vvEPA
United States
Environmental Protection
Agency
Office of Water Regulations
and Standards (WH-522)
Industrial Technology Division
August 1988
820R88100
Office of Water
Analytical Methods
for the
National Sewage Sludge
Survey
A.RY
-i'OTECTION SGENW
M. I 08317.
-------
ANALYTICAL METHODS FOR
THE NATIONAL SEWAGE SLUDGE SURVEY
Prepared for:
W. A. Telliard, Chief
Energy and Mining Industry Branch
USEPA Office of Water Regulations and Standards
401 M Street, SW
Washington, DC 20460
Under EPA Contract No. 68-01-6990
Publication Date: August 1, 1988
-------
INTRODUCTION
This document is a compilation of the analytical methods that the USEPA Office of Water
Regulations and Standards (OWRS) will use in the National Sewage Sludge Survey.
These methods have been compiled from three sources other than OWRS, they are:
1) "Methods for Chemical Analysis of Water and Wastes," USEPA, EMSL,
Cincinnati, OH 45268, EPA-600/4-79-020 (Revised March 1983).
Note: This document is currently available from National Technical
Information Service, Springfield, VA 22161, PB84-128677.
2) "Test Methods for Evaluating Solid Waste," USEPA, OSW, Washington, DC
20460, SW-846, (November 1986).
Note: This document is currently available from the Superintendent of
Documents, U.S. Government Printing Office, Washington, DC 20402.
3) Method 8290 is included in draft form and was developed by :
USEPA
Environmental Monitoring Systems Laboratory
Las Vegas, Nevada 09193-3478
Questions concerning this document should be addressed to:
W.A. Telliard
USEPA Office of Water Regulations and Standards
Sample Control Center
P.O. Box 1407
Alexandria, VA 22313
703/557-5040
-------
ANALYTICAL METHODS FOR
THE NATIONAL SEWAGE SLUDGE SURVEY
TABLE OF CONTENTS-
CATEGORY FRACTION TECHNIQUE
Organics
METHOD MODIFICATION PAGE
Metals
Classicals
VOA
ABN
PEST/HERB
PCDD/PCDF
Furnace
ICP
Antimony
Arsenic
Selenium
Thallium
Mercury
ICP-22 Ele.
Residue
Cyanide
Fluoride
TKN
Nitrate
Nitrite
Phosphorous
GCMS
GCMS
GC
GCMS
Digestion
Digestion
GFAA
GFAA
GFAA
GFAA
CVAA
ICPAES
Grav.-TOT
Spectro.
Electrode
Block Digest.
Cd Reduct.
Cd Reduct.
CrHoOr Reduct.
1624C
1625C
1618
8290
3050
3050
204.2
206.2
270.2
279.2
245.5
200.7
160.3
335.2
340.2
351.2
353.2
353.2
365.2
<95°C-Sb
HC1 Reflux
MSA
MSA
MSA
MSA
+42 Ele. Screen
3
31
81
115
249
249
259
263
267
271
275
281
295
299
309
313
319
319
327
-------
EPA METHOD 1624C
VOLATILE ORGANIC COMPOUNDS BY ISOTOPE DILUTION GCMS
EPA METHOD 1625C
SEMIVOLATILE ORGANIC COMPOUNDS BY ISOTOPE DILUTION GCMS
-------
Introduction
Methods 1624 and 1625 were developed by the Industrial
Technology Division (ITD) within EPA's Office of Water
Regulations and Standards to provide improved precision and
accuracy of analysis of pollutants in aqueous and solid
matrices. The ITD is responsible for development and
promulgation of nationwide standards setting limits on
pollutant levels in industrial discharges.
Methods 1624 and 1625 are isotope dilution, gas
chromatography-mass spectrometry methods for analysis of the
volatile and semi-volatile, organic "priority" pollutants, and
other organic pollutants amenable to gas chromatography-mass
spectrometry. Isotope dilution is a technique which employs
stable, isotopically labeled analogs of the compounds of
interest as internal standards in the analysis.
Questions concerning the Methods or their application should
be addressed to:
W. A. Tel Hard
USEPA
Office of Water Regulations and Standards
401 M Street SW
Washington, DC 20460
202-382-7131
OR
USEPA OWRS
Sample Control Center
P.O. Box 1407
Alexandria, Virginia 22313
703-557-5040
Publication date: March 1988
-------
METHOD 1624 15 February 1988 Revision C
Volatile Organic Compounds by Isotope Dilution GCMS
1 SCOPE AND APPLICATION
1.1 This method is designed to determine the
volatile toxic organic pollutants
associated with the 1976 Consent Decree;
the Resource Conservation and Recovery
Act; the Comprehensive Environmental
Response, Compensation and Liabilities
Act; and other compounds amenable to purge
and trap gas chromatography-mass
spectrometry (GCMS).
Table 1
VOLATILE ORGANIC COMPOUNDS DETERMINED BY CALIBRATED GCMS USING ISOTOPE
DILUTION AND INTERNAL STANDARD TECHNIQUES
Compound
acetone
acrolein
acrylonitrile
benzene
bromodi ch loromethane
bromoform
bromomethane
carbon tetrachloride
chlorobenzene
chloroethane
2-chloroethylvinyl ether
chloroform
ch loromethane
di bromoch loromethane
1 , 1 -dich loroethane
1,2-dichtoroethane
1 ,1-dichloroethene
trans-1,2-dichlorethene
1 , 2-d ich I oropropane
trans- 1 ,3-di ch loropropene
diethyl ether
p-dioxane
ethyl benzene
methylene chloride
methyl ethyl ketone
1 , 1 ,2,2- tetrachloroethane
tetrachlorethene
toluene
1,1, 1-tri chloroethane
1,1, 2- trich loroethane
trichloroethene
vinyl chloride
Storet
81552
34210
34215
34030
32101
32104
34413
32102
34301
34311
34576
32106
344 tt
32105
34496
32103
34501
34546
34541
34699
81576
81582
34371
34423
81595
34516
34475
34010
34506
34511
39180
39175
Pollutant
CAS Registry EPA-EGD
67-64-1
107-02-8
107-13-1
71-43-2
75-27-4
75-25-2
74-83-9
56-23-5
108-90-7
75-00-3
110-75-8
67-66-3
74-87-3
124-48-1
75-34-3
107-06-2
75-35-4
156-60-5
78-87-5
10061-02-6
60-29-7
123-91-1
100-41-4
75-09-2
78-93-3
79-34-5
127-18-4
108-88-3
71-55-6
79-00-5
79-01-6
75-01-4
516 V
002 V
003 V
004 V
048 V
047 V
046 V
006 V
007 V
016 V
019 V
023 V
045 V
051 V
013 V
010 V
029 V
030 V
032 V
033 V
515 V
527 V
038 V
044 V
514 V
015 V
085 V
086 V
011 V
014 V
087 V
088 V
NPDES
001 V
002 V
003 V
012 V
005 V
020 V
006 V
007 V
009 V
010 V
011 V
021 V
008 V
014 V
015 V
016 V
026 V
017 V
019 V
022 V
023 V
024 V
025 V
027 V
028 V
029 V
031 V
Labeled Compound
Analog CAS Registry
d6
d-
H
C
13c
d,
1?3c
d
ds
13C
d_
^
d,
d4
d2
d.
d4
d10
da
dio
d°
d7
C-
d8
13*3
r
13C.,
666-52-4
33984-05-3
53807-26-4
1076-43-3
93952-10-4
72802-81-4
1111-88-2
32488-50-9
3114-55-4
19199-91-8
31717-44-9
1111-89-3
93951-99-6
56912-77-7
17070-07-0
22280-73-5
42366-47-2
93952-08-0
93951-86-1
2679-89-2
17647-74-4
25837-05-2
1665-00-5
53389-26-7
33685-54-0
32488-49-6
2037-26-5
2747-58-2
93952-09-1
93952-00-2
6745-35-3
EPA-
EGD
616 V
202 V
203 V
204 V
248 V
247 V
246 V
206 V
207 V
216 V
223 V
245 V
251 V
213 V
210 V
229 V
230 V
232 V
233 V
615 V
627 V
238 V
244 V
614 V
215 V
285 V
286 V
211 V
214 V
287 V
288 V
-------
1.2 The chemical compounds listed in tables 1
and 2 may be determined in waters, soils,
and municipal sludges by this method. The
method is designed to meet the survey
requirements of the Environmental
Protection Agency.
Table 2
VOLATILE ORGANIC COMPOUNDS TO BE
DETERMINED BY REVERSE SEARCH AND
QUANT I TATION USING KNOWN RETENTION TIMES,
RESPONSE FACTORS, REFERENCE COMPOUNDS, AND
MASS SPECTRA
interferences rather than instrumental
limitations. The levels in table 3 typify
the minimum quantity that can be detected
with no interferences present.
1.4 The GCMS portions of this method are for
use only by analysts experienced with GCMS
or under the close supervision of such
qualified persons. Laboratories unfamil-
iar with analyses of environmental samples
by GCMS should run the performance tests
in reference 1 before beginning.
2 SUMMARY OF METHOD
EGO
No.
532
533
534
535
536
537
538
539
540
541
542
543
544
545
546
547
548
549
550
551
552
553
554
951
952
Compound CAS Registry
allyl alcohol*
carbon disulfide
2-chloro-1,3-butadiene
(chloroprene)
chloroacetonitrile*
3 - ch I oropropene
crotonaldehyde*
1,2-dibromoethane (EDB)
dibromomethane
trans- 1,4-
dichloro-2-butene
1 ,3-dichloropropane
cis-1 ,3-dichloropropene
ethyl cyanide*
ethyl methacrylate
2-hexanone
iodome thane
isobutyl alcohol*
methacrylonitrile
methyl methacrylate
4-methyl-2-pentanone
1,1,1,2- tetrach loroethane
t r i ch I orof I uoromethane
1,2,3-trichloropropane
vinyl acetate
m-xylene
o- •» p-xylene
107-18-6
75-15-0
126-99-8
107-14-2
107-05-1
123-73-9
106-93-4
74-95-3
110-57-6
142-28-9
10061-01-5
107-12-0
97-63-2
591-78-6
74-88-4
78-83-1
126-98-7
78-83-1
108-10-1
630-20-6
75-69-4
96-18-4
108-05-4
108-38-3
* determined at a purge temperature of
75 - 85 °C
1.3 The detection limit of this method is
usually dependent on the level of
2.1 The percent solids content of the sample
is determined. If the solids content is
known or determined to be less than one
percent, stable isotopically labeled
analogs of the compounds of interest are
added to a 5 ml sample and the sample is
purged with an inert gas at 20 - 25 °C in
a chamber designed for soil or water
samples.
If the solids content is greater than one
percent, five mL of reagent water and the
labeled compounds are added to a 5 gram
aliquot of sample and the mixture is
purged at 40 °C. Compounds that will not
purge at 20 - 25 °C or at 40 °C are purged
at 75 - 85 "C. In the purging process,
the volatile compounds are transferred
from the aqueous phase into the gaseous
phase where they are passed into a sorbent
column and trapped. After purging is
completed, the trap is backflushed and
heated rapidly to desorb the compounds
into a gas chromatograph (GC). The
compounds are separated by the GC and
detected by a mass spectrometer (MS)
(references 2 and 3). The labeled
compounds serve to correct the variability
of the analytical technique.
2.2 Identification of a pollutant (qualitative
analysis) is performed in one of three
ways: (1) For compounds listed in table 1
and other compounds for which authentic
standards are available, the GCMS system
is calibrated and the mass spectrum and
retention time for each standard are
-------
2.3
stored in a user created library. A
compound is identified when its retention
time and mass spectrum agree with the
library retention time and spectrum. (2)
For compounds listed in table 2 and other
compounds for which standards are not
available, a compound is identified when
the retention time and mass spectrum agree
with those specified in this method. (3)
For chromatographic peaks which are not
identified by (1) and (2) above, the
background corrected spectrum at the peak
maxinun is compared with spectra in the
EPA/NIH Mass Spectral File (reference 4).
Tentative identification is established
when the spectrum agrees.
Quantitative analysis is performed in one
of four ways by GCMS using extracted ion
current profile (EICP) areas: (1) For
compounds listed in table 1 and other
compounds for which standards and labeled
analogs are available, the GCMS system is
calibrated and the compound concentration
is determined using an isotope dilution
technique. (2) For compounds listed in
table 1 and for other compounds for which
authentic standards but no labeled
2.4
3.1
compounds are available, the GCMS system
is calibrated and the compound
concentration is determined using an
internal standard technique. (3) For
compounds listed in table 2 and other
compounds for which standards are not
available, compound concentrations are
determined using known response factors.
(4) For compounds for which neither
standards nor known response factors are
available, compound concentration is
determined using the sum of the EICP areas
relative to the sum of the EICP areas of
the nearest eluted internal standard.
Quality is assured through reproducible
calibration and testing of the purge and
trap and GCMS systems.
CONTAMINATION AND INTERFERENCES
Impurities in the purge gas, organic
compounds out-gassing from the plumbing
upstream of the trap, and solvent vapors
in the laboratory account for the majority
of contamination problems. The analytical
system is demonstrated to be free from
interferences under conditions of the
Table 3
GAS CHROMATOGRAPHY OF PURGEABLE ORGANIC COMPOUNDS
EGD
No.
(1)
245
345
246
346
288
388
216
316
244
344
546
616
716
Compound
chloromethane-d-
chloromethane
bromomethane-d.
bromomethane
vinyl chloride-d.
vinyl chloride
chloroethane-d.
chloroethane
methylene chloride-d-
methylene chloride
iodomethane
acetone-d,
o
acetone
Retention time
Mean EGD
(sec) Ref
147
148
243
246
301
304
378
386
512
517
498
554
565
181
245
181
246
181
288
181
216
181
244
181
181
616
Relative (2)
0.141 -
0.922 -
0.233 -
0.898 -
0.286 -
0.946 -
0.373 -
0.999 -
0.582 -
0.999 -
0.68
0.628 -
0.984 -
0.270
1.210
0.423
1.195
0.501
1.023
0.620
1.060
0.813
1.017
0.889
1.019
Mini-
mum
level
(ug/L)
50
50
50
50
50
10
50
50
10
10
50
50
Method Detection
Limit (4)
Low High
solids solids
(ug/kg) (ug/kg)
207* 13
148* 11
190* 11
789* 24
566* 280*
3561* 322*
-------
202
302
203
303
533
552
543
229
329
536
532
181
213
313
615
715
230
330
614
714
223
323
535
210
310
539
548
547
211
311
627
727
206
306
554
248
348
534
537
232
332
542
287
387
541
204
304
251
351
214
314
233
333
acrolein-d^
acrolein
acrylomtrile-dj
acrylonitrile
carbon disulfide
trichlorof luoromethane
ethyl cyanide
1 , 1-dichloroethene-d-
1,1-dichloroethene
3-chloropropene
allyl alcohol
bromoch loromethane (I.S.)
1 , 1 -di ch loroethane-dj
1,1-dichloroethane
diethyl ether-d.-
di ethyl ether
trans-1 ,2-dichloroethene-d_
trans-1 ,2-dichloroethene
methyl ethyl ketone-d,
methyl ethyl ketone
chloroform- C.
chloroform
chloroacetonitrile
1,2-dichloroethane-d^
1,2-dichloroethane
dibromomethane
methacrylonitrile
isobutyl alcohol
1 , 1 , 1 - 1 r i ch I oroethane- Cp
1,1, 1-trichloroethane
p-dioxane-d_
o
p-dioxane
carbon tetrachloride- C.
carbon tetrachloride
vinyl acetate
bromodichloromethane- C1
bromodi ch 1 oromethane
2-chloro-1,3-butadiene
crotonaldehyde
1 ,2-dichloropropane-d,
1 , 2-di ch I oropropane
cis-1,3-dichloropropene
trichloroethene- C^
trichloroethene
1 ,3-dichloropropane
benzene-d^
benzene
chlorodibromomethane- C^
chlorodibromomethane
1,1,2-trichloroethane- Cp
1,1,2-trichloroethane
trans-1 ,3-dichloropropene-d,
trans-1 ,3-dichloropropene
564
566
606
612
631
663
672
696
696
696
703
730
778
786
804
820
821
821
840
848
861
861
884
901
910
910
921
962
989
999
982
1001
1018
1018
1031
1045
1045
1084
1098
1123
1134
1138
1172
1187
1196
1200
1212
1222
1222
1224
1224
1226
1226
181
202
181
203
181
181
181
181
229
181
181
181
181
213
181
615
181
230
181
614
181
223
181
181
210
181
181
181
181
211
181
627
182
206
182
182
248
182
182
182
232
182
182
287
182
182
204
182
251
182
214
182
233
0.641 -
0.984 -
0.735 -
0.985 -
0.86
0.91
0.92
0.903 -
0.999 -
0.95
0.96
1.000 -
1.031 -
0.999 -
1.067 -
1.010 -
1.056 -
0.996 -
0.646 -
0.992 -
1.092 -
0.961 -
1.21
1.187 -
0.973 -
1.25
1.26
1.32
1.293 -
0.989 -
1.262 -
1.008 -
0.754 -
0.938 -
0.79
0.766 -
0.978 -
0.83
0.84
0.830 -
0.984 •
0.87
0.897 -
0.991 -
0.92
0.888 •
1.002 -
0.915 -
0.989 -
0.922 -
0.975 -
0.922 -
0.993 -
0.903(5)
1.018(5)
0.926
1.030
0.976
1.011
1.000
1.119
1.014
1.254
1.048
1.228
1.011
1.202
1.055
1.322
1.009
1.416
1.032
1.598
1.044
1.448(5)
1.040(5)
0.805
1.005
0.825
1.013
0.880
1.018
0.917
1.037
0.952
1.026
0.949
1.030
0.953
1.027
0.959
1.016
50
50
50
50
10
10
10
10
10
50
50
10
10
50
50
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
377*
360*
31
16
63
41
241*
21
23
16
--
87
28
29
41
23
15
26
(6)«
18
9
5
1
12
3
80*
2
3
4
140*
9
3
5
2
8
2
1
(6)*
-------
019
538
182
549
247
347
551
550
553
215
315
545
285
385
540
183
544
286
386
207
307
238
338
185
951
952
2-chloroethyt vinyl ether
1,2-dibromoethane
2-bromo-1-chloropropane U.S.)
methyl methacrylate
bromoform- C.
bromoform
1,1,1,2- tetrach loroethane
4-methyl -2-pentanone
1 ,2,3-trichloropropane
1,1,2,2-tetrachloroethane-d2
1 , 1 ,2,2- tetrachloroethane
2-hexanone
tetrachloroethene- C2
tetrach I oroethene
trans-1,4-dichloro-2-butene
1,4-dichlorobutane (int std)
ethyl methacrylate
toluene-dg
toluene
ch lorobenzene-dc
chlorobenzene
ethylbenzene-d..
ethylbenzene
bromof I uorobenzene
m-xylene
o- * p-xylene
1278
1279
1306
1379
1386
1386
1408
1435
1520
1525
1525
1525
1528
1528
1551
1555
1594
1603
1619
1679
1679
1802
1820
1985
2348
2446
182
182
182
182
182
247
182
183
183
183
215
183
183
285
183
183
183
183
286
183
207
183
238
183
183
183
0.983 -
0.98
1.000 -
1.06
1.048 -
0.992 -
1.08
0.92
0.98
0.969 -
0.890 -
0.98
0.966 -
0.997 -
1.00
1.000 -
1.03
1.016 -
1.001 -
1.066 -
0.914 -
1.144 -
0.981 -
1.255 -
1.51
1.57
1.026
1.000
1.087
1.003
0.996
1.016
0.996
1.003
1.000
1.054
1.019
1.135
1.019
1.293
1.018
1.290
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
122 21
91 7
20 6
106 10
27 4
21 58*
28 4
(1) Reference numbers beginning with 0, 1, 5, or 9 indicate a pollutant quantified by the internal standard
method; reference numbers beginning with 2 or 6 indicate a labeled compound quantified by the internal standard
method; reference numbers beginning with 3 or 7 indicate a pollutant quantified by isotope dilution.
(2) The retention time limits in this column are based on data from four wastewater laboratories. The single
values for retention times in this column are based on data from one wastewater laboratory.
(3) This is a minimum level at which the analytical system shall give recognizable mass spectra (background
corrected) and acceptable calibration points when calibrated using reagent water. The concentration in the
aqueous or solid phase is determined using the equations in section 13.
(4) Method detection limits determined in digested sludge (low solids) and in filter cake or compost (high
sol ids).
(5) Specification derived from related compound.
(6) An unknown interference in the particular sludge studied precluded measurement of the Method Detection
Limit (HDL) for this compound.
*Background levels of these compounds were present in the sludge resulting in higher than expected MOL's. The
HDL for these compounds is expected to be approximately 20 ug/kg (100 - 200 for the gases and water soluble
compounds) for the low solids method and 5-10 ug/kg (25 - 50 for the gases and water soluble compounds) for
the high solids method, with no interferences present.
Column: 2.4 m (8 ft) x 2 mm i.d. glass, packed with one percent SP-1000 coated on 60/80 Carbopak B.
Carrier gas: helium at 40 ml/min.
Temperature program: 3 min at 45 °C, 8 "C per min to 240 °C, hold at 240 "C for 15 minutes.
-------
analysis by analyzing reagent water blanks
initially and with each sample batch
(samples analyzed on the same 8 hr shift),
as described in section 8.5.
3.2 Samples can be contaminated by diffusion
of volatile organic compounds (particu-
larly methylene chloride) through the
bottle seal during shipment and storage.
A field blank prepared from reagent water
and carried through the sampling and
handling protocol serves as a check on
such contamination.
3.3 Contamination by carry-over can occur when
high level and low level samples are
analyzed sequentially. To reduce carry-
over, the purging device (figure 1 for
samples containing less than one percent
solids; figure 2 for samples containing
one percent solids or greater) is cleaned
or replaced with a clean purging device
after each sample is analyzed. When an
unusually concentrated sample is
encountered, it is followed by analysis of
a reagent water blank to check for carry-
over. Purging devices are cleaned by
washing with soap solution, rinsing with
tap and distilled water, and drying in an
oven at 100-125 °C. The trap and other
parts of the system are also subject to
contamination; therefore, frequent bakeout
and purging of the entire system may be
required.
3.4 Interferences resulting from samples will
vary considerably from source to source,
depending on the diversity of the site
being sampled.
4 SAFETY
4.1 The toxicity or carcinogenic}ty of each
compound or reagent used in this method
has not been precisely determined;
however, each chemical compound should be
treated as a potential health hazard.
Exposure to these compounds should be
reduced to the lowest possible level. 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 data handling sheets
should also be made available to all
personnel involved in these analyses.
Additional information on laboratory
safety can be found in references 5 - 7.
4.2 The following compounds covered by this
method have been tentatively classified as
known or suspected human or mammalian car-
cinogens: benzene, carbon tetrachloride,
chloroform, and vinyl chloride. Primary
standards of these toxic compounds should
be prepared in a hood, and a N10SH/MESA
approved toxic gas respirator should be
worn when high concentrations are handled.
5 APPARATUS AND MATERIALS
5.1 Sample bottles for discrete sampling
5.1.1 Bottle--25 to 40 ml with screw cap (Pierce
13075, or equivalent). Detergent wash,
rinse with tap and distilled water, and
dry at >105 °C for one hr minimum before
use.
5.1.2 Septum--Teflon-faced silicone (Pierce
12722, or equivalent), cleaned as above
and baked at 100 - 200 °C for one hour
minimum.
5.2 Purge and trap device--consists of purging
device, trap, and desorber.
5.2.1 Purging devices for water and soil samples
5.2.1.1 Purging device for water samples—designed
to accept 5 ml samples with water column
at least 3 cm deep. The volume of the
gaseous head space between the water and
trap shall be less than 15 ml. The purge
gas shall be introduced less than 5 mm
from the base of the water column and
shall pass through the water as bubbles
with a diameter less than 3 mm. The
purging device shown in figure 1 meets
these criteria.
-------
OPTIONAL
FOAM TRAP
INLET 1/4 IN OD
10 MM GLASS FRIT
MEDIUM POROSITY
SAMPLE INLET
2 WAY SYRINGE VALVE
17 CM 20 GAUGE SYRINGE NEEDLE
6 MM O 0 RUBBER SEPTUM
INLET 1>4 IN O D
1/16 IN OO
"STAINLESS STEEL
13X
MOLECULAR SIEVE
PURGE GAS FILTER
PURGE GAS
1 FLOW CONTROL
FIGURE 1 Purging Device for Waters
PURGE INLET FITTING
SAMPLE OUTLET FITTING
3' x 6 MM O D GLASS TUBING
FIGURES Purging Device for Soils or Waters
5.2.1.2 Purging device for solid samples—designed
to accept 5 grams of solids plus 5 mL of
water. The volume of the gaseous head
space between the water and trap shall be
less than 25 ml. The purge gas shall be
introduced less than 5 mm from the base of
the sample and shall pass through the
water as bubbles with a diameter less than
3 mm. The purging device shall be capable
of operating at ambient temperature (20 -
25 °C) and of being controlled at
temperatures of 40 ± 2 °C and 80 ± 5 "C
while the sample is being purged. The
purging device shown in figure 2 meets
these criteria.
5.2.2 Trap--25 to 30 cm x 2.5 mm i.d. minimum,
containing the following:
5.2.2.1 Methyl silicone packing—one t 0.2 cm, 3
percent OV-1 on 60/80 mesh Chromosorb U,
or equivalent.
5.2.2.2 Porous potymer--15 ± 1.0 cm, Tenax GC
(2,6-diphenylene oxide polymer), 60/80
mesh, chromatographic grade, or
equivalent.
5.2.2.3 Silica gel--8 ± 1.0 cm, Davison Chemical,
35/60 mesh, grade 15, or equivalent. The
trap shown in figure 3 meets these
specifications.
5.2.4 Desorbei—shall heat the trap to 175 i 5
°C in 45 seconds or less. The polymer
section of the trap shall not exceed a
temperature of 180 °C and the remaining
sections shall not exceed 220 °C during
desorb, and no portion of the trap shall
exceed 225 °C during bakeout. The
desorber shown in figure 3 meets these
specifications.
5.2.5 The purge and trap device may be a
separate unit or coupled to a GC as shown
in figures 4 and 5.
-------
PACKING DETAIL
__*- 5 MM GLASS WOOL
7 7 CM SILICA GEL
CONSTRUCTION DETAIL
COMPRESSION
FITTING NUT
AND FERRULES
14 FT 7D/FOOT
RESISTANCE WIRE
WRAPPED SOLID
15 CM TENAX GC
•- i CM 3°; 0V i
~~S- 5 MM GLASS WOOL
FIGURES Trap Construction and Packings
5.3 Gas chromatograph—shall be linearly
temperature programmable with initial and
final holds, shall contain a glass jet
separator as the MS interface, and shall
produce results which meet the calibration
(sect i on 7), quaIi ty assurance < sect i on
8), and performance tests (section 11) of
this method.
CARRIER GAS P UCKJID INJECTlON PO*"!
FLOW CONTROL \X I I— COLUMN OVEN
PRESSURE K W
'RESSUHE
REGULATOR
OPTIONAL a PORT COLUMN
SELECTION VALVE
CONFIRMATORY COLUMN
TO DETECTOR
ANALYTICAL COLUMN
PURGE GAS L3 ^.
