1
December 1987
-------
5.0
25
4.0
t
Non-Clogging Support Bushing
1 Inch Blade at 30° to Horizontal
9.0
Figure 1. Extractor.
1330 - 4
Revision 1
December 1987
-------
o
I
X
Ul
I
o
oc
oi
Ł
.1
1330 - 5
Revision 1
December 1987
-------
X
0)
E
0.
r>
«
.1
iZ
i 5
1330 - 6
Revision 1
December 1987
-------
Combined
Weight
.33 kg (.73 Ib)
' 15.25 cm \
(6")
(3.15cm)
11.25")
Sample
Eiastomeric
r
Sample Holder*
T / /
7.1 cm
(2.8")
3.3cm |^g.
(1.3") «^
* Elutomeric sample holder fabricated of material firm enough to support the sample.
Figure 4. Compaction tester.
1330 - 7
Revision 1
December 1987
-------
METHOD 1330
EXTRACTION PROCEDURE FOR OIL WASTE
welgr, filter
memora-ie ana
pref1leer
7.2
Accetic Je
filter holoer.
e f 1 lte"s
phase cettlc:
e«f>fifu9« If
Does waste
appear to contain
< 5X solias?
1330 - 8
Revision 1
December 1987
-------
METHOD 1330
(Continued)
7.8
Discard Solid
Area > 3.1 cmZ/gnt
or passes tnr<
9.S mm slaves
Area < 3.1 cm2/gn
s^ or particle site
Is surface^x* 9.5mm sieve
area or particle
slit o< trie
material?
7.10.1
Material Is In
s Ingl* Place
Cut or
cast
cyllnoer fro"!
«aste material
for Structural
Integrity Proc
7. tO.2
7.9
Prepare
materlal
for extraction
by crushing
cutting or
gr ma Ing
Astembla
tcater; oroo
hammer 15 times
7.10.3
Memove
•olid ••tcrlal:
Meign; transfer
to Extractor
1330 - 9
Revision 1
December 1987
-------
TABLE 2. ERA-APPROVED FILTRATION MEDIA
Supplier
Filter to be used
for aqueous systems
Filter to be used
for organic systems
Coarse prefilter
Gel man
Nuclepore
Millipore
Medium prefilters
Nuclepore
Millipore
Fine prefilters
Gel man
Nuclepore
Millipore
Fine filters (0.45 urn)
Gel man
Pall
Nuclepore
Millipore
Selas
61631, 61635
210907, 211707
AP25 035 00,
AP25 127 50
210905, 211705
AP20 035 00,
AP20 124 50
64798, 64803
210903, 211703
AP15 035 00,
AP15 124 50
L
60173, 60177
NX04750, NX14225
142218
HAWP 047 00,
HAWP 142 50
83485-02,
83486-02
61631, 61635
210907, 211707
AP25 035 00,
AP25 127 50
210905, 211705
AP20 035 00,
AP20 124 50
64798, 64803
210903, 211703
APIS 035 00,
AP15 124 50
60540 or 66149,
60544 or 66151
1422183
FHUP 047 00,
FHLP 142 50
83485-02,
83486-02
aSusceptible to decomposition by certain polar organic solvents.
1310 - 9
Revision 1
December 1987
-------
TABLE 3. PRECISIONS OF EXTRACTION-ANALYSIS
PROCEDURES FOR SEVERAL ELEMENTS
Element
Arsenic
Barium
Cadmium
Chromium
Mercury
Sample Matrix
1.
2.
3.
1.
2.
3.
1.
2.
3.
4.
5.
1.
2.
3.
4.
5.
1.
2.
3.
Auto fluff
Barrel sludge
Lumber treatment
company sediment
Lead smelting emission
control dust
Auto fluff
Barrel sludge
Lead smelting emission
control dust
Wastewater treatment
sludge from
electroplating
Auto fluff
Barrel sludge
Oil refinery
tertiary pond sludge
Wastewater treatment
sludge from
electroplating
Paint primer
Paint primer filter
Lumber treatment
company sediment
Oil refinery
tertiary pond sludge
Barrel sludge
Wastewater treatment
sludge from
electroplating
Lead smelting emission
control dust
Analysis
Method
7060
7060
7060
6010
7081
7081
3010/7130
3010/7130
7131
7131
7131
3010/7190
7191
7191
7191
7191
7470
7470
7470
Laboratory
Replicates
1.8, 1.5 ug/L
0.9, 2.6 ug/L
28, 42 mg/L
0.12, 0.12 mg/L
791, 780 ug/L
422, 380 ug/L
120, 120 mg/L
360, 290 mg/L
470, 610 ug/L
1100, 890 ug/L
3.2, 1.9 ug/L
1.1, 1.2 mg/L
61, 43 ug/L
--
0.81, 0.89 mg/L
--
0.15, 0.09 ug/L
1.4, 0.4 ug/L
0.4, 0.4 ug/L
1310 - 10
Revision 1
December 1987
-------
TABLE 3 (Continued)
Element
Sample Matrix
Analysis
Method
Laboratory
Replicates
Lead
Nickel
Chromium(VI)
1. Lead smelting emission
control dust
2. Auto fluff
3. Incinerator ash
4. Barrel sludge
5. Oil refinery
tertiary pond sludge
1. Sludge
2. Wastewater treatment
sludge from
electroplating
1. Wastewater treatment
sludge from
electroplating
3010/7420 940, 920 mg/L
7421
7421
7421
7421
7521
3010/7520
7196
1540, 1490 ug/L
1000, 974 ug/L
2550, 2800 ug/L
31, 29 ug/L
2260, 1720 ug/L
130, 140 mg/L
18, 19 ug/L
1310 - 11
Revision 1
December 1987
-------
FIGURE 1.
EXTRACTOR
Non-Clogging Support Bushing
1 Inch Bladt
-------
FIGURE 2.
ROTARY EXTRACTOR
1310 - 13
Revision 1
December 1987
-------
FIGURE 3.
EPRI EXTRACTOR
j
1310 - 14
Revision 1
December 1987
-------
FIGURE 4.
COMPACTION TESTER
Combined
• Weight
.33 kg (.73 Ib)
Elastomeric
Semple HoWtf *
T / /K
T
7.1 em
(2.8")
I
3.3 em
(1.3") '
I rf
^
* Elanomcric sample hoMtr fabricated of mattrial firm enough to support the temple.
1310 - 15
Revision 1
December 1987
-------
METHOD 1310
EXTRACTION PROCEDURE (EP) TOXICITY TEST METHOD
AND STRUCTURAL INTEGRITY TEST
filter
mcmorane ana
prefliter
7.Z
Assemole
filter rtoioer.
memorsnes. end
prefliters
pfi»*e cettlc
c«ntrlfuo«
75]
Filter
out liguia
ph»ie and
refrloerite It
7.6 |
trelgh vet solid
prose
Does
•ooetr to contain
<.5X sollos?
Olculcte
percent solid*
1310 - 16
Revision 1
December 1987
-------
METHOD 1310
(Continued)
7.a |
Discard Solid
Arc* > 3.1 cmZ/gm
or passes througn
9.S Him sieves
is surface
eree or particle
Size Of the
materiel?
Art* < 3.1 cmZ/gm
or particle size
> 9.5mm sieve
7.10.1
Materiel ic in
single piece
Cut or
cast
cylinder from
M*«te niteriei
for Structure]
Integrity Proc.
7.10.2
7.9
Preeere
' etaterial
for extraction
by crushing.
cutting or
grinding
Assemble
tester: crop
he»»er is tines
7.10.3
MeMove
solid (Mterial:
weign; transfer
to Extractor
o
1310 - 17
Revision 1
December 1987
-------
METHOD 1310
(Continued)
7. Ill
'Calculate
•mount of
liqule «no acid
to u«e for
••traction
7. 12|
Place
materiel into
extractor; mae
deionizea voter
7. 13
AllO"
• elurrie<
to etena.
•et up filter
eoperotuc
filter
Uee lOOg
of neterlel
for eittrectlon
proceourc
7. 16
Combine
' liquids
from section
7.S eno 7.is
to eneiyze for
contaminant*
Agitate
for 24 noura
•no monitor pH
of aolutlon
7.13
7.17
Obtain
analytical
method from
Table 1
Calibrate ana
ad)u>t pM meter
7. 16 I
Compare extract
concentration to max
contamination limits
In Table 1. to
determine CP toxlclty
7.14|
At and of
••traction
period add
delonited water
( Stop J
o
1310 - 18
Revision 1
December 1987
-------
METHOD 1312
SYNTHETIC PRECIPITATION LEACHING PROCEDURE
1.0 SCOPE AND APPLICATION
1.1 Method 1312 is designed to determine the mobility of both organic
and inorganic analytes present in samples of soils and wastes.
2.0 SUMMARY OF METHOD
2.1 For liquid samples (i.e., those containing less than 0.5 % dry
solid material), the sample, after filtration through a 0.6 to 0.8 jum glass
fiber filter, is defined as the 1312 extract.
2.2 For samples containing greater than 0.5 % solids, the liquid phase,
if any, is separated from the solid phase and stored for later analysis; the
particle size of the solid phase is reduced, if necessary. 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 region of the
country where the sample site is located if the sample is a soil. If the sample
is a waste or wastewater, the extraction fluid employed is a pH 4.2 solution.
A special extractor vessel is used when testing for volatile analytes (see Table
1 for a list of volatile compounds). Following extraction, the liquid extract
is separated from the sample by 0.6 to 0.8 urn glass fiber filter.
2.3 If compatible (i.e., multiple phases will not form on combination),
the initial liquid phase of the waste is added to the liquid extract, and these
are analyzed together. If incompatible, the liquids are analyzed separately and
the results are mathematically combined to yield a volume-weighted average
concentration.
3.0 INTERFERENCES
3.1 Potential interferences that may be encountered during analysis are
discussed in the individual analytical methods.
4.0 APPARATUS AND MATERIALS
4.1 Agitation apparatus: The agitation apparatus must be capable of
rotating the extraction vessel in an end-over-end fashion (see Figure 1) at 30
+ 2 rpm. Suitable devices known to EPA are identified in Table 2.
4.2 Extraction Vessels
4.2.1 Zero Headspace Extraction Vessel (ZHE). This device is for
use only when the sample is being tested for the mobility of volatile
analytes (i.e., those listed in Table 1). The ZHE (depicted in Figure 2)
allows for liquid/solid separation within the device and effectively
precludes headspace. This type of vessel allows for initial liquid/solid
separation, extraction, and final extract filtration without opening the
vessel (see Step 4.3.1). These vessels shall have an internal volume of
1312 - 1 Revision 0
November 1992
-------
500-600 ml and be equipped to accommodate a 90-110 mm filter. The devices
contain VITON*1 0-rings which should be replaced frequently. Suitable ZHE
devices known to EPA are identified in Table 3.
For the ZHE to be acceptable for use, the piston within the ZHE
should be able to be moved with approximately 15 psi or less. If it takes
more pressure to move the piston, the 0-rings in the device should be
replaced. If this does not solve the problem, the ZHE is unacceptable for
1312 analyses and the manufacturer should be contacted.
The ZHE should be checked for leaks after every extraction. If the
device contains a built-in pressure gauge, pressurize the device to 50
psi, allow it to stand unattended for 1 hour, and recheck the pressure.
If the device does not have a built-in pressure gauge, pressurize the
device to 50 psi, submerge it in water, and check for the presence of air
bubbles escaping from any of the fittings. If pressure is lost, check all
fittings and inspect and replace 0-rings, if necessary. Retest the
device. If leakage problems cannot be solved, the manufacturer should be
contacted.
Some ZHEs use gas pressure to actuate the ZHE piston, while others
use mechanical pressure (see Table 3). Whereas the volatiles procedure
(see Step 7.3) refers to pounds-per-square-inch (psi), for the
mechanically actuated piston, the pressure applied is measured in torque-
inch-pounds. Refer to the manufacturer's instructions as to the proper
conversion.
4.2.2 Bottle Extraction Vessel. When the sample is being
evaluated using the nonvolatile extraction, a jar with sufficient capacity
to hold the sample and the extraction fluid is needed. Headspace is
allowed in this vessel.
The extraction bottles may be constructed from various materials,
depending on the analytes to be analyzed and the nature of the waste (see
Step 4.3.3). It is recommended that borosilicate glass bottles be used
instead of other types of glass, especially when inorganics are of
concern. Plastic bottles, other than polytetrafluoroethylene, shall not
be used if organics are to be investigated. Bottles are available from a
number of laboratory suppliers. When this type of extraction vessel is
used, the filtration device discussed in Step 4.3.2 is used for initial
liquid/solid separation and final extract filtration.
4.3 Filtration Devices: It is recommended that all filtrations be
performed in a hood.
4.3.1 Zero-Headspace Extraction Vessel (ZHE): When the sample
is evaluated for volatiles, the zero-headspace extraction vessel described
in Step 4.2.1 is used for filtration. The device shall be capable of
supporting and keeping in place the glass fiber filter and be able to
withstand the pressure needed to accomplish separation (50. psi).
1VITON® is a trademark of Du Pont.
1312 - 2 Revision 0
November 1992
-------
NOTE: When it is suspected that the glass fiber filter has been ruptured, an
in-line glass fiber filter may be used to filter the material within the
ZHE.
4.3.2 Filter Holder: When the sample is evaluated for other than
volatile analytes, a filter holder capable of supporting a glass fiber
filter and able to withstand the pressure needed to accomplish separation
may be used. Suitable filter holders range from simple vacuum units to
relatively complex systems capable of exerting pressures of up to 50 psi
or more. The type of filter holder used depends on the properties of the
material to be filtered (see Step 4.3.3). These devices shall have a
minimum internal volume of 300 ml and be equipped to accommodate a minimum
filter size of 47 mm (filter holders having an internal capacity of 1.5 L
or greater, and equipped to accommodate a 142 mm diameter filter, are
recommended). Vacuum filtration can only be used for wastes with low
solids content (<10 %) and for highly granular, liquid-containing wastes.
All other types of wastes should be filtered using positive pressure
filtration. Suitable filter holders known to EPA are shown in Table 4.
4.3.3 Materials of Construction: Extraction vessels and
filtration devices shall be made of inert materials which will not leach
or absorb sample components. Glass, polytetrafluoroethylene (PTFE), or
type 316 stainless steel equipment may be used when evaluating the
mobility of both organic and inorganic components. Devices made of high-
density polyethylene (HOPE), polypropylene (PP), or polyvinyl chloride
(PVC) may be used only when evaluating the mobility of metals.
Borosilicate glass bottles are recommended for use over other types of
glass bottles, especially when inorganics are analytes of concern.
4.4 Filters: Filters shall be made of borosilicate glass fiber, shall
contain no binder materials, and shall have an effective pore size of 0.6 to
0.8-/im or equivalent. Filters known to EPA which meet these specifications are
identified in Table 5. Pre-filters must not be used. When evaluating the
mobility of metals, filters shall be acid-washed prior to use by rinsing with IN
nitric acid followed by three consecutive rinses with deionized distilled water
(a minimum of 1-L per rinse is recommended). Glass fiber filters are fragile and
should be handled with care.
4.5 pH Meters: The meter should be accurate to + 0.05 units at 25°C.
4.6 ZHE Extract Collection Devices: TEDLAR*2 bags or glass, stainless
steel or PTFE gas-tight syringes are used to collect the initial liquid phase and
the final extract when using the ZHE device. These devices listed are
recommended for use under the following conditions:
4.6.1 If a waste contains an aqueous liquid phase or if a waste
does not contain a significant amount of nonaqueous liquid (i.e.. <1 % of
total waste), the TEDLAR* bag or a 600 ml syringe should be used to collect
and combine the initial liquid and solid extract.
2TEDLAR* is a registered trademark of Du Pont.
1312 - 3 Revision 0
November 1992
-------
4.6.2 If a waste contains a significant amount of nonaqueous
liquid in the initiaj liquid phase (i.e., >1 % of total waste), the
syringe or the TEDLAR* bag may be used for both the initial solid/liquid
separation and the final extract filtration. However, analysts should use
one or the other, not both.
4.6.3 If the waste contains no initial liquid phase (is 100 %
solid)eor has no significant solid phase (is 100 % liquid), either the
TEDLAR* bag or the syringe may be used. If the syringe is used, discard
the first 5 ml of liquid expressed from the device. The remaining
aliquots are used for analysis.
4.7 ZHE Extraction Fluid Transfer Devices: Any device capable of
transferring the extraction fluid into the ZHE without changing the nature of the
extraction fluid is acceptable (e.g., a positive displacement or peristaltic
pump, a gas-tight syringe, pressure filtration unit (see Step 4.3.2), or other
ZHE device).
4.8 Laboratory Balance: Any laboratory balance accurate to within +
0.01 grams may be used (all weight measurements are to be within +0.1 grams).
4.9 Beaker or Erlenmeyer flask, glass, 500 mL.
4.10 Watchglass, appropriate diameter to cover beaker or Erlenmeyer
flask.
4.11 Magnetic stirrer.
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 Reagent Water. Reagent water is defined as water in which an
interferant is not observed at or above the method's detection limit of the
analyte(s) of interest. For nonvolatile extractions, ASTM Type II water or
equivalent meets the definition of reagent water. For volatile extractions, it
is recommended that reagent water be generated by any of the following methods.
Reagent water should be monitored periodically for impurities.
5.2.1 Reagent water for volatile extractions may be generated
by passing tap water through a carbon filter bed containing about 500
grams of activated carbon (Calgon Corp., Filtrasorb-300 or equivalent).
5.2.2 A water purification system (Millipore Super-Q or
equivalent) may also be used to generate reagent water for volatile
extractions.
1312 - 4 Revision 0
November 1992
-------
5.2.3 Reagent water for volatile extractions may also be prepared
by boiling water for 15 minutes. Subsequently, while maintaining the
water temperature at 90 ± 5 degrees C, bubble a contaminant-free inert gas
(e.g. nitrogen) through the water for 1 hour. While still hot, transfer
the water to a narrow mouth screw-cap bottle under zero-headspace and seal
with a Teflon-lined septum and cap.
5.3 Sulfuric acid/nitric acid (60/40 weight percent mixture) H2S04/HN03.
Cautiously mix 60 g of concentrated sulfuric acid with 40 g of concentrated
nitric acid.
5.4 Extraction fluids.
5.4.1 Extraction fluid #1: This fluid is made by adding the
60/40 weight percent mixture of sulfuric and nitric acids to reagent water
(Step 5.2) until the pH is 4.20 ± 0.05. The fluid is used to determine
the Teachability of soil from a site that is east of the Mississippi
River, and the Teachability of wastes and wastewaters.
NOTE: Solutions are unbuffered and exact pH may not be attained.
5.4.2 Extraction fluid #2: This fluid is made by adding the
60/40 weight percent mixture of sulfuric and nitric acids to reagent water
(Step 5.2) until the pH is 5.00 ± 0.05. The fluid is used to determine
the Teachability of soil from a site that is west of the Mississippi
River.
5.4.3 Extraction fluid #3: This fluid is reagent water (Step
5.2) and is used to determine cyanide and volatiles Teachability.
NOTE: These extraction fluids shouTd be monitored frequentTy for impurities.
The pH should be checked prior to use to ensure that these fluids are made
up accurately. If impurities are found or the pH is not within the above
specifications, the fluid shall be discarded and fresh extraction fluid
prepared.
5.5 Analytical standards shall be prepared according to the appropriate
analyticaT method.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 ATT sampTes shall be collected using an appropriate sampling plan.
6.2 There may be requirements on the minimal size of the field sample
depending upon the physical state or states of the waste and the analytes of
concern. An aliquot is needed for the preliminary evaluations of the percent
solids and the particle size. An aliquot may be needed to conduct the
nonvolatile analyte extraction procedure (see Step 1.4 concerning the use of this
extract for volatile organics). If volatile organics are of concern, another
aliquot may be needed. Quality control measures may require additional aliquots.
Further, it is always wise to colTect more sampTe just in case something goes
wrong with the initiaT attempt to conduct the test.
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6.3 Preservatives shall not be added to samples before extraction.
6.4 Samples may be refrigerated unless refrigeration results in
irreversible physical change to the waste. If precipitation occurs, the entire
sample (including precipitate) should be extracted.
6.5 When the sample is to be evaluated for volatile analytes, care
shall be taken to minimize the loss of volatiles. Samples shall be collected and
stored in a manner intended to prevent the loss of volatile analytes (e.g..
samples should be collected in Teflon-lined septum capped vials and stored at
4°C. Samples should be opened only immediately prior to extraction).
6.6 1312 extracts should be prepared for analysis and analyzed as soon
as possible following extraction. Extracts or portions of extracts for metallic
analyte determinations must be acidified with nitric acid to a pH < 2, unless
precipitation occurs (see Step 7.2.14 if precipitation occurs). Extracts should
be preserved for other analytes according to the guidance given in the individual
analysis methods. Extracts or portions of extracts for organic analyte
determinations shall not be allowed to come into contact with the atmosphere
(i.e., no headspace) to prevent losses. See Section 8.0 (Quality Control) for
acceptable sample and extract holding times.
7.0 PROCEDURE
7.1 Preliminary Evaluations
Perform preliminary 1312 evaluations on a minimum 100 gram aliquot of
sample. This aliquot may not actually undergo 1312 extraction. These
preliminary evaluations include: (1) determination of the percent solids (Step
7.1.1); (2) determination of whether the waste contains insignificant solids and
is, therefore, its own extract after filtration (Step 7.1.2); and (3)
determination of whether the solid portion of the waste requires particle size
reduction (Section 7.1.3).
7.1.1 Preliminary determination of percent solids: Percent
solids is defined as that fraction of a waste sample (as a percentage of
the total sample) from which no liquid may be forced out by an applied
pressure, as described below.
7.1.1.1 If the sample will obviously yield no free
liquid when subjected to pressure filtration (i.e.. is 100%
solids), weigh out a representative subsample (100 g minimum) and
proceed to Step 7.1.3.
7.1.1.2 If the sample is liquid or multiphasic,
liquid/solid separation to make a preliminary determination of
percent solids is required. This involves the filtration device
discussed in Step 4.3.2, and is outlined in Steps 7.1.1.3 through
7.1.1.9.
7.1.1.3 Pre-weigh the filter and the container that will
receive the filtrate.
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7.1.1.4 Assemble filter holder and filter following the
manufacturer's instructions. Place the filter on the support
screen and secure.
7.1.1.5 Weigh out a subsample of the waste (100 gram
minimum) and record the weight.
7.1.1.6 Allow slurries to stand to permit the solid phase
to settle. Samples that settle slowly may be centrifuged prior to
filtration. Centrifugation is to be used only as an aid to
filtration. If used, the liquid should be decanted and filtered
followed by filtration of the solid portion of the waste through
the same filtration system.
7.1.1.7 Quantitatively transfer the sample to the filter
holder (liquid and solid phases). Spread the sample evenly over
the surface of the filter. If filtration of the waste at 4eC
reduces the amount of expressed liquid over what would be expressed
at room temperature, then allow the sample to warm up to room
temperature in the device before filtering.
NOTE: If sample material (>1 % of original sample weight) has obviously adhered
to the container used to transfer the sample to the filtration apparatus,
determine the weight of this residue and subtract it from the sample
weight determined in Step 7.1.1.5 to determine the weight of the sample
that will be filtered.
Gradually apply vacuum or gentle pressure of 1-10 psi, until air
or pressurizing gas moves through the filter. If this point is not
reached under 10 psi, and if no additional liquid has passed through the
filter in any 2-minute interval, slowly increase the pressure in 10 psi
increments to a maximum of 50 psi. After each incremental increase of 10
psi, if the pressurizing gas has not moved through the filter, and if no
additional liquid has passed through the filter in any 2-minute interval,
proceed to the next 10-psi increment. When the pressurizing gas begins to
move through the filter, or when liquid flow has ceased at 50 psi (i.e..
filtration does not result in any additional filtrate within any 2-minute
period), stop the filtration.
NOTE: Instantaneous application of high pressure can degrade the glass fiber
filter and may cause premature plugging.
7.1.1.8 The material in the filter holder is defined as
the solid phase of the sample, and the filtrate is defined as the
liquid phase.
NOTE: Some samples, such as oily wastes and some paint wastes, will obviously
contain some material that appears to be a liquid, but even after applying
vacuum or pressure filtration, as outlined in Step 7.1.1.7, this material
may not filter. If this is the case, the material within the filtration
device is defined as a solid. Do not replace the original filter with a
fresh filter under any circumstances. Use only one filter.