CLOW CONTROL A V
• I/ MOLECULAR
SIEVE FILTER
PURGING
DEVICE
NOTE
ALL LINES BETWEEN TRAP
ANO GC SHOULD BE HEATED
TO HO C
FIGURE 4 Schematic of Purge and Trap
Device-Purge Mode
CARRIER GAS
FLOW CONTROL
LIQUID INJECTION PORTS
COLUMN OVEN
OPTIONAL 4 PORT COLUMN
SELECTION VALVE
CONFIRMATORY COLUMN
TO DETECTOR
IALVTICAL COLUMN
PURGE GAS a ',
FLOW CONTROL 4 V
I3X MOLECULAR
SIEVE FILTEP
PURGING
DEVICE
NOTE
ALL LINES BETWEEN TRAP
AND GC SHOULD 86 HEATED
'oaoc
FIGURE 5 Schematic of Purge and Trap
Device-Desorb Mode
5.3.1 Column--2.8 ± 0.4 m x 2 t 0.5 mm i.d.
glass, packed with one percent SP-1000 on
Carbopak B, 60/80 mesh, or equivalent.
5.4 Mass spectrometer--70 eV electron impact
ionization; shall repetitively scan from
20 to 250 amu every 2-3 seconds, and
produce a unit resolution (valleys between
m/z 174-176 less than 10 percent of the
height of the m/z 175 peak), background
corrected mass spectrum from 50 ng 4-
bromofluorobenzene (BFB) injected into the
GC. The BFB spectrum shall meet the mass-
intensity criteria in table 4. All
portions of the GC column, transfer lines,
and separator which connect the GC column
to the ion source shall remain at or above
the column temperature during analysis to
preclude condensation of less volatile
compounds.
Table 4
BFB MASS-INTENSITY SPECIFICATIONS
M/z
Intensity Required
50 15 to 40 percent of m/z 95
75 30 to 60 percent of m/z 95
95 base peak, 100 percent
96 5 to 9 percent of m/z 95
173 less than 2 percent of m/z 174
174 greater than 50 percent of m/z 95
175 5 to 9 percent of m/z
176 95 to 101 percent of m/z 174
177 5 to 9 percent of m/z 176
10
-------
5.5 Data system--shall collect and record MS
data, store mass-intensity data in
spectral libraries, process GCMS data and
generate reports, and shall calculate and
record response factors.
5.5.1 Data acquisition--mass spectra shall be
collected continuously throughout the
analysis and stored on a mass storage
device.
5.5.2 Mass spectral libraries—user created
libraries containing mass spectra obtained
from analysis of authentic standards shall
be employed to reverse search GCMS runs
for the compounds of interest (section
7.2).
5.5.3 Data processing—the data system shall be
used to search, locate, identify, and
quantify the compounds of interest in each
GCMS analysis. Software routines shall be
employed to compute retention times and
EICP areas. Displays of spectra, mass
chromatograms, and library comparisons are
required to verify results.
5.5.4 Response factors and multipoint calibra-
tions—the data system shall be used to
record and maintain lists of response
factors (response ratios for isotope dilu-
tion) and generate multi-point calibration
curves (section 7). Computations of rela-
tive standard deviation (coefficient of
variation) are useful for testing calibra-
tion linearity. Statistics on initial and
on-going performance shall be maintained
(sections 8 and 11).
5.6 Syringes--5 mL glass hypodermic, with
Luer-lok tips.
5.7 Micro syringes--10, 25, and 100 uL.
5.8 Syringe valves--2-way, with Luer ends
(Teflon or Kel-F).
5.9 Syringe--5 ml, gas-tight, with shut-off
valve.
5.10
8ottles--15
liner.
ml.
screw-cap with Teflon
5.11 Balances
5.11.1 Analytical, capable of weighing 0.1 mg.
5.11.2 Top loading, capable of weighing 10 mg.
5.12 Equipment for determining percent moisture
5.12.1 Oven, capable of being temperature
controlled at 110 ± 5 "C.
5.12.2 Oessicator.
5.12.3 Beakers--50 - 100 mL.
6 REAGENTS AND STANDARDS
6.1 Reagent watei—water in which the
compounds of interest and interfering
compounds are not detected by this method
(section 11.7). It may be generated by
any of the following methods:
6.1.1 Activated carbon—pass tap water through a
carbon bed (Calgon Filtrasorb-300, or
equivalent).
6.1.2 Water purifier--pass tap water through a
purifier (Millipore Super Q, or
equivalent).
6.1.3 Boil and purge—heat tap water to 90-100
°C and bubble contaminant free inert gas
through it for approximately one hour.
While still hot, transfer the water to
screw-cap bottles and seal with a Teflon-
lined cap.
6.2 Sodium thiosulfate—ACS granular.
6.3 Methanol--pesticide quality or equivalent.
6.4 Standard solutions--purchased as solutions
or mixtures with certification to their
purity, concentration, and authenticity,
or prepared from materials of known purity
and composition. If compound purity is 96
percent or greater, the weight may be used
without correction to calculate the
concentration of the standard.
6.5 Preparation of stock solutions—prepare in
methanol using liquid or gaseous standards
11
-------
per the steps below. Observe the safety
precautions given in section 4.
6.5.1 Place approximately 9.8 ml of metHanoi in
a 10 mL ground glass stoppered volumetric
flask. Allow the flask to stand unstop-
pered for approximately 10 minutes or un-
til all methanol wetted surfaces have
dried.
In each case, weigh the flask, immediately
add the compound, then immediately reweigh
to prevent evaporation losses from
affecting the measurement.
6.5.1.1 Liquids—using a 100 uL syringe, permit 2
drops of liquid to fall into the methanol
without contacting the neck of the flask.
Alternatively, inject a known volume of
the compound into the methanol in the
flask using a micro-syringe.
that can be used to determine the accuracy
of calibration standards are available
from the US Environmental Protection
Agency, Environmental Monitoring and Sup-
port Laboratory, Cincinnati, Ohio.
6.6 Labeled compound spiking solution--from
stock standard solutions prepared as
above, or from mixtures, prepare the spik-
ing solution to contain a concentration
such that a 5-10 uL spike into each 5 mL
sample, blank, or aqueous standard ana-
lyzed will result in a concentration of 20
ug/L of each labeled compound. For the
gases and for the water soluble compounds
(acrolein, acrylonitrile, acetone, diethyl
ether, and MEK), a concentration of 100
ug/L may be used. Include the internal
standards (section 7.5) in this solution
so that a concentration of 20 ug/L in each
sample, blank, or aqueous standard will be
produced.
6.5.1.2 Gases (chloromethane, bromomethane,
chloroethane, vinyl chtoride)--fiit a
valved 5 mL gas-tight syringe with the
compound.
Lower the needle to approximately 5 mm
above the methanol meniscus. Slowly
introduce the compound above the surface
of the meniscus. The gas will dissolve
rapidly in the methanol.
6.5.2 Fill the flask to volume, stopper, then
mix by inverting several times. Calculate
the concentration in mg/mL (ug/uL) from
the weight gain (or density if a known
volume was injected).
6.5.3 Transfer the stock solution to a Teflon
sealed screw-cap bottle.
6.7 Secondary standards—using stock solu-
tions, prepare a secondary standard in
methanol to contain each pollutant at a
concentration of 500 ug/mL. For the gases
and water soluble compounds (section 6.6),
a concentration of 2.5 mg/mL may be used.
6.7.1 Aqueous calibration standards--using a 25
uL syringe, add 20 uL of the secondary
standard (section 6.7) to 50, 100, 200,
500, and 1000 mL of reagent water to
produce concentrations of 200, 100, 50,
20, and 10 ug/L, respectively. If the
higher concentration standard for the
gases and water soluble compounds was
chosen (section 6.6), these compounds will
be at concentrations of 1000, 500, 250,
100, and 50 ug/L in the aqueous
calibration standards.
Store, with minimal headspace, in the dark
at -10 to -20 "C.
6.5.4 Prepare fresh standards weekly for the
gases and 2-chloroethylvinyl ether. All
other standards are replaced after one
month, or sooner if comparison with check
standards indicate a change in concentra-
tion. Quality control check standards
6.7.2 Aqueous performance standard--an aqueous
standard containing all pollutants,
internal standards, labeled compounds, and
BFB is prepared daily, and analyzed each
shift to demonstrate performance (section
11). This standard shall contain either
20 or 100 ug/L of the labeled and
pollutant gases and water soluble
compounds, 10 ug/L BFB, and 20 ug/L of all
12
-------
other pollutants, labeled compounds, and
internal standards. It may be the nominal
20 ug/L aqueous calibration standard
(section 6.7.1).
6.7.3 A methanolic standard containing all
pollutants and internal standards is
prepared to demonstrate recovery of these
compounds when syringe injection and purge
and trap analyses are compared.
This standard shall contain either 100
ug/mL or 500 ug/mL of the gases and water
soluble compounds, and 100 ug/mL of the
remaining pollutants and internal
standards (consistent with the amounts in
the aqueous performance standard in
6.7.2).
6.7.4 Other standards which may be needed are
those for test of BFB performance (section
7.1) and for collection of mass spectra
for storage in spectral libraries (section
7.2).
7 CALIBRATION
Calibration of the GCMS system is
performed by purging the compounds of
interest and their labeled analogs from
reagent water at the temperature to be
used for analysis of samples.
7.1 Assemble the gas chromatographic apparatus
and establish operating conditions given
in table 3. By injecting standards into
the GC, demonstrate that the analytical
system meets the minimum levels in table 3
for the compounds for which calibration is
to be performed, and the mass-intensity
criteria in table 4 for 50 ng BFB.
7.2 Mass spectral libraries—detection and
identification of the compounds of
interest are dependent upon the spectra
stored in user created libraries.
7.2.1 For the compounds in table 1 and other
compounds for which the GCMS is to be
calibrated, obtain a mass spectrum of each
pollutant and labeled compound and each
internal standard by analyzing an
authentic standard either singly or as
part of a mixture in which there is no
interference between closely eluted
components. That only a single compound
is present is determined by examination of
the spectrum. Fragments not attributable
to the compound under study indicate the
presence of an interfering compound.
Adjust the analytical conditions and scan
rate (for this test only) to produce an
undistorted spectrum at the GC peak
maximum. An undistorted spectrum will
usually be obtained if five complete
spectra are collected across the upper
half of the GC peak. Software algorithms
designed to "enhance" the spectrum may
eliminate distortion, but may also
eliminate authentic m/z's or introduce
other distortion.
7.2.3 The authentic reference spectrum is
obtained under BFB tuning conditions
(section 7.1 and table 4) to normalize it
to spectra from other instruments.
7.2.4 The spectrum is edited by saving the 5
most intense mass spectral peaks and all
other mass spectral peaks greater than 10
percent of the base peak. The spectrum
may be further edited to remove common
interfering masses. If 5 mass spectral
peaks cannot be obtained under the scan
conditions given in section 5.4, the mass
spectrometer may be scanned to an m/z
lower than 20 to gain additional spectral
information. The spectrum obtained is
stored for reverse search and for compound
confirmation.
7.2.5 For the compounds in table 2 and other
compounds for which the mass spectra,
quantitation m/z's, and retention times
are known but the instrument is not to be
calibrated, add the retention time and
reference compound (table 3); the response
factor and the quantitation m/z (table 5);
and spectrum (Appendix A) to the reverse
search library. Edit the spectrum per
section 7.2.4, if necessary.
7.3 Assemble the purge and trap device. Pack
the trap as shown in figure 3 and
13
-------
Table 5
VOLATILE ORGANIC COMPOUND CHARACTERISTIC M/Z'S
Compound
acetone
acrolein
acrylom'trile
allyl alcohol
benzene
2-bromo-1-chloropropane <3)
bromoch I oromethane (3)
bromod i ch I oromethane
bromoform
bromomethane
carbon disulfide
carbon tetrachloride
2-chloro-1,3-butadiene
chloroacetonitri le
chlorobenzene
chloroethane
2-chloroethyl vinyl ether
chloroform
ch I oromethane
3-chloropropene
crotonatdehyde
di bromoch loromethane
1 ,2-dibromoethane
di bromomethane
1,4-dichlorobutane (3)
trans- 1,4-dichloro-2-butene
1,1-dichloroethane
1,2-dichloroethane
1 , 1 -dichloroethene
trans- 1 , 2-di ch I orethene
1 , 2 - d i ch I oropropane
1,3-dichtoropropane
cis-1 ,3-dichloropropene
trans-1,3-dichloropropene
di ethyl ether
p-dioxane
ethyl cyanide
ethyl methacrylate
ethylbenzene
2-hexanone
iodomethane
isobutyl alcohol
methylene chloride
methyl ethyl ketone
methyl methacrylate
4-methyl-2-pentanone
Labeled
analog
d6
d4
dj
d6
13C
13C
d_
13c
"5
d5
d7
13c7
"3
13c
Cvj
d4
d2
d2
d6
d4
d10
d8
d10
d2
dj
Primary
m/z's
58/64
56/60
53/56
57
78/84
77
128
83/86
173/176
96/99
76
47/48
53
75
112/117
64/71
106/113
85/86
50/52
76
70
129/130
107
93
55
75
63/66
62/67
61/65
61/65
63/67
76
75
75/79
74/84
88/96
54
69
106/116
58
142
74
84/88
72/75
69
58
Reference
compound
(1)
181
181
182
181
181
182
182
181
183
182
182
181
183
183
181
181
182
183
Response factor at
purge temp, of
20 °C 80 "C
(2)
1.93
0.29
(2)
0.43
(2)
0.86
1.35
0.093
0.89
0.29
(2)
0.69
0.076
4.55
(2)
0.23
0.15
0.20
2.02
0.50
1.12
0.63
0.090
0.68
1.91
0.14
0.88
0.41
1.26
0.52
0.33
2.55
0.22
0.79
0.29
-------
methacryloni tri le
1,1,1, 2- tetrach I oroethane
1,1,2,2-tetrachloroethane d-
tetrachlorethene C_
toluene d_
1,1,1-trichloroethane d,
13 ^
1,1.2-trichloroethane C,
trichloroethene C,
t r i ch I orof I uoromethane
1,2,3-trichloropropane
vinyl acetate
vinyl chloride d.
m-xylene
o- + p-xylene
67
131
83/84
166/172
92/99
97/102
83/84
95/136
101
75
86
62/65
106
106
181
182
181
183
182
183
183
0.25
0.20
2.31
0.89
0.054
1.69
3.33
0.79
0.25
2.19
0.72
0.19
-
~
(1) 181 « bromochloromethane 182 = 2-bromo-1-chloropropane
(2) not detected at a purge temperature of 25 °C
(3) internal standard
183 = 1,4-dichlorobutane
condition overnight at 170 - 180 °C by
back-flushing with an inert gas at a flow
rate of 20 - 30 mL/min. Condition traps
daily for a minimum of 10 minutes prior to
use.
The exact value must be determined by
experience for each instrument. It is
used to match the calibration range of the
instrument to the analytical range and
detection limits required.
7.3.1 Analyze the aqueous performance standard
(section 6.7.2) according to the purge and
trap procedure in section 10. Compute the
area at the primary m/z (table 5) for each
compound. Compare these areas to those
obtained by injecting one uL of the
metHanoiic standard (section 6.7.3) to
determine compound recovery. The recovery
shall be greater than 20 percent for the
water soluble compounds (section 6.6), and
60 - 110 percent for all other compounds.
This recovery is demonstrated initially
for each purge and trap GCMS system. The
test is repeated only if the purge and
trap or GCMS systems are modified in any
way that might result in a change in
recovery.
7.3.2 Demonstrate that 100 ng toluene (or
toluene-d.) produces an area at m/z 91 (or
99) approximately one-tenth that required
to exceed the linear range of the system.
7.4 Calibration by isotope dilution—the iso-
tope dilution approach is used for the
purgeable organic compounds when appropri-
ate labeled compounds are available and
when interferences do not preclude the
analysis. If labeled compounds are not
available, or interferences are present,
the internal standard method (section 7.5)
is used. A calibration curve encompassing
the concentration range of interest is
prepared for each compound determined.
The relative response (RR) vs concentra-
tion (ug/L) is plotted or computed using a
linear regression. An example of a cali-
bration curve for toluene using toluene-dg
is given in figure 6. Also shown are the
± 10 percent error limits (dotted lines).
Relative response is determined according
to the procedures described below. A min-
imum of five data points are required for
calibration (section 7.4.4).
15
-------
10-
ifs
O
> 1.0-
0.1-
2 10 20 50 100 200
CONCENTRATION (ug/L)
FIGURES Relative Response Calibration Curve for
Toluene. The Dotted Lines Enclose a +/- 10 Percent
Error Window
7.4.1 The relative response (RR) of pollutant to
labeled compound is determined from iso-
tope ratio values calculated from acquired
data. Three isotope ratios are used in
this process:
R = (area at-m^/z)
(area at
If either of the areas is zero, it is as-
signed a value of one in the calculations;
that is, if:
area of m./z = 50721, and
area of nu/z = 0, then
R = 50721 = 50720
1
The m/z's are always selected such that R
> R . When there is a difference in re-
tention times (RT) between the pollutant
and labeled compounds, special precautions
are required to determine the isotope ra-
tios.
R , R , and Rffl are defined as follows:
R = [area m./z (at RT.)]
1
[area m-/z (at RT.)]
R = the isotope ratio measured in the
pure pollutant (figure 7A).
R = the isotope ratio of pure labeled
compound (figure 78).
Rm = the isotope ratio measured in the an-
alytical mixture of the pollutant and la-
beled compounds (figure 7C).
[area m./z (at
[area uu/z (at RT-)]
7.4.3 An example of the above calculations can
be taken from the data plotted in figure 7
for toluene and toluene-d.. For these
data,
168900
The correct way to calculate RR is:
7.4.2
RR
(Rm-Rx)(Ry*1)
If R is not between 2R and 0.5R , the
method does not apply and the sample is
analyzed by the internal standard method
(section 7.5).
In most cases, the retention times of the
pollutant and labeled compound are the
same and isotope ratios (R's) can be cal-
culated from the EICP areas, where:
V
1 = 0.00001640
60960
R = 96868 = 1.174
ffl '
82508
The RR for the above data is then calcu-
lated using the equation given in section
7.4.1. For the example, RR = 1.174. Not
all labeled compounds elute before their
pollutant analogs.
16
-------
(A)
AREA=168920
M.
• M/Z 98
• M/Z 92
(B)
AREA=60960
• M/Z 98
• M/2 92
(C)
M/Z 92 96868
M/Z 98 " 82508
• M/Z 98
• M/Z 92
FIGURE 7 Extracted Ion Current Profiles for (A)
Toluene, (B) Toluene-ds, and (C) a Mixture of
Toluene and Toluene-ds
7.4.4 To calibrate the analytical system by
isotope dilution, analyze a 5 ml aliquot
of each of the aqueous calibration
standards (section 6.7.1) spiked with an
appropriate constant amount of the labeled
compound spiking solution (section 6.6),
using the purge and trap procedure in
section 10. Compute the RR at each
concentration.
7.4.5 Linearity-if the ratio of relative
response to concentration for any compound
is constant (less than 20 percent
coefficient of variation) over the 5 point
calibration range, an averaged relative
response/concentration ratio may be used
for that compound; otherwise, the complete
calibration curve for that compound shall
be used over the 5 point calibration
range.
7.5 Calibration by internal standard--used
when criteria for isotope dilution
(section 7.4) cannot be met. The method
is applied to pollutants having no labeled
analog and to the labeled compounds.
The internal standards used for volatiles
analyses are bromochloro methane, 2-
bromo-1-chloropropane, and 1,4-dichlorobu-
tane. Concentrations of the labeled com-
pounds and pollutants without labeled
analogs are computed relative to the near-
est eluted internal standard, as shown in
tables 3 and 5.
7.5.1 Response factors--calibration requires the
determination of response factors (RF)
which are defined by the following
equation:
RF = (A x C. ). where
(Ais x V
A is the EJCP area at the characteristic
m/z for the compound in the daily stan-
dard.
A. is the EICP area at the characteristic
m/z for the internal standard.
C. is the concentration (ug/L) of the in-
ternal standard.
C is the concentration of the pollutant
in the daily standard.
7.5.2 The response factor is determined at 10,
20, 50, 100, and 200 ug/L for the
pollutants (optionally at five times these
concentrations for gases and water soluble
pollutants--see section 6.7), in a way
analogous to that for calibration by
isotope dilution (section 7.4.4). The RF
is plotted against concentration for each
compound in the standard (C ) to produce a
s
calibration curve.
7.5.3 Linearity--if the response factor (RF) for
any compound is constant (less than 35
percent coefficient of variation) over the
5 point calibration range, an averaged
response factor may be used for that
compound; otherwise, the complete
calibration curve for that compound shall
be used over the 5 point range.
7.6 Combined calibration--by adding the
isotopicatly labeled compounds and
17
-------
internal standards (section 6.6) to the
aqueous calibration standards (section
6.7.1), a single set of analyses can be
used to produce calibration curves for the
isotope dilution and internal standard
methods. These curves are verified each
shift (section 11.5) by purging the
aqueous performance standard (section
6.7.2).
Recalibration is required only if
calibration and on-going performance
(section 11.5) criteria cannot be met.
7.7 Elevated purge temperature calibration--
samples containing greater than one
percent solids are analyzed at a
temperature of 40 t 2 °C (section 10).
For these samples, the analytical system
may be calibrated using a purge
temperature of 40 ± 2. °C in order to more
closely approximate the behavior of the
compounds of interest in high solids
samples.
8 QUALITY ASSURANCE/QUALITY CONTROL
costs of measurements, provided all
performance specifications are met. Each
time a modification is made to the method,
the analyst is required to repeat the
procedure in section 8.2 to demonstrate
method performance.
8.1.3 Analyses of blanks are required to
demonstrate freedom from contamination and
that the compounds of interest and
interfering compounds have not been
carried over from a previous analysis
(section 3). The procedures and criteria
for analysis of a blank are described in
sections 8.5.
8.1.4 The laboratory shall spike all samples
with labeled compounds to monitor method
performance. This test is described in
section 8.3.
When results of these spikes indicate
atypical method performance for samples,
the samples are diluted to bring method
performance within acceptable limits
(section H.2).
8.1 Each laboratory that uses this method is
required to operate a formal quality
assurance program (reference 8). The
minimum requirements of this program
consist of an initial demonstration of
laboratory capability, analysis of samples
spiked with labeled compounds to evaluate
and document data quality, and analysis of
standards and blanks as tests of continued
performance. Laboratory performance is
compared to established performance
criteria to determine if the results of
analyses meet the performance
characteristics of the method.
8.1.1 The analyst shall make an initial
demonstration of the ability to generate
acceptable accuracy and precision with
this method.
This ability is established as described
in section 8.2.
8.1.2 The analyst is permitted to modify this
method to improve separations or lower the
8.1.5 The laboratory shall, on an on-going
basis, demonstrate through the analysis of
the aqueous performance standard (section
6.7.2) that the analysis system is in
control. This procedure is described in
sections 11.1 and 11.5.
8.1.6 The laboratory shall maintain records to
define the quality of data that is
generated. Development of accuracy
statements is described in sections 8.4
and 11.5.2.
8.2 Initial precision and accuracy—to
establish the ability to generate
acceptable precision and accuracy, the
analyst shall perform the following
operations for compounds to be calibrated:
8.2.1 Analyze two sets of four 5-mL aliquots (8
aliquots total) of the aqueous performance
standard (section 6.7.2) according to the
method beginning in section 10.
18
-------
8.2.2 Using results of the first set of four
analyses in section 8.2.1, compute the
average recovery (X) in ug/L and the
standard deviation of the recovery (s) in
ug/L for each compound, by isotope
dilution for pollutants with a labeled
analog, and by internal standard for
labeled compounds and pollutants with no
labeled analog.
8.2.3 For each compound, compare s and X with
the corresponding limits for initial
precision and accuracy found in table 6.
If s and X for all compounds meet the
acceptance criteria, system performance is
acceptable and analysis of blanks and
samples may begin. If, however, any
individual s exceeds the precision limit
or any individual X falls outside the
range for accuracy, system performance is
unacceptable for that compound.
NOTE: The large number of compounds in
table 6 present a substantial probability
that one or more will fail one of the
acceptance criteria when all compounds are
analyzed. To determine if the analytical
system is out of control, or if the
failure can be attributed to probability,
proceed as follows:
8.2.4 Using the results of the second set of
four analyses, compute s and X for only
those compounds which failed the test of
the first set of four analyses (section
8.2.3). If these compounds now pass,
system performance is acceptable for all
compounds and analysis of blanks and
samples may begin. If, however, any of
the same compounds fail again, the
analysis system is not performing properly
for the compound (s) in question. In this
event, correct the problem and repeat the
entire test (section 8.2.1).
8.3 The laboratory shall spike all samples
with labeled compounds to assess method
performance on the sample matrix.
8.3.1 Spike and analyze each sample according to
the method beginning in section 10.
8.3.2 Compute the percent recovery (P) of the
labeled compounds using the internal
standard method (section 7.5).
8.3.3 Compare the percent recovery for each
compound with the corresponding labeled
compound recovery limit in table 6. If
the recovery of any compound falls outside
its warning limit, method performance is
unacceptable for that compound in that
sample.
Therefore, the sample matrix is complex
and the sample is to be diluted and
reanalyzed, per section 14.2.
8.4 As part of the QA program for the
laboratory, method accuracy for wastewater
samples shall be assessed and records
shall be maintained. After the analysis
of five wastewater samples for which the
labeled compounds pass the tests in
section 8.3.3, compute the average percent
recovery (P) and the standard deviation of
the percent recovery (s ) for the labeled
compounds only. Express the accuracy
assessment as a percent recovery interval
For example, if
the accuracy
70 - 110%.
Update the accuracy assessment for each
compound on a regular basis (e.g. after
each 5-10 new accuracy measurements).
8.5 Blanks--reagent water blanks are analyzed
to demonstrate freedom from carry-over
(section 3) and contamination.
8.5.1 The level at which the purge and trap
system will carry greater than 5 ug/L of a
pollutant of interest (tables 1 and 2)
into a succeeding blank shall be
determined by analyzing successively
larger concentrations of these compounds.
When a sample contains this concentration
or more, a blank shall be analyzed
immediately following this sample to
demonstrate no carry-over at the 5 ug/L
level.