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7.1.1.9 Determine the weight of the liquid phase by
subtracting the weight of the filtrate container (see Step 7.1.1.3)
from the total weight of the filtrate-filled container. Determine
the weight of the solid phase of the sample by subtracting the
weight of the liquid phase from the weight of the total sample, as
determined in Step 7.1.1.5 or 7.1.1.7.
Record the weight of the liquid and solid phases.
Calculate the percent solids as follows:
Weight of solid (Step 7.1.1.9)
Percent solids = x 100
Total weight of waste (Step 7.1.1.5 or 7.1.1.7)
7.1.2 If the percent solids determined in Step 7.1.1.9 is equal
to or greater than 0.5%, then proceed either to Step 7.1.3 to determine
whether the solid material requires particle size reduction or to Step
7.1.2.1 if it is noticed that a small amount of the filtrate is entrained
in wetting of the filter. If the percent solids determined in Step
7.1.1.9 is less than 0.5%, then proceed to Step 7.2.9 if the nonvolatile
1312 analysis is to be performed, and to Section 7.3 with a fresh portion
of the waste if the volatile 1312 analysis is to be performed.
7.1.2.1 Remove the solid phase and filter from the
filtration apparatus.
7.1.2.2 Dry the filter and solid phase at 100 ± 20"C
until two successive weighings yield the same value within + 1 %.
Record the final weight.
Note: Caution should be taken to ensure that the subject solid will not flash
upon heating. It is recommended that the drying oven be vented to a hood
or other appropriate device.
7.1.2.3 Calculate the percent dry solids as follows:
Percent (Weight of dry sample + filter) - tared weight of filter
dry solids = x 100
Initial weight of sample (Step 7.1.1.5 or 7.1.1.7)
7.1.2.4 If the percent dry solids is less than 0.5%,
then proceed to Step 7.2.9 if the nonvolatile 1312 analysis is to
be performed, and to Step 7.3 if the volatile 1312 analysis is to
be performed. If the percent dry solids is greater than or equal
to 0.5%, and if the nonvolatile 1312 analysis is to be performed,
return to the beginning of this Section (7.1) and, with a fresh
portion of sample, determine whether particle size reduction is
necessary (Step 7.1.3).
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7.1.3 Determination of whether the sample requires particle-size
reduction (particle-size is reduced during this step): Using the solid
portion of the sample, evaluate the solid for particle size. Particle-
size reduction is required, unless the solid has a surface area per gram
of material equal to or greater than 3.1 cm2, or is smaller than 1 cm in
its narrowest dimension (i.e.. is capable of passing through a 9.5 mm
(0.375 inch) standard sieve). If the surface area is smaller or the
particle size larger than described above, prepare the solid portion of
the sample for extraction by crushing, cutting, or grinding the waste to
a surface area or particle size as described above. If the solids are
prepared for organic volatiles extraction, special precautions must be
taken (see Step 7.3.6).
Note: Surface area criteria are meant for filamentous (e.g., paper, cloth, and
similar) waste materials. Actual measurement of surface area is not
required, nor is it recommended. For materials that do not obviously meet
the criteria, sample-specific methods would need to be developed and
employed to measure the surface area. Such methodology is currently not
available.
7.1.4 Determination of appropriate extraction fluid:
7.1.4.1 For soils, if the sample is from a site that is
east of the Mississippi River, extraction fluid #1 should be used.
If the sample is from a site that is west of the Mississippi River,
extraction fluid #2 should be used.
7.1.4.2 For wastes and wastewater, extraction fluid #1
should be used.
7.1.4.3 For cyanide-containing wastes and/or soils,
extraction fluid #3 (reagent water) must be used because leaching
of cyanide-containing samples under acidic conditions may result
in the formation of hydrogen cyanide gas.
7.1.5 If the aliquot of the sample used for the preliminary
evaluation (Steps 7.1.1 - 7.1.4) was determined to be 100% solid at Step
7.1.1.1, then it can be used for the Section 7.2 extraction (assuming at
least 100 grams remain), and the Section 7.3 extraction (assuming at least
25 grams remain). If the aliquot was subjected to the procedure in Step
7.1.1.7, then another aliquot shall be used for the volatile extraction
procedure in Section 7.3. The aliquot of the waste subjected to the
procedure in Step 7.1.1.7 might be appropriate for use for the Section 7.2
extraction if an adequate amount of solid (as determined by Step 7.1.1.9)
was obtained. The amount of solid necessary is dependent upon whether a
sufficient amount of extract will be produced to support the analyses. If
an adequate amount of solid remains, proceed to Step 7.2.10 of the
nonvolatile 1312 extraction.
7.2 Procedure when Volatiles are not Involved
A minimum sample size of 100 grams (solid and liquid phases) is
recommended. In some cases, a larger sample size may be appropriate, depending
on the solids content of the waste sample (percent solids, See Step 7.1.1),
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whether the initial liquid phase of the waste will be miscible with the aqueous
extract of the solid, and whether inorganics, semivolatile organics, pesticides,
and herbicides are all analytes of concern. Enough solids should be generated
for extraction such that the volume of 1312 extract will be sufficient to support
all of the analyses required. If the amount of extract generated by a single
1312 extraction will not be sufficient to perform all of the analyses, more than
one extraction may be performed and the extracts from each combined and aliquoted
for analysis.
7.2.1 If the sample will obviously yield no liquid when subjected
to pressure filtration (i.e.. is 100 % solid, see Step 7.1.1), weigh out
a subsample of the sample (100 gram minimum) and proceed to Step 7.2.9.
7.2.2 If the sample is liquid or multiphasic, liquid/solid
separation is required. This involves the filtration device described in
Step 4.3.2 and is outlined in Steps 7.2.3 to 7.2.8.
7.2.3 Pre-weigh the container that will receive the filtrate.
7.2.4 Assemble the filter holder and filter following the
manufacturer's instructions. Place the filter on the support screen and
secure. Acid wash the filter if evaluating the mobility of metals (see
Step 4.4).
Note: Acid washed filters may be used for all nonvolatile extractions even when
metals are not of concern.
7.2.5 Weigh out a subsample of the sample (100 gram minimum) and
record the weight. If the waste contains <0.5 % dry solids (Step 7.1.2),
the liquid portion of the waste, after filtration, is defined as the 1312
extract. Therefore, enough of the sample should be filtered so that the
amount of filtered liquid will support all of the analyses required of the
1312 extract. For wastes containing >0.5 % dry solids (Steps 7.1.1 or
7.1.2), use the percent solids information obtained in Step 7.1.1 to
determine the optimum sample size (100 gram minimum) for filtration.
Enough solids should be generated by filtration to support the analyses to
be performed on the 1312 extract.
7.2.6 Allow slurries to stand to permit the solid phase to settle.
Samples that settle slowly may be centrifuged prior to filtration. Use
centrifugation only as an aid to filtration. If the sample is
centrifuged, the liquid should be decanted and filtered followed by
filtration of the solid portion of the waste through the same filtration
system.
7.2.7 Quantitatively transfer the sample (liquid and solid phases)
to the filter holder (see Step 4.3.2). Spread the waste sample evenly
over the surface of the filter. If filtration of the waste at 4°C reduces
the amount of expressed liquid over what would be expressed at room
temperature, then allow the sample to warm up to room temperature in the
device before filtering.
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NOTE: If waste material (>1 % of the original sample weight) has obviously
adhered to the container used to transfer the sample to the filtration
apparatus, determine the weight of this residue and subtract it from the
sample weight determined in Step 7.2.5, to determine the weight of the
waste sample that will be filtered.
Gradually apply vacuum or gentle pressure of 1-10 psi, until air
or pressurizing gas moves through the filter. If this point if not
reached under 10 psi, and if no additional liquid has passed through the
filter in any 2-minute interval, slowly increase the pressure in 10-psi
increments to maximum of 50 psi. After each incremental increase of 10
psi, if the pressurizing gas has not moved through the filter, and if no
additional liquid has passed through the filter in any 2-minute interval,
proceed to the next 10-psi increment. When the pressurizing gas begins to
move through the filter, or when the liquid flow has ceased at 50 psi
(i.e., filtration does not result in any additional filtrate within a
2-minute period), stop the filtration.
NOTE: Instantaneous application of high pressure can degrade the glass fiber
filter and may cause premature plugging.
7.2.8 The material in the filter holder is defined as the solid
phase of the sample, and the filtrate is defined as the liquid phase.
Weigh the filtrate. The liquid phase may now be either analyzed (see
Steps 7.2.12) or stored at 4°C until time of analysis.
NOTE: Some wastes, such as oily wastes and some paint wastes, will obviously
contain some material which appears to be a liquid. Even after applying
vacuum or pressure filtration, as outlined in Step 7.2.7, this material
may not filter. If this is the case, the material within the filtration
device is defined as a solid, and is carried through the extraction as a
solid. Do not replace the original filter with a fresh filter under any
circumstances. Use only one filter.
7.2.9 If the sample contains <0.5% dry solids (see Step 7.1.2),
proceed to Step 7.2.13. If the sample contains >0.5 % dry solids (see
Step 7.1.1 or 7.1.2), and if particle-size reduction of the solid was
needed in Step 7.1.3, proceed to Step 7.2.10. If the sample as received
passes a 9.5 mm sieve, quantitatively transfer the solid material into the
extractor bottle along with the filter used to separate the initial liquid
from the solid phase, and proceed to Step 7.2.11.
7.2.10 Prepare the solid portion of the sample for extraction by
crushing, cutting, or grinding the waste to a surface area or particle-
size as described in Step 7.1.3. When the surface area or particle-size
has been appropriately altered, quantitatively transfer the solid material
into an extractor bottle. Include the filter used to separate the initial
liquid from the solid phase.
NOTE: Sieving of the waste is not normally required. Surface area requirements
are meant for filamentous (e.g.. paper, cloth) and similar waste
materials. Actual measurement of surface area is not recommended. If
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sieving is necessary, a Teflon-coated sieve should be used to avoid
contamination of the sample.
7.2.11 Determine the amount of extraction fluid to add to the
extractor vessel as follows:
20 x % solids (Step 7.1.1) x weight of waste
filtered (Step 7.2.5 or 7.2.7)
Weight of =
extraction fluid
100
Slowly add this amount of appropriate extraction fluid (see Step
7.1.4) to the extractor vessel. Close the extractor bottle tightly (it is
recommended that Teflon tape be used to ensure a tight seal), secure in
rotary extractor device, and rotate at 30 ± 2 rpm for 18 + 2 hours.
Ambient temperature (i .e.. temperature of room in which extraction takes
place) shall be maintained at 23 + 2°C during the extraction period.
NOTE: As agitation continues, pressure may build up within the extractor bottle
for some types of sample (e.g., limed or calcium carbonate-containing
sample may evolve gases such as carbon dioxide). To relieve excess
pressure, the extractor bottle may be periodically opened (e.g., after 15
minutes, 30 minutes, and 1 hour) and vented into a hood.
7.2.12 Following the 18 ± 2 hour extraction, separate the material
in the extractor vessel into its component liquid and solid phases by
filtering through a new glass fiber filter, as outlined in Step 7.2.7.
For final filtration of the 1312 extract, the glass fiber filter may be
changed, if necessary, to facilitate filtration. Filter(s) shall be
acid-washed (see Step 4.4) if evaluating the mobility of metals.
7.2.13 Prepare the 1312 extract as follows:
7.2.13.1 If the sample contained no initial liquid phase,
the filtered liquid material obtained from Step 7.2.12 is defined
as the 1312 extract. Proceed to Step 7.2.14.
7.2.13.2 If compatible (e.g.. multiple phases will not
result on combination), combine the filtered liquid resulting from
Step 7.2.12 with the initial liquid phase of the sample obtained
in Step 7.2.7. This combined liquid is defined as the 1312
extract. Proceed to Step 7.2.14.
7.2.13.3 If the initial liquid phase of the waste, as
obtained from Step 7.2.7, is not or may not be compatible with the
filtered liquid resulting from Step 7.2.12, do not combine these
liquids. Analyze these liquids, collectively defined as the 1312
extract, and combine the results mathematically, as described in
Step 7.2.14.