8.5.2 With each sample lot (samples analyzed on
the same 8 hr shift), a blank shall be
from P - 2s to P + 2s
P = 90X and s = 10X,
interval is expressed as
19
-------
Table 6
ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS
Acceptance criteria at 20 ug/L or as noted
Initial precision
and accuracy
Section 8.2.3
Comoound
acetone*
acroleln*
ecrylonitrile*
benzene
bromod i ch 1 oromethane
bromoform
bromomethane
carbon tetrachloride
ch I orobenzene
chloroethane
2-chloroethylvinyl ether
chloroform
ch I oromethane
dibromoch I oromethane
1,1 -di chloroethane
1,2-di chloroethane
1,1-dichloroethene
trans-1,2-dichlorethene
1,2-di chloropropane
cis-1,3-dichloropropene
trans-1,3-dichloropropene
diethyl ether*
p-dioxane
ethyl benzene
methylene chloride
methyl ethyl ketone*
1 ,1 ,2,2-tetrachloroethane
tetrachlorethene
toluene
1,1,1-trichloroethane
1,1,2-trichloroethane
trichloroethene
vinyl chloride
8 (IK9/L)
51.0
72.0
16.0
9.0
8.2
7.0
25.0
6.9
8.2
15.0
36.0
7.9
26.0
7.9
6.7
7.7
12.0
7.4
19.0
22.0
15.0
44.0
7.2
9.6
9.7
57.0
9.6
6.6
6.3
5.9
7.1
8.9
228.0
X (ug/L)
77 •
32 •
70 •
13 •
7 •
7 •
d -
16 -
14 •
d -
d -
12 -
d -
11 -
11 -
12 -
d -
11 -
d -
d -
d -
75 -
13 -
16 -
d -
66 -
11 -
15 -
15 -
11 •
12 -
17 •
d •
153
168
132
28
32
35
54
25
30
47
70
26
56
29
31
30
50
32
47
51
40
146
27
29
50
159
30
29
29
33
30
30
59
Labeled
compound
recovery
Sec 8.3
and 14.2
P (X)
35 -
37 •
ns •
ns •
ns •
ns •
ns •
42 -
ns -
ns -
ns -
18 -
ns •
16 -
23 -
12 -
ns -
15 -
ns -
ns -
ns -
44 -
ns -
ns •
ns -
36 -
5 -
31 -
4 -
12 -
21 -
35 -
ns -
165
163
204
196
99
214
414
165
205
308
554
172
410
185
191
192
315
195
343
381
284
156
239
203
316
164
199
181
193
200
184
196
452
On- go ing
accuracy
Sec 11.5
R (ug/L)
55 •
7 -
58 •
4 •
4 •
6 -
d -
12 •
4 -
d -
d •
8 -
d -
8 -
9 -
8 -
d •
8 -
d -
d •
d -
55 -
11 -
5 -
d -
42 -
7 -
11 -
6 •
8 •
9 -
12 -
d -
145
190
144
33
34
36
61
30
35
51
79
30
64
32
33
33
52
34
51
56
44
14
29
35
50
158
34
32
33
35
32
34
65
* Acceptance criteria at 100 ug/L
d = detected; result must be greater than zero.
ns = no specification; limit would be below detection limit.
20
-------
analyzed immediately after analysis of the
aqueous performance standard (section
11.1) to demonstrate freedom from
contamination. If any of the compounds of
interest (tables 1 and 2) or any
potentially interfering compound is found
in a blank at greater than 10 ug/L
(assuming a response factor of 1 relative
to the nearest eluted internal standard
for compounds not listed in tables 1 and
2), analysis of samples is halted until
the source of contamination is eliminated
and a blank shows no evidence of
contamination at this level.
8.6 The specifications contained in this
method can be met if the apparatus used is
calibrated properly, then maintained in a
calibrated state. The standards used for
calibration (section 7), calibration
verification (section 11.5) and for
initial (section 8.2) and on-going
(section 11.5) precision and accuracy
should be identical, so that the most
precise results will be obtained. The
GCMS instrument in particular will provide
the most reproducible results if dedicated
to the settings and conditions required
for the analyses of volatiles by this
method.
8.7 Depending on specific program require-
ments, field replicates may be collected
to determine the precision of the sampling
technique, and spiked samples may be re-
quired to determine the accuracy of the
analysis when the internal method is used.
9 SAMPLE COLLECTION, PRESERVATION, AND
HANDLING
9.2 Samples are maintained at 0 - 4 "C from
the time of collection until analysis. If
an aqueous sample contains residual
chlorine, add sodium thiosulfate
preservative (10 mg/40 mL) to the empty
sample bottles just prior to shipment to
the sample site. EPA Methods 330.4 and
330.5 may be used for measurement of
residual chlorine (reference 9). If
preservative has been added, shake the
bottle vigorously for one minute
Immediately after filling.
9.3 For aqueous samples, experimental evidence
indicates that some aromatic compounds,
notably benzene, toluene, and ethyl
benzene are susceptible to rapid
biological degradation under certain
environmental conditions. Refrigeration
alone may not be adequate to preserve
these compounds in wastewaters for more
than seven days.
For this reason, a separate sample should
be collected, acidified, and analyzed when
these aromatics are to be determined.
Collect about 500 mL of sample in a clean
container. Adjust the pH of the sample to
about 2 by adding HCl (1+1) while
stirring. Check pH with narrow range (1.4
to 2.8) pH paper. Fill a sample container
as described in section 9.1. If residual
chlorine is present, add sodium
thiosulfate to a separate sample container
and fill as in section 9.1.
9.4 All samples shall be analyzed within 14
days of collection.
10 PURGE, TRAP, AND GCMS ANALYSIS
9.1 Grab samples are collected in glass
containers having a total volume greater
than 20 mL. For aqueous samples which
pour freely, fill sample bottles so that
no air bubbles pass through the sample as
the bottle is filled and seal each bottle
so that no air bubbles are entrapped.
Maintain the hermetic seal on the sample
bottle until time of analysis.
Samples containing less than one percent
solids are analyzed directly as aqueous
samples (section 10.4). Samples con-
taining one percent solids or greater are
analyzed as solid samples (section 10.5).
10.1 Determination of percent solids
10.1.1 Weigh 5
beaker.
10 g of sample into a tared
21
-------
10.1.2 Dry overnight (12 hours minimum) at 110 ±
5 °C, and cool in a dessicator.
10.1.3 Determine percent solids as follows:
% solids = weight of sample dry x 100
weight of sample wet
10.2 Remove standards and samples from cold
storage and bring to 20 - 25 "C.
10.3 Adjust the purge gas flow rate to 40 t 4
mL/min.
10.4 Samples containing less than one percent
solids
10.4.1 Mix the sample by shaking vigorously.
Remove the plunger from a 5 mL syringe and
attach a closed syringe valve. Open the
sample bottle and carefully pour the
sample into the syringe barrel until it
overflows. Replace the plunger and
compress the sample. Open the syringe
valve and vent any residual air while
adjusting the sample volume to 5.0 t 0.1
mL. Because this process of taking an
aliquot destroys the validity of the
sample for future analysis, fill a second
syringe at this time to protect against
possible loss of data.
10.4.2 Add an appropriate amount of the labeled
compound spiking solution (section 6.6)
through the valve bore, then close the
valve.
10.4.3 Attach the syringe valve assembly to the
syringe valve on the purging device. Open
both syringe valves and inject the sample
into the purging chamber. Purge the
sample per section 10.6.
10.5 Weighing of samples containing one percent
solids or greater.
10.5.1 Mix the sample thoroughly using a clean
spatula.
10.5.2 Weigh 5+1 grams of sample into a purging
vessel (figure 2).
Record the weight to three significant
figures.
10.5.3 Add 5.0 ± 0.1 mL of reagent water to the
vessel.
10.5.4 Using a metal spatula, break up any lumps
of sample to disperse the sample in the
water.
10.5.5 Add an appropriate amount of the labeled
compound spiking solution (section 6.6) to
the sample in the purge vessel. Place a
cap on the purging vessel and and shake
vigorously to further disperse the sample.
Attach the purge vessel to the purging
device.
10.6 Purge the sample for 11.0 ± 0.1 minutes at
20 - 25 °C for samples containing less
than one percent solids. Purge samples
containing one percent solids or greater
at 40 ± 2 "C. If the compounds in table 2
that do not purge at 20 - 40 °C are to be
determined, a purge temperature of 80 ± 5
°C is used.
10.7 After the 11 minute purge time, attach the
trap to the chromatograph and set the
purge and trap apparatus to the desorb
mode (figure 5). Desorb the trapped
compounds into the GC column by heating
the trap to 170 - 180 °C while
backflushing with carrier gas at 20 - 60
mL/min for four minutes. Start MS data
acquisition upon start of the desorb
cycle, and start the GC column temperature
program 3 minutes later. Table 3
summarizes the recommended operating
conditions for the gas chromatograph.
Included in this table are retention times
and minimum levels that can be achieved
under these conditions. An example of the
separations achieved by the column listed
is shown in figure 9. Other columns may
be used provided the requirements in
section 8 are met. If the priority
pollutant gases produce GC peaks so broad
that the precision and recovery
specifications (section 8.2) cannot be
met, the column may be cooled to ambient
22
-------
or subambient temperatures to sharpen
these peaks.
10.8 After desorbing the sample for four 11.5
minutes, recondition the trap by purging
with purge gas while maintaining the trap
temperature at 170 - 180 °C. After
approximately seven minutes, turn off the
trap heater to stop the gas flow through
the trap. When cool, the trap is ready
for the next sample.
10.9 While analysis of the desorbed compounds
proceeds, remove and clean the purge
device. Rinse with tap water, clean with
detergent and water, rinse with tap and
distilled water, and dry for one hour
minimum in an oven at a temperature 11.5.1
greater than 150 °C.
11 SYSTEM PERFORMANCE
than 10 percent of the taller of the two
peaks.
Calibration verification and on-going
precision and accuracy -- compute the
concentration of each pollutant (table 1)
by isotope dilution (section 7.4) for
those compounds which have labeled
analogs. Compute the concentration of
each pollutant (table 1) which has no
labeled analog by the internal standard
method (section 7.5). Compute the
concentration of the labeled compounds by
the internal standard method. These
concentrations are computed based on the
calibration data determined in section 7.
For each pollutant and labeled compound,
compare the concentration with the
corresponding limit for on-going accuracy
in table 6.
11.1 At the beginning of each 8 hr shift during
which analyses are performed, system
calibration and performance shall be
verified for the pollutants and labeled
compounds (table 1). For these tests,
analysis of the aqueous performance
standard (section 6.7.2) shall be used to
verify all performance criteria.
Adjustment and/or recalibration (per
section 7) shall be performed until all
performance criteria are met. Only after
all performance criteria are met may
blanks and samples be analyzed.
11.2 BFB spectrum validity—the criteria in
table 4 shall be met.
11.3 Retention times—the absolute retention
times of the internal standards shall be
as follows: bromochloromethane: 653 - 782
seconds; 2-bromo-1-chloropropane: 1270 -
1369 seconds; 1,4-dichlorobutane: 1510 -
1605 seconds. The relative retention
times of all pollutants and labeled
compounds shall fall within the limits
given in table 3.
11.5.1.1
11.5.1.2
11.4 GC resolution—the valley height between
toluene and toluene-d. (at m/z 91 and 99
o
plotted on the same graph) shall be less
If all compounds meet the acceptance
criteria, system performance is acceptable
and analysis of blanks and samples may
continue. If any individual value falls
outside the range given, system
performance is unacceptable for that
compound.
NOTE: The large number of compounds in
table 6 present a substantial probability
that one or more will fail the acceptance
criteria when all compounds are analyzed.
To determine if the analytical system is
out of control, or if the failure may be
attributed to probability, proceed as
follows:
Analyze a second aliquot of the aqueous
performance standard (section 6.7.2).
Compute the concentration for only those
compounds which failed the first test
(section 11.5.1). If these compounds now
pass, system performance is acceptable for
all compounds and analyses of blanks and
samples may proceed. If, however, any of
the compounds fail again, the measurement
system is not performing properly for
these compounds. In this event, locate
and correct the problem or recalibrate the
23
-------
system (section 7), and repeat the entire
test (section 11.1) for all compounds.
12.1 Labeled compounds and pollutants having no
labeled analog (tables 1 and 2):
11.5.2 Add results which pass the specification
in 11.5.1.2 to initial (section 8.2) and
previous on-going data. Update QC charts
to form a graphic representation of
laboratory performance (figure 8).
oo
y
(-
5
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i t i t i i t i i
TOLUENE-D.
* • .
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123456789 10
ANALYSIS NUMBER
lU Q 110
£2 100<>
TOLUENE
--+3s
-3s
6/1 6/1 6/1 6/1 6/2 6/2 6/3 6/3 6/4 6/5
DATE ANALYZED
FIGURES Quality Control Charts Showing Area
(top graph) and Relative Response of Toluene to
Toluene-ds (lower graph) Plotted as Function of
Time or Analysis Number
Develop a statement of accuracy for each
pollutant and labeled compound by
calculating the average percent recovery
(R) and the standard deviation of percent
recovery (sr). Express the accuracy as a
recovery interval from R - 2s to R + 2s .
For example, if R = 95X and sp = 5X, the
accuracy is 85 - 105 percent.
12 QUALITATIVE DETERMINATION
Identification is accomplished by
comparison of data from analysis of a
sample or blank with data stored in the
mass spectral libraries. For compounds
for which the relative retention times and
mass spectra are known, identification is
confirmed per sections 12.1 and 12.2. For
unidentified GC peaks, the spectrum is
compared to spectra in the EPA/NIH mass
spectral file per section 12.3.
12.1.1 The signals for all characteristic m/z's
stored in the spectral library (section
7.2.4) shall be present and shall maximize
within the same two consecutive scans.
12.1.2 Either (1) the background corrected EICP
areas, or (2) the corrected relative
intensities of the mass spectral peaks at
the GC peak maximum shall agree within a
factor of two (0.5 to 2 times) for all
masses stored in the library.
12.1.3 For the compounds for which the system has
been calibrated (table 1), the relative
retention time shall be within the windows
specified in table 3.
12.1.4 For the compounds for which the system has
not been calibrated but the relative
retention times and mass spectra are known
(table 2), the retention time relative to
the internal standard specified in table 3
shall be within ± 20 scans or t 60
seconds, whichever is greater, based on
the nominal relative retention time
specified in table 3.
12.2 Pollutants having a labeled analog (table
1):
12.2.1 The signals for all characteristic m/z's
stored in the spectral library (section
7.2.4) shall be present and shall maximize
within the same two consecutive scans.
12.2.2 Either (1) the background corrected EICP
areas, or (2) the corrected relative
intensities of the mass spectral peaks at
the GC peak maximum shall agree within a
factor of two for all masses stored in the
spectral library.
13.2.3 The relative retention time between the
pollutant and its labeled analog shall be
within the windows specified in table 3.
12.3 Unidentified GC peaks
-------
12.3.1 The signals for m/z's specific to a GC
peak shall all maximize within the same
two consecutive scans.
12.3.2 Either (1) the background corrected EICP
areas, or (2) the corrected relative
intensities of the mass spectral peaks at
the GC peak maximum shall agree within a
factor of two with the masses stored in
the EPA/NIH Mass Spectral File.
12.4 M/z's present in the experimental mass
spectrum that are not present in the
reference mass spectrum shall be accounted
for by contaminant or background ions. If
the experimental mass spectrum is
contaminated, or if identification is
ambiguous, an experienced spectrometrist
(section 1.4) is to determine the presence
or absence of the compound.
13 QUANTITATIVE DETERMINATION
13.1 Isotope dilution -- by adding a known
amount of a labeled compound to every
sample prior to purging, correction for
recovery of the pollutant can be made be-
cause the pollutant and its labeled analog
exhibit the same effects upon purging,
desorption, and gas chromatography.
Relative response (RR) values for sample
mixtures are used in conjunction with
calibration curves described in section
7.4 to determine concentrations directly,
so long as labeled compound spiking levels
are constant. For the toluene example
given in figure 7 (section 7.4.3), RR
would be equal to 1.174. For this RR
value, the toluene calibration curve given
in figure 6 indicates a concentration of
31.8 ug/L.
13.2 Internal standard—calculate the concen-
tration of each pollutant using the
response factor determined from
calibration data (section 7.5) for the
compounds which were calibrated (table 1),
or from table 5 for compounds which were
not calibrated (table 2), using the
following equation:
Concentration = (A_ x
where the terms are as defined in section
7.5.1.
13.3 The concentration of the pollutant in the
solid phase of the sample is computed
using the concentration of the pollutant
detected in the aqueous solution, as
follows:
Concentration in solid (ug/kg) =
O.OOS L x aqueous cone (ug/L)
weight of solids (g)
where "X solids" is from section 10.1.3.
13.4 If the EICP area at the quantisation m/z
exceeds the calibration range of the
system, samples are diluted by successive
factors of 10 until the area is within the
calibration range.
13.4.1 F6r aqueous samples, bring 0.50 mL, 0.050
ml, 0.0050 mL etc. to 5 mL volume with
reagent water and analyze per section
10.4.
13.4.2 For samples containing high solids,
substitute 0.50 or 0.050 gram in section
10.5.2 to achieve a factor of 10 or 100
dilution, respectively.
13.4.3 For dilution of high solids samples
greater than a factor of 100, add 5 grams
of sample to 10 mL methanol in a
calibrated 15 - 25 mL centrifuge tube.
Cap and shake vigorously for 15 - 20
seconds to disperse the sample in the
methanol. Centrifuge to settle suspended
particles, if necessary.
13.4.3.1 Remove 0.1 percent of the volume of the
supernate with a 15 - 25 uL syringe. This
volume will be in the range of 10 - 15 uL.
Add this volume to 5 mL reagent water in a
5 mL syringe and analyze per section
10.4.1.
25
-------
13.4.3.2 For further dilutions, remove 1 mL of the
supernate (14.4.3) and dilute to 10 ml,
100 mL, 1000 mL etc. in reagent water.
Remove a volume of this sample/reagent
water mixture equivalent to the volume
determined in step 13.4.3.1, add to 5 mL
reagent water in a 5 mL syringe, and
analyze per section 10.4.1.
13.5 For GC peaks which are to be identified
(per section 12.3), the sample is diluted
by successive factors of 10 when any peak
in the unconnected mass spectrum at the GC
peak maximum is saturated.
outside the range given in table 6. If
the recovery remains outside of the range
for this diluted sample, the aqueous
performance standard shall be analyzed
(section 11) and calibration verified
(section 11.5). If the recovery for the
labeled compound in the aqueous
performance standard is outside the range
given in table 6, the analytical system is
out of control. In this case, the
instrument shall be repaired, the
performance specifications in section 11
shall be met, and the analysis of the
undiluted sample shall be repeated.
13.6 Report results for all pollutants, labeled
compounds, and tentatively identified
compounds found in all standards, blanks,
and samples, in ug/L for samples
containing less than one percent solids
and in ug/kg for samples in which the
undiluted sample contains one percent
solids or greater, to three significant
figures. Results for samples which have
been diluted are reported at the least
dilute level at which the area at the
quantisation m/z is within the calibration
range (section 13.4) or at which no m/z in
the spectrum is saturated (section 13.5).
For compounds having a labeled analog,
results are reported at the least dilute
level at which the area at the
quantitat ion m/z is within the calibration
range (section 13.4) and the labeled
compound recovery is within the normal
range for the method (section 14.2).
14 ANALYSIS OF COMPLEX SAMPLES
14.1 Some samples may contain high levels
(>1000 ug/kg) of the compounds of interest
and of interfering compounds. Some
samples will foam excessively when purged;
others will overload the trap/or GC
column.
If the recovery for the aqueous
performance standard is within the range
given in table 6, the method does not work
on the sample being analyzed and the
result may not be reported for regulatory
compliance purposes.
14.3 Reverse search computer programs can
misinterpret the spectrum of chromato-
graphically unresolved pollutant and
labeled compound pairs with overlapping
spectra when a high level of the pollutant
is present. Examine each chromatogram for
peaks greater than the height of the
internal standard peaks. These peaks can
obscure the compounds of interest.
15 METHOD PERFORMANCE
15.1 The specifications for this method were
taken from the interlaboratory validation
of-EPA Method 624 (reference 10). Method
1624 has been shown to yield slightly
better performance on treated effluents
than method 624. Results of initial tests
of this method at a purge temperature of
80 °C can be found in reference 11 and
results of initial tests of this method on
municipal sludge can be found in reference
12.
14.2 Dilute 0.5 mL of samples containing less
than one percent solids or 0.5 gram of
samples containing one percent solids or
greater with 4.5 mL of reagent water and
analyze this diluted sample when the
recovery of any labeled compound is
15.2 A chromatogram of the 20 ug/L aqueous
performance standards (sections 6.7.2 and
11.1) is shown in figure 9.
26
-------
MASS CHROMATOGRAM DATA: UOAI01945 II
89/81/84 23:95:86 CALI: UOAID1945 #1
SAMPLE: UO.S.OPR-86820,88,U,NA:NA,HAS
CONOS.: 16248,3.811,2111,3845,45-24888,15e248,20H./mNS
RANGE: G 1.1288 LABEL: N 8, 4.8 QUAN: A 8, 1.8 J
SCANS 1 TO 1286
8 BASE: U 28, 3
188. B-i
47
251
222976.
46.514
256.575
406
13:48
eee
28:38
886
27:28
1608
34:18
1208 SCAN
41:66 TIME
FIGURE 9 Chromatogram of Aqueous
Performance Standard
27
-------
REFERENCES
1. "Performance Tests for the Evaluation of
Computerized Gas Chromatography/Mass
Spectrometry Equipment and Laboratories,"
USEPA, EMSL Cincinnati, OH 45268, EPA-
600/4-80-025 (April 1980).
2. Bellar, T. A. and Lichtenberg, J. J.,
"Journal American Water Works Assoc-
iation," 66, 739 (1974).
3. Bellar, T. A. and Lichtenberg, J. J.,
"Semi-automated Headspace Analysis of
Drinking Waters and Industrial Waters for
Purgeable Volatile Organic Compounds," in
Measurement of Organic Pollutants in Water
and Wastewater. C. E. VanHall, ed.,
American Society for Testing Materials,
Philadelphia, PA, Special Technical
Publication 686, (1978).
4. National Standard Reference Data System,
"Mass Spectral Tape Format", US National
Bureau of Standards (1979 and later
attachments).
5. "Working with Carcinogens," DHEW, PHS,
NIOSH, Publication 77-206 (1977).
6. "OSHA Safety and Health Standards, General
Industry," 29 CFR 1910, OSHA 2206, (1976).
7. "Safety in Academic Chemistry Laborato-
ries," American Chemical Society Publica-
tion, Committee on Chemical Safety (1979).
8. "Handbook of Analytical Quality Control in
Water and Wastewater Laboratories," USEPA,
EMSL Cincinnati, OH 45268, EPA-4-79-019
(March 1979).
9. "Methods 330.4 and 330.5 for Total
Residual Chlorine," USEPA, EMSL Cincin-
nati, OH 45268, EPA-4-79-020 (March 1979).
Purge and Trap Volatiles Analysis", S-
CUBED Division of Maxwell Laboratories,
Inc., Prepared for W. A. Telliard,
Industrial Technology Division (WH-552),
USEPA, 401 M St SW, Washington DC 20460
(July 1986).
12. Colby, Bruce N. and Ryan, Philip W.,
"Initial Evaluation of Methods 1634 and
1635 for the Analysis of Municipal
Wastewater Treatment Sludges by Isotope
Dilution GCMS", Pacific Analytical Inc.,
Prepared for W. A. Telliard, Industrial
Technology Division (WH-552), USEPA, 401 M
St SW, Washington DC 20460 (July 1986).
10. "Method 624--Purgeables", 40 CFR Part 136
(49 FR 43234), 26 October 1984.
11. "Narrative for SAS 106: Development of an
Isotope Dilution GC/MS Method for Hot
28
-------
Appendix A: Mass Spectra in the Form of Mass/intensity Lists
532 allyI alcohol
m/z int. m/z int.
42 30 43 39
56 58 57 1000
533 carbon disulfide
m/z int. m/z int.
44 282 46 10
534 2-chloro-1,3-butadiene (chloroprene)
m/z int. m/z int.
48 21 49 91
54 41 61 30
87 12 88 452
535 chloroacetonitrile
m/z int. m/z int.
47 135 48 1000
74 43 75 884
536 3-chloropropene
m/z int. m/z int.
35 39 36 40
49 176 51 64
76 1000 77 74
537 crotonaldehyde
m/z int. m/z
35 26 40
50 40 51
69 511 70
538 1,2-dibromoethane (EDS)
m/z int. m/z int.
79 50 80 13
105 32 106 29
186 13 188 27
539 dibromomethane
m/z int. m/z int.
43 99 44 101
91 142 92 61
172 375 173 14
540 trans-1,4-dichloro-2-butene
m/z int. m/z int.
49 166 50 171
62 286 64 91
90 93 91 129
541 1,3-dichloropropane
m/z int. m/z int.
40 15 42 44
61 18 62 22
77 46 78 310
542 cis-1,3-dichloropropene
m/z int. m/z int.
37 262 38 269
77 328 110 254
int.
28
20
1000
m/z
42
52
71
int.
339
21
43
m/z
43
53
int.
48
31
m/z
44
55
int.
335
55
m/z
49
68
int,
27
24
543 ethyl cyanide
m/z int. m/z
44 115 50
55 193
int.
34
m/z
44
58
m/z
64
ne>
m/z
50
62
89
m/z
49
76
m/z
40
52
78
m/z
42
52
71
m/z
31
107
190
m/z
45
93
174
m/z
51
75
124
m/z
47
63
79
m/z
39
112
m/z
51
int.
232
300
int.
14
int.
223
54
22
int.
88
39
int.
44
31
324
int.
339
21
43
int.
51
1000
13
int.
30
1000
719
int.
289
1000
138
int.
19
131
12
int.
998
161
int.
166
m/z
45
61
m/z
76
m/z
51
63
90
m/z
50
77
m/z
42
61
m/z
43
53
m/z
82
108
m/z
79
94
175
m/z
52
77
126
m/z
48
65
m/z
49
m/z
52
int.
12
15
int.
1000
int.
246
11
137
int.
294
278
int.
206
29
int.
48
31
int.
15
38
int.
184
64
12
int.
85
323
86
int.
20
38
int.
596
int.
190
53
m/z
77
m/z
52
64
m/z
51
m/z
47
73
int.
13
int.
27
int.
241
16
int.
12
int.
40
22
m/z
93
109
m/z
80
95
176
m/z
53
88
128
m/z
49
75
m/z
51
m/z
53
int.
54
922
int.
35
875
342
int.
878
246
12
int.
193
47
int.
189
int.
127
m/z
55
m/z
78
m/z
53
73
m/z
73
m/z
58
75
m/z
95
110
m/z
81
160
m/z
54
89
m/z
51
76
m/z
75
m/z
54
int.
59
int.
82
int.
1000
21
int.
22
int.
35
138
int.
42
19
int.
175
18
int.
273
415
int.
55
1000
int.
1000
int.
1000
-------
544 ethyl methacrylate
M/Z int. M/Z int. M/Z int.
42 127 43 48 45 155
69 1000 70 83 71 25
96 17 99 93 113 11
545 2-hexanone (methyl butyl ketone)
m/z int. M/Z int. M/Z int.
42 61 43 1000 44 24
59 21 71 36 85 37
546 iodomethane
M/Z
44
142
int.
57
1000
M/Z
127
143
int.
328
12
M/Z
128
int.
17
M/Z
139
int.
39
M/Z
140
int.
34
M/Z
141
int
120
int.
21
446
400
M/Z
41
53
67
int.
26
19
1000
547 isobutyl alcohol
M/Z int. M/Z
34 21 35
43 1000 44
59 25 73
548 Methacrylonitrile
m/z int. M/Z
38 24 39
51 214 52
65 55 66
549 methyl methacrylate
m/z int. M/Z
42 127 43
59 124 68
98 20 99
550 4-methyl-2-pentanone (methyl isobutyl ketone; MIBK)
m/z int. m/z int. M/Z int.
42 69 43 1000 44 54
57 205 58 346 59 20
100 94
int.
13
42
12
M/Z
36
45
74
int.
13
21
63
M/Z
37
55
int.
11
40
M/Z
39
56
int.
10
37
M/Z
42
57
int
575
21
int.
52
28
89
M/Z
45
69
100
int.
48
1000
442
m/z
53
70
101
int.
30
51
22
m/z
55
82
int.
100
26
M/Z
56
85
int.
49
45
551 1.1,1,2-tetrachloroethane
m/z int. m/z int.
47 144 49 163
84 31 95 416
121 236 131 1000
552 trichlorofluoromethane
M/Z int. M/Z int.
44 95 47 153
68 53 82 40
105 102 117 16
553 1,2,3-trichloropropane
m/z int. m/z int.
49 285 51 87
76 38 77 302
99 103 110 265
554 vinyl acetate
m/z int. M/Z int.