7.2.14 Following collection of the 1312 extract, the pH of the
extract should be recorded. Immediately aliquot and preserve the extract
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for analysis. Metals aliquots must be acidified with nitric acid to pH <
2. If precipitation is observed upon addition of nitric acid to a small
aliquot of the extract, then the remaining portion of the extract for
metals analyses shall not be acidified and the extract shall be analyzed
as soon as possible. All other aliquots must be stored under
refrigeration (4°C) until analyzed. The 1312 extract shall be prepared
and analyzed according to appropriate analytical methods. 1312 extracts
to be analyzed for metals shall be acid digested except in those instances
where digestion causes loss of metallic analytes. If an analysis of the
undigested extract shows that the concentration of any regulated metallic
analyte exceeds the regulatory level, then the waste is hazardous and
digestion of the extract is not necessary. However, data on undigested
extracts alone cannot be used to demonstrate that the waste is not
hazardous. If the individual phases are to be analyzed separately,
determine the volume of the individual phases (to ± 0.5 %), conduct the
appropriate analyses, and combine the results mathematically by using a
simple volume-weighted average:
(V,) (C,) + (V2) (C2)
Final Analyte Concentration =
where:
V, = The volume of the first phase (L).
C, = The concentration of the analyte of concern in the first phase (mg/L).
V2 = The volume of the second phase (L).
C2 = The concentration of the analyte of concern in the second phase
(mg/L).
7.2.15 Compare the analyte concentrations in the 1312 extract with
the levels identified in the appropriate regulations. Refer to Section
8.0 for quality assurance requirements.
7.3 Procedure when Volatiles are Involved
Use the ZHE device to obtain 1312 extract for analysis of volatile
compounds only. Extract resulting from the use of the ZHE shall not be used to
evaluate the mobility of non-volatile analytes (e.g., metals, pesticides, etc.).
The ZHE device has approximately a 500 ml internal capacity. The ZHE can
thus accommodate a maximum of 25 grams of solid (defined as that fraction of a
sample from which no additional liquid may be forced out by an applied pressure
of 50 psi), due to the need to add an amount of extraction fluid equal to 20
times the weight of the solid phase.
Charge the ZHE with sample only once and do not open the device until the
final extract (of the solid) has been collected. Repeated filling of the ZHE to
obtain 25 grams of solid is not permitted.
Do not allow the sample, the initial liquid phase, or the extract to be
exposed to the atmosphere for any more time than is absolutely necessary. Any
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manipulation of these materials should be done when cold (4°C) to minimize loss
of volatiles.
7.3.1 Pre-weigh the (evacuated) filtrate collection container
(see Step 4.6) and set aside. If using a TEDLAR* bag, express all liquid
from the ZHE device into the bag, whether for the initial or final
liquid/solid separation, and take an aliquot from the liquid in the bag
for analysis. The containers listed in Step 4.6 are recommended for use
under the conditions stated in Steps 4.6.1-4.6.3.
7.3.2 Place the ZHE piston within the body of the ZHE (it may be
helpful first to moisten the piston 0-rings slightly with extraction
fluid). Adjust the piston within the ZHE body to a height that will
minimize the distance the piston will have to move once the ZHE is charged
with sample (based upon sample size requirements determined from Step 7.3,
Step 7.1.1 and/or 7.1.2). Secure the gas inlet/outlet flange (bottom
flange) onto the ZHE body in accordance with the manufacturer's
instructions. Secure the glass fiber filter between the support screens
and set aside. Set liquid inlet/outlet flange (top flange) aside.
7.3.3 If the sample is 100% solid (see Step 7.1.1), weigh out
a subsample (25 gram maximum) of the waste, record weight, and proceed to
Step 7.3.5.
7.3.4 If the sample contains <0.5% dry solids (Step 7.1.2), the
liquid portion of waste, after filtration, is defined as the 1312 extract.
Filter enough of the sample so that the amount of filtered liquid will
support all of the volatile analyses required. For samples containing
>0.5% dry solids (Steps 7.1.1 and/or 7.1.2), use the percent solids
information obtained in Step 7.1.1 to determine the optimum sample size to
charge into the ZHE. The recommended sample size is as follows:
7.3.4.1 For samples containing <5% solids (see Step
7.1.1), weigh out a 500 gram subsample of waste and record the
weight.
7.3.4.2 For wastes containing >5% solids (see Step
7.1.1), determine the amount of waste to charge into the ZHE as
follows:
25
Weight of waste to charge ZHE = x 100
percent solids (Step 7.1.1)
Weigh out a subsample of the waste of the appropriate size and
record the weight.
7.3.5 If particle-size reduction of the solid portion of the
sample was required in Step 7.1.3, proceed to Step 7.3.6. If particle-
size reduction was not required in Step 7.1.3, proceed to Step 7.3.7.
7.3.6 Prepare the sample for extraction by crushing, cutting, or
grinding the solid portion of the waste to a surface area or particle size
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as described in Step 7.1.3.1. Wastes and appropriate reduction equipment
should be refrigerated, if possible, to 4°C prior to particle-size
reduction. The means used to effect particle-size reduction must not
generate heat in and of itself. If reduction of the solid phase of the
waste is necessary, exposure of the waste to the atmosphere should be
avoided to the extent possible.
NOTE: Sieving of the waste is not recommended due to the possibility that
volatiles may be lost. The use of an appropriately graduated ruler is
recommended as an acceptable alternative. Surface area requirements are
meant for filamentous (e.g., paper, cloth) and similar waste materials.
Actual measurement of surface area is not recommended.
When the surface area or particle-size has been appropriately
altered, proceed to Step 7.3.7.
7.3.7 Waste slurries need not be allowed to stand to permit the
solid phase to settle. Do not centrifuge samples prior to filtration.
7.3.8 Quantitatively transfer the entire sample (liquid and solid
phases) quickly to the ZHE. Secure the filter and support screens into
the top flange of the device and secure the top flange to the ZHE body in
accordance with the manufacturer's instructions. Tighten all ZHE fittings
and place the device in the vertical position (gas inlet/outlet flange on
the bottom). Do not attach the extraction collection device to the top
plate.
Note: If sample material (>1% of original sample weight) has obviously adhered
to the container used to transfer the sample to the ZHE, determine the
weight of this residue and subtract it from the sample weight determined
in Step 7.3.4 to determine the weight of the waste sample that will be
filtered.
Attach a gas line to the gas inlet/outlet valve (bottom flange)
and, with the liquid inlet/outlet valve (top flange) open, begin applying
gentle pressure of 1-10 psi (or more if necessary) to force all headspace
slowly out of the ZHE device into a hood. At the first appearance of
liquid from the liquid inlet/outlet valve, quickly close the valve and
discontinue pressure. If filtration of the waste at 4°C reduces the
amount of expressed liquid over what would be expressed at room
temperature, then allow the sample to warm up to room temperature in the
device before filtering. If the waste is 100 % solid (see Step 7.1.1),
slowly increase the pressure to a maximum of 50 psi to force most of the
headspace out of the device and proceed to Step 7.3.12.
7.3.9 Attach the evacuated pre-weighed filtrate collection
container to the liquid inlet/outlet valve and open the valve. Begin
applying gentle pressure of 1-10 psi to force the liquid phase of the
sample into the filtrate collection container. If no additional liquid
has passed through the filter in any 2-minute interval, slowly increase
the pressure in 10-psi increments to a maximum of 50 psi. After each
incremental increase of 10 psi, if no additional liquid has passed through
the filter in any 2-minute interval, proceed to the next 10-psi increment.
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When liquid flow has ceased such that continued pressure filtration at 50
psi does not result in any additional filtrate within a 2-minute period,
stop the filtration. Close the liquid inlet/outlet valve, discontinue
pressure to the piston, and disconnect and weigh the filtrate collection
container.
NOTE: Instantaneous application of high pressure can degrade the glass fiber
filter and may cause premature plugging.
7.3.10 The material in the ZHE is defined as the solid phase of
the sample and the filtrate is defined as the liquid phase.
NOTE: Some samples, such as oily wastes and some paint wastes, will obviously
contain some material which appears to be a liquid. Even after applying
pressure filtration, this material will not filter. If this is the case,
the material within the filtration device is defined as a solid, and is
carried through the 1312 extraction as a solid.
If the original waste contained <0.5 % dry solids (see Step 7.1.2),
this filtrate is defined as the 1312 extract and is analyzed directly.
Proceed to Step 7.3.15.
7.3.11 The liquid phase may now be either analyzed immediately
(see Steps 7.3.13 through 7.3.15) or stored at 4'C under minimal headspace
conditions until time of analysis. Determine the weight of extraction
fluid #3 to add to the ZHE as follows:
20 x % solids (Step 7.1.1) x weight
of waste filtered (Step 7.3.4 or 7.3.8)
Weight of extraction fluid =
100
7.3.12 The following steps detail how to add the appropriate
amount of extraction fluid to the solid material within the ZHE and
agitation of the ZHE vessel. Extraction fluid #3 is used in all cases
(see Step 5.7).
7.3.12.1 With the ZHE in the vertical position, attach a
line from the extraction fluid reservoir to the liquid inlet/outlet
valve. The line used shall contain fresh extraction fluid and
should be preflushed with fluid to eliminate any air pockets in the
line. Release gas pressure on the ZHE piston (from the gas
inlet/outlet valve), open the liquid inlet/outlet valve, and begin
transferring extraction fluid (by pumping or similar means) into
the ZHE. Continue pumping extraction fluid into the ZHE until the
appropriate amount of fluid has been introduced into the device.
7.3.12.2 After the extraction fluid has been added,
immediately close the liquid inlet/outlet valve and disconnect the
extraction fluid line. Check the ZHE to ensure that all valves are
in their closed positions. Manually rotate the device in an
end-over-end fashion 2 or 3 times. Reposition the ZHE in the
1312 - 16 Revision 0
November 1992
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vertical position with the liquid inlet/outlet valve on top.
Pressurize the ZHE to 5-10 psi (if necessary) and slowly open the
liquid inlet/outlet valve to bleed out any headspace (into a hood)
that may have been introduced due to the addition of extraction
fluid. This bleeding shall be done quickly and shall be stopped
at the first appearance of liquid from the valve. Re-pressurize
the ZHE with 5-10 psi and check all ZHE fittings to ensure that
they are closed.
7.3.12.3 Place the ZHE in the rotary extractor apparatus
(if it is not already there) and rotate at 30 + 2 rpm for 18+2
hours. Ambient temperature (i.e., temperature of room in which
extraction occurs) shall be maintained at 23 ± 2°C during
agitation.
7.3.13 Following the 18+2 hour agitation period, check the
pressure behind the ZHE piston by quickly opening and closing the gas
inlet/outlet valve and noting the escape of gas. If the pressure has not
been maintained (i.e., no gas release observed), the ZHE is leaking.
Check the ZHE for leaking as specified in Step 4.2.1, and perform the
extraction again with a new sample of waste. If the pressure within the
device has been maintained, the material in the extractor vessel is once
again separated into its component liquid and solid phases. If the waste
contained an initial liquid phase, the liquid may be filtered directly
into the same filtrate collection container (i.e., TEDLAR* bag) holding the
initial liquid phase of the waste. A separate filtrate collection
container must be used if combining would create multiple phases, or there
is not enough volume left within the filtrate collection container.
Filter through the glass fiber filter, using the ZHE device as discussed
in Step 7.3.9. All extracts shall be filtered and collected if the TEDLAR*
bag is used, if the extract is multiphasic, or if the waste contained an
initial liquid phase (see Steps 4.6 and 7.3.1).
NOTE: An in-line glass fiber filter may be used to filter the material within
the ZHE if it is suspected that the glass fiber filter has been ruptured
7.3.14 If the original sample contained no initial liquid phase,
the filtered liquid material obtained from Step 7.3.13 is defined as the
1312 extract. If the sample contained an initial liquid phase, the
filtered liquid material obtained from Step 7.3.13 and the initial liquid
phase (Step 7.3.9) are collectively defined as the 1312 extract.
7.3.15 Following collection of the 1312 extract, immediately
prepare the extract for analysis and store with minimal headspace at 4°C
until analyzed. Analyze the 1312 extract according to the appropriate
analytical methods. If the individual phases are to be analyzed
separately (i.e., are not miscible), determine the volume of the
individual phases (to 0.5%), conduct the appropriate analyses, and combine
the results mathematically by using a simple volume- weighted average:
(V,) (C,) + (V2) (C2)
Final Analyte
Concentration V1 + V2
1312 - 17 Revision 0
November 1992
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where:
V, = The volume of the first phases (L).
C1 = The concentration of the analyte of concern in the first phase (mg/L).
V2 = The volume of the second phase (L).
C2 = The concentration of the analyte of concern in the second phase
(mg/L).
7.3.16 Compare the analyte concentrations in the 1312 extract with
the levels identified in the appropriate regulations. Refer to Section
8.0 for quality assurance requirements.
8.0 QUALITY CONTROL
8.1 A minimum of one blank (using the same extraction fluid as used for
the samples) for every 20 extractions that have been conducted in an extraction
vessel.
8.2 A matrix spike shall be performed for each waste type (e.g.,
wastewater treatment sludge, contaminated soil, etc.) unless the result exceeds
the regulatory level and the data is being used solely to demonstrate that the
waste property exceeds the regulatory level. A minimum of one matrix spike must
be analyzed for each analytical batch. As a minimum, follow the matrix spike
addition guidance provided in each analytical method.
8.2.1 Matrix spikes are to be added after filtration of the 1312
extract and before preservation. Matrix spikes should not be added prior
to 1312 extraction of the sample.
8.2.2 In most cases, matrix spike levels should be added at a
concentration equivalent to the corresponding regulatory level. If the
analyte concentration is less than one half the regulatory level, the
spike concentration may be as low as one half of the analyte
concentration, but may not be less than five times the method detection
limit. In order to avoid differences in matrix-effects, the matrix spikes
must be added to the same nominal volume of 1312 extract as that which was
analyzed for the unspiked sample.
8.2.3 The purpose of the matrix spike is to monitor the
performance of the analytical methods used, and to determine whether
matrix interferences exist. Use of other internal calibration methods,
modification of the analytical methods, or use of alternate analytical
methods may be needed to accurately measure the analyte concentration in
the 1312 extract when the recovery of the matrix spike is below the
expected analytical method performance.
8.2.4 Matrix spike recoveries are calculated by the following
formula:
%R (% Recovery) = 100 (Xs - Xu) / K
where:
Xs = measured value for the spiked sample
Xu = measured value for the unspiked sample, and
K = known value of the spike in the sample.
1312 - 18 Revision 0
November 1992
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K = known value of the spike in the sample.
8.3 All quality control measures described in the appropriate analytical
methods shall be followed.
8.4 The use of internal calibration quantitation methods shall be
employed for a metallic contaminant if: (1) Recovery of the contaminant from the
1312 extract is not at least 50% and the concentration does not exceed the
appropriate regulatory level, and (2) The concentration of the contaminant
measured in the extract is within 20% of the appropriate regulatory level.
8.4.1. The method of standard additions shall be employed as the
internal calibration quantitation method for each metallic contaminant.
8.4.2 The method of standard additions requires preparing
calibration standards in the sample matrix rather than reagent water or
blank solution. It requires taking four identical aliquots of the
solution and adding known amounts of standard to three of these aliquots.
The forth aliquot is the unknown. Preferably, the first addition should
be prepared so that the resulting concentration is approximately 50% of
the expected concentration of the sample. The second and third additions
should be prepared so that the concentrations are approximately 100% and
150% of the expected concentration of the sample. All four aliquots are
maintained at the same final volume by adding reagent water or a blank
solution, and may need dilution adjustment to maintain the signals in the
linear range of the instrument technique. All four aliquots are analyzed.
8.4.3 Prepare a plot, or subject data to linear regression, of
instrument signals or external-calibration-derived concentrations as the
dependant variable (y-axis) versus concentrations of the additions of
standards as the independent variable (x-axis). Solve for the intercept
of the abscissa (the independent variable, x-axis) which is the concentra-
tion in the unknown.
8.4.4 Alternately, subtract the instrumental signal or external-
calibration-derived concentration of the unknown (unspiked) sample from
the instrumental signals or external-calibration-derived concentrations of
the standard additions. Plot or subject to linear regression of the
corrected instrument signals or external-calibration-derived concentra-
tions as the dependant variable versus the independent variable. Derive
concentrations for the unknowns using the internal calibration curve as if
it were an external calibration curve.
8.5 Samples must undergo 1312 extraction within the following time
periods:
1312 - 19 Revision 0
November 1992
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SAMPLE MAXIMUM HOLDING TIMES (davsl
Volatiles
Semi-
volatiles
Mercury
Metals,
except
mercury
From: Field
Collec-
tion
To: 1312
extrac-
tion
14
14
28
180
From: 1312
extrac-
tion
To: Prepara-
tive
extrac-
tion
NA
7
NA
NA
From: Prepara-
tive
extrac-
tion
To: determi-
native
analysis
14
40
28
180
Total
Elapsed
Time
28
61
56
360
NA = Not Applicable
If sample holding times are exceeded, the values obtained will be considered
minimal concentrations. Exceeding the holding time is not acceptable in
establishing that a waste does not exceed the regulatory level. Exceeding the
holding time will not invalidate characterization if the waste exceeds the
regulatory level.
9.0 METHOD PERFORMANCE
9.1 Precision results for semi-volatiles and metals: An eastern soil
with high organic content and a western soil with low organic content were used
for the semi-volatile and metal leaching experiments. Both types of soil were
analyzed prior to contaminant spiking. The results are shown in Table 6. The
concentrations of contaminants leached from the soils were consistently
reproducible, as shown by the low relative standard deviations (RSDs) of the
recoveries (generally less than 10 % for most of the compounds).
9.2 Precision results for volatiles: Four different soils were spiked
and tested for the extraction of volatiles. Soils One and Two were from western
and eastern Superfund sites. Soils Three and Four were mixtures of a western
soil with low organic content and two different municipal sludges. The results
are shown in Table 7. Extract concentrations of volatile organics from the
eastern soil were lower than from the western soil. Replicate Teachings of Soils
Three and Four showed lower precision than the leachates from the Superfund
soils.
1312 - 20
Revision 0
November 1992
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10.0 REFERENCES
1.0 Environmental Monitoring Systems Laboratory, "Performance Testing of
Method 1312; QA Support for RCRA Testing: Project Report". EPA/600/4-
89/022. EPA Contract 68-03-3249 to Lockheed Engineering and Sciences
Company, June 1989.
2.0 Research Triangle Institute, "Interlaboratory Comparison of Methods 1310,
1311, and 1312 for Lead in Soil". U.S. EPA Contract 68-01-7075, November
1988.
1312 - 21 Revision 0
November 1992
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Table 1. Volatile Analytes1
Compound CAS No.
Acetone 67-64-1
Benzene 71-43-2
n-Butyl alcohol 71-36-3
Carbon disulfide 75-15-0
Carbon tetrachloride 56-23-5
Chlorobenzene 108-90-7
Chloroform 67-66-3
1,2-Dichloroethane 107-06-2
1,1-Dichloroethylene 75-35-4
Ethyl acetate 141-78-6
Ethyl benzene 100-41-4
Ethyl ether 60-29-7
Isobutanol 78-83-1
Methanol 67-56-1
Methylene chloride 75-09-2
Methyl ethyl ketone 78-93-3
Methyl isobutyl ketone 108-10-1
Tetrachloroethylene 127-18-4
Toluene 108-88-3
1,1,1,-Trichloroethane 71-55-6
Trichloroethylene 79-01-6
Trichlorofluoromethane 75-69-4
l,l,2-Trichloro-l,2,2-trifluoroethane 76-13-1
Vinyl chloride 75-01-4
Xylene 1330-20-7
1 When testing for any or all of these analytes, the zero-headspace extractor
vessel shall be used instead of the bottle extractor.
1312 - 22 Revision 0
November 1992
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Table 2. Suitable Rotary Agitation Apparatus1
Company
Location
Model No.
Analytical Testing and
Consulting Services,
Inc.
Associated Design and
Manufacturing Company
Environmental Machine and
Design, Inc.
IRA Machine Shop and
Laboratory
Lars Lande Manufacturing
Millipore Corp.
Warrington, PA
(215) 343-4490
Alexandria, VA
(703) 549-5999
Lynchburg, VA
(804) 845-6424
Santurce, PR
(809) 752-4004
4-vessel extractor (DC20S);
8-vessel extractor (DC20);
12-vessel extractor (DC20B)
2-vessel
4-vessel
6-vessel
8-vessel
12-vessel
24-vessel
(3740-2);
(3740-4);
(3740-6);
(3740-8);
(3740-12);
(3740-24)
8-vessel (08-00-00)
4-vessel (04-00-00)
8-vessel (011001)
Whitmore Lake, MI 10-vessel (10VRE)
(313) 449-4116 5-vessel (5VRE)
Bedford, MA
(800) 225-3384
4-ZHE or
4 1-liter
bottle extractor
(YT300RAHW)
1 Any device that rotates the extraction vessel in an end-over-end fashion at 30
+2 rpm is acceptable.
1312 - 23
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November 1992
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Table 3. Suitable Zero-Headspace Extractor Vessels1
Company
Location
Model No.
Analytical Testing &
Consulting Services, Inc.
Associated Design and
Manufacturing Company
Lars Lande Manufacturing2
Millipore Corporation
Environmental Machine
and Design, Inc.
Warrington, PA
(215) 343-4490
Alexandria, VA
(703) 549-5999
Whitmore Lake, MI
(313) 449-4116
Bedford, MA
(800) 225-3384
Lynchburg, VA
(804) 845-6424
C102, Mechanical
Pressure Device
3745-ZHE, Gas
Pressure Device
ZHE-11, Gas
Pressure Device
YT30090HW, Gas
Pressure Device
VOLA-TOX1, Gas
Pressure Device
1 Any device that meets the specifications listed in Step 4.2.1 of the method is
suitable.
2 This device uses a 110 mm filter.
1312 - 24
Revision 0
November 1992
-------
Table 4. Suitable Filter Holders1
Company
Nucleopore Corporation
Micro Filtration
Systems
Millipore Corporation
Location
Pleasanton, CA
(800) 882-7711
Dublin, CA
(800) 334-7132
(415) 828-6010
Bedford, MA
(800) 225-3384
Model/
Catalogue #
425910
410400
302400
311400
YT30142HW
XX1004700
Size
142 mm
47 mm
142 mm
47 mm
142 mm
47 mm
1 Any device capable of separating the liquid from the solid phase of the waste
is suitable, providing that it is chemically compatible with the waste and the
constituents to be analyzed. Plastic devices (not listed above) may be used when
only inorganic analytes are of concern. The 142 mm size filter holder is
recommended.
Table 5. Suitable Filter Media1
Company
Millipore Corporation
Nucleopore Corporation
Whatman Laboratory
Products, Inc.
Micro Filtration
Systems
Location Model
Bedford, MA AP40
(800) 225-3384
Pleasanton, CA 211625
(415) 463-2530
Clifton, NJ GFF
(201) 773-5800
Dublin, CA GF75
(800) 334-7132
(415) 828-6010
Pore
Size
(Mm)
0.7
0.7
0.7
0.7
1 Any filter that meets the specifications in Step 4.4 of the Method is suitable.
1312 - 25 Revision 0
November 1992
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TABLE 6 - METHOD 1312 PRECISION RESULTS FOR SEMI-VOLATILES AND METALS
Eastern Soil (vH 4.2)
FORTIFIED ANALYTES
bis(2-chloroethyl) -
ether
2-Chlorophenol
1,4-Dichlorobenzene
1 , 2-Dichlorobenzene
2-Methylphenol
Nitrobenzene
2 ,4-Dimethylphenol
Hexachlorobutadiene
Acenaphthene
2 , 4-Dinitrophenol
2 ,4-Dinitrotoluene
Hexachlorobenzene
gamma BHC (Lindane)
beta BHC
METALS
Lead
Cadmium
Amount
Spiked
1040
1620
2000
8920
3940
1010
1460
6300
3640
1300
1900
1840
7440
640
5000
1000
Amount
Recovered*
(A«g)
834
1010
344
1010
1860
812
200
95
210
896**
1150
3.7
230
35
70
387
% RSD
12.5
6.8
12.3
8.0
7.7
10.0
18.4
12.9
8.1
6.1
5.4
12.0
16.3
13.3
4.3
2.3
Western Soil (pH 5.0)
Amount
Recovered*
(Mg)
616
525
272
1520
1130
457
18
280
310**
23**
585
10
1240
65.3
10
91
% RSD
14.2
54.9
34.6
28.4
32.6
21.3
87.6
22.8
7.7
15.7
54.4
173.2
55.2
51.7
51.7
71.3
* = Triplicate analyses.