36 5 42 103
M/Z
60
96
133
M/Z
49
84
119
m/z
61
83
111
M/Z
43
int.
303
152
955
int.
43
28
14
int.
300
23
28
int.
1000
951 M-xylene
M/Z
65
951 o-
M/Z
51
int.
62
+ p-xylene
int.
88
M/Z
77
M/Z
77
int.
124
int.
131
m/z
91
m/z
91
int.
1000
int.
1000
M/Z
105
M/Z
105
int.
245
int.
229
m/z
106
m/z
106
int
580
int
515
M/Z
55
85
114
M/Z
55
100
M/Z
139
M/Z
37
55
m/z
42
62
68
m/z
53
70
101
m/z
53
67
m/z
61
97
135
m/z
51
101
m/z
62
96
112
M/Z
44
m/z
105
M/Z
105
int.
32
14
119
int.
12
56
int.
39
int.
11
40
int.
100
24
51
int.
30
51
22
int.
11
12
int.
330
270
301
int.
21
1000
int.
107
29
164
int.
70
int.
245
int.
229
M/Z
58
86
M/Z
57
int.
39
169
int.
130
m/z
49
63
int.
19
59
m/z
55
69
m/z
62
98
m/z
52
102
m/z
63
97
114
m/z
45
int.
15
10
int.
98
84
int.
14
10
int.
98
166
25
int.
8
M/Z
68
87
M/Z
58
M/Z
50
64
m/z
56
85
m/z
82
117
M/Z
66
103
m/z
75
m/z
86
M/Z
M/z
int.
60
21
int.
382
int.
60
136
int.
1?
9
int.
45
804
int.
162
671
int.
1000
20
int.
57
int.
int.
30
-------
Method 1625, Revision C 15 February 1988 Draft
Semivolatile Organic Compounds by Isotope Dilution GCMS
1 SCOPE AND APPLICATION
1.1 This method is designed to determine the 1.3
semivolatile toxic organic pollutants
associated with the 1976 Consent Decree;
the Resource Conservation and Recovery
Act; the Comprehensive Environmental
Response, Compensation and Liabilities
Act; and other compounds amenable to
extraction and analysis by capillary 1.4
column gas chromatography-mass
spectrometry (GCMS).
1.2 The chemical compounds listed in tables 1
through 4 may be determined in waters,
soils, and municipal sludges by this
method. The method is designed to meet
the survey requirements of the
Environmental Protection Agency (EPA).
The detection limit of this method is
usually dependent on the level of
interferences rather than instrumental
limitations. The limits in tables 5 and 6
typify the minimum quantity that can be
detected with no interferences present.
The GCMS portions of this method are for
use only by analysts experienced with GCMS
or under the close supervision of such
qualified persons. Laboratories unfamil-
iar with analyses of environmental samples
by GCMS should run the performance tests
in reference 1 before beginning.
Table 1
BASE/NEUTRAL EXTRACTABLE COMPOUNDS
DETERMINED BY CALIBRATED GCMS USING ISOTOPE DILUTION AND INTERNAL STANDARD TECHNIQUES
Compound
Pollutant
Storet CAS Registry EPA-EGD
NPDES
Labeled Compound
Analog CAS Registry EPA-EGD
acenaphthene
acenaphthylene
anthracene
benzidine
benzo(a)anthracene
benzo( b ) f I uoranthene
benzo< k ) f I uoranthene
benzo(a)pyrene
benzo(ghi )perytene
biphenyl (Appendix C)
bis(2-chloroethyl) ether
bis(2-chloroethoxy)methane
bis(2-chloroisopropyl) ether
bis(2-ethylhexyl) phthalate
4-bromophenyl phenyl ether
butyl benzyl phthalate
n-C10 (Appendix C)
34205
34200
34220
39120
34526
34230
34242
34247
34521
81513
34273
34278
34283
39100
34636
34292
77427
83-32-9
208-96-8
120-12-7
92-87-5
56-55-3
205-99-2
207-08-9
50-32-8
191-24-2
92-52-4
111-44-4
111-91-1
108-60-1
117-81-7
101-55-3
85-68-7
124-18-5
001 B
077 B
078 B
005 B
072 B
074 B
075 B
073 B
079 B
512 8
018 B
043 B
042 B
066 B
041 B
067 B
517 8
001 B
002 B
003 B
004 B
005 B
007 B
009 B
006 B
008 B
011 B
010 B
012 B
013 B
014 B
015 B
d10
da
d10
d8
d12
d12
d12
d12
d12
d10
d8
d8
d12
d4
d5
d4
d22
15067-20-2
93951-97-4
1719-06-8
92890-63-6
1718-53-2
93951-98-5
93952-01-3
63466-71-7
93951-66-7
1486-01-7
93952-02-4
93966-78-0
93951-67-8
93951-87-2
93951-83-8
93951-88-3
16416-29-8
201 B
277 B
278 B
205 B
272 B
274 B
275 B
273 B
279 B
612 B
218 B
243 B
242 B
266 B
241 8
267 B
617 B
31
-------
Storet
Pollutant
EPA-EGD
NPDES
Labeled Compound
Analog CAS Registry
EPA-EGD
n-C12 (Appendix C)
n-C14 (Appendix C)
n-C16 (Appendix C)
n-C18 (Appendix C)
n-C20 (Appendix C)
n-C22 (Appendix C)
n-C24 (Appendix C)
n-C26 (Appendix C)
n-C28 (Appendix C)
n-C30 (Appendix C)
carbazole (4c)
2-chloronaphthalene
4-chlorophenyl phenyl ether
chrysene
p-cymene (Appendix C)
dibenzo( a, h) anthracene
dibenzofuran (Appendix C & 4c)
dibenzothiophene (Synfuel)
di-n-butyl phthalate
1,2-dichlorobenzene
1 ,3-dichlorobenzene
1,4-dichlorobenzene
3,3' -dichlorobenzidine
diethyl phthalate
2, 4 -dime thy I phenol
dimethyl phthalate
2,4-dinitrotoluene
2,6-dinitrotoluene
di-n-octyl phthalate
diphenylamine (Appendix C)
diphenyl ether (Appendix C)
1 ,2-diphenylhydrazine
f luoranthene
f luorene
hexach 1 orobenzenc
hexach I orobutadi ene
hexach loroethane
hexachlorocyclopentadiene
ideno(1 ,2,3-cd)pyrene
isophorone
naphthalene
beta-naphthylamine (Appendix C)
nitrobenzene
N-nitrosodimethylamine
N-nitrosodi-n-proplyamine
M - n i t rosod i pheny I ami ne
phenanthrene
phenol
alpha-picoline (Synfuel)
pyrene
styrene (Appendix C)
alpha- terpineol (Appendix C)
1,2,3-trichlorobcnzene (4c)
1,2,4-trichl orobenzene
77588
77691
77757
77804
77830
77859
77886
77901
78116
78117
77571
34581
34641
34320
77356
34556
81302
77639
39110
34536
34566
34571
34631
34336
34606
34341
34611
34626
34596
77579
77587
34346
34376
34381
39700
34391
34396
34386
34403
34408
34696
82553
34447
34438
34428
34433
34461
34694
77088
34469
77128
77493
77613
34551
112-40-3
629-59-4
544-76-3
593-45-3
112-95-8
629-97-0
646-31-1
630-01-3
630-02-4
638-68-6
86-74-8
91-58-7
7005-72-3
218-01-9
99-87-6
53-70-3
132-64-9
132-65-0
84-74-2
95-50-1
541-73-1
106-46-7
91-94-1
84-66-2
105-67-9
131-11-3
121-14-2
606-20-2
117-84-0
122-39-4
101-84-8
122-66-7
206-44-0
86-73-7
118-74-1
87-68-3
67-72-1
77-47-4
193-39-5
78-59-1
91-20-3
91-59-8
98-95-3
62-75-9
621-64-7
86-30-6
85-01-8
108-95-2
109-06-8
129-00-0
100-42-5
98-55-5
87-61-6
120-82-1
506 B
518 B
519 B
520 B
521 B
522 B
523 B
524 B
525 B
526 B
528 B
020 B
040 B
076 B
513 B
082 B
505 B
504 B
068 B
025 B
026 B
027 B
028 B
070 B
034 A
071 B
035 B
036 B
069 8
507 B
508 B
037 B
039 B
080 B
009 B
052 B
012 B
053 B
083 B
054 B
055 B
502 B
056 B
061 B
063 B
062 B
081 B
065 A
503 B
084 B
510 B
509 B
529 B
008 B
016 B
017 B
018 B
019 B
026 B
020 B
021 B
022 B
023 B
024 B
003 A
025 B
027 B
028 B
029 B
030 B
031 B
032 B
033 B
034 B
036 B
035 B
037 B
038 B
039 B
040 B
041 B
042 B
043 B
044 B
010 A
045 B
046 B
d26
"34
d42
d50
d62
d8
d5
d12
d14
d14
d8
d8
d4
d4
d4
d4
d6
d4
d4
cL
d4
d10
do
do
d10
13*10
lO'"
13 6
f*
^ C
C4
d8
d7
d5
d6
d14
d6
d10
d5
d7
d10
d.
"3
16416-30-1
15716-08-2
62369-67-9
16416-32-3
93952-07-9
38537-24-5
93951-84-9
93951-85-0
1719-03-5
93952-03-5
13250-98-1
93952-04-6
33262-29-2
93952-11-5
2199-69-1
2199-70-4
3855-82-1
93951-91:8
93952-12-6
93951-75-8
93951-89-4
93951-68-9
93951-90-7
93952-13-7
37055-51-9
93952-05-7
93951-92-9
93951-69-0
81103-79-9
93952-14-8
93951-70-3
93952-15-9
93951-71-4
93952-16-0
1146-65-2
93951-94-1
4165-60-0
17829-05-9
93951-96-3
93951-95-2
1517-22-2
4165-62-2
93951-93-0
1718-52-1
5161-29-5
93952-06-8
3907-98-0
2199-72-6
606 B
618 B
619 B
620 B
621 B
622 B
623 B
624 B
625 B
626 B
628 B
220 B
240 B
276 B
613 B
282 B
605 B
604 B
268 8
225 8
226 B
227 B
228 B
270 B
234 A
271 B
235 B
236 B
269 B
607 B
608 B
237 B
231 B
280 B
209 B
252 B
212 B
253 B
254 B
255 B
602 B
256 B
261 B
263 B
262 B
281 B
265 A
603 B
284 B
610 B
609 B
629 B
208 B
32
-------
Table 2
ACID EXTRACTABLE COMPOUNDS
DETERMINED BY CALIBRATED GCMS USING ISOTOPE DILUTION AND INTERNAL STANDARD TECHNIQUES
Compound
Storet
Pollutant Labeled Compound
CAS Registry EPA-EGO NPDES Analog CAS Registry EPA-EGO
4-chloro-3-methylphenol
2-chlorophenol
2,4-dichlorophenot
2,4-dinitrophenol
2-methyl-4,6-dinitrophenol
2-nitrophenol
4-nitrophenol
pentachlorophenol
2,3,
2.4,
2.4,
6-trichlorophenol (4c)
5-trichlorophenol (4c)
6-trichlorophenol
34452 59-50-7
34586 95-57-8
34601 120-83-2
34616 51-28-5
34657 534-52-1
34591 88-75-5
34646 100-02-7
39032 87-86-5
77688 933-75-5
95-95-4
34621 88-06-2
Table 3
022 A
024 A
031 A
059 A
060 A
057 A
058 A
064 A
530 A
531 A
021 A
BASE/NEUTRAL EXTRACTABLE COMPOUNDS TO BE DETERMINED BY
EGO
No.
ibb
556
557
558
559
560
561
562
563
564
565
566
567
568
569
570
571
572
573
574
575
USING KNOWN RETENTION
Compound
acetophenone
4-aminobiphenyl
aniline
o-anisidine
aramite
benzanthrone
1,3-benzenediol
(resorcinol)
benzenethiol
2,3-benzof luorene
benzyl alcohol
2-bromochlorobenzene
3-bromochlorobenzene
4-chloro-2-nitroaniline
5-chloro-o-toluidine
4-chloroani line
3-chloronitrobenzene
o-cresol
crotoxyphos
2,6-di-tert-butyl-
p-benzoquinone
2,4-diaminotoluene
1,2-dibromo-3-
chloropropane
TIMES, RESPONSE FACTORS
CAS
Registry
98-86-2
92-67-1
62-53-3
90-04-0
140-57-8
82-05-3
108-46-3
108-98-5
243-17-4
100-51-6
694-80-4
108-37-2
89-63-4
95-79-4
106-47-8
121-73-3
95-48-7
7700-17-6
719-22-2
95-80-7
96-12-8
008 A d2 93951-72-5 222 A
001 A d4 93951-73-6 224 A
002 A dj 93951-74-7 231 A
005 A dj 93951-77-0 259 A
004 A d2 93951-76-9 260 A
006 A d^ 93951-75-1 257 A
007 A d4 93951-79-2 258 A
009 A 13C, 85380-74-1 264 A
O
d2 93951-81-6 630 A
d2 93951-82-
7 631 A
011 A d2 93951-80-5 221 A
REVERSE SEARCH AND QUANT I TAT I ON
, REFERENCE COMPOUND, AND MASS SPECTRA
EGD
No.
576
577
578
579
580
581
582
583
584
585
586
587
588
589
590
591
592
593
Compound
2,6-dichloro-4-
nitroaniline
1,3-dichloro-2-propanol
2,3-dichloroani line
2,3-dichlcronitro-
benzene3209-22-1
1,2:3,4-diepoxybutane
3,3'-dimethoxybenzidine
dimethyl sulfone
p-dimethylamino-
azobenzene
7, 12-dimethylbenz-
(a)anthracene
N,N-dimethylformamide
3,6-dimethylphenanthrene
1,4-dinitrobenzene
diphenyldisulfide
ethyl methanesulfonate
ethyl eneth i ourea
ethynylestradiol
3-methyl ether
hexach I oropropene
2- isopropylnaphthalene
CAS
Registry
99-30-9
96-23-1
608-27-5
1464-53-5
119-90-4
67-71-0
60-11-7
57-97-6
68-12-2
1576-67-6
100-25-4
882-33-7
62-50-0
96-45-7
72-33-3
1888-71-7
2027-17-0
33
-------
EGD
Mo.
Compound
CAS
Registry
594 isosafrole 120-58-1
595 longifolene 475-20-7
596 malachite green 569-64-2
597 methapyrilene 91-80-5
598 methyl methanesulfonate 66-27-3
599 2-methylbenzothioazole 120-75-2
900 3-methylcholanthrene 56-49-5
901 4,4'-methylene-
bis(2-chloroaniline) 101-14-4
902 4,5-methylene-
phenanthrene 203*64-5
903 ,1-methylfluorene 1730-37-6
904 2-methylnaphthalene 91-57-6
905 1-methylphenanthrene 832-69-9
906 2-(methylthio)-
benzothiazole 615-22-5
907 1,5-naphthalenediamine 2243-62-1
908 1,4-naphthoquinone 130-15-4
909 alpha-naphthylamine 134-32-7
910 5-nitro-o-toluidine 99-55-8
911 2-nitroaniline 88-74-4
912 3-nitroaniline 99-09-2
913 4-nitroaniline 100-01-6
914 4-nitrobiphenyl 92-93-3
915 N-nitrosodi-n-butylamine 924-16-3
916 N-nitrosodiethylamine 55-18-5
917 N-m'trosomethyl-
ethylamine 10595-95-6
918 N-nitrosomethyl-
phenylamine 614-00-6
919 N-nitrosomorpholine 59-89-2
920 N-nitrosopiperidine 100-75-4
921 pentachlorobenzene 608-93-5
922 pentachloroethane 76-01-7
923 pentamethylbenzene 700-12-9
924 perylene 198-55-0
925 phenacetin 62-44-2
926 phenothiazine 92-84-2
927 1-phenylnaphthalene 605-02-7
928 2-phenylnaphthalene 612-94-2
929 pronamide 23950-58-5
930 pyridine 110-86-1
931 safrole 94-59-7
932 squalene 7683-64-9
933 1,2.4,5-tetra-
chlorobenzene 95-94-3
934 thianaphthene
(2,3-benzothiophene) 95-15-8
935 thioacetamide 62-55-5
936
937
938
939
940
941
942
thioxanthone
o-toluidine
1 ,2,3-trimethoxybenzene
2,4,5-trimethylamline
triphenylene
t r i propy t eneg I yco I
methyl ether
1.3,5-trithiane
492-22-8
95-53-4
634-36-6
137-17-7
217-59-4
20324-33-8
291-21-4
Table 4
ACID EXTRACTABLE COMPOUNDS
TO BE DETERMINED BY REVERSE SEARCH
AND QUANTI TAT ION USING
KNOWN RETENTION TIMES,
RESPONSE FACTORS,
REFERENCE COMPOUND,
AND MASS SPECTRA
EGD
No.
943
944
945
946
947
948
Compound
benzoic acid
p-cresol
3,5-dibromo-
4-hydroxybenzonitrile
2,6-dichloroph'enol
hexanoic acid
2,3,4,6-tetrachlorophenot
CAS
Registry
65-85-0
106-44-5
1689-84-5
87-65-0
142-62-1
58-90-2
2 SUMMARY OF METHOD
2.1 The percent solids content of a sample is
determined. Stable isotopically labeled
analogs of the compounds of interest are
added to the sample. If the solids
content is less than one percent, a one
liter sample is extracted at pH 12 - 13,
then at pH <2 with methylene chloride
using continuous extraction techniques.
If the solids content is 30 percent
percent or less, the sample is diluted to
one percent solids with reagent water,
homogenized ultrasonical ly, and extracted
at pH 12-13, then at pH <2 with methylene
chloride using continuous extraction
techniques. If the solids content is
greater than 30 percent, the sample is
extracted using ultrasonic techniques.
-------
Table 5
GAS CHROMATOGRAPHY OF BASE/NEUTRAL EXTRACTABLE COMPOUNDS
EGO
No.
(1)
164
930
261
361
585
580
603
703
917
598
610
710
916
577
589
582
562
922
557
613
713
265
365
218
318
617
717
226
326
227
327
225
325
935
564
242
342
571
263
363
555
212
312
937
919
575
256
Compound
2.2'-difluorobiphenyl (int std)
pyridine
N-nitrosodimethylamine-d, (5)
N-nitrosodimethylamine (5)
N.N-dimethylformaimde
1 ,2:3,4-diepoxybutane
alpha picotine-d.
alpha picoline
N-nitrosomethylethylamine
methyl methanesulfonate
styrene-d_
styrene
N-nitrosodiethylamine
1,3-dichloro-2-propanot
ethyl methanesulfonate
dimethyl sulfone
benzenethiol
pentachloroethane
aniline
p-cymene-d..
p-cymene
phenol-d-
phenol
bis(2-chloroethyl) ether-dg
bis(2-chloroethyl) ether
n-decane-d-.
n-decane
1 ,3-dichlorobenzene-d^
1 , 3-di ch I orobenzene
1 ,4-dichlorobenzene-d.
1,4-dichl orobenzene
1,2-dichlorobenzene-d^
1 , 2 - d i ch I or obenz ene
thioacetamide
benzyl alcohol
bis(2-chloroisopropyl) ether-d^
bis(2-chloroisopropyt ) ether
o-cresol
N-nitrosodi-n-propylamine-d., (5)
N-nitrosodi-n-propylam'ine (5)
acetophenone
hexachloroethane- C
hexachloroethane
o-toluidine
N-nitrosomorpholine
1 ,2-dibromo-3-chloropropane
nitrobenzene-d-
Retention
Mean
(sec)
1163
378
378
385
407
409
417
426
451
511
546
549
570
589
637
649
667
680
694
742
755
696
700
696
704
698
720
722
724
737
740
758
760
768
785
788
799
814
817
830
818
819
823
830
834
839
845
time
EGO
Ref
164
164
164
261
164
164
164
603
164
164
164
610
164
164
164
164
164
164
164
164
613
164
265
164
218
164
617
164
226
164
227
164
225
164
164
164
242
164
164
263
164
164
212
164
164
164
164
Relative (2)
1.000 - 1.000
0.325
0.286 - 0.364
1.006 - 1.028
0.350
0.352
0.326 - 0.393
1.006 - 1.028
0.338
0.439
0.450 - 0.488
1.002 - 1.009
0.490
0.506
0.548
0.558
0.574
0.585
0.597
0.624 - 0.652
1.008 - 1.023
0.584 - 0.613
0.995 - 1.010
0.584 - 0.607
1.007 • 1.016
0.585 - 0.615
1.022 - 1.038
0.605 - 0.636
0.998 - 1.008
0.601 - 0.666
0.997 - 1.009
0.632 - 0.667
0.995 - 1.008
0.660
0.675
0.664 - 0.691
1.010 - 1.016
0.700
0.689 - 0.716
1.008 - 1.023
0.703
0.690 - 0.717
0.999 - 1.001
0.714
0.717
0.721
0.706 - 0.727
Mini
mum
Lev
el (3)
(uq/mL)
10
50
50
50
50
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
20
20
10
10
10
Method Detection
Limit (4)
low high
solids solids
(ug/kg) (ug/kg)
16 27
25 87
149* 17
426* 912"
2501* 757<
32 22
299* 1188-
46 26
35 20
63 16
24 39
46 47
58 55
35
-------
EGO
No.
(1)
356
566
565
941
254
354
942
920
234
334
243
343
208
308
558
255
355
934
609
709
606
706
629
729
252
352
918
592
569
570
915
923
561
931
939
904
599
568
938
933
253
353
594
594
578
574
220
320
518
612
712
608
708
Compound
nitrobenzene
3 - bromoch I orobenzene
2-bromoch lorobenzene
tripropylene glycol methyl ether
isophorone-dg
i sophorone
1,3,5-trithiane
N-nitrosopiperidine
2,4-dimethylphenol-d_
2,4-dimethylphenol
bis(2-chloroethoxy) methane-d. (5)
bis(2-chloroethoxy) methane (5)
1,2,4-trichlorobenzene-d,
1 ,2, 4- t rich lorobenzene
o-anisidine
naphthalene-d_
naphthalene
thianapthene
a I pha- terpi neol -d.
alpha-terpineol
n-dodecane-dj^
n-dodecane
1,2,3-trichlorobenzene-cL (5)
1,2,3-trichlorobenzene (5)
hexach 1 orobutadi ene- C^
hexach I orobutadi ene
N-nitrosomethylphenylamine
hexach loropropene
4-chloroaniline
3-chloronitrobenzene
N-ni trosodi -n-butylamine
pentame thy I benzene
1 ,3-benzenediol
safrole
2,4,5-trimethylani I ine
2 -methyl naphtha I ene
2-methylbenzothiazole
5-chloro-o-toluidine
1 ,2,3-trimethoxybenzene
1 , 2 , 4 , 5 - tet rach lorobenzene
13
hexachlorocyclopentadiene- C^
hexach I orocyc I opentadi ene
isosafrole (cis or trans)
isosafrole (cis or trans)
2,3-dichloroaniline
2,4-diaminotoluene
2-chloronaphthalene-d_
2-chloronaphthalene
n-tetradecane
biphenyl-d1Q
biphenyt
diphenyl ether-d1Q
diphenyl ether
Retention
Mean
(sec)
849
854
880
881
881
889
889
895
921
924
933
939
955
958
962
963
967
971
973
975
953
981
1000
1003
1005
1006
1006
1013
1016
1018
1063
1083
1088
1090
1091
1098
1099
1101
1128
1141
1147
1142
1147
1190
11oO
1187
1185
1200
1203
1195
1205
1211
1216
time
EGO
Ref
256
164
164
164
164
254
164
164
164
234
164
243
164
208
164
164
255
164
164
609
164
606
164
629
164
252
164
164
164
164
164
164
164
164
164
164
164
164
164
164
164
253
164
164
164
164
164
220
164
164
612
164
608
Relative (2)
1.002 - 1.007
0.734
0.757
0.758
0.747 - 0.767
0.999 - 1.017
0.764
0.770
0.781 - 0.803
0.999 - 1.003
0.792 - 0.807
1.000 - 1.013
0.813 - 0.830
1.000 - 1.005
0.827
0.819 - 0.836
1.001 - 1.006
0.835
0.829 - 0.844
0.998 - 1.008
0.730 - 0.908
0.986 - 1.051
0.852 - 0.868
1.000 - 1.005
0.856 - 0.871
0.999 - 1.002
0.865
0.871
0.874
0.875
0.914
0.931
0.936
0.937
0.938
0.944
0.945
0.947
0.970
0.981
0.976 - 0.986
0.999 - 1.001
0.986
1.023
0.997
1.021
1.014 - 1.024
0.997 - 1.007
1.034
1.016 - 1.027
1.001 - 1.006
1.036 - 1.047
0.997 - 1.009
Mini
mum
Lev
el (3)
(ug/mL)
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
10
Method Detection
Limit (4)
low high
solids solids
(uq/kq) (uq/kq)
39 28
8 5
26 13
26 23
49 24
62 42
nd nd
860* 3885*
260* 164*
46 22
nd nd
80 59
256 3533
67 55
44 12
36
-------
Mini Method Detection
mum Limit (4)
EGD
No.
(1)
579
911
908
595
277
377
593
587
576
271
371
573
236
336
912
201
301
605
705
921
909
235
335
602
702
590
280
380
240
340
270
370
906
567
910
913
619
719
237
337
607
707
262
362
241
341
925
903
209
309
556
929
281
Compound
2,3-dichloronitrobenzene
2-nitroanitine
1 ,4-naphthoquinone
longifolene
acenaphthylene-dg
acenaphthylene
2- isopropylnaphthalene
1 ,4-dinitrobenzene
2,6-dichloro-4-nitroaniline
dimethyl phthalate-d^
dimethyl phthalate
2,6-di-t-butyl-p-benzoquinone
2,6-dinitrotoluene-d,
2,6-dinitrotoluene
3-nitroaniline
acenaphthene-d1Q
acenaphthene
dibenzofuran-dg
dibenzofuran
pentach I orobenzene
alpha-naphthy'lamine
2,4-dinitrotoluene-d,
2,4-dinitrotoluene
beta-naphthylamine-d^
beta-naphthylamine
ethylenethiourea
f luorene-d.-
f luorene
4-chlorophenyl phenyl ether-d.
4-chlorophenyl phenyl ether
diethyl phthalate-d4
di ethyl phthalate
2-(methylthio)benzothiazole
4-chloro-2-nitroani line
5-nitro-o-toluidine
4-nitroaniline
n-hexadecane-dj^
n-hexadecane
1 ,2-diphenylhydrazine- ,8
1,2-diphenylhydrazine (6)
diphenylamine-d.jg
d i phenyl ami ne
N-nitrosodiphenylamine-d,
N-nitrosodiphenylamine (7)
4-bromophenyl phenyl ether-d- (5)
4-bromophenyl phenyl ether (5)
phenacetin
1-methy If luorene
hexachlorobenzene- C,
o
hexachlorobenzene
4 - ami nobi phenyl
pronamide
phenanthrene-d.,.