** = Duplicate analyses; one value was rejected as an outlier at the 90%
confidence level using the Dixon Q test.
1312 - 26
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November 1992
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TABLE 7 - METHOD 1312 PRECISION RESULTS FOR VOLATILES
Soil
No.
1
(Western)
Avg.
Compound Name
Acetone
Acrylonitrile
Benzene
n- Butyl Alcohol
(1-Butanol)
Carbon disulfide
Carbon tetrachloride
Chlorobenzene
Chloroform
1, 2-Dichloroethane
1 , 1-Dichloroethane
Ethyl acetate
Ethylbenzene
Ethyl ether
Isobutanol (4-Methyl
-1-propanol)
Methylene chloride
Soil
No. 2
(Eastern)
Avg.
%Rec.* %RSD %Rec.^
44.0
52.5
47.8
55.5
21.4
40.6
64.4
61.3
73.4
31.4
76.4
56.2
48.0
0.0
47.5
12
68
8
2
16
18
6
8
4
14
9
9
16
ND
30
.4
.4
.29
.91
.4
.6
.76
.04
.59
.5
.65
.22
.4
.3
43
50
34
49
12
22
41
54
68
22
75
23
55
0
42
.8
.5
.8
.2
.9
.3
.5
.8
.7
.9
.4
.2
.1
.0
.2
* %RSD
2.
70.
16.
14.
49.
29.
13.
16.
11.
39.
4.
11.
9.
ND
42.
25
0
3
6
5
1
1
4
3
3
02
5
72
9
Soil No. 3
(Western and
Sludge)
Avg.
%Rec.** %RSD
116.0
49.3
49.8
65.5
36.5
36.2
44.2
61.8
58.3
32.0
23.0
37.5
37.3
61.8
52.0
11.5
44.9
36.7
37.2
51.5
41.4
32.0
29.1
33.3
54.4
119.8
36.1
31.2
37.7
37.4
Soil No. 4
(Western and
Sludge)
Avg.
%Rec.
21.3
51.8
33.4
73.0
21.3
24.0
33.0
45.8
41.2
16.8
11.0
27.2
42.0
76.0
37.3
*** %RSD
71.4
4.6
41.1
13.9
31.5
34.0
24.9
38.6
37.8
26.4
115.5
28.6
17.6
12.2
16.6
Methyl ethyl ketone
(2-Butanone)
Methyl isobutyl
ketone
1,1,1,2-Tetrachloro-
ethane
1,1,2,2-Tetrachloro-
ethane
Tetrachloroethene
Toluene
1,1,1-Trichloro-
ethane
1,1,2-Trichloro-
e thane
Trichloroethene
Trichloro-
fluoromethane
1,1,2-Trichloro-
trifluoroethane
Vinyl chloride
56.7
81.1
69.0
85.3
45.1
59.2
47.2
76.2
54.5
5
10
6
7
12
8
16
5
11
.94
.3
.73
.04
.7
.06
.0
.72
.1
61
88
41
58
15
49
33
67
39
.9
.9
.1
.9
.2
.3
.8
.3
.4
3
2
11
4
17
10
22
8
19
.94
.99
.3
.15
.4
.5
.8
.43
.5
73.
58.
50.
64.
26.
45.
40.
61.
38.
7
3
8
0
2
7
7
7
8
31.3
32.6
31.5
25.7
44.0
35.2
40.6
28.0
40.9
40.6
39.8
36.8
53.6
18.6
31.4
26.2
46,4
25.6
39.0
40.3
23.8
15.8
24.2
37.2
38.8
25.4
34.1
20.7 24.5
18.1 26.7
10.2 20.3
12.6
6.95
7.17
60.1
58.0
72.8
28.5
21.5
25.0
34.0
67.8
61.0
19.8 33.9
15.3
11.8
24.8
25.4
* Triplicate analyses
** Six replicate analyses
**•* Five replicate analyses
I
131V 27
Revision 0
November 1992
-------
Motor
(30 jf 2 rpm
Extraction Vessel Holder
_JUU
Figure 1. Rotary Agitation Apparatus
1312 -I
Revision 0
November 1992
-------
Liquid Inlet/Outlet Valve
Top Flange
^•^^•i
Support Screen •V,
7
Support Screen'
Viton O-Rings
Bottom Flange
Pressurized Gas
Inlet/Outlet Valve
Pressure
Gauge
Figure 2. Zero-Headspace Extractor (ZHE)
1312 - 29
Revision 0
November 199
-------
METHOD 1312
SYNTHETIC PRECIPITATION LEACHING PROCEDURE
721* sample
100% solid?
7 3 Assemble filter
holder, weigh out
subsample. allow
so 1 ids to set tie,
transfer subsample
to f11 ter holder,
filter, determine %
sol ids
3 - 6 Begin again
with larger
subsample
7 4 Dry filter and
solid phase, record
weight, calculate %
dry solids
745 Begin again
with new subsample
7 4 4 Discard
solid and 1 iquid
phases, will use
new liquid phase as
extract
1312 - 30
Revision 0
November 1992
-------
METHOD 1312
SYNTHETIC PRECIPITATION LEACHING PROCEDURE (continued)
;
r.d
5 D«t«rmin«
p*rtiel«-ii
uetion due
if
.
appropriate
••UtcUon fluid to
7 6
optim
for
im tamplc »n*
f il tration
7621 Mia* solid
pha*« of tampl* to
i*tti« prior to
f li'.ration
1312 - 31
Revision 0
November 1992
-------
METHOD 1312
SYNTHETIC PRECIPITATION LEACHING PROCEDURE (continued)
7 6
Quant i tj
2 2
tivviy
t ransf er
aopropr la
of sample
KoLdec
• amount
to filUr
apply
pressure to filter
until Liquid flow
ceases
7 6 2
filtrate
filtcat
9 tor* unt
ex t raci
3 W.igh
analyza
• now or
iL tim. of
inalyjn
quantitativ«ly
tran*f*r «olid> and
filter lo entractor
7 6 3 Add
appropriate amount
of extraction fluid
to extractor,
ext ract for 18
hours
7 6 4
After
en traction , f il ter
liquid and solid
phases
1312 - 32
Revision 0
November 1992
-------
METHOD 1312
SYNTHETIC PRECIPITATION LEACHING PROCEDURE (continued)
7 6 5 Filtered
material is def-red
as extract
765 filtered
liquid from Steps
7 6 ^ and 7 6 2 3
are ^ef ined a j
extract
766-767
Record pH of
extract, preserve,
ana 1 y ze by
appropriate methods
768 Compa r e
contaminan t
concentrations in
ex t ract to
appropriate
thresholds
STOP
1312 - 33
Revision 0
November 1992
-------
METHOD 1312
SYNTHETIC PRECIPITATION LEACHING PROCEDURE (continued)
7 7 Assemble ZHE
device for volatile
analysis
7?4 Determine
optimum sample > ize
>0 5%
7 7 3 Heigh
subsampl e
776 Cool sample
and reduction
equipment. reduce
particle size
without generating
heat
774 Filtrate is
defined as ex tract
777 Do not
centrifuge wastes
prior to filtration
1312 - 34
Revision 0
November 1992
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METHOD 1312
SYNTHETIC PRECIPITATION LEACHING PROCEDURE (continued)
1 7 3 Transfer
sample to ZHE
1 1 9 Attach
f11 t rate collection
container apply
pressure un ti1
1iquia flow ceases
1 1 10 Filtrate is
aefined as extract
7711
extraction fluid 13
to add to ZHE
7 7 13 pump
ax traction fluid
into ZHE
7 7 14 R«mov« any
headsoac*,
repr.ssunz. ZHE
7
for
' 15 Rotat. ZHE
18 hour, it 22C
1 7 16 Reoeat
procedure «i th -.e
samp Ie
7 7 16 Separate
phases
1 7 17 Defin.
extract
7 7 18 Analyze by
appr oprlate
methods. combine
results if
appropriate
7 7 19 Compar*
contaminant
concentrations to
appropriate
thresholds
STOP
1312 - 35
Revision 0
November 1992
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METHOD 9045B
SOIL AND WASTE oH
1.0 SCOPE AND APPLICATION
1.1 Method 9045 is an electrometric procedure for measuring pH in soils
and waste samples. Wastes may be solids, sludges, or non-aqueous liquids. If
water is present, it must constitute less than 20% of the total volume of the
sample.
2.0 SUMMARY OF METHOD
2.1 The sample is mixed with reagent water, and the pH of the resulting
aqueous solution is measured.
3.0 INTERFERENCES
3.1 Samples with very low or very high pH may give incorrect readings
on the meter. For samples with a true pH of >10, the measured pH may be
incorrectly low. This error can be minimized by using a low-sodium-error
electrode. Strong acid solutions, with a true pH of <1, may give incorrectly
high pH measurements.
3.2 Temperature fluctuations will cause measurement errors.
3.3 Errors will occur when the electrodes become coated. If an
electrode becomes coated with an oily material that will not rinse free, the
electrode can either (1) be cleaned with an ultrasonic bath, or (2) be washed
with detergent, rinsed several times with water, placed in 1:10 HC1 so that the
lower third of the electrode is submerged, and then thoroughly rinsed with water.
4.0 APPARATUS AND MATERIALS
4.1 pH Meter with means for temperature compensation.
4.2 Electrodes:
4.2.1 Calomel electrode.
4.2.2 Glass electrode.
4.2.3 A combination electrode can be employed instead of calomel
or glass.
4.3 Beaker: 50-mL.
4.4 Thermometer.
4.5 Analytical balance: capable of weighing 0.1 g.
9045B - 1 Revision 2
November 1992
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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 Reagent water. All reference to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Primary standard buffer salts are available from the National
Institute of Standards and Technology (NIST) and should be used in situations
where extreme accuracy is necessary. Preparation of reference solutions from
these salts requires some special precautions and handling, such as low-
conductivity dilution water, drying ovens, and carbon-dioxide-free purge gas.
These solutions should be replaced at least once each month.
5.4 Secondary standard buffers may be prepared from NIST salts or
purchased as solutions from commercial vendors. These commercially available
solutions, which have been validated by comparison with NIST standards, are
recommended for routine use.
6.0 SAMPLE PRESERVATION AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine of this manual.
6.2 Samples should be analyzed as soon as possible.
7.0 PROCEDURE
7.1 Calibration:
7.1.1 Because of the wide variety of pH meters and accessories,
detailed operating procedures cannot be incorporated into this method.
Each analyst must be acquainted with the operation of each system and
familiar with all instrument functions. Special attention to care of the
electrodes is recommended.
7.1.2 Each instrument/electrode system must be calibrated at a
minimum of two points that bracket the expected pH of the samples and are
approximately three pH units or more apart. Repeat adjustments on
successive portions of the two buffer solutions until readings are within
0.05 pH units of the buffer solution value.
7.2 Sample preparation and pH measurement of soils:
7.2.1 To 20 g of soil in a 50-mL beaker, add 20 mL of reagent
water and stir the suspension several times during the next 30 minutes.
9045B - 2 Revision 2
November 1992
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7.2.2 Let the soil suspension stand for about 1 hour to allow
most of the suspended clay to settle out from the suspension or filter off
the aqueous phase for pH measurement.
7.2.3 Adjust the electrodes in the clamps of the electrode holder
so that, upon lowering the electrodes into the beaker, the glass electrode
will be immersed just deep enough into the clear supernatant solution to
establish a good electrical contact through the ground-glass joint or the
fiber-capillary hole. Insert the electrodes into the sample solution in
this manner. For combination electrodes, immerse just below the
suspension.
7.2.4 If the sample temperature differs by more than 2*C from the
buffer solution, the measured pH values must be corrected.
7.2.5 Report the results as "soil pH measured in water at _°C"
where eC is the temperature at which the test was conducted.
7.3 Sample preparation and pH measurement of waste materials:
7.3.1 To 20 g of waste sample in a 50-mL beaker, add 20 ml of
reagent water and stir the suspension several times during the next 30
minutes.
7.3.2 Let the waste suspension stand for about 15 minutes to
allow most of the suspended waste to settle out from the suspension or
filter off aqueous phase for pH measurement.
NOTE: If the waste is hydroscopic and absorbs all the reagent water,
begin the experiment again using 20 g of waste and 40 mL of
reagent water.
NOTE: If the supernatant is multiphasic, decant the oily phase and
measure the pH of the aqueous phase. The electrode may need to be
cleaned (Step 3.3) if it becomes coated with an oily material.
7.3.3 Adjust the electrodes in the clamps of the electrode holder
so that, upon lowering the electrodes into the beaker, the glass electrode
will be immersed just deep enough into the clear supernatant to establish
good electrical contact through the ground-glass joint or the fiber-
capillary hole. Insert the electrode into the sample solution in this
manner. For combination electrodes, immerse just below the suspension.
7.3.4 If the sample temperature differs by more than 2°C from the
buffer solution, the measured pH values must be corrected.
7.3.5 Report the results as "waste pH measured in water at _°C"
where °C is the temperature at which the test was conducted.
8.0 QUALITY CONTROL
8.1 Duplicate samples and check standards should be analyzed with each
analytical batch.
9045B - 3 Revision 2
November 1992
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8.2 Electrodes must be thoroughly rinsed between samples.
9.0 METHOD PERFORMANCE
9.1 No data provided.
10.0 REFERENCES
1. Black, Charles Allen; Methods of Soil Analysis; American Society of
Agronomy: Madison, WI, 1973.
2. National Institute of Standards and Technology, Standard Reference
Material Catalog, 1986-87, Special Publication 260.
9045B - 4 Revision 2
November 1992
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METHOD 9045B
SOIL AND WASTE pH
START
721 Add 20 mL
water to 20 9
soil, stir
during next 30
minutes
7 2 2 Let soil
su»pension
stand for 1
hour or fi1ter
7 1 Calibrate
each
ins trument/
elect rode
sys tern
Inser t
elect rodes
into sample
solution
731 Add 20 mL
water to 20 g
was te, stir
during next 30
minutes
732 Let waste
suspension
stand for 15
minu tea or
filter
Correct
easured pH
va1ues
Repor t
r esulIs and
tempera ture
Repeat
experiment
with 20 g
waste and 40
mL water
Decan t oily
phase,
measure pH of
aqueous phase
Aqueous
Phase
9045B - 5
Revision 2
November 1992
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ERRATA FOR SW-846 PROPOSED UPDATE II
METHOD 9040A
THIS PACKET CONTAINS AN OFFICIAL ERRATA SHEET FOR
METHOD 9040A, PROPOSED UPDATE II, OF
TEST METHODS FOR EVALUATING SOLID WASTE, PHYSICAL/CHEMICAL
METHODS, SW-846, 3RD EDITION
Incorporate this errata for Proposed Update II Method
9040A by either:
1. Placing the errata sheet at the beginning of
the method, or
2. Manually editing the method to show the
text change.
If you have any problems including this errata in your copy of the
Proposed Update II for SW-846, please telephone the Methods Information
Communication Exchange (MICE) at (703) 821-4789.
If you have any problems or questions concerning your SW-846
subscription, please telephone the U.S. Government Printing Office (GPO) at
(202) 783-3238.
Recycled/Recyclable
Printed with Soy/Canola Ink on paper that
contains at least 50% recycled fiber
-------
ERRATA FOR PROPOSED UPDATE II METHOD 9040A
In Step 7.1.2., replace the following text:
(For corrosivity characterization, the calibration of the pH meter should
include a buffer of pH 2 for acidic wastes and a pH 12 buffer for caustic
wastes; also for corrosivity characterization, the sample must be measured
at 25'C ± TC if the pH of the waste is above 12.0.)
with:
(For corrosivity characterization, the calibration of the pH meter should
include a buffer of pH 2 for acidic wastes and a pH 12 buffer for caustic
wastes.)
9040A ERRATA - 1 November 1992
-------
METHOD 9096
LIQUID RELEASE TEST (LRT) PROCEDURE
1.0 SCOPE AND APPLICATION
1.1 The Liquid Release Test (LRT) is a laboratory test designed to
determine whether or not liquids will be released from sorbents when they are
subjected to overburden pressures in a landfill.
1.2 Any liquid-loaded sorbent that fails the EPA Paint Filter Free
Liquids Test (PFT) (SW-846 Method 9095), may be assumed to release liquids in
this test. Analysts should ensure that the material in question will pass the
PFT before performing the LRT.
2.0 SUMMARY OF METHOD
2.1 A representative sample of the liquid-loaded sorbent, standing 10
cm high in the device, is placed between twin stainless steel screens and two
stainless-steel grids, in a device capable of simulating landfill overburden
pressures. An absorptive filter paper is placed on the side of each stainless-
steel grid opposite the sample (i.e., the stainless-steel screen separates the
sample and the filter paper, while the stainless-steel grid provides a small air
gap to prevent wicking of liquid from the sample onto the filter paper). A
compressive force of 50 psi is applied to the top of the sample. Release of
liquid is indicated when a visible wet spot is observed on either filter paper.
3.0 INTERFERENCES
3.1 When testing sorbents are loaded with volatile liquids (e.g.,
solvents), any released liquid migrating to the filter paper may rapidly
evaporate. For this reason, filter papers should be examined immediately after
the test has been conducted.
3.2 It is necessary to thoroughly clean and dry the stainless-steel
screens prior to testing to prevent false positive or false negative results.
Material caught in screen holes may impede liquid transmission through the screen
causing false negative results. A stiff bristled brush, like those used to clean
testing sieves, may be used to dislodge material from holes in the screens. The
screens should be ultrasonically cleaned with a laboratory detergent, rinsed with
deionized water, rinsed with acetone, and thoroughly dried.
When sorbents containing oily substances are tested, it may be necessary
to use solvents (e.g., methanol or methylene chloride) to remove any oily residue
from the screens and from the sample holder surfaces.
3.3 When placing the 76 mm screen on top of the loaded sample it is
important to ensure that no sorbent is present on top of the screen to contact
the filter paper and cause false positive results. In addition, some sorbent
residue may adhere to container sidewalls and contact the filter as the sample
compresses under load, causing wet spots on the edges of the filter. This type
of false positive may be avoided by carefully centering the 76 mm filter paper
in the device prior to initiating the test.
9096 - 1 Revision 0
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3.4 Visual examination of the sample may indicate that a release is
certain (e.g.. free standing liquid or a sample that flows like a liquid),
raising concern over unnecessary clean-up of the LRT device. An optional 5
minute Pre-Test, described in Appendix A of this procedure, may be used to
determine whether or not an LRT must be performed.
4.0 APPARATUS AND MATERIALS
4.1 LRT Device (LRTD): A device capable of applying 50 psi of pressure
continuously to the top of a confined, cylindrical sample (see Figure 1). The
pressure is applied by a piston on the top of the sample. All device components
contacting the sample (i.e.. sample-holder, screens, and piston) should be
resistant to attack by substances being tested. The LRTD consists of two basic
components, described below.
4.1.1 Sample holder: A rigid-wall cylinder, with a bottom plate,
capable of holding a 10 cm high by 76 mm diameter sample.
4.1.2 Pressure Application Device: In the LRTD (Figure 1),
pressure is applied to the sample by a pressure rod pushing against a
piston that lies directly over the sample. The rod may be pushed against
the piston at a set pressure using pneumatic, mechanical, or hydraulic
pressure. Pneumatic pressure application devices should be equipped with
a pressure gauge accurate to within ±1 psi, to indicate when the desired
pressure has been attained and whether or not it is adequately maintained
during the test. Other types of pressure application devices (e.g..
mechanical or hydraulic) may be used if they can apply the specified
pressure continuously over the ten minute testing time. The pressure
application device must be calibrated by the manufacturer, using a load
cell or similar device placed under the piston, to ensure that 50 + 1 psi
is applied to the top of the sample. The pressure application device
should be sufficiently rugged to deliver consistent pressure to the sample
with repeated use.
4.2 Stainless-Steel Screens: To separate the sample from the filter,
thereby preventing false positive results from particles falling on the filter
paper. The screens are made of stainless steel and have hole diameters of 0.012
inches with 2025 holes per square inch. Two diameters of screens are used: a
larger (90 mm) screen beneath the sample and a smaller (76 mm) screen that is
placed on top of the sample in the sample-holding cylinder.
4.3 Stainless-Steel Grids: To provide an air gap between the
stainless-steel screen and filter paper, preventing false positive results from
capillary action. The grids are made of 1/32" diameter, woven, stainless steel
wire cut to two diameters, 90 mm and 76 mm.
4.4 Filter Papers: To detect released liquid. Two sizes, one 90 mm
and one 76 mm, are placed on the side of the screen opposite the sample. The
76 mm diameter filter paper has the outer 6 mm cut away except 3 conical points
used for centering the paper (see Figure 2). Blue, seed-germination filter paper
manufactured by Schleicher and Schuell (Catalog Number 33900) is suitable. Other
colored, absorptive papers may be used as long as they provide sufficient wet/dry
contrast for the operator to clearly see a wet spot.
9096 - 2 Revision 0
November 1992
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4.5 Spatula: To assist in loading and removing the sample.
4.6 Rubber or wooden mallet: To tap the sides of the device to settle
and level the sample.
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 Reagent water. All references to water in this method refer to
reagent water, as defined in Chapter One.
5.3 Acetone.
6.0 SAMPLE COLLECTION, PRESERVATION AND HANDLING
6.1 All samples should be collected using a sampling plan that
addresses the considerations discussed in "Test Methods for Evaluating Solid
Wastes (SW-846)." The sampling plan should be designed to detect and sample any
pockets of liquids that may be present in a container (i.e.. in the bottom or top
of the container).
6.2 Preservatives should not be added to samples.
6.3 Samples should be tested as soon as possible after collection, but
in no case after more than three days after collection. If samples must be
stored, they can be stored in sealed containers and maintained under dark, cool
conditions (temperature ranging between 35° and 72° F). Samples should not be
frozen.
7.0 PROCEDURE
The procedure below was developed for the original LRTD, manufactured by
Associated Design and Manufacturing Company (ADM). Procedures for other LRTDs,
along with evidence for equivalency to the ADM device, should be supplied by the
manufacturer.
7.1 Disassemble the LRTD and make sure that all parts are clean and
dry.
7.2 Invert the sample-holding cylinder and place the large stainless-
steel screen, the large stainless-steel grid, then a 90 mm filter paper on the
cylinder base (bottom-plate side).
7.3 Secure the bottom plate (plate with a hole in the center and four
holes located on the outer circumference) to the flange on the bottom of the
sample-holding cylinder using four knob screws.
9096 - 3 Revision 0
November 1992
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7.4 Turn the sample holder assembly to the right-side-up position
(bottom-plate-side down). Fill the sample holder with a representative sample
until the sample height measures 10 cm (up to the etched line in the cylinder).
7.5 Tap the sides of the sample holder with a rubber or wooden mallet
to remove air pockets and to settle and level the sample.
7.6 Repeat filling, and tapping until a sample height of 10 cm is
maintained after tapping.
7.7 Smooth the top of the sample with a spatula to create a horizontal
surface.
7.8 Place the small stainless-steel screen, then the small stainless-
steel grid on top of the sample.
NOTE: Prior to placing the stainless-steel grid on top of the screen,
make sure that no sorbent material is on the grid side of the
stainless-steel screen.
7.9 Place the 76 mm filter paper on top of the small stainless-steel
grid, making sure the filter paper is centered in the device.
7.10 Using the piston handle (screwed into the top of the piston) lower
the piston into the sample holder until it sits on top of the filter paper.
Unscrew and remove the handle.
7.11 Place the loaded sample holder into position on the baseplate and
lock into place with two toggle clamps.
7.12 Place the pressure application device on top of the sample-holder.
Rotate the device to lock it into place and insert the safety key.
7.13 Connect air lines.
7.14 Initiate rod movement and pressure application by pulling the air-
valve lever toward the operator and note time on data sheet. The pressure gauge
at the top of the pressure application device should read as specified in the
factory calibration record for the particular device. If not, adjust regulator
to attain the specified pressure.
NOTE: After pressure application, the air lines can be disconnected, the
toggle clamps can be released, and the LRTD can be set aside for
10 minutes while other LRTDs are pressurized. LRTD pressures
should be checked every 3 minutes to ensure that the specified
pressure is being maintained. If the specified pressure is not
being maintained to within + psi, the LRTD must be reconnected to
the air lines and pressure applied throughout the 10 minute test.
7.15 After 10 minutes place the LRTD on the baseplate, reconnect air
lines and toggle clamps, and turn off pressure (retract the rod) by pushing the
air-valve lever away from the operator. Note time on data sheet.