Retention
Mean
(sec)
1214
1218
1224
1225
1265
1247
1254
1255
1259
1269
1273
1273
1283
1300
1297
1298
1304
1331
1335
1340
1358
1359
1364
1368
1371
1381
1395
1401
1406
1409
1409
1414
1415
1421
1422
1430
1447
1469
1433
1439
1437
1439
1447
1464
1495
1498
1512
1514
1521
1522
1551
1578
1578
time
EGO
Ref
164
164
164
164
164
277
164
164
164
164
271
164
164
236
164
164
201
164
605
164
164
164
235
164
602
164
164
281
164
240
164
270
164
164
164
164
164
619
164
237
164
607
164
262
164
241
164
164
164
209
164
164
164
Relative (2)
1.044
1.047
1.052
1.053
1.080 - 1.095
1.000 - 1.004
1.078
1.079
1.083
1.083 - 1.102
0.998 - 1.005
1.095
1.090 - 1.112
1.001 - 1.005
1.115
1.107 - 1.125
0.999 - 1.009
1.134 - 1.155
0.998 - 1.007
1.152
1.168
1.152 - 1.181
1.000 - 1.002
1.163 - 1.189
0.996 - 1.007
1.187
1.185 - 1.214
0.999 - 1.008
1.194 - 1.223
0.990 • 1.015
1.197 - 1.229
0.996 - 1.006
1.217
1.222
1.223
1.230
1.010 - 1.478
1.013 - 1.020
1.216 - 1.248
0.999 - 1.009
1.213 - 1.249
1.000 - 1.007
1.225 - 1.252
1.000 - 1.002
1.271 - 1.307
0.990 - 1.015
1.300
1.302
1.288 - 1.327
0.999 - 1.001
1.334
1.357
1.334 - 1.380
Lev
el (3)
(ug/mL)
10
10
10
10
10
10
10
10
10
10
10
10
50
50
10
10
10
10
10
10
10
10
20
20
20
20
20
20
10
10
10
10
10
low high
solids solids
(ug/kg) (ug/kg)
57 1
62 2
55 4
64 5
77 21
65 20
49 3
69 6
73 5
52 1
116* 64
48 2
58 5
55 2
55 1
51 i
17
48
37
-------
EGO
No.
520
381
278
378
604
704
588
914
927
628
728
621
721
907
902
905
268
368
928
586
597
926
239
339
572
936
284
384
205
305
522
559
559
583
563
623
723
932
267
367
276
376
901
272
372
581
228
328
940
560
266
366
524
Compound
n-octadecane
phenanthrene
anthracene-d.g
anthracene
dibenzothiophene-d-
dibenzothiophene
diphenyldisulfide
4-nitrobiphenyl
1 - pheny I naph thai ene
carbazole-d- (5)
carbazole (5)
n-eicosane-d,p
n-eicosane
1,5-naphthalenediamine
4, 5-methylenephenanthrene
1 -methylphenanthrene
di-n-butyl phthalate-d.
di-n-butyl phthalate
2-phenylnaphthalene
3,6-dimethylphenanthrene
methapyrilene
phenothiazine
f luoranthene-d.Q
f tuoranthene
crotoxyphos
thioxanthone
pyrene-d1Q
pyrene
benzidine-d-
benzidine
n-docosane
aramite
aramite
p-di methyl ami noazobenzene
2,3-benzof luorene
n-tetracosane-d^0
n-tetracosane
squalene
butylbenzyl phthalate-d, (5)
butylbenzyl phthalate (5)
chrysene-d.p
chrysene
4,4'methylenebis(2-chloroaniline)
benzo(a)anthracene-d.2
benzo(a)anthracene
3,3'-dimethoxybenzidine
3,3'-dichlorobenzidine-d,
3,3'-dichlorobenzidine
triphenylene
benzanthrone
bis(2-ethylhexyl) phthalate-d^
bis(2-ethylhexyl) phthalate
n-hexacosane
Retention
Mean
(sec)
1580
1583
1588
1592
1559
1564
1623
1639
1643
1645
1650
1655
1677
1676
1690
1697
1719
1723
1733
1763
1781
1796
1813
1817
1822
1836
1844
1852
1854
1853
1889
1901
1916
1922
1932
1997
2025
2039
2058
2060
2081
2083
2083
2082
2090
2090
2088
2086
2088
2106
2123
2124
2147
time
EGO
Ref
164
281
164
278
164
604
164
164
164
164
628
164
621
164
164
164
164
268
164
164
164
164
164
239
164
164
164
284
164
205
164
164
164
164
164
164
612
164
164
267
164
276
164
164
272
164
164
228
164
164
164
266
164
Relative (2)
1.359
1.000 - 1.005
1.342 - 1.388
0.998 - 1.006
1.314 - 1.361
1.000 - 1.006
1.396
1.409
1.413
1.388 - 1.439
1.000 - 1.006
1.184 - 1.662
1.010 - 1.021
1.441
1.453
1.459
1.446 - 1.510
1.000 - 1.003
1.490
1.516
1.531
1.544
1.522 - 1.596
1.000 - 1.004
1.567
1.579
1.523 - 1.644
1.001 - 1.003
1.549 - 1.632
1.000 - 1.002
1.624
1.635
1.647
1.653
1.661
1.671 - 1.764
1.012 - 1.015
1.753
1.715 - 1.824
1.000 - 1.002
1.743 - 1.837
1.000 - 1.004
1.791
1.735 - 1.846
0.999 - 1.007
1.797
1.744 - 1.848
1.000 - 1.001
1.795
1.811
1.771 - 1.880
1.000 - 1.002
1.846
Mini
mum
Lev
el (3)
(ug/mL)
10
10
10
10
10
10
20
20
10
10
10
10
10
10
10
10
50
50
10
10
10
10
10
10
10
10
10
50
50
10
10
10
Method Detection
Limit (4)
low high
solids solids
(ug/kg) (ug/kg)
134*
42
52
72
47
83
64
54
40
nd
432*
--
60
51
61
62
553*
609*
844*
22
21
71
24
229*
80
22
48
nd
447*
--
65
48
47
111
1310*
886*
38
-------
EGO
No.
(1) Concound
Retention time
Mean EGO
(sec) Ref Relative (2)
Mini
mum
Lev
el (3)
(uq/mL)
Method Detection
Limit (4)
low high
solids solids
(ua/kg) (ug/kg)
J12
312
591 ethynylestradiol 3-methyl ether
269 di-n-octyl phthalate-d4
369 di-n-octyl phthalate
525 n-octacosane
584 7,12-dimethylbenz(a)anthracene
274 benzo(b)fluoranthene-d,
374 benzo(b)fluoranthene
275 benzo(k)fluoranthene-d,
375 benzo(k)fluoranthene
924 perylene
273 benzo(a)pyrene-d.|2
373 benzo(a)pyrene
626 n-triacontane-d,,
. ot
726 n-tnacontane
596 malachite green
900 3-methylcholanthrene
083 indeno(1,2,3-cd)pyrene
282 dibenzo(a,h)anthracene-d.4 (5)
382 dibenzo(a,h)anthracene (5)
279 benzo(ghi)perylene-d12
379 benzo(ghi)perylene
2209
2239
2240
2272
2284
2281
2293
2287
2293
2349
2351
2350
2384
2429
2382
2439
2650
2649
2660
2741
2750
164
164
269
164
164
164
274
164
275
164
164
273
164
626
164
164
164
164
282
164
279
1.899
1.867 - 1.982
1.000 - 1.002
1.954
1.964
1.902 - 2.025
1.000 - 1.005
1.906 - 2.033
1.000 - 1.005
2.020
1.954 - 2.088
1.000 - 1.004
1.972 - 2.127
1.011 - 1.028
2.048
2.097
2.279
2.107 - 2.445
1.000 - 1.007
2.187 - 2.524
1.001 - 1.006
10
10
10
10
10
10
10
10
10
10
10
20
20
20
20
20
72
492*
54
95
52
252*
67
49
44
62
1810*
30
20
15
658*
263*
125
nd
(1) Reference numbers beginning with 0, 1, 5, or 9 indicate a pollutant quantified by the internal standard
method; reference numbers beginning with 2 or 6 indicate a labeled compound quantified by the internal standard
method; reference numbers beginning with 3 or 7 indicate a pollutant quantified by isotope dilution.
(2) Single values in this column are based on single laboratory data.
(3) This is a minimum level at which the analytical system shall give recognizable mass spectra (background
corrected) and acceptable calibration points. The concentration in the aqueous or solid phase is determined
using the equations in section 14.
(4) Method detection limits determined in digested sludge (low solids) and in filter cake or compost (high
solids).
(5) Specification derived from related compound.
(6) Detected as azobenzene
(7) Detected as diphenylamine
nd = not detected when spiked into the sludge tested
*Background levels of these compounds were present in the sludge tested, resulting in higher than expected
MDL's. The MOL for these compounds is expected to be approximately 50 ug/kg with no interferences present.
Column: 30 +/- 2 m x 0.25 +/- 0.02 mm i.d. 94% methyl, 4X phenyl, 1X vinyl bonded phase fused silica capillary
Temperature program: 5 min at 30°C; 30 - 280°C at 8°C per min; isothermal at 280°C until benzo(ghi)perylene
elutes
Gas velocity: 30 +/- 5 cm/sec at 30"C
39
-------
Table 6
GAS CHROMATOGRAPHY OF ACID EXTRACTABLE COMPOUNDS
EGO
No.
(1)
164
224
324
947
944
257
357
231
331
943
946
222
322
221
321
631
731
530
259
359
258
358
948
260
360
945
264
364
Compound
2,2'-difluorobiphenyl (int std)
2-chlorophenol-d^
2-chlorophenol
hexanoic acid
p-cresol
2-nitrophenot-d^
2-nitrophenol
2,4-dichlorophenol-cU
2,4-dichlorophenol
benzoic acid
2,6-dichlorophenol
4-chloro-3-methylphenol-d-
4-chloro-3-methylphenol
2,4,6- trichlorophenol-d.
2,4,6-trichlorophenol,
2,4,5-trichlorophenol-d2 (5)
2,4,5- trichlorophenol
2,3,6-trichlorophenol
2,4-dinitrophenol-d,
2,4-dinitrophenol
4-ni trophenol -d.
4-nitrophenol
2 , 3 , 4 , 6- tet rach I oropheno t
2-methyl-4,6-dinitrophenol-d_
2-methy I -4, 6-dini trophenot
3,5-dibromo-4-hydroxybenzonitrile
pentachlorophenol- C,
pen tach I oropheno I
Retention time
Mean EGO
(sec) Ref
1163
701
705
746
834
898
900
944
947
971
981
1086
1091
1162
1165
1167
1170
1195
1323
1325
1349
1354
1371
1433
1435
1481
1559
1561
164
164
224
164
164
164
257
164
231
164
164
164
222
164
221
164
631
164
164
259
164
258
164
164
260
164
164
264
Relative (2)
1.000 - 1.000
0.587 - 0.618
0.997 - 1.010
0.641
0.717
0.761 - 0.783
0.994 - 1.009
0.802 - 0.822
0.997 - 1.006
0.835
0.844
0.930 - 0.943
0.998 - 1.003
0.994 - 1.005
0.998 - 1.004
0.998 - 1.009
0.998 - 1.004
1.028
1.127 - 1.149
1.000 - 1.005
1.147 - 1.175
0.997 - 1.006
1.179
1.216 - 1.249
1.000 - 1.002
1.273
1.320 - 1.363
0.998 - 1.002
Mini-
mum
Level
(3)
(ug/mL)
10
10
10
20
20
10
10
10
10
10
10
10
10
10
50
50
50
50
20
20
50
50
Method Detection
Limit (4)
low high
solids solids
(ug/kg) (ug/kg)
18 10
39 44
24 116
41 62
46 111
32 55
58 37
565 642
287 11
385 83
51 207
(1) Reference numbers beginning with 0, 1, 5, or 9 indicate a pollutant quantified by the internal standard
method; reference numbers beginning with 2 or 6 indicate a labeled compound quantified by the internal standard
method; reference numbers beginning with 3 or 7 indicate a pollutant quantified by isotope dilution.
(2) Single values in this column are based on single laboratory data.
(3) This is a minimum level at which the analytical system shall give recognizable mass spectra (background
corrected) and acceptable calibration points. The concentration in the aqueous or solid phase is determined
using the equations in section 14.
(4) Method detection limits determined in digested sludge (low solids) and in filter cake or compost (high
sol ids).
•Background levels of these compounds were present in the sludge resulting in higher than expected MDL's. The
MDL for these compounds is expected to be approximately 50 ug/kg with no interferences present.
(5) Specification derived from related compound.
Column: 30 +/- 2 m x 0.25 +/• 0.02 mm i.d. 94X methyl, 4X phenyl, 1X vinyl bonded phase fused silica capillary
Temperature program: 5 min at 30°C; 30 - 250°C or until pentachlorophenol elutes
Gas velocity: 30 +/- 5 cm/sec at 30°C
-------
Each extract is dried over sodium sulfate,
concentrated to a volume of five mt,
cleaned up using gel permeation
chromatography (GPC), if necessary, and
concentrated to one mL. An internal
standard is added to the extract, and a
one uL aliquot of the extract is injected
into the gas chromatograph (GC). The
compounds are separated by GC and detected
by a mass spectrometer (MS). The labeled
compounds serve to correct the variability
of the analytical technique.
2.2 Identification of a pollutant (qualitative
analysis) is performed in one of three
ways: (1) for compounds listed in tables
1 and 2, and for other compounds for which
authentic standards are available, the
GCMS system is calibrated and the mass
spectrum and retention time for each
standard are stored in a user created
library. A compound is identified when
its retention time and mass spectrum agree
with the library retention time and
spectrum. (2) For compounds listed in
tables 3 and 4, and for other compounds
for which standards are not available, a
compound is identified when the retention
time and mass spectrum agree with those
specified in this method. (3) For
chromatographic peaks which are not
identified by (1) and (2) above, the
background corrected spectrum at the peak
maximum is compared with spectra in the
EPA/NIH Mass Spectral File (reference 2).
Tentative identification is established
when the spectrum agrees.
2.3 Quantitative analysis is performed in one
of four ways by GCMS using extracted ion
current profile (EICP) areas: (1) For
compounds listed in tables 1 and 2, and
for other compounds for which standards
and labeled analogs are available, the
GCMS system is calibrated and the compound
concentration is determined using an
isotope dilution technique. (2) For
compounds listed in tables 1 and 2, and
for other compounds for which authentic
standards but no labeled compounds are
available, the GCMS system is calibrated
and the compound concentration is
determined using an internal standard
technique. (3) For compounds listed in
tables 3 and 4, and for other compounds
for which standards are not available,
compound concentrations are determined
using known response factors. (4) For
compounds for which neither standards nor
known response factors are available,
compound concentration is determined using
the sum of the EICP areas relative to the
sum of the EICP areas of the internal
standard.
2.4 Quality is assured through reproducible
calibration and testing of the extraction
and GCMS systems.
3 CONTAMINATION AND INTERFERENCES
3.1 Solvents, reagents, glassware, and other
sample processing hardware may yield
artifacts and/or elevated baselines
causing misinterpretation of chromatograms
and spectra. All materials used in the
analysis shall be demonstrated to be free
from interferences under the conditions of
analysis by running method blanks
initially and with each sample tot
(samples started through the extraction
process on a given 8 hr shift, to a
maximum of 20). Specific selection of
reagents and purification of solvents by
distillation in all-glass systems may be
required. Glassware and, where possible,
reagents are cleaned by solvent rinse and
baking at 450°C for one hour minimum.
3.2 Interferences coextracted from samples
will vary considerably from source to
source, depending on the diversity of the
site being sampled.
4 SAFETY
4.1 The toxicity or careinogenicity of each
compound or reagent used in this method
has not been precisely determined;
however, each chemical compound should be
treated as a potential health hazard.
Exposure to these compounds should be
reduced to the lowest possible level. 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 data handling sheets
should also be made available to all
personnel involved in these analyses.
Additional information on laboratory
safety can be found in references 3-5.
4.2 The following compounds covered by this
method have been tentatively classified as
known or suspected human or mammalian
carcinogens: benzo(a)anthracene, 3,3'-
dichlorobenzidine, benzo(a)pyrene.
dibenzo(a,h)anthracene, N-nitrosodimethy-
lamine, and beta-naphthylamine. Primary
standards of these compounds shall be
prepared in a hood, and a N10SH/MESA
approved toxic gas respirator should be
worn when high concentrations are handled.
5 APPARATUS AND MATERIALS
5.1 Sampling equipment for discrete or
composite sampling.
5.1.1 Sample Bottles and Caps
5.1.1.1 Liquid Samples (waters, sludges and
similar materials that contain less than
five percent solids)--Sample bottle, amber
glass, 1.1 liters minimum, with screw cap.
5.1.1.2 Solid samples (soils, sediments, sludges,
filter cake, compost, and similar
materials that contain more than five
percent solids)--Sample bottle, wide
mouth, amber glass, 500 mL minimum.
5.1.1.3 If amber bottles are not available,
samples shall be protected from light.
5.1.1.A Bottle caps--threaded to fit sample
bottles. Caps shall be lined with Teflon.
5.1.1.5 Cleaning
5.1.1.5.1 Bottles are detergent water washed, then
solvent rinsed or baked at 450 "C for one
hour minimum before use.
5.1.1.5.2 Liners are detergent water washed, then
reagent water (section 6.5.1) and solvent
rinsed, and baked -at approx 200 °C for one
hour minimum prior to use.
5.1.2 Compositing equipment—automatic or manual
compositing system incorporating glass
containers cleaned per bottle cleaning
procedure above. Sample containers are
kept at 0 - 4 "C during sampling. Glass
or Teflon tubing only shall be used. If
the sampler uses a peristaltic pump, a
minimum length of compressible silicone
rubber tubing may be used in the pump
only. Before use, the tubing shall be
thoroughly rinsed with methanol, followed
by repeated rinsings with reagent water
(section 6.5.1) to minimize sample
contamination. An integrating flow meter
is used to collect proportional composite
samples.
5.2 Equipment for determining percent moisture
5.2.1 Oven, capable of being temperature
controlled at 110 +/- 5 °C.
5.2.2 Dessicator
5.3 Sonic disruptor--375 watt with pulsing
capability and 3/4 in. disrupter horn
(Ultrasonics, Inc, Model 375C, or
equivalent).
5.4 Extraction apparatus
5.4.1 Continuous liquid-liquid extractor--Tefton
or glass connecting joints and stopcocks
without lubrication, 1.5 - 2 liter
capacity (Hershberg-Wolf Extractor, Ace
Glass 6841-10, or equivalent).
5.4.2 Beakers
5.4.2.1 1.5 - 2 liter, calibrated to one liter
5.4.2.2 400 - 500 mL
5.4.2.3 Spatulas--stainless steel
5.4.3 Filtration apparatus
-------
5.4.3.1 Glass funnel--125 - 250 mL
5.4.3.2 Filter paper for above (Whatman 41, or
equivalent)
5.5 Drying column—15 to 20 mm i.d. Pyrex
chromatographic col urn equipped with
coarse glass frit or glass wool plug.
5.6 Kuderna-Oanish (K-D) apparatus
5.6.1 Concentrator tube--10mL, graduated (Kontes
K-570050-1025, or equivalent) with
calibration verified. Ground glass
stopper (size 19/22 joint) is used to
prevent evaporation of extracts.
5.6.2 Evaporation flask--500 ml (Kontes K-
570001-0500, or equivalent), attached to
concentrator tube with springs (Kontes K-
662750-0012).
5.6.3 Snyder column--three ball macro (Kontes K-
503000-0232, or equivalent).
5.6.4 Snyder column--two ball micro (Kontes K-
469002-0219, or equivalent).
5.6.5 Boiling chips--approx 10/40 mesh,
extracted with methylene chloride and
baked at 450 °C for one hr minimum.
5.7 Water bath—heated, with concentric ring
cover, capable of temperature control (+/-
2 °C), installed in a fume hood.
5.8 Sample vials — amber glass, 2 - 5 mL with
Teflon-lined screw cap.
5.9 Balances
5.9.1 Analytical —capable of weighing 0.1 mg.
5.9.2 Top loading—capable of weighing 10 mg.
5.10 Automated gel permeation chromatograph
(Analytical Biochemical Labs, Inc.,
Columbia, MO, Model GPC Autoprep 1002, or
equivalent)
5.10.1 Column—600 - 700 mm x 25 mm i.d., packed
with 70 g of SX-3 Bio-beads (Bio-Rad
Laboratories, Richmond, CA)
5.10.2 UV detectors -- 254-mu, preparative or
semi-prep flow cell:
5.10.2.1 Schmadzu, 5 mm path length
5.10.2.2 Beckman-Altex 152W, 8 uL micro-prep flow
cell, 2 mm path
5.10.2.3 Pharmacia UV-1, 3 mm flow cell
5.10.2.4 LDC Milton-Roy UV-3, monitor #1203
5.11 Gas chromatograph—shall have split less or
on-column injection port for capillary
column, temperature program with 30 °C
hold, and shall meet all of the
performance specifications in section 12.
5.11.1 Column--30 ±5 m x 0.25 ± 0.02 mm i.d. 5%
phenyl, 94X methyl, 1X vinyl silicone
bonded phase fused silica capillary column
(J & W DB-5, or equivalent).
5.12 Mass spectrometer--70 eV electron impact
ionization, shall repetitively scan from
35 to 450 amu in 0.95 - 1.00 second, and
shall produce a unit resolution (valleys
between m/z 441-442 less than 10 percent
of the height of the 441 peak), background
corrected mass spectrum from 50 ng
decafluorotriphenylphosphine (DFTPP) in-
troduced through the GC inlet. The
spectrum shall meet the mass-intensity
criteria in table 7 (reference 6). The
mass spectrometer shall be interfaced to
the GC such that the end of the capillary
column terminates within one centimeter of
the ion source but does not intercept the
electron or ion beams. All portions of
the column which connect the GC to the ion
source shall remain at or above the column
temperature during analysis to preclude
condensation of less volatile compounds.
-------
Table 7
DFTPP MASS-INTENSITY SPECIFICATIONS*
Mass Intensity required
51 8-82 percent of m/z 198
68 less than 2 percent of m/z 69
69 11-91 percent of m/z 198
70 less than 2 percent of m/z 69
127 32 - 59 percent of m/z 198
197 less than 1 percent of m/z 198
198 base peak, 100 percent abundance
199 4-9 percent of m/z 198
275 11-30 percent of m/z 198
441 44-110 percent of m/z 443
442 30-86 percent of m/z 198
443 14 - 24 percent of m/z 442
•Reference 6
5.13 Data system--shall collect and record MS
data, store mass- intensity data in
spectral libraries, process GCMS data,
generate reports, and shall compute and
record response factors.
5.13.1 Data acquisition--mass spectra shall be
collected continuously throughout the
analysis and stored on a mass storage
device.
5.13.2 Mass spectral libraries—user created
libraries containing mass spectra obtained
from analysis of authentic standards shall
be employed to reverse search GCMS runs
for the compounds of interest (section
7.2).
5.13.3 Data processing—the data system shall be
used to search, locate, identify, and
quantify the compounds of interest in each
GCMS analysis. Software routines shall be
employed to compute retention times and
peak areas. Displays of spectra, mass
chromatograms, and library comparisons are
required to verify results.
5.13.4 Response factors and multipoint
calibrations—the data system shall be
used to record and maintain lists of
response factors (response ratios for
isotope dilution) and multi-point
calibration curves (section 7).
Computations of relative standard
deviation (coefficient of variation) are
used for testing calibration linearity.
Statistics on initial (section 8.2) and
on-going (section 12.7) performance shall
be computed and maintained.
6 REAGENTS AND STANDARDS
6.1 Reagents for adjusting sample pH
6.1.1 Sodium hydroxide--reagent grade, 6N in
reagent water.
6.1.2 Sulfuric acid—reagent
reagent water.
grade, 6N in
6.2 Sodium sulfate—reagent grade, granular
anhydrous, rinsed with methylene chloride
(20 mL/g), baked at 450 °C for one hour
minimum, cooled in a dessicator, and
stored in a pre-cleaned glass bottle with
screw cap which prevents moisture fr.om
entering.
6.3 Methylene chloride—distilled in glass
(Burdick and Jackson, or equivalent).
6.4 GPC calibration solution — containing 300
mg/mL corn oil, 15 mg/mL bis(2-ethylhexyl)
phthalate, 1.4 mg/mL pentachlorophenol,
0.1 mg/mL perylene, and 0.5 mg/mL sulfur
6.5 Reference matrices
6.5.1 Reagent water—water in which the
compounds of interest and interfering
compounds are not detected by this method.
6.5.2 High solids reference matrix—playground
sand or similar material in which the
compounds of interest and interfering
compounds are not detected by this method.
6.6 Standard solutions—purchased as solutions
or mixtures with certification to their
purity, concentration, and authenticity,
or prepared from materials of known purity
and composition. If compound purity is 96
percent or greater, the weight may be used
44
-------
without correction to compute the
concentration of the standard. When not
being used, standards are stored in the
dark at -20 to -10 °C in screw-capped
vials with Teflon-lined lids. A mark is
placed on the vial at the level of the
solution so that solvent evaporation loss
can be detected. The vials are brought to
room temperature prior to use. Any
precipitate is redissotved and solvent is
added if solvent loss has occurred.
6.7 Preparation of stock solutions--prepare in
methylene chloride, benzene, p-dioxane, or
a mixture of these solvents per the steps
below. Observe the safety precautions in
section 4. The large number of labeled
and unlabeled acid and base/neutral
compounds used for combined calibration
(section 7) and calibration verification
(12.5) require high concentrations (approx
40 mg/mL) when individual stock solutions
are prepared, so that dilutions' of
mixtures will permit calibration with.all
compounds in a single set of solutions.
•The working range for most compounds is
10-200 ug/mL. Compounds with a reduced MS
response may be prepared at higher
concentrations.
6.7.1 Dissolve an appropriate amount of assayed
reference material in a suitable solvent.
For example, weigh 400 mg naphthalene in a
10 ml ground glass stoppered volumetric
flask and fill to the mark with benzene.
After the naphthalene is completely
dissolved, transfer the solution to a 15
ml vial with Teflon-lined cap.
6.7.2 Stock standard solutions should be checked
for signs of degradation prior to the
preparation of calibration or performance
test standards. Quality control check
samples that can be used to determine the
accuracy of calibration standards are
available from the US Environmental
Protection Agency, Environmental Monitor-
ing and Support 'Laboratory, Cincinnati,
Ohio 45268.
6.7.3 Stock standard solutions shall be replaced
after six months, or sooner if comparison
with quality control check standards
indicates a change in concentration.
6.8 Labeled compound spiking sotut ion--from
stock standard solutions prepared as
above, or from mixtures, prepare the
spiking solution at a concentration of 200
ug/mL, or at a concentration appropriate
to the MS response of each compound.
6.9 Secondary standard--using stock solutions
(section 6.7), prepare a secondary
standard containing all of the compounds
in tables 1 and 2 at a concentration of
400 ug/mL, or higher concentration
appropriate to the MS response of the
compound.
6.10 Internal standard solution—prepare 2,2'-
difluorobiphenyl (DFB) at a concentration
of 10 mg/mL in benzene.
6.11 OFTPP solution--prepare at 50 ug/mL in
acetone.
6.12 Solutions for obtaining authentic mass
spectra (section 7.2)--prepare mixtures of
compounds at concentrations which will
assure authentic spectra are obtained for
storage in libraries.
6.13 Calibration solutions—combine 0.5 mL of
the solution in section 6.8 with 25, 50,
125, 250, and 500 uL of the solution in
section 6.9 and bring to 1.00 mL total
volume each. This will produce
calibration solutions of nominal 10, 20,
50, 100 and 200 ug/mL of the pollutants
and a constant nominal 100 ug/mL of the
labeled compounds. Spike each solution
with 10 uL of the internal standard
solution (section 6.10). These solutions
permit the relative response (labeled to
unlabeled) to be measured as a function of
concentration (section 7.4).
6.14 Precision and recovery standard--used for
determination of initial (section 8.2) and
on-going (section 12.7) precision and
recovery. This solution shall contain the
pollutants and labeled compounds at a
nominal concentration of 100 ug/mL.