9096 - 4 Revision 0
November 1992
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7.16 When the air gauge reaches 0 psi, disconnect the air lines and
remove the pressure-application device by removing the safety key, rotating the
device, and lifting it away from the sample holder.
7.17 Screw the piston handle into the top of the piston.
7.18 Lift out the piston.
7.19 Remove the filter paper and immediately examine it for wet spots
(wet area on the filter paper). The presence of a wet spot(s) indicates a
positive test (i.e., liquid release). Note results on data sheet.
7.20 Release toggle clamps and remove sample holder from baseplate.
Invert sample holder onto suitable surface and remove the knob screws holding the
bottom plate.
7.21 Remove the bottom plate and immediately examine the filter paper
for wet spots as described in Step 7.19. Note results on data sheet. Wet
spot(s) on either filter indicates a positive test.
8.0 QUALITY CONTROL
8.1 Duplicate samples should be analyzed every twenty samples or every
analytical batch, whichever is more frequent.
9.0 METHOD PERFORMANCE
9.1 Precision and accuracy data are not available at this time.
10.0 REFERENCES
1. Hoffman, P., G. Kingsbury, B. Lesnik, M. Meyers, "Background Document for
the Liquid Release Test (LRT) Procedure"; document submitted to the Environmental
Protection Agency by Research Triangle Institute: Research Triangle Park, NC
under Contract No. 68-01-7075, Work Assignment 76 and Contract No. 68-WO-0032,
Work Assignment 12.
9096 - 5 Revision 0
November 1992
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APPENDIX A
1.0 SCOPE AND APPLICATION
1.1 The LRT Pre-Test is an optional, 5 minute laboratory test designed
to determine whether or not liquids will be definitely released from sorbents
before applying the LRT. This test is performed to prevent unnecessary cleanup
and possible damage to the LRT device.
1.2 This test is purely optional and completely up to the discretion
of the operator as to when it should be used.
2.0 SUMMARY OF METHOD
A representative sample will be loaded into a glass grid that is placed on
a glass plate already stained with 2 dyes (one water soluble and one oil
soluble). A second glass plate will be placed on top and a 2 Ib. weight placed
on top for 5 minutes. At the end of 5 minutes the base of the glass grid is
examined for any dye running along the edges, this would indicate a liquid
release.
3.0 INTERFERENCES
A liquid release can be detected at lower Liquid Loading Levels with
extremely clean glassware. The glass plates and glass grid should be cleaned
with a laboratory detergent, rinsed with Deionized water, rinsed with acetone,
and thoroughly dried.
4.0 APPARATUS AND MATERIALS
4.1 Glass Plate: 2 glass plates measuring 7.5 cm x 7.5 cm.
4.2 Glass Grid: See Figure 3.
4.3 Paint Brush: Two small paint brushes for applying dyes.
4.4 Spatula: To assist in loading the sample.
4.5 Weight: 2.7 kg weight to apply pressure to the sample.
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 Methylene Blue dye in methanol.
5.3 Anthraquinone dye in toluene.
9096 - 6
Revision 0
November 1992
-------
6.0 SAMPLE COLLECTION, PRESERVATION AND HANDLING
See LRT Procedure.
7.0 PROCEDURE
7.1 Paint one strip, approximately 1 cm wide, of methylene blue dye
across the center of a clean and dry glass plate (see Figure 4). The dye is
allowed to dry.
7.2 Paint one strip, approximately 1 cm wide, of anthraquinone dye
across the center of the same glass plate (see Figure 4). This strip should be
adjacent to and parallel with the methylene blue strip. The dye is allowed to
dry.
7.3 Place the glass grid in the center of the dye-painted glass plate.
7.4 Place a small amount of sample into the glass-grid holes, pressing
down gently until the holes are filled to slightly above the grid top.
7.5 Place a second, clean and dry, glass plate on top of the sample and
grid.
7.6 Place a 2.7 kg weight on top of the glass for 5 minutes.
7.7 After 5 minutes remove the weight and examine the base of the grid
extending beyond the sample holes for any indication of dyed liquid. The entire
assembly may be turned upside down for observation. Any indication of liquid
constitutes a release and the LRT does not need to be performed.
8.0 QUALITY CONTROL
8.1 Refer to Chapter One for specific quality control procedures.
9.0 METHOD PERFORMANCE
9.1 Precision and accuracy data are not available at this time.
10.0 REFERENCES
1. Research Triangle Institute. "Background Document for the Liquid Release
Test: Single Laboratory Evaluation and 1988 Collaborative Study".
Submitted to the Environmental Protection Agency under Contract No. 68-01-
7075, Work Assignment 76 and Contract No. 68-WO-0032, Work Assignment 12.
September 18, 1991.
9096 - 7 Revision 0
November 1992
-------
FIGURE 1.
LRT DEVICE
Pressure
Application
Device
50 psi
Sample-HoldIng Cylinder
Filter
Separator Plate
Separator Plate
ter
Bottom Plate
9096 - 8
Revision 0
November 1992
-------
FIGURE 2.
76 MM DIAMETER FILTER PAPER
120'
9096 - 9
Revision 0
November 1992
-------
FIGURE 3.
GLASS GRID SPECIFICATIONS.
0.25 inch
glass rod
1.7cm
4.0 cm
1
9.7 cm
9096 - 10
Revision 0
November 1992
-------
FIGURE 4.
POSITIONING OF DYE ON GLASS PLATE
Methylene Blue
Anthraquinone
7.5 cm
7.5 cm
9096 - 11
Revision 0
November 1992
-------
METHOD 9096
LIQUID RELEASE TEST (LRT) PROCEDURE
START
7
6 Add more
sample
c
STOP
9096 - 12
Revision 0
November 1992
-------
METHOD 9040A
DH ELECTROMETRIC MEASUREMENT
1.0 SCOPE AND APPLICATION
1.1 Method 9040 Is used to measure the pH of aqueous wastes and those
multiphase wastes where the aqueous phase constitutes at least 20% of the total
volume of the waste.
1.2 The corrosivity of concentrated acids and bases, or of concentrated
acids and bases mixed with inert substances, cannot be measured. The pH
measurement requires some water content.
2.0 SUMMARY
2.1 The pH of the sample is determined electrometrically using either
a glass electrode in combination with a reference potential or a combination
electrode. The measuring device is calibrated using a series of standard
solutions of known pH.
3.0 INTERFERENCES
3.1 The glass electrode, in general, is not subject to solution
interferences from color, turbidity, colloidal matter, oxidants, reductants, or
high salinity.
3.2 Sodium error at pH levels >10 can be reduced or eliminated by using
a low-sodium-error electrode.
3.3 Coatings of oily material or particulate matter can impair
electrode response. These coatings can usually be removed by gentle wiping or
detergent washing, followed by rinsing with distilled water. An additional
treatment with hydrochloric acid (1:10) may be necessary to remove any remaining
film.
3.4 Temperature effects on the electrometric determination of pH arise
from two sources. The first is caused by the change in electrode output at
various temperatures. This interference can be controlled with instruments
having temperature compensation or by calibrating the electrode-instrument system
at the temperature of the samples. The second source of temperature effects is
the change of pH due to changes in the sample as the temperature changes. This'
error is sample-dependent and cannot be controlled. It should, therefore, be
noted by reporting both the pH and temperature at the time of analysis.
4.0 APPARATUS AND MATERIALS
4.1 pH meter: Laboratory or field model. Many instruments are commer-
cially available with various specifications and optional equipment.
4.2 Glass electrode.
9040A - 1 Revision 1
November 1992
-------
4.3 Reference electrode: A silver-silver chloride or other reference
electrode of constant potential may be used.
NOTE: Combination electrodes incorporating both measuring and referenced
functions are convenient to use and are available with solid, gel-type
filling materials that require minimal maintenance.
4.4 Magnetic stirrer and Teflon-coated stirring bar.
4.5 Thermometer or temperature sensor for automatic compensation.
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 Primary standard buffer salts are available from the National
Institute of Standards and Technology (NIST) and should be used in situations
where extreme accuracy is necessary. Preparation of reference solutions from
these salts requires some special precautions and handling, such as low-
conductivity dilution water, drying ovens, and carbon-dioxide-free purge gas.
These solutions should be replaced at least once each month.
5.3 Secondary standard buffers may be prepared from NIST salts or
purchased as solutions from commercial vendors. These commercially available
solutions have been validated by comparison with NIST standards and are
recommended for routine use.
6.0 SAMPLE COLLECTION, PRESERVATION, AND HANDLING
6.1 All samples must be collected using a sampling plan that addresses
the considerations discussed in Chapter Nine of this manual.
6.2 Samples should be analyzed as soon as possible.
7.0 PROCEDURE
7.1 Calibration:
7.1.1 Because of the wide variety of pH meters and accessories,
detailed operating procedures cannot be incorporated into this method.
Each analyst must be acquainted with the operation of each system and
familiar with all instrument functions. Special attention to care of the
electrodes is recommended.
7.1.2 Each instrument/electrode system must be calibrated at a
minimum of two points that bracket the expected pH of the samples and are
approximately three pH units or more apart. (For corrosivity characteri-
zation, the calibration of the pH meter should include a buffer of pH 2
for acidic wastes and a pH 12 buffer for caustic wastes; also for
9040A - 2 Revision 1
November 1992
-------
corrosivity characterization, the sample must be measured at 25'C ± 1*C if
the pH of the waste is above 12.0.) Various instrument designs may
involve use of a dial (to "balance" or "standardize") or a slope
adjustment, as outlined in the manufacturer's instructions. Repeat
adjustments on successive portions of the two buffer solutions until
readings are within 0.05 pH units of the buffer solution value.
7.2 Place the sample or buffer solution in a clean glass beaker using
a sufficient volume to cover the sensing elements of the electrodes and to give
adequate clearance for the magnetic stirring bar. If field measurements are
being made, the electrodes may be immersed directly into the sample stream to an
adequate depth and moved in a manner to ensure sufficient sample movement across
the electrode-sensing element as indicated by drift-free readings (<0.1 pH).
7.3 If the sample temperature differs by more than 2*C from the buffer
solution, the measured pH values must be corrected. Instruments are equipped
with automatic or manual compensators that electronically adjust for temperature
differences. Refer to manufacturer's instructions.
7.4 Thoroughly rinse and gently wipe the electrodes prior to measuring
pH of samples. Immerse the electrodes into the sample beaker or sample stream
and gently stir at a constant rate to provide homogeneity and suspension of
solids. Note and record sample pH and temperature. Repeat measurement on
successive volumes of sample until values differ by <0.1 pH units. Two or three
volume changes are usually sufficient.
8.0 QUALITY CONTROL
8.1 Duplicate samples and check standards should be analyzed with each
analytical batch.
8.2 Electrodes must be thoroughly rinsed between samples.
9.0 METHOD PERFORMANCE
9.1 Forty-four analysts in twenty laboratories analyzed six synthetic
water samples containing exact increments of hydrogen-hydroxyl ions, with the
following results:
Accuracy as
Standard Deviation Bias Bias
pH Units pH Units % oH Units
3.5
3.5
7.1
7.2
8.0
8.0
0.10
0.11
0.20
0.18
0.13
0.12
-0.29
-0.00
+1.01
-0.03
-0.12
+0.16
-0.01
+0.07
-0.002
-0.01
+0.01
10.0 REFERENCES
1. National Institute of Standards and Technology,
Material Catalog 1986-87, Special Publication 260.
9040A - 3
Standard Reference
Revision 1
November 1992
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METHOD 9040A
pH ELECTROMETRIC MEASUREMENT
Start
7 1 Calibrate pH
me ter
7 2 Place sample or
buffer solution in
glass beaker
7 3 Co r rect
measured pH va1ues
7 4 Immerse
electrodes and
measure pH of
sample
7 4 Note and record
pH and temperature,
repeat 2 or 3 times
with different
vo1umes
Stop
9040A - 4
Revision 1
November 1992
-------
METHOD 9096
APPENDIX A
START
7 1 Paint methylene
blue strip on
glass, dry
7 ^ Paint
anthraquinone atrip
on glass parallel'
to first strip, dry
7 3 Place grid in
center of glass
plate
7 4 Fill holes of
grid with sample
7 S Place second
glass plate on top
of sample
7 6 Apply
glass for
weight on
S minutes
7 7 Remove weight
and check for wet
spot(s)
STOP
9096 - 13
Revision 0
November 1992
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