-------
6.15 Stability of solutions—all standard
solutions (sections 6.8 - 6.14) shall be
analyzed within 48 hours of preparation
and on a monthly basis thereafter for
signs of degradation. Standards will
remain acceptable if the peak area at the
quantisation mass relative to the DFB
internal standard remains within t 15
percent of the area obtained in the
initial analysis of the standard.
7 CALIBRATION
7.1 Assemble the GCMS and establish the
operating conditions in table 5. Analyze
standards per the procedure in section 11
to demonstrate that the analytical system
meets the minimum levels in tables 5 and
6, and the mass-intensity criteria in
table 7 for 50 ng DFTPP.
7.2 Mass spectral libraries—detection and
identification of compounds of interest
are dependent upon spectra stored in user
created libraries.
7.2.1 Obtain a mass spectrum of each pollutant,
labeled compound, and the internal
standard by analyzing an authentic
standard either singly or as part of a
mixture in which there is no interference
between closely eluted components. That
only a single compound is present is
determined by examination of the spectrum.
Fragments not attributable to the compound
under study indicate the presence of an
interfering compound.
7.2.2 Adjust the analytical conditions and scan
rate (for this test only) to produce an
undistorted spectrum at the GC peak
maximum. An undistorted spectrum will
usually be obtained if five complete
spectra are collected across the upper
half of the GC peak. Software algorithms
designed to "enhance" the spectrum may
eliminate distortion, but may also
eliminate authentic masses or introduce
other distortion.
7.2.3 The authentic reference spectrum is
obtained under DFTPP tuning conditions
(section 7.1 and table 7) to normalize it
to spectra from other instruments.
7.2.4 The spectrum is edited by saving the 5
most intense mass spectral peaks and all
other mass spectral peaks greater than 10
percent of the base peak. The spectrum
may be further edited to remove common
Table 8
BASE/NEUTRAL EXTRACT ABLE COMPOUND CHARACTERISTIC M/Z'S
AND RESPONSE FACTORS
Compound
acenaphthene
acenaphthylene
acetophenone
4-aminobiphenyl
ani I ine
o-anisidine
anthracene
aramite
benzanthrone
1,3-benzenediol
benzenethiol
benzidine
benzo(a)anthracene
benzo( b) f I uoranthene
benzo( k ) f luoranthene
Labeled
analog
d10
d8
d10
d8
d12
d12
«*1,
Primary
m/z
154/164
152/160
105
169
93
108
178/188
185
230
110
110
184/192
228/240
252/264
252/264
Response
Factor (1)
0.79
0.81
1.04
0.43
0.19
0.15
0.78
0.18
-------
Comoound
benzo(a)pyrene
benzo(ghi )perytene
2,3-benzof luorene
benzoic acid
benzyl alcohol
biphenyl
bis(2-chloroethyl) ether
bis(2-chloroethoxy)methane
bis(2-chloroisopropyl) ether
bis(2-ethylhexyl> phthalate
2 - bromoch I orobenzene
3 - bromoch I orobenzene
4-bromophenyl phenyl ether
butyl benzyl phthalate
n-C10
n-C12
n-C14
n-C16
n-C18
n-C20
n-C22
n-C24
n-C26
n-C28
n-C30
carbazole
4-chloro-2-nitroaniline
5-chloro-o-toluidine
4-chloroani line
2-chlorsnaphthalene
3 - ch 1 oron i t robenzene
4-chlorophenyl phenyl ether
3-chloropropionitri le
chrysene
o-cresol
crotoxyphos
p-cymene
2,6-di-tert-butyl-p-benzoquinone
di-n-butyl phthalate
2 , 4 - d i ami noto I uene
dibenzo(a,h)anthracene
dibenzofuran
d i benzot h i ophene
1,2-dibromo-3-chloropropane
2,6-dichloro-4-nitroani I ine
1 ,3-dichloro-2-propanol
2,3-dichloroani 1 ine
1,2-dichlorobenzene
1,3-dichlorobenzene
1 , 4 -d i ch I orobenzene
3,3'-dichlorobenzidine
2,2'-difluorobiphenyl (int std)
2,3-dichloronitrobenzene
1 , 2 : 3 , 4 - di epoxybutane
Labeled
analog
d12
d12
d10
4
d6
4
d4
*5
<
d22
d26
"34
d42
"50
d62
d8
d7
d5
d12
d14
d4
d14
d8
d8
d4
d4
d4
d6
Primary
m/z
252/264
276/288
216
105
79
154/164
93/101
93/99
121/131
149/153
111
192
248/253
149/153
55/66
55/66
55
55/66
55
55/66
55
55/66
55
55
55/66
167/175
172
106
127
162/169
157
204/209
54
228/240
108
127
119/130
220
149/153
122
278/292
168/176
184/192
157
124
79
161
146/152
146/152
146/152
252/258
190
191
55
Response
Factor (1)
0.35
0.16
0.47
0.33
0.40
0.20
0.50
0.73
0.18
0.42
0.59
0.017
0.078
0.059
0.22
0.019
0.68
0.47
0.11
0.27
-------
Labeled
Compound analog
diethyl phthalate d^
3,3'-dimethoxybenzidine
dimethyl phthalate d^
dimethyl sul forte
p-di methyl ami noazobenzene
7, 12-dimethylbenz(a)anthracene
N.N-dimethylformamide
3,6-dimethylphenanthrene
2,4-dimethylphenol d-
1,4-dinitrobenzene
2,4-dinitrotoluene d-
2,6-dinitrotoluene d.
di-n-octyl phthalate d.
diphenylamine d^Q
diphenyl ether d^
diphenyldi sul fide
1,2-diphenylhydrazine (2) d1Q
ethyl methanesulfonate
ethylenethiourea
ethynylestradiol 3-methyl ether
fluoranthene d.-
f luorene d.Q
hexach I orobenzene C^
hexach I orobutadi ene C^
hexach I oroe thane C
hexachlorocyclopentadiene C^
hexach I oropropene
indeno(1,2,3-cd)pyrene
isophorone d-
2-isopropylnaphthalene
isosafrole
longifolene
malachite green
methapyrilene
methyl methanesulfonate
2-methylbenzothiazole
3-methylcholanthrene
4,4'-methylenebis(2-chloroaniline)
4,5-methylenephenanthrene
1 -methyl f luorene
2-methylnaphthalene
1-methylphenanthrene
2-(methylthio)benzothiazole
naphthalene dg
1 ,5-naphthalenediamine
1,4-naphthoquinone
alpha-naphthylamine
heta-naphthylamine d_
5-nitro-o-toluidine
2-nitroani line
3-nitroani line
4-nitroaniline
nitrobenzene d_
4-nitrobiphenyl
H-nitrosodi-n-butylamine
N-nitrosodi-n-propylamine d..
Primary
m/z
149/153
244
163/167.
79
120
256
73
206
122/125
168
165/168
165/167
149/153
169/179
170/180
218
77/82
109
102
227
202/212
166/176
284/292
225/231
201/204
237/241
213
276
82/88
170
162
161
330
97
80
149
268
231
190
180
142
192
181
128/136
158
158
143
143/150
152
138
138
138
128/128
199
84
70/84
Response
Factor (1)
0.19
0.40
0.23
0.58
0.51
0.72
0.24
0.25
0.28
0.22
0.28
0.23
0.32
0.33
0.14
0.43
0.20
0.59
0.59
0.21
0.44
0.37
0.99
0.65
0.42
0.085
0.021
0.89
0.31
0.39
0.27
0.11
0.35
0.47
48
-------
Labeled
Compound analog
N-nitrosodiethylamine
N-nitrosodimethylanrine d.
N-nitrosodiphenylamine (3) d.
M-nitrosomethylethylamine
N-ni trosomethylphenylaraine
N-nt trosomorphol ine
N-nitrosopiperidine
pentach lorobenzene
pentach I oroethane
pentamethylbenzene
perylene
phenacetin
phenanthrene d...
phenol dj
phenothiazine
1 -phenylnaphthalene
2-phenylnaphthalene
alpha-picoline d-,
pronamide
pyrene d1Q
pyridine
safrole
squa I ene
styrene d.
alpha-terpineol d.
1,2,4,5-tetrachlorobenzene
thianaphthene
thioacetamide
thioxanthone
o-toluidine
1,2,3-trichlorobenzene d.
1,2,4-trichlorobenzene d.
1 ,2,3-trimethoxybenzene
2,4,5-trimethylani line
triphenylene
tripropylene glycol methyl ether
1,3,5-trithiane
Primary
m/z
102
74/80
169/175
88
106
56
114
248
117
148
252
108
178/188
94/71
199
204
204
93/100
173
202/212
79
162
69
104/109
59/62
216
134
75
212
106
180/183
180/183
168
120
228
59
138
Response
Factor (1)
0.45
0.33
0.024
0.49
0.41
0.25
0.20
0.42
0.30
0.38
0.15
0.48
0.73
0.31
0.68
0.45
0.042
0.43
1.52
0.28
0.23
1.04
0.48
0.28
1.32
0.092
0.15
(1) referenced to 2,2'-difluorobiphenyl
(2) detected as azobenzene
(3) detected as diphenylamine
-------
Interfering masses. If 5 mass spectral
peaks cannot be obtained under the scan
conditions given in section 5.12, the mass
spectrometer may be scanned to an m/z
lower than 35 to gain additional spectral
information. The spectrum obtained is
stored for reverse search and for compound
confirmation.
7.2.5 For the compounds in tables 3 and 4 and
for other compounds for which the mass
spectra, quantisation m/z's, and retention
times are known but the instrument is not
to be calibrated, add the retention time
and reference compound (tables 5 and 6);
the response factor and the quantitation
m/z (tables 8 and 9); and spectrum
(Appendix A) to the reverse search
library. Edit the spectrum per section
7.2.4, if necessary.
Table 9
ACID EXTRACTABLE COMPOUND CHARACTERISTIC M/Z'S
Compound
Labeled Primary Response
analog m/z Factor (1)
benzoic acid
4-chloro-3-methylphenol
2-chlorophenol
p-cresol
3,5-dibromo-
4-hydroxybenzonitrile
2,4-dichlorophenol
2,6-dichlorophenol
2,4-dini trophenol
hexanoic acid
2-methy I -4 , 6-di ni trophenol
2-ni trophenol
4-ni trophenol
pentachlorophenol
2,3,4,6-tetrachlorophenol
2,3,6-trichlorophenol
2,4,5-trichlorophenol
2,4,6-trichl orophenol
d2
d4
"3
dj
d2
d4
13d*
C6
d2
d2
d2
105
107/109
128/132
108
277
162/167
162
184/187
60
198/200
139/143
139/143
266/272
232
196/200
196/200
196/200
0.16
0.61
0.12
0.42
0.62
0.17
(1) referenced to 2,2'-difluorobiphenyl
7.3 Analytical range--demonstrate that 20 ng
anthracene or phenanthrene produces an
area at m/z 178 approx one-tenth that
required to exceed the linear range of the
system. The exact value must be
determined by experience for each
instrument. It is used to match the
calibration range of the instrument to the
analytical range and detection limits
required, and to diagnose instrument
sensitivity problems (section 15.3). The
20 ug/mL calibration standard (section
6.13) can be used to demonstrate this
performance.
7.3.1 Polar compound detection—demonstrate that
unlabeled pentachlorophenol and benzidine
are detectable at the 50 ug/mL level (per
all criteria in section 13). The 50 ug/mL
calibration standard (section 6.13) can be
used to demonstrate this performance.
7.4 Calibration with isotope dilution--isotope
dilution is used when 1) labeled compounds
are available, 2) interferences do not
preclude its use, and 3) the quantitation
m/z (tables 8 and 9) extracted ion current
profile (EICP) area for the compound is in
the calibration range. Alternate labeled
compounds and quantitation m/z's may be
used based on availability. If any of the
above conditions preclude isotope
dilution, the internal standard method
(section 7.5) is used.
7.4.1 A calibration curve encompassing the
concentration range is prepared for each
compound to be determined. The relative
response (pollutant to labeled) vs
concentration in standard solutions is
plotted or computed using a linear
regression. The example in Figure 1 shows
a calibration curve for phenol using
phenol-d, as the isotopic diluent. Also
shown are the t 10 percent error limits
(dotted lines). Relative Response (RR) is
determined according to the procedure::
described below. A minimum of five data
points are employed for calibration.
7.4.2 The relative response of a pollutant to
its labeled analog is determined from
isotope ratio values computed from
50
-------
10-
cfl
O
a.
in
> 10-
0 1-
2 10 20 50 100 200
CONCENTRATION (ug/mL)
FIGURE 1 Relative Response Calibration Curve
for Phenol. The Dotted Lines Enclose a ± TO Per-
cont Error Window.
RTOl
Cares nyz (at RT,)]
as measured in the mixture of the
pollutant and labeled compounds (figure
2), and RR = R.
FIGURE 2 Extracted Ion Current Profiles for
Chromatographically Resolved Labeled (m2/z)
and Unlabeled (m,/z) Pairs.
7.4.3
acquired data. Three isotope ratios are
used in this process:
R = the isotope ratio measured for the
pure pollutant.
R = the isotope ratio measured for the
labeled compound.
R = the isotope ratio of an analytical
m
mixture of pollutant and labeled
compounds.
The m/z's are selected such that R > R .
If Rm is not between 2R and 0.5RX, the
method does not apply and the sample is
analyzed by the internal standard method.
Capillary columns usually separate the
pollutant- labeled pair, with the labeled
compound eluted first (figure 2). For
this case,
R = [area m./z (at RTQ]
1
V
7.4.4 Special precautions are taken when the
pollutant-labeled pair is not separated,
or when another labeled compound with
interfering spectral masses overlaps the
•pollutant (a case which can occur with
isomeric compounds). In this case, it is
necessary to determine the respective
contributions of the pollutant and labeled
compounds to the respective EICP areas.
If the peaks are separated well enough to
permit the data system or operator to
remove the contributions of the compounds
to each other, the equations in section
7.4.3 apply. This usually occurs when the
height of the valley between the two GC
peaks at the same m/z is less than 10
percent of the height of the shorter of
the two peaks. If significant GC and
spectral overlap occur, RR is computed
using the following equation:
RR = (R - R KR
[area m_/z (at
(R
where R is measured as shown in figure
3A, R is measured as shown in figure 38,
and R is measured as shown in figure 3C.
m
For the example.
R = 46100 = 9.644
4780
51
-------
R = 2650 = 0.0608
V 43600
f?m = 49200 = 1.019
48300
RR = 1.114.
(3A)
AREA = 46100
AREA = 4780
(section 7.4) cannot be met. The internal
standard to be used for both acid and
base/neutral analyses is 2,2'-difluorobi-
phenyl. The internal standard method is
also applied to determination of compounds
having no labeled analog, and to
measurement of labeled compounds for
intra-laboratory statistics (sections 8.4
and 12.7.4).
7.5.1 Response factors--calibration requires the
determination of response factors (RF)
which are defined by the following
equation:
AREA = 43600
AREA =48300
<3C)
AREA = 49200
FIGURE 3 Extracted Ion Current Profiles for (3A)
Unlabeled Compound, (3B) Labeled Com-
pound, and (3C) Equal Mixture of Unlabeled
and Labeled Compounds.
7.4.5 To calibrate the analytical system by
isotope dilution, analyze a 1.0 uL aliquot
of each of the calibration standards
(section 6.13) using the procedure in
section 11. Compute the RR at each
concentration.
7.4.6 Linearity--if the ratio of relative
response to concentration for any compound
is constant (less than 20 percent
coefficient of variation) over the 5 point
calibration range, an averaged relative
response/concentration ratio may be used
for that compound; otherwise, the complete
calibration curve for that compound shall
be used over the 5 point calibration
range.
7.5 Calibration by internal standard--used
when criteria for isotope dilution
7.5.1.1
7.5.1.2
7.6
-------
pollutants, labeled compounds, and the
internal standard, a single set of
analyses can be used to produce
calibration curves for the isotope
dilution and internal standard methods.
These curves are verified each shift
(section 12.5) by analyzing the 100 ug/mL
calibration standard (section 6.13).
Recall bration is required only if
calibration verification (section 12.5)
criteria cannot be met.
8 QUALITY ASSURANCE/QUALITY CONTROL
8.1.3 Analyses of blanks are required to
demonstrate freedom from contamination.
The procedures and criteria for analysis
of a blank are described in section 8.5.
8.1.4 The laboratory 'shall spike all samples
with labeled compounds to monitor method
performance. This test is described in
section 8.3. When results of these spikes
indicate atypical method performance for
samples, the samples are diluted to bring
method performance within acceptable
limits (section 15).
8.1 Each laboratory that uses this method is
required to operate a formal quality
assurance program (reference 7). The
minimum requirements of this program
consist of an initial demonstration of
laboratory capability, analysis of samples
spiked with labeled compounds to evaluate
and document data quality, and analysis of
standards and blanks as tests of continued
performance. Laboratory performance is
compared to established performance
criteria to determine if the results of
analyses meet the performance
characteristics of the method. If the
method is to be applied routinely to
samples containing high solids with very
little moisture (e.g., soils, filter cake,
compost), the high solids reference matrix
(section 6.5.2) is substituted for the
reagent water (6.5.1) in all performance
tests, and the high solids method (section
10) is used for these tests.
8.1.1 The analyst shall make an initial
demonstration of the ability to generate
acceptable accuracy and precision with
this method. This ability is established
as described in section 8.2.
8.1.2 The analyst is permitted to modify this
method to improve separations or lower the
costs of measurements, provided all
performance specifications are met. Each
time a modification is made to the method,
the analyst is required to repeat the
procedure in section 8.2 to demonstrate
method performance.
8.1.5 The laboratory shall, on an on-going
basis, demonstrate through calibration
verification and the analysis of the
precision and recovery standard (section
6.14) that the analysis system is in
control. These procedures are described
in sections 12.1, 12.5, and 12.7.
8.1.6 The laboratory shall maintain records to
define the quality of data that is
generated. Development of accuracy
statements is described in section 8.4.
8.2 Initial precision and accuracy--to
establish the ability to generate
acceptable precision and accuracy, the
analyst shall perform the following
operations:
8.2.1 For low solids (aqueous samples), extract,
concentrate, and analyze two sets of four
one-liter aliquots (8 aliquots total) of
the precision and recovery standard
(section 6.14) according to the procedure
in section 10. For high solids samples,
two sets of four 30 gram aliquots of the
high solids reference matrix are used.
8.2.2 Using results of the first set of four
analyses, compute the average recovery (X)
in ug/mL and the standard deviation of the
recovery (s) in ug/mL for each compound,
by isotope dilution for pollutants with a
labeled analog, and by internal standard
for labeled compounds and pollutants with
no labeled analog.
53
-------
8.2.3 For each compound, compare s and X with
the corresponding limits for initial
precision and accuracy in table 10. If s
and X for all compounds meet the
acceptance criteria, system performance is
acceptable and analysis of blanks and
samples may begin. If, however, any
individual s exceeds the precision limit
or any individual X falls outside the
range for accuracy, system performance is
unacceptable for that compound. NOTE: The
large number of compounds in table 10
present a substantial probability that one
or more will fail the acceptance criteria
when all compounds are analyzed. To
determine if the analytical system is out
of control, or if the failure can be
attributed to probability, proceed as
follows:
8.2.4 Using the results of the second set of
four analyses, compute s and X for only
those compounds which failed the test of
the first set of four analyses (section
8.2.3). If these compounds now pass,
system performance is acceptable for all
compounds and analysis of blanks and
Table 10
samples may begin. If, however, any of
the same compounds fail again, the
analysis system is not performing properly
for these compounds. In this event,
correct the problem and repeat the entire
test (section 8.2.1).
8.3 The laboratory shall spike all samples
with labeled compounds to assess method
performance on the sample matrix.
8.3.1 Analyze each sample according to the
method beginning in section 10.
8.3.2 Compute the percent recovery (P) of the
labeled compounds using the internal
standard method (section 7.5).
8.3.3 Compare the labeled compound recovery for
each compound with the corresponding
limits in table 10. If the recovery of
any compound falls outside its warning
limit, method performance is unacceptable
for that compound in that sample.
Therefore, the sample is complex. Water
samples are diluted, and smaller amounts
of soils, sludges, and sediments are
reanalyzed per section 15.
ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS
EGD
No.
(1)
301
201
377
277
378
278
305
205
372
272
374
274
375
275
373
Acceptance criteria
Initial
precision
and accuracy
Section 8.2.3
(ug/L)
Compound
acenaphthene
acenaphthene-d.Q
acenaphthylene
acenaphthylene-d.
anthracene
anthracene-d1Q
benzidine
benzidine-dp
benzo( a ) anth racene
benzo(a)anthracene-d.2
benzo(b)f luoranthene
benzo(b) f luoranthene-d. -
benzo( k ) f I uorant hene
benzo( k ) f I uoranthene-d. 2
benzo(a)pyrene
s
21
38
38
31
41
49
119
269
20
41
183
168
26
114
26
X
79 -
38 -
69 -
39 -
58 -
31 -
16 -
ns(2)
65 -
25 -
32 -
11 -
59 -
15 -
62 -
134
147
186
146
174
194
518
ns
168
298
545
577
143
514
195
Labeled
compound
recovery
Sec 8.3
and 14.2
P
20
23
14
ns
12
ns
ns
(X)
- 270
- 239
- 419
- ns
- 605
ns
ns
Cal ibra-
tion
ver i f i -
cation
Sec 12.5
(ug/mL)
80 -
71 -
60 -
66 -
60 -
58 -
34 -
ns •
70 -
28 -
61 -
14 -
13 -
13 -
78 -
125
141
166
152
168
171
296
ns
142
357
164
ns
ns
ns
129
On -go ing
accuracy
Sec 12.7
R (ug/L)
72 -
30 -
61 -
33 -
50 -
23 -
11 -
ns -
62 -
22 -
20 -
ns -
53 -
ns -
59 -
144
180
207
168
199
242
672
ns
176
329
ns
ns
155
685
206
-------
EGO
NO.
(1)
273
379
279
712
612
318
218
343
243
342
242
366
266
341
241
367
267
717
617
706
606
518
719
619
520
721
621
522
723
623
524
525
726
626
728
628
320
220
322
222
324
224
340
240
376
276
713
613
382
282
Comoound
benzo(a)pyrene-d12
benzo(ghi )perylene
benzo( gh i )pery I ene-d.. _
biphenyl (Appendix C)
biphenyl-d1Q
bis(2-chloroethyl) ether
bis(2-chloroethyl) ether-dg
bis(2-chloroethoxy)methane
bis(2-chloroethoxy)methane (3)
bis(2-chloroisopropyl) ether
bis(2-chloroisopropyl)ether-d.-
bis(2-ethylhexyl) phthalate
bis(2-ethylhexyl) phthalate-d.
4-bromophenyl phenyl ether
4-bromophenylphenyl ether-d5(3)
butyl benzyl phthalate
butyl benzyl phthalate-d, (3)
n-C10 (Appendix C)
n-ClO-d22
n-C12 (Appendix C)
n-C12-d26
n-C14 (Appendix C) (3)
n-d6 (Appendix C)
n-C16-dj4
n-C18 (Appendix C) (3)
n-C20 (Appendix C)
n-C20-d42
n-C22 (Appendix C) (3)
n-C24 (Appendix C)
n-C24-d5Q
n-C26 (Appendix C) (3)
n-C28 (Appendix C) (3)
n-C30 (Appendix C)
n-C30-d.2
carbazote (4c)
carbazole-d- (3)
2-chloronaphthalene
2-chloronaphthalene-d^
4-chloro-3-methylphenol
4-chloro-3-methy I phenol -d-
2-chlorophenol
2-chtorophenol-d^
4-chlorophenyl phenyl ether
4-chlorophenyl phenyl ether-d-
chrysene
chrysene-d.j2
p-cymene (Appendix C)
p-cymene-d..
dibenzo( a, h) anthracene
dibenzo(a,h)anthracene-d. , (3)
Acceptance criteria
Initial
precision
and accuracy
Section 8.2.3
(ug/L)
s X
24
21
45
41
43
34
33
27
33
17
27
31
29
44
52
31
29
51
70
74
53
109
33
46
39
59
34
31
11
28
35
35
32
41
38
31
100
41
37
111
13
24
42
52
51
69
18
67
55
45
35
72
29
75
28
55
29
43
29
81
35
69
32
44
40
19
32
24
ns
35
ns
ns
80
37
42
53
34
45
80
27
35
35
61
27
36
48
46
30
76
30
79
36
75
40
59
33
76
ns
23
29
- 181
- 160
- 268
- 148
- 165
- 196
- 196
- 153
- 196
- 138
- 149
- 220
- 205
- 140
- 161
- 233
- 205
- 195
- 298
- 369
- 331
- ns
- 162
- 162
- 131
- 263
- 172
- 152
- 139
- 211
- 193
- 193
- 200
- 242
- 165
- 130
- 357
- 168
- 131
- 174
- 135
- 162
- 166
- 161
- 186
- 219
- 140
- 359
- 299
- 268
Labeled.
compound
recovery
Sec 8.3
and 14.2
P (X)
21 -
14 -
ns -
15 -
15 -
20 -
18 -
19 -
18 -
ns -
ns -
18 -
19 -
15 -
13 -
29 -
15 -
ns -
23 -
19 -
13 -
ns -
14 -
290
529
ns
372
372
260
364
325
364
ns
ns
308
306
376
479
215
324
613
255
325
512
ns
529
Calibra-
tion
verifi-
cation
Sec 12.5
(uq/mD
12 -
69 -
13 -
58 -
52 -
61 -
52 -
44 -
52 -
67 -
44 -
76 -
43 -
52 -
57 -
22 -
43 -
42 -
44 -
60 -
41 -
37 -
72 -
54 -
40 -
54 -
62 -
40 -
65 -
50 -
26 -
26 -
66 -
24 -
44 -
69 -
58 -
72 -
85 -
68 -
78 -
55 -
71 •
57 -
70 -
24 -
79 -
66 -
13 -
13 -
ns
145
ns
171
192
164
194
228
194
148
229
131
232
193
175
450
232
235
227
166
242
268
138
186
249
184
162
249
154
199
392
392
152
423
227
145
171
139
115
147
129
180
142
175
142
411
127
152
761
ns
On-going
accuracy
Sec 12.7
R (ug/U)
32 -
58 -
25 -
62 -
17 •
50 -
25 -
39 -
25 -
77 -
30 -
64 -
28 -
35 -
29 -
35 -
28 -
19 -
ns -
29 -
ns -
ns -
71 -
28 -
35 -
46 -
29 -
39 -
78 -
25 -
31 -
31 -
56 -
23 -
31 -
40 -
35 -
24 -
62 -
14 -
76 -
33 -
63 -
29 -
48 -
23 -
72 -
ns -
19 -
25 -
194
168
303
176
267
213
222
166
222
145
169
232
224
172
212
170
224
237
504
424
408
ns
181
202
167
301
198
195
142
229
212
212
215
274
188
156
442
204
159
314
138
176
194
212
221
290
147
468
340
303
55
-------
EGD
No.
(1)
705
605
704
604
368
268
325
225
326
226
327
227
328
228
331
231
370
270
334
234
371
271
359
259
335
235
336
236
369
269
707
607
708
608
337
237
339
239
380
280
309
209
352
252
312
212
353
253
083
354
254
Compound
dibenzofuran (Appendix C)
dibenzofuran-dg
dibenzothiophene (Synfuel)
dibenzothiophene-dg
di-n-butyl phthalate
di-n-butyl phthalate-d^
1,2-dichlorobenzene
1,2-dichlorobenzene-d,
1 ,3-dichlorobenzene
1 ,3-dichlorobenzene-d^
1 , 4 - d i ch I orobenzene
1,4-dichlorobenzene-d^
3,3'-dichlorobenzidine
3,3'-dichlorobenzidine-d^
2,4-dichlorophenol
2,4-dichlorophenol-oL
di ethyl phthalate
diethyl phthalate-d^
2, 4 -dimethyl phenol
2,4-dimethylphenol-d,
dimethyl phthalate
dimethyl phthalate-d.
2,4-dinitrophenol
2,4-dinitrophenol-dj
2,4-dinitrotoluene
2,4-dinitrotoluene-dj
2,6-dinitrotoluene
2,6-dinitrotoluene-d,
di-n-octyl phthalate
di-n-octyl phthalate-d^
diphenylamine (Appendix C)
diphenytamine-d.-
diphenyl ether (Appendix C)
diphenyl ether-d...
1 ,2-diphenylhydrazine
1 ,2-diphenylhydrazine-d.g
f luoranthene
f tuoranthene-d^Q
f luorene
f luorene-d.-
hexach I orobenzene
hexachlorobenzene- C^
hexach lorobutadiene
hexachlorobutadiene- C^
hexach I oroethane
hexach 1 oroethane- C
hexach I orocyc I open tad i ene
hexachlorocyclopentadiene- C^
ideno(1,2,3-cd)pyrene (3)
isophorone
isophorone-d_
Acceptance criteria
Initial
precision
and accuracy
Section 8.2.3
(ug/L)
s X
20
31
31
31
15
23
17
35
43
48
42
48
26
80
12
28
44
78
13
22
36
108
18
66
18
37
30
59
16
46
45
42
19
37
73
35
33
35
29
43
16
81
56
63
227
77
15
60
55
25
23
85 -
47 -
79 -
48 -
76 -
23 -
73 -
14 -
63 -
13 -
61 -
15 -
68 -
ns -
85 -
38 -
75 -
ns -
62 -
15 -
74 -
ns -
72 -
22 -
75 -
22 -
80 -
44 -
77 -
12 -
58 -
27 -
82 -
36 -
49 -
31 -
71 -
36 -
81 -
51 -
90 -
36 -
51 -
ns -
21 -
ns -
69 -
ns -
23 -
76 -
49 -
136
136
150
130
165
195
146
212
201
203
194
193
174
562
131
164
196
260
153
228
188
640
134
308
158
245
141
184
161
383
205
206
136
155
308
173
177
161
132
131
124
228
251
316
ns
400
144
ns
299
156
133
Labeled
compound
recovery
Sec 8.3
and'14.2
P (X)
28 -
29 -
13 -
ns -
ns -
ns -
ns -
24 -
ns -
ns -
ns -
ns -
10 -
17 -
ns -
11 -
19 -
17 -
20 -
27 -
13 -
ns -
ns -
ns -
33 -
220
215
346
494
550
474
ns
260
ns
449
ns
ns
514
442
ns
488
281
316
278
238
595
ns
ns
ns
193
Calibra-
tion
verifi-
cation
Sec 12.5
(uq/mL)
73 -
66 -
72 -
69 -
71 -
52 -
74 -
61 -
65 -
52 -
62 -
65 -
77 -
18 -
67 -
64 -
74 -
47 -
67 -
58 -
73 -
50 -
75 -
39 -
79 -
53 -
55 -
36 -
71 -
21 -
57 -
59 -
83 -
77 -
75 -
58 -
67 -
47 -
74 -
61 -
78 -
38 -
74 -
68 -
71 -
47 -
77 -
47 -
13 -
70 -
52 -
136
150
140
145
142
192
135
164
154
192
161
153
130
558
149
157
135
211
150
172
137
201
133
256
127
187
183
278
140
467
176
169
120
129
134
174
149
215
135
164
128
265
135
148
141
212
129
211
761
142
194
On- go ing
accuracy
Sec 12.7
R (ug/L)
79 -
39 -
70 -
40 -
74 -
22 -
70 -
11 -
55 -
ns -
53 -
11 -
64 -
ns -
83 -
34 -
65 -
ns -
60 -
14 -
67 -
ns -
68 -
17 -
72 -
19 -
70 -
31 -
74 -
10 -
51 -
21 -
77 -
29 -
40 -
26 -
64 -
30 -
70 -
38 -
85 -
23 -
43 -
ns -
13 -
ns -
67 -
ns -
19 -
70 -
44 -
146
160
168
156
169
209
152
247
225
260
219
245
185
ns
135
182
222
ns
156
242
207
ns
141
378
164
275
159
250
166
433
231
249
144
186
360
200
194
187
151
172
132
321
287
413
ns
563
148
ns
340
168
147
56
-------
EGO
No.
(1)
360
260
355
255
702
602
356
256
357
257
358
258
361
261
363
263
362
262
364
264
381
281
365
265
703
603
384
284
710
610
709
609
729
629
308
208
530
731
631
321
221
Compound
2-methyl-4,6-dinitrophenol
2-methyl-4,6-dinitrophenol-d-
naphthalene
naphtha I ene-dg
beta-naphthylamine (Appendix C)
beta-naphthylamine-d-
nitrobenzene
nitrobenzene-dc
5
2-nitrophenot
2-nitrophenol-d,
4-nitrophenol
4-nitrophenol-d,
N - n i t rosod i methy I ami ne
N-nitrosodimethylamine-d, (3)
M-nitrosodi-n-propylamine
N-nitrosodi-n-propylamine (3)
N-nitrosodipheny(amine
N-nitrosodiphenylamine-d,
pentachlorophenol
pentachlorophenol- C,
phenanthrene
phenanthrene-d^Q
phenol
phenol -d-
alpha-picoline (Synfuel)
alpha-picoline-d^
pyrene
pyrene-d1Q
styrene (Appendix C)
styrene-dj
alpha-terpineol (Appendix C)
alpha-terpineol-oL
1,2,3-trichlorobenzene (4c)
1,2.3-trichlorobenzene-dj (3)
1,2,4-trichlorobenzene
1,2,4-trichlorobenzene-d,
2,3,6-trichlorophenol (4c) (3)
2,4,5-trichlorophenol (4c)
2,4,5-trichlorophenol-d2 (3)
2,4,6-trichlorophenol
2,4,6-trichlorophenol-d-
Acceptance criteria
Initial
precision
and accuracy
Section 8.2.3
(ug/L)
s X
19
64
20
39
49
33
25
28
15
23
42
188
49
33
45
37
45
37
21
49
13
40
36
161
38
138
19
29
42
49
44
48
69
57
19
57
30
30
47
57
47
77
36
80
23
10
ns
69
18
78
41
62
14
10
ns
65
54
65
54
76
37
93
45
77
21
59
11
76
32
53
ns
42
22
15
15
82
15
58
58
43
59
43
- 133
- 247
- 139
- 157
- ns
ns
- 161
- 265
- 140
- 145
- 146
- 398
- ns
- ns
- 142
- 126
- 142
- 126
- 140
- 212
- 119
- 130
- 127
- 210
- 149
- 380
- 152
- 176
- 221
- 281
- 234
- 292
- 229
- 212
- 136
- 212
- 137
- 137
- 183
- 205
- 183
Labeled
compound
recovery
Sec 8.3
and 14.2
P (X)
16
14
ns
ns
27
ns
ns
26
26
18
24
ns
ns
18
ns
ns
ns
ns
21
21
- 527
- 305
- ns
- ns
- 217
- ns
- ns
- 256
- 256
- 412
- 241
- ns
- ns
- 303
- ns
- 672
- 592
- 592
- 363
- 363
Calibra-
tion
verifi-
cation
Sec 12.5
(ug/mL)
69 -
56 -
73 -
71 -
39 -
44 -
85 -
46 -
77 -
61 -
55 -
35 -
39 -
44 -
68 -
59 -
68 -
59 -
77 -
42 -
75 -
67 -
65 -
48 -
60 -
31 -
76 -
48 -
65 -
44 -
54 -
20 -
60 -
61 -
78 -
61 -
56 -
56 -
69 -
81 -
69 -
145
177
137
141
256
230
115
219
129
163
183
287
256
230
148
170
148
170
130
237
133
149
155
208
165
324
132
210
153
228
186
502
167
163
128
163
180
180
144
123
144
On- go ing
accuracy
Sec 12.7
R (uq/L)
72
28
75
22
ns
ns
65
15
75
37
51
ns
ns
ns
53
40
53
40
71
29
87
34
62
ns
50
ns
72
28
48
ns
38
18
11
10
77
10
51
51
34
48
34
- 142
- 307
- 149
- 192
- ns
- ns
- 169
- 314
- 145
- 158
- 175
- ns
- ns
- ns
- 173
- 166
- 173
- 166
- 150
- 254
- 126
- 168
- 154
- ns
- 174
- 608
- 159
- 196
- 244
- 348
- 258
- 339
- 297
- 282
- 144
- 282
- 153
- 153
- 226
- 244
- 226
(1) Reference numbers beginning with 0, 1 or 5 indicate a pollutant quantified by the internal standard method;
reference numbers beginning with 2 or 6 indicate a labeled compound quantified by the internal standard method;
reference numbers beginning with 3 or 7 indicate a pollutant quantified by isotope dilution.
(2) ns = no specification: limit is outside the range that can be measured reliably.
(3) This compound is to be determined by internal standard; specification is derived from related compound.
57
-------
8.4 As part of the QA program for the
laboratory, method accuracy for samples
shall be assessed and records shall be
maintained. After the analysis of five
samples or a given matrix type (water,
soil, sludge, sediment) for which the
labeled compounds pass the tests in
section 8.3, compute the average percent
recovery (P) and the standard deviation of
the percent recovery (s ) for the labeled
compounds only. Express the accuracy
assessment as a percent recovery interval
from P -2s to P + 2s for each matrix.
For example, if P = 90S and s = 10X for
five analyses of compost, trie accuracy
interval is expressed as 70 - 110%.
Update the accuracy assessment for each
compound in each matrix on a regular basis
Ce.g. after each 5-10 new accuracy
measurements).
8.5 Blanks--reagent water and high solids
reference matrix blanks -are analyzed to
demonstrate freedom from contamination.
calibrated properly, then maintained in a
calibrated state. The standards used for
calibration (section 7), calibration
verification (section 12.5), and for
initial (section 8.2) and on-going
(section 12.7) precision and recovery
should be identical, so that the most
precise results will be obtained. The
GCMS instrument in particular will provide
the most reproducible results if dedicated
to the settings and conditions required
for the analyses of semi-volatrles by this
method.
8.7 Depending on specific program require-
ments, field replicates may be collected
to determine the precision of the sampling
technique, and spiked samples may be
required to determine the accuracy of the
analysis when the internal standard method
is used.
9 SAMPLE COLLECTION, PRESERVATION, AND
HANDLING
8.5.1 Extract and concentrate a one liter
reagent water blank or a high solids
reference matrix blank with each sample
lot (samples started through the
extraction process on the same 8 hr shift,
to a maximum of 20 samples). Analyze the
blank immediately after analysis of the
precision and recovery standard (section
6.14) to demonstrate freedom from
contamination.
8.5.2 If any of the compounds of interest
(tables 1 thru 4) or any potentially
interfering compound is found in an
aqueous blank at greater than 10 ug/L, or
in a high solids reference matrix blank at
greater than 100 ug/kg (assuming a
response factor of 1 relative to the
internal standard for compounds not listed
in tables 1 thru 4), analysis of samples
is halted until the source of
contamination is eliminated and a blank
shows-no evidence of contamination at this
level.
8.6 The specifications contained in this
method can be met if the apparatus used is
9.1 Collect samples in glass containers
following conventional sampling practices
(reference 8). Aqueous samples which flow
freely are collected in refrigerated
bottles using automatic sampling
equipment. Solid samples are collected as
grab samples using wide mouth jars.
9.2 Maintain samples at 0 - 4 °C from the time
of collection until extraction. If
residual chlorine is present in aqueous
samples, add 80 mg sodium thiosulfate per
liter of water. EPA methods 330.4 and
330.5 may be used to measure residual
chlorine (reference 9).
9.3 Begin sample extraction within seven days
of collection, and analyze all extracts
within 40 days of extraction.
10 SAMPLE EXTRACTION, CONCENTRATION, AND
CLEANUP
Samples containing one percent solids or
less are extracted directly using
continuous liquid/liquid extraction
techniques (section 10.2.1 and figure 4).
58
-------
10.2.2.3
10.2.2.4
Samples containing one to 30 percent 10.2.2.2
solids are diluted to the one percent
level with reagent water (section 10.2.2)
and extracted using continuous
liquid/liquid extraction techniques.
Samples containing greater than 30 percent
solids are extracted using ultrasonic
techniques (section 10.2.5)
10.1 Determination of percent solids
10.1.1 Weigh 5 - 10 g of sample into a tared
beaker.
10.1.2 Dry overnight (12 hours minimum) at 110 ±
5 °C, and cool in a dessicator.
10.1.3 Determine percent solids as follows:
% solids = weight of dry sample x 100
weight of wet sample
10.2 Preparation of samples for extraction 10.2.2.5
10.2.1 Samples containing one percent solids or
less—extract sample directly using 10.2.2.6
continuous liquid/liquid extraction
techniques.
10.2.1.1 Measure 1.00 ± 0.01 liter of sample into a 10.2.2.7
clean 1.5 - 2.0 liter beaker.
10.2.1.2 Dilute aliquot—for samples which are
expected to be difficult to extract, 10.2.2.8
concentrate, or clean-up, measure an
additional 100.0 ± 1.0 ml into a clean 1.5
- 2.0 liter beaker and dilute to a final
volume of 1.00 ± 0.1 liter with reagent
water.
10.2.1.3 Spike 0.5 mL of the labeled compound
spiking solution (section 6.8) into the
sample aliquots. Proceed to preparation
of the QC aliquots for low solids samples
(section 10.2.3).
Using the percent solids found in 10.1.3,
determine the weight of sample required to
produce one liter of solution containing
one percent solids as follows:
sample weight
1000
X solids
grams
Place the weight determined in 10.2.2.2 in
a clean 1.5 - 2.0 liter beaker. Discard
all sticks, rocks, leaves and other
foreign material prior to weighing.
Dilute aliquot--for samples which are
expected to be difficult to extract,
concentrate, or clean-up, weigh an amount
of sample equal to one-tenth the amount
determined in 10.2.2.2 into a second clean
1.5 - 2.0 liter beaker. When diluted to
1.0 liter, this dilute aliquot will
contain 0.1 percent solids.
Bring the sample aliquot(s) above to 100 -
200 mL volume with reagent water.
Spike 0.5 ml of the labeled compound
spiking solution (section 6.8) into each
sample aliquot.
Using a clean metal spatula, break any
solid portions of the sample into small
pieces.
Place the 3/4 in. horn on the ultrasonic
probe approx 1/2 in. below the surface of
each sample aliquot and pulse at 50
percent for three minutes at full power.
If necessary, remove the probe from the
solution and break any large pieces using
the metal spatula or a stirring rod and
repeat the sonication.
Clean the probe with methylene
chloride:acetone (1:1) between samples to
preclude cross-contamination.
10.2.2 Samples containing one to 30 percent
solids
10.2.2.1 Mix sample thoroughly.
10.2.2.9 Bring the sample volume to 1.0 ± 0.1 liter
with reagent water.
10.2.3 Preparation of QC aliquots for samples
containing low solids (<30 percent).
59
-------
[10.2.3.1 J
[10.2.1.3, 10.2.3.2]
[10.2.3.3]
[10.2.4]
[10.3.2]
[10.3.4]
H0.5]
[10.6]
[11.3]
[11.4]
STANDARD
1 L REAGENT
WATER
SPIKE
1.0mL
OF STANDARDS
STIR AND
EQUILIBRATE
STANDARD OR BLANK
EXTRACT BASE/
NEUTRAL
ORGANIC I AQUEOUS
EXTRACT ACID
CONCENTRATE
TO 2-4 ml
CONCENTRATE
TO 2-4 mL
CONCENTRATE
TOLOmL
ADD INTERNAL
STANDARD
INJECT
BLANK
1 L REAGENT
WATER
SPIKE 500 pL
OF 200 pg/mL
ISOTOPES
STIR AND
EQUILIBRATE
SAMPLE
1 L ALIQUOT
SPIKE 500 pL
OF 200 pg/mL
ISOTOPES
STIR AND
EQUILIBRATE
EXTRACT BASE/
NEUTRAL
ORGANIC
AQUEOUS
EXTRACT ACID
CONCENTRATE
TO 1.0 mL
CONCENTRATE
TOLOmL
ADD INTERNAL
STANDARD
ADD INTERNAL
STANDARD
INJECT
INJECT
FIGURE 4 Flow Chart for Extraction/Concentration of Low Solids Precision and Recovery Standard, Blank, and
Sample by Method 1635. Numbers in Brackets [ ] Refer to Section Numbers in the Method.
60
-------
10.2.3.1 For each sample or sample lot (to a
maximum of 20) to be extracted at the same
time, place three 1.0 ± 0.01 liter
aliquots of reagent water in clean 1.5 -
2.0 liter beakers.
10.2.3.2 Spike 0.5 mL of the labeled compound
spiking solution (section 6.8) into one
reagent water aliquot. This aliquot will
serve as the blank.
10.2.3.3 Spike 1.0 ml of the precision and recovery
standard (section 6.14) into the two
remaining reagent water aliquots.
10.2.4 Stir and equilibrate all sample and QC
solutions for 1 - 2 hours. Extract the
samples and QC aliquots per section 10.3.
10.2.5 Samples containing 30 percent solids or
greater
10.2.5.1 Mix the sample thoroughly
10.2.5.2 Weigh 30 t 0.3 grams into a clean 400 •
500 ml beaker. Discard all sticks, rocks,
leaves and other foreign material prior to
weighing.
10.2.5.3 Dilute aliquot—for samples which are
expected to be difficult to extract,
concentrate, or clean-up, weigh 3 t 0.03
grams into a clean 400 - 500 ml beaker.
10.2.5.4 Spike 0.5 ml of the labeled compound
spiking solution (section 6.8) into each
sample aliquot.
10.2.5.5 QC aliquots--for each sample or sample lot
(to a maximum of 20) to be extracted at
the same time, place three 30 t 0.3 gram
aliquots of the high solids reference
matrix in clean 400 - 500 mL beakers.
10.2.5.6 Spike 0.5 ml of the labeled compound
spiking solution (section 6.8) into one
high solids reference matrix aliquot.
This aliquot will serve as the blank.
10.2.5.7 Spike 1.0 mL of the precision and recovery
standard (section 6.14) into the two
remaining high solids reference matrix
aliquots. Extract, concentrate, and clean
up the high solids samples per sections
10.4 through 10.8.
10.3 Continuous extraction of low solids
(aqueous) samples--piace 100 - 150 mL
methylene chloride in each continuous
extractor and 200 - 300 mL in each
distilling flask.
10.3.1 Pour the sample(s), blank, and standard
aliquots into the extractors. Rinse the
glass containers with 50 - 100 mL
methylene chloride and add to the
respective extractors. Include all solids
in the extraction process.
10.3.2 Base/neutral extraction—adjust the pH of
the waters in the extractors to 12 - 13
with 6N NaOH while monitoring with a pH
meter. Begin the extraction by heating
the flask until the methylene chloride is
boiling. When properly adjusted, 1 - 2
drops of methylene chloride per second
will fall from the condenser tip into the
water. Test and adjust the pH of the
waters- during the first to second hour and
during the fifth to tenth hour of
extraction. Extract for 24 - 48 hours.
10.3.3 Remove the distilling flask, estimate and
record the volume of extract (to the
nearest 100 mL), and pour the contents
through a drying column containing 7 to 10
cm anhydrous sodium sutfate. Rinse the
distilling flask with 30 - 50 mL of
methylene chloride and pour through the
drying column. Collect the solution in a
500 mL K-D evaporator flask equipped with
a 10 mL concentrator tube. Seal, label as
the base/neutral fraction, and concentrate
per sections 10.5 to 10.6.
10.3.4 Acid extraction—adjust the pH of the
waters in the extractors to 2 or less
using 6N sulfuric acid. Charge clean
distilling flasks with 300 - 400 mL of
methylene chloride. Test and adjust the
pH of the waters during the first 1 - 2 hr
and during the fifth to tenth hr of
extraction. Extract for 24 - 48 hours.
61
-------
Repeat section 10.3.3, except label as the
acid fraction.
10.4 Ultrasonic extraction of high solids
samples
10.4.1 Add 60 grams of anhydrous sodium sulfate
the sample and QC aliquot(s) (section
10.2.5) and mix thoroughly.
10.4.2 Add 100 ± 10 mL of acetone:methylene
chloride (1:1) to the sample and mix
thoroughly.
10.4.3 Place the 3/4 in. horn on the ultrasonic
probe approx 1/2 in. below the surface of
the solvent but above the solids layer and
pulse at 50 percent for three minutes at
full power. If necessary, remove the
probe from the solution and break any
large pieces using the metal spatula or a
stirring rod and repeat the sonication.
10.4.4 Decant the extracts through Whatman 41
filter paper using glass funnels and
collect in 500 - 1000 mL graduated
cylinders.
10.4.5 Repeat the extraction steps (10.4.2 -
10.4.4) twice more for each sample and QC
aliquot. On the final extraction, swirl
the sample or QC aliquot, pour into its
respective glass funnel, and rinse with
acetone:methylene chloride. Record the
total extract volume.
10.4.6 Pour each extract through a drying column
containing 7 to 10 cm of anhydrous sodium
sulfate. Rinse the graduated cylinder
with 30 - 50 mL of methylene chloride and
pour through the drying column. Collect
each extract in a 500 mL K-D evaporator
flask equipped with a 10 mL concentrator
tube. Seal and label as the high solids
semi-volatile fraction. Concentrate and
clean up the samples and QC aliquots per
sections 10.5 through 10.8.
10.5 Macro concentration--concentrate the
extracts in separate 500 mL K-D flasks
equipped with 10 mL concentrator tubes.
10.5.1 Add 1 to 2 clean boiling chips to the
flask and attach a three-ball macro Snyder
column. Prewet the column by adding
approx one mL of methylene chloride
through the top. Place the K-D apparatus
in a hot water bath so that the entire
tower rounded surface of the flask is
bathed with steam. Adjust the vertical
position of the apparatus and the water
temperature as required to complete the
concentration in 15 to 20 minutes. At the
proper rate of distillation, the balls of
the column will actively chatter but the
chambers will not flood. When the liquid
has reached an apparent volume of 1 mL,
remove the K-D apparatus from the bath and
allow the solvent to drain and cool for at
least 10 minutes. Remove the Snyder column
and rinse the flask and its lowers joint
into the concentrator tube with 1 - 2 mL
of methylene chloride. A 5 mL syringe is
recommended for this operation.
10.5.2 For performance standards (sections 8.2
and 12.7) and for blanks (section 8.5),
combine the acid and base/neutral extracts
for each at this point. Do not combine
the acid and base/neutral extracts for
aqueous samples.
10.6 Micro-concentration--Add a clean boiling
chip and attach a two-ball micro Snyder
column to the concentrator tube. Prewet
the column by adding approx 0.5 mL
methylene chloride through the top. Place
the apparatus in the hot water bath.
Adjust the vertical position and the water
temperature as required to complete the
concentration in 5 - 10 minutes. At the
proper rate of distillation, the balls of
the column will actively chatter but the
chambers will not flood. When the liquid
reaches an apparent volume of approx 0.5
mL, remove the apparatus from the water
bath and allow to drain and cool for at
least 10 minutes. Remove the micro Snyder
column and rinse its lower joint into the
concentrator tube with approx 0.2 mL of
methylene chloride. Adjust the final
volume to 5.0 mL if the extract is to be
cleaned up by GPC, or to 1.0 mL if it has
62
-------
been cleaned up or does not require clean-
up.
10.7 Transfer the concentrated extract to a
clean screw-cap vial. Seal the vial with a
Teflon-lined lid, and mark the level on
the vial. Label with the sample number and
fraction, and store in the dark at -20 to
-10 °C until ready for analysis.
10.8 GPC setup and calibration
10.8.1 Column packing
10.8.1.1 Place 75 ± 5 g of SX-3 Bio-beads in a 400
- 500 ml beaker.
10.8.1.2 Cover the beads and allow to swell
overnight (12 hours minimum).
10.8.1.3 Transfer the swelled beads to the column
and pump solvent through the column, from
bottom to top, at 4.5 - 5.5 mL/min prior
to connecting the column to the detector.
10.8.1.4 After purging the column with solvent for
1 - 2 hours, adjust the column head
pressure to 7 - 10 psig, and purge for 4 -
5 hours to remove air from the column.
Maintain a head pressure of 7 - 10 psig.
Connect the column to the detector.
10.8.2 Column calibration
10.8.2.1 Load 5 mL of the calibration solution
(section 6.4) into the sample loop.
10.8.2.2 Inject the calibration solution and record
the signal from the detector. The elution
pattern wilt be corn oil, bis(2-
ethylhexyl) phthatate, pentachlorophenol,
perylene, and sulfur.
10.8.2.3 Set the "dump time" to allow >85% removal
of the corn oil and >85% collection of the
phthalate.
10.8.2.4 Set the "collect time" to the peak minimum
between perylene and sulfur.
10.8.2.5 Verify the calibration with the
calibration solution after every 20
extracts. Calibration is verified if the
recovery of the pentachlorophenol is
greater than 85X. If calibration is not
verified, the system shall be recalibrated
using the calibration solution, and the
previous 20 samples shall be re-extracted
and cleaned up using the calibrated GPC
system.
10.9 Extract cleanup
10.9.1 Filter the extract or load through the
filter holder to remove particulates.
Load the 5.0 mL extract onto the column.
The maximum capacity of the column is 0.5
- 1.0 gram. If necessary, split the
extract into multiple aliquots to prevent
column overload.
10.9.2 Elute the extract using the calibration
data determined in 10.8.2. Collect the
eluate in a clean 400 - 500 mL beaker.
10.9.3 Concentrate the cleaned up extract per
section 10.5.
10.9.4 Rinse the sample loading tube thoroughly
with methylene chloride between extracts
to prepare for the next sample.
10.9.5 If a particularly dirty extract is
encountered, a 5.0 mL methylene chloride
blank shall be run through the system to
check for carry-over.
10.9.6 Reconcentrate the extract to one mL and
transfer to a screw-cap vial per sections
10.6 and 10.7.
11 GCMS ANALYSIS
11.1 Establish the operating conditions given
in tables 5 or 6 for analysis of the
base/neutral or acid extracts, respec-
tively. For analysis of combined extracts
(section 10.5.2 and 10.9.6), use the
operating conditions in table 5.
11.2 Bring the concentrated extract (section
10.7) or standard (sections 6.13 - 6.14)
to room temperature and verify that any
precipitate has redissolved. Verify the
63
-------
level on the extract (sections 6.6 and
10.7) and bring to the mark with solvent
if required.
11.3 Add the internal standard solution
(section 6.10) to the extract (use 1.0 uL
of solution per 0.1 ml of extract)
immediately prior to injection to minimize
the possibility of loss by evaporation,
adsorption, or reaction. Mix thoroughly.
11.4 Inject a volume of the standard solution
or extract such that 100 ng of the
internal standard will be injected, using
on-column or splitless injection. For 1
ml extracts, this volume will be 1.0 uL.
Start the GC column initial isothermal
hold upon injection. Start MS data
collection after the solvent peak elutes.
Stop data collection after the
benzo(ghi)perylene or pentachlorophenol
peak elutes for the base/neutral (or semi-
volatile) or acid fraction, respectively.
Return the column to the initial
temperature for analysis of the next
sample.
12 SYSTEM AND LABORATORY PERFORMANCE
12.1 At the beginning of each 8 hr shift during
which analyses are performed, GCMS system
performance and calibration are verified
for all pollutants and labeled compounds.
For these tests, analysis of the 100 ug/mL
calibration standard (section 6.13) shall
be used to verify all performance
criteria. Adjustment and/or recalibration
(per section 7) shall be performed until
all performance criteria are met. Only
after all performance criteria are met may
samples, blanks, and precision and
recovery standards be analyzed.
12.2 DFTPP spectrum validity-inject 1 uL of
the DFTPP solution (section 6.11) either
separately or within a few seconds of
injection of the standard (section 12.1)
analyzed at the beginning of each shift.
The criteria in table 7 shall be met.
12.3 Retention times—the absolute retention
time of 2,2'-difluorobiphenyl shall be
within the range of 1078 to 1248 seconds
and the relative retention times of all
pollutants and labeled compounds shall
fall within the limits given in tables 5
and 6.
12.4 GC resolution--the valley height between
anthracene and phenanthrene at m/z 178 (or
the analogs at m/z 188) shall not exceed
10 percent of the taller of the two peaks.
12.5 Calibration verification--compute the
concentration of each pollutant (tables 1
and 2) by isotope dilution (section 7.4)
for those compounds which have labeled
analogs. Compute the concentration of
each pollutant which has no labeled analog
by the internal standard method (section
7.5). Compute the concentration of the
labeled compounds by the internal standard
method. These concentrations are computed
based on the calibration data determined
in section 7.
12.5.1 For each pollutant and labeled compound
being tested, compare the concentration
with the calibration verification limit in
table 10. If all compounds meet the
acceptance criteria, calibration has been
verified and analysis of blanks, samples,
and precision and recovery standards may
proceed. If, however, any compound fails,
the measurement system is not performing
properly for that compound. In this
event, prepare a fresh calibration
standard or correct the problem causing
the failure and repeat the test (section
12.1), or recalibrate (section 7).
12.6 Multiple peaks--each compound injected
shall give a single, distinct GC peak.
12.7 On-going precision and accuracy.
12.7.1 Analyze the extract of one of the pair of
precision and recovery standards (section
10) prior to analysis of samples from the
same lot.
12.7.2 Compute the concentration of each
pollutant (tables 1 and 2) by isotope
dilution (section 7.4) for those compounds
64
-------
which have labeled analogs. Compute the
concentration of each pollutant which has
no labeled analog by the internal standard
method (section 7.5). Compute the concen-
tration of the labeled compounds by the
internal standard method.
12.7.3 For each pollutant and labeled compound,
compare the concentration with the limits
for on-going accuracy in table 10. If all
compounds meet the acceptance criteria,
system performance is acceptable and
analysis of blanks and samples may
proceed. If, however, any individual
concentration falls outside of the range
given, system performance is unacceptable
for that compound.
for each pollutant and labeled compound in
each matrix type by calculating the
average percent recovery (R) and the
standard deviation of percent recovery
Express the accuracy as a recovery
For
= 5X, the
accuracy is 85 - 105X.
(sr).
interval from R - 2s to R + 2s
example, if R = 9SX and
QC
ANTHRACENE-D,,
•
•
.
--3s
a 123456789 10
ANALYSIS NUMBER
NOTE: The large number of compounds in
table 10 present a substantial probability
that one or more will fail when all
compounds are analyzed. To determine if
the extraction/concentration system is out
of control or if the failure is caused by
probability, proceed as follows:
12.7.3.1 Analyze the second aliquot of the pair of
precision and recovery standards (section
10).
12.7.3.2 Compute the concentration of only those
pollutants or labeled compounds that
failed the previous test (section 12.7.3).
If these compounds now pass, the
extraction/concentration processes are in
control and analysis of blanks and samples
may proceed. If, however, any of the same
compounds fail again, the extrac-
tion/concentration processes are not being
performed properly for these compounds.
In this event, correct the problem, re-
extract the sample lot (section 10) and
repeat the on-going precision and recovery
test (section 12.7).
12.7.4 Add results which pass the specifications
in section 12.7.3 to initial and previous
on-going data for each compound in each
matrix. Update QC charts to form a
graphic representation of continued
laboratory performance (Figure 5).
Develop a statement of laboratory accuracy
E Z
5 "
oc 5
100-*'
090
ANTHRACENE
r • ~ •*""","
• , •
( •
•3s
-3s
6/1 6/1 6/1 5/1 6/2 6/2 6/3 6/3 6/4 6/5
DATE ANALYZED
FIGURE 5 Quality Control Charts Showing Area
(top graph) and Relative Response of
Anthracene to Anthracene-d,0 (lower graph)
Plotted as a Function of Time or Analysis
Number.
13 QUALITATIVE DETERMINATION
Identification is accomplished by
comparison of data from analysis of a
sample or blank with data stored in the
mass spectral libraries. For compounds
for which the relative retention times and
mass spectra are known, identification is
confirmed per sections 13.1 and 13.2. For
unidentified GC peaks, the spectrum is
compared to spectra in the EPA/NIH mass
spectral file per section 13.3.
13.1 Labeled compounds and pollutants having no
labeled analog (tables 1 thru 4):
13.1.1 The signals for all characteristic m/z's
stored in the spectral library (section
7.2.4) shall be present and shall maximize
within the same two consecutive scans.
65
-------
13.1.2 Either <1) the background corrected EICP
areas, or (2) the corrected relative
intensities of the mass spectral peaks at
the GC peak maximum shall agree within a
factor of two (0.5 to 2 times) for all
masses stored in the library.
13.1.3 For the compounds for which the system has
been calibrated (tables 1 and 2), the
retention time shall be within the windows
specified in tables 5 and 6, or within t
15 scans or ± 15 seconds (whichever is
greater) for compounds for which no window
is specified.
intensities of the mass spectral peaks at
the GC peak maximum shall agree within a
factor of two with the masses stored in
the EPA/NIH Mass Spectral File.
13.4 M/z's present in the experimental mass
spectrum that are not present in the
reference mass spectrum shall be accounted
for by contaminant or background ions. If
the experimental mass spectrum is
contaminated, or if identification is
ambiguous, an experienced spectrometrist
(section 1.4) is to determine the presence
or absence of the compound.
13.1.4 For the compounds for which the system has
not been calibrated but the relative
retention times and mass spectra are known
(tables 3 and 4), the retention time
relative to the 2,2'-difluorobiphenyl
internal standard shall be within t 30
scans or ± 30 seconds (whichever is
greater) based on the nominal retention
time specified in tables 5 and 6.
13.2 Pollutants having a labeled analog (tables
1 and 2):
13.2.1 The signals for all characteristic m/z's
stored in the spectral library (section
7.2.4) shall be present and shall maximize
within the same two consecutive scans.
13.2.2 Either (1) the background corrected EICP
areas, or (2) the corrected relative
intensities of the mass spectral peaks at
the GC peak maximum shall agree within a
factor of two for all masses stored in the
spectral library.
13.2.3 The relative retention time between the
pollutant and its labeled analog shall be
within the windows specified in tables 5
and 6.
13.3 Unidentified GC peaks
13.3.1 The signals for masses specific to a GC
peak shall all maximize within t 1 scan.
13.3.2 Either (1) the background corrected EICP
areas, or (2) the corrected relative
14 QUANTITATIVE DETERMINATION
14.1 Isotope dilution—by adding a known amount
of a labeled compound to every sample
prior to extraction, correction for
recovery of the pollutant can be made
because the pollutant and its labeled
analog exhibit the same effects upon
extraction, concentration, and gas
chromatography. Relative response (RR)
values for sample mixtures are used in
conjunction with calibration curves
described in section 7.4 to determine
concentrations directly, so long as
labeled compound spiking levels are
constant. For the phenol example given in
figure 1 (section 7.4.1), RR would be
equal to 1.114. For this RR value, the
phenol calibration curve given in figure 1
indicates a concentration of 27 ug/mL in
the sample extract (C x).
14.2 Internal standard--compute the concentra-
tion in the extract using the response
factor determined from calibration data
(section 7.5) and the following equation:
(A-s x RF)
where C is the concentration of the
compound in the extract, and the other
terms are as defined in section 7.5.1.
14.3 The concentration of the pollutant in the
solid phase of the sample is computed
using the concentration of the pollutant
66
-------
in the extract and the weight of the
solids (section 10), as follows:
Concentration in solid (ug/kg) =
is the extract volume in mL, and
where
U is the sample weight in kg.
14.4 If the EICP area at the quant i tat ion m/z
for any compound exceeds the calibration
range of the system, the extract of the
dilute aliquot (section 10) is analyzed by
isotope dilution. If further dilution is
required and the sample holding time has
not been exceeded, a smaller sample
aliquot is extracted per section 14.4.1 -
14.4.3. If the sample holding time has
been exceeded, the sample extract is
diluted by successive factors of 10,
internal standard is added to give a
concentration of 100 ug/mL in the diluted
extract, and the diluted extract, is
analyzed by the internal standard method.
14.4.1 For samples containing one percent solids
or less for which the holding time has not
been exceeded, dilute 10 ml, 1.0 mL, 0.1
ml etc. of sample to one liter with
reagent water and extract per section
10.2.1.
14.4.2 For samples containing 1-30 percent
solids for which the holding time has not
been exceeded, extract an amount of sample
equal to 1/100 the amount determined in
10.2.2.2. Extract per section 10.2.2.
14.4.3 For samples containing 30 percent solids
or greater for which the holding time has
not been exceeded, extract 0.30 i 0.003 g
of sample per section 10.2.5.
14.5 For GC peaks which are to be identified
(per section 13.3), the sample is diluted
by successive factors of 10 when any peak
in the uncorrected mass spectrum at the GC
peak maximum is saturated.
14.6 Results are reported for all pollutants,
labeled compounds, and tentatively
identified compounds found in all
standards, blanks, and samples, in units
of ug/L for aqueous samples or in ug/kg
dry weight of solids for samples
containing one percent solids or greater
(soils, sediments, filter cake, compost),
to three significant figures. Results for
samples which have been diluted are
reported at the least dilute level at
which the area at the quant i tat ion m/z is
within the calibration range (section
14.4) or at which no m/z in the spectrum
is saturated (section 14.5). For
compounds having a labeled analog, results
are reported at the least dilute level at
which the area at the quantitation m/z is
within the calibration range (section
14.4) and the labeled compound recovery is
within the normal range for the method
(section 15.4).
15 ANALYSIS OF COMPLEX SAMPLES
15.1 Some samples may contain high levels
(>1000 ug/L) of the compounds of interest,
interfering compounds, and/or polymeric
materials. Some samples will not
concentrate to one mL (section 10.6);
others will overload the GC column and/or
mass spectrometer.
15.2 Analyze the dilute aliquot (section 10)
when the sample will not concentrate to
1.0 mL. If a dilute aliquot was not
extracted, and the sample holding time
(section 9.3) has not been exceeded,
dilute an aliquot of an aqueous sample
with reagent water, or weigh a dilute
aliquot of a high solids sample and re-
extract (section 10); otherwise, dilute
the extract (section 14.4) and analyze by
the internal standard method (section
14.2).
15.3 Recovery of internal standard—the EICP
area of the internal standard should be
within a factor of two of the area in the
shift standard (section 12.1). If the
absolute areas of the labeled compounds
are within a factor of two of the
respective areas in the shift standard,
and the internal standard area is less
67
-------
than one-half of its respective area, then
internal standard loss in the extract has
occurred. In this rase, use one of the
labeled compounds (preferably a
polynuclear aromatic hydrocarbon) to
compute the concentration of a pollutant
with no labeled analog.
15.4 Recovery of . labeled compounds--in most
samples, labeled compound recoveries will
be similar to those from reagent water or
from the high solids reference matrix
(section 12.7). If the labeled compound
recovery is outside the limits given in
table 10, the extract from the dilute
aliquot (section 10) is analyzed as in
section U.4. If the recoveries of all
labeled compounds and the internal
standard are low (per the criteria above),
then a loss in instrument sensitivity is
the most likely cause. In this case, the
100 ug/mL calibration standard (section
12.1) shall be analyzed and calibration
verified (section 12.5). If a loss in
sensitivity has occurred, the instrument
shall be repaired, the performance
specifications in section 12 shall be met,
and the extract reanalyzed. If a loss in
instrument sensitivity has not occurred,
the method does not work on the sample
being analyzed and the result may not be
reported for regulatory compliance
purposes.
16 METHOD PERFORMANCE
16.1 Interlaboratory performance for this
method is detailed in reference 10.
Reference mass spectra, retention times,
and response factors are from references
11 and 12. Results of initial tests of
this method on municipal sludge can be
found in reference 13.
16.2 A chromatogram of the 100 ug/mL
acid/base/neutral calibration standard
(section 6.13) is shown in figure 6.
68
-------
RIC DATA: ABHiouee #1
83/13--S4 5::4:08 CALI: HBtllDliee »1
SAMPLE: AB,G,VER,00108.de,C.NA:NH,NA$
CONOS.: 1625A,3811,0.251111,5638,30-28086,156230,30CIVSJ
RANGE: G 1,3288 LABEL: N 2, 3.0 QUAN: A 2, 2,0 J
SCANS
1 TO 3288
8 BASE: U 20, 3
RIC
U
71577S.
see
7:55
ieee
15:50
1500
23:45
2608
31:40
2500
39:35
sees
47:30
SCAN
TIME
FIGURE 6 Chromatooram of Combined Acid/Base/Neutral Standard.
69
-------
REFERENCES
"Performance Tests for the Evaluation of
Computerized Gas Chromatography/Mass
Spectrometry Equipment and Laboratories"
USEPA, EMSL Cincinnati, Ohio 45268, EPA-
600/4-80-025 (April 1980).
National Standard Reference Data System,
"Mass Spectral Tape Format", US National
Bureau of Standards (1979 and later
attachments).
"Working with Carcinogens," DHEU, PHS,
CDC, NIOSH, Publication 77-206, (Aug
1977).
"OSHA Safety and Health Standards, General
Industry" OSHA 2206, 29 CFR 1910 (Jan
1976).
"Safety in Academic Chemistry
Laboratories," ACS Committee on Chemical
Safety (1979).
"Interlaboratory Validation of U. S.
Environmental Protection Agency Method
1625A, Addendum Report", SRI
International, Prepared for Analysis and
Evaluation Division (WH-557), USEPA, 401 M
St SW, Washington DC 20460 (January
1985).
Division, Washington, DC 20460 (June 15,
1984).
11 "Narrative for Episode 1036: Paragraph
4(c) Mass Spectra, Retention Times, and
Response Factors", U S Testing Co, Inc,
Prepared for W. A. Telliard, Industrial
Technology Division (WH-552), USEPA, 401 M
St SW, Washington DC 20460 (October 1985).
12 "Narrative for SAS 109: Analysis of
Extractable Organic Pollutant Standards by
Isotope Dilution GC/MS", S-CU8ED Division
of Maxwell Laboratories, Inc., Prepared
for W. A. Telliard, Industrial Technology
Division (WH-552), USEPA, 401 M St SW,
Washington DC 20460 (July 1986).
13 Colby, Bruce N. and Ryan, Philip W.,
"Initial Evaluation of Methods 1634 and
1635 for the analysis of Municipal
Wasteuater Treatment Sludges by Isotope,
Dilution GCMS", Pacific Analytical Inc.,
Prepared for W. A. Telliard, Industrial
Technology Division (WH-552), USEPA, 401 M
St SW, Washington DC 20460 (July 1986).
7 "Handbook, of Analytical Quality Control in
Water and Wastewater Laboratories," USEPA,
EMSL, Cincinnati, OH 45268, EPA-600/4-79-
019 (March 1979).
8 "Standard Practice for Sampling Water,"
ASTM Annual Book of Standards, ASTM,
Philadelphia, PA, 76 (1980).
9 "Methods 330.4 and 330.5 for Total
Residual Chlorine," USEPA, EMSL,
Cincinnati, OH 45268, EPA 600/4-70-020
(March 1979).
10 "Inter-laboratory Validation of US
Environmental Protection Agency Method
1625," USEPA, Effluent Guidelines
70
-------
Appendix A: Mass Spectra in the Form of Mass/Intensity Lists
555
m/z
42
61
75
105
556
m/z
51
139
557
m/z
40
51
63
91
558
m/z
40
53
65
80
108
•559
m/z
41
77
163
319
560
m/z
74
101
202
561
m/z
40
51
62
71
111
562
m/z
45
77
563
m/z
74
108
216
acetophenone
int.
21
13
36
1000
m/z
43
62
76
106
int.
245
26
62
87
m/z
49
63
77
120
int.
19
422
941
479
m/z
50
65
78
121
int.
221
31
11
38
m/z
51
73
89
int.
524
13
12
m/Z
52
74
91
int.
75
64
22
4-aminobiphenyl
int.
55
65
aniline
int.
65
47
59
10
m/z
63
141
m/z
41
52
64
92
int.
65
132
int.
66
54
33
136
m/z
72
167
m/z
42
53
65
93
int.
82
163
int.
16
12
226
1000
m/z
83
168
m/z
46
54
66
94
int.
73
280
int.
11
40
461
73
m/z
85
169
m/z
47
61
74
int.
163
1000
int.
75
17
11
m/z
115
170
m/z
50
62
78
int.
142
216
int.
40
28
14
o-anisidine
int.
22
286
142
915
1000
arami te
int.
606
155
143
270
m/z
41
54
66
81
109
m/z
57
91
175
334
int.
43
39
20
41
55
int.
758
339
182
137
m/z
42
61
76
92
122
m/z
59
105
185
int.
10
12
13
47
123
int.
328
153
1000
m/z
50
62
77
93
844
m/z
63
107
187
int.
60
25
36
14
124
int.
782
239
328
m/z
51
63
68
94
56
m/z
65
121
191
int.
106
43
32
18
int.
285
107
346
m/z
52
64
79
105
m/z
74
123
197
int.
202
24
25
18
int.
113
120
191
benzanthrone
int.
69
278
762
m/z
75
150
203
int.
71
58
126
m/z
87
174
230
int.
97
67
1000
m/z
88
199
231
int.
160
63
177
m/z
99
200
int.
69
350
m/z
100
201
int.
215
236
1,3-benzenediol
int.
64
54
27
16
51
m/z
41
52
63
81
int.
19
29
74
201
m/z
52
53
64
82
int.
42
184
61
251
m/z
43
54
65
95
int.
36
89
13
13
m/z
49
55
68
109
int.
11
97
56
11
m/z
50
61
69
110
int.
43
15
119
1000
benzenethiol
int.
128
161
m/z
50
84
int.
149
259
m/z
51
109
int.
205
316
m/z
65
110
int.
175
1000
m/z
66
111
int.
505
102
m/z
69
int.
114
2,3- benzof I uorene
int.
52
491
1000
m/z
81
187
217
int.
69
75
166
m/z
94
189
int.
143
90
m/z
95
213
int.
253
233
m/z
106
214
int.
60
60
m/z
107
215
int.
205
987
71
-------
943
01/Z
45
75
564
m/z
40
61
75
89
108
565
m/z
49
76
566
m/z
49
76
567
m/z
49
63
76
126
568
m/z
50
79
143
569
m/z
41
91
129
570
m/z
50
85
571
m/z
50
89
944
m/z
50
80
572
m/z
40
105
benzole acid
int.
29
25
m/z
50
76
int.
221
81
m/z
51
77
int.
413
778
m/z
52
78
int.
45
76
m/z
66
105
int.
11
1000
m/z
74
122
int.
53
868
benzyl alcohol
int.
17
11
13
65
737
m/z
59
62
76
90
109
int.
16
31
18
64
43
m/z
50
63
77
91
int.
155
70
565
125
m/z
51
64
78
105
int.
319
12
116
38
m/z
52
65
79
106
int.
78
75
1000
18
m/z
53
74
80
107
int.
84
35
73
523
2 - bromoch I orobenzene
int.
237
202
m/z
50
111
int.
890
961
m/z
51
113
int.
183
287
m/z
73
190
int.
158
638
m/z
74
192
int.
506
809
m/z
75
194
int.
1000
193
3 - bromoch I orobenzene
int.
201
197
m/z
50
111
int.
834
1000
m/z
51
113
int.
174
301
m/z
73
190
int.
169
625
m/z
74
192
int.
509
802
m/z
75
194
int.
914
191
4-chloro-2-nitroaniline
int.
119
1000
127
766
m/z
50
64
78
128
int.
174
315
152
234
m/z
51
65
90
142
int.
260
192
724
211
m/z
52
73
91
172
int.
531
290
253
915
m/z
61
74
101
174
int.
205
105
232
289
m/z
62
75
114
int.
394
156
312
5-chloro-o-toluidine
int.
115
140
313
m/z
51
89
int.
261
152
m/z
52
106
int.
257
1000
m/z
53
140
int.
137
599
m/z
77
141
int.
420
964
m/z
78
142
int.
134
265
4-chloroaniline
int.
60
63
292
m/z
62
92
int.
55
186
m/z
63
99
int.
147
67
m/z
64
100
int.
135
115
m/z
65
127
int.
329
1000
m/z
73
128
int.
51
81
3-chloronit r obenzene
int.
619
101
o-cresol
int.
102
114
p-cresol
int.
136
145
m/z
51
99
m/z
51
90
m/z
51
90
int.
189
258
int.
181
231
int.
224
122
m/z
73
111
m/z
53
107
m/z
52
107
int.
144
851
int.
144
783
int.
106
822
m/z
74
113
m/z
77
108
m/z
53
108
int.
330
266
int.
358
1000
int.
196
1000
m/z
75
157
m/z
79
m/z
77
int.
1000
424
int.
380
int.
420
m/z
76
159
m/z
80
m/i
79
int.
169
137
int.
159
int.
308
crotoxyphos
int.
633
484
m/z
44
109
int.
448
21
m/z
67
127
int.
42
1000
m/z
77
166
int.
70
180
m/z
79
193
int.
41
401
m/z
104
194
int.
100
20
72
-------
573 2,6-di-t-butyl-p-benzoquinone
m/z
51
77
135
220
574
m/z
40
67
105
575
m/z
42
77
106
159
945
m/z
53
170
576
m/z
41
65
133
577
m/z
40
49
78
578
m/z
52
73
163
579
m/z
49
74
110
161
946
m/z
49
126
580
m/z
40
57
int. m/z int. m/z
392 53 586 55
376 79 308 91
538 136 240 149
410
2,4-diaminototuene
int. m/z int. m/z
70 42 55 51
50 77 147 78
134 106 67 121
1 ,2-dibromo-3-chloropropane
int. m/z int. m/z
38 59 341 51
331 81 43 93
17 119 -74 121
204 187 10
int.
325
456
429
int.
76
69
958
int.
104
117
66
m/z
57
95
163
m/z
52
93
122
m/z
61
95
155
int.
668
322
292
int.
70
63
1000
int.
38
106
635
m/z
65
107
177
m/z
53
94
123
m/z
75
97
157
int.
416
248
1000
int.
51
224
79
int.
1000
12
784
m/z
67
121
205
m/z
61
104
m/z
76
105
158
int.
927
255
203
int.
91
128
int.
75
67
20
3,5-dibromo-4-hydroxybenzonitrile
int. m/z int. m/z
148 61 193 62
141 275 489 277
2,6-dichloro-4-ni troani I ine
int. m/z int. m/z
206 52 1000 61
137 89 218 90
218 160 401 176
1,3-dichtoro-2-propanol
int. m/z int. m/z
14 42 55 43
113 50 15 51
11 79 1000 80
2,3-dichloroani line
int. m/z int. m/z
138 61 151 62
130 90 460 99
626 165 101
2,3-dichloronitrobenzene
int. m/z int. m/z
220 50 257 61
976 75 743 84
204 111 303 133
190 163 121 191
2,6-dichlorophenol
int. m/z int. m/z
111 62 160 "^?
260 162 1000 164
1 , 2:3, 4-di epoxybutane
int. m/z int. m/z
37 41 29 42
155 58 16 85
int.
222
1000
int.
523
443
431
int.
503
37
25
int.
265
202
int.
150
351
701
411
int.
714
613
int.
83
13
m/z
88
279
m/z
62
97
178
m/z
44
57
81
m/z
63
125
m/z
62
85
135
193
m/z
73
166
m/z
43
int.
632
451
int.
828
458
134
int.
22
10
310
int.
455
108
int.
120
166
435
263
in*.
132
101
int.
60
m/z
117
m/z
63
124
206
m/z
47
61
m/z
64
126
m/z
63
86
145
m/z
" 98
m/z
55
int.
137
int.
588
954
378
int.
\2
12
int.
142
149
int.
173
125
580
int.
293
int.
1000
m/z
168
m/z
73
126
m/z
58
75
m/z
65
161
m/z
73
109
147
m/z
99
m/z
56
int.
152
int.
470
401
int.
15
14
int.
105
1000
int.
336
1000
368
int.
117
int.
67
73
-------
581
m/z
65
122
245
582
m/z
44
63
96
583
m/z
42
104
584
m/z
101
125
237
252
585
m/z
40
57
586
tn/z
76
190
587
m/z
50
76
588
m/z
50
110
589
m/z
42
64
97
590
m/z
41
73
591
m/z
41
160
310
592
m/z
47
141
3,3' -dimethoxybenzidine
int. m/z int. m/z
44 79 222 85
115 158 154 186
152
dimethyl sulfone
int. m/z int. m/z
10 45 94 46
69 64 22 65
23
p-dimethylaminoazobenzene
int. m/z int. m/z
483 51 181 77
142 105 190 120
int.
69
144
int.
29
19
int.
447
1000
m/z
93
201
m/z
47
79
m/z
78
148
int.
84
552
int.
18
1000
int.
120
160
m/z
107
229
m/z
48
81
m/z
79
225
int.
46
162
int.
69
36
int.
147
676
m/z
115
244
m/z
62
94
m/z
91
int.
110
1000
int.
14
528
int.
109
7, 12-dimethylbenzo(a)anthracene
int. m/z int. m/z
24 112 34 113
46 126 81 127
23 239 313 240
68 253 33 255
N,N-dimethylformamide
int. m/z int. m/z
58 41 79 42
17 58 83 72
3,6-dimethylphenanthrene
int. m/z int. m/z
113 89 129 94
193 191 430 205
1,4-dinitrobenzene
int. m/z int. m/z
1000 51 131 63
664 92 240 122
diphenyldisulf ide
int. m/z int. m/z
153 51 293 65
132 154 191 185
ethyl methanesulfonate
int. m/z int. m/z
16 43 72 45
22 65 93 79
206 109 579 111
ethylenethiourea
int. m/z int. m/z
46 42 126 45
151 102 1000
ethynylestradiol 3-methyl
int. m/z int. m/z
155 53 101 91
115 173 199 174
516
hexach I oropropene
int. m/z int. m/z
131 71 333 106
206 143 196 211
int.
112
60
230
84
int.
497
89
int.
179
246
int.
228
166
int.
671
117
int.
208
1000
18
int.
97
ether
int.
157
313
int.
334
631
m/z
114
128
241
256
m/z
43
73
m/z
101
206
m/z
64
168
m/z
59
218
m/z
48
80
123
m/z
46
m/z
115
227
m/z
108
213
int.
38
76
433
1000
int.
115
994
int.
142
1000
int.
218
399
int.
282
418
int.
40
127
15
int.
42
int.
143
1000
int.
200
1000
m/z
119
215
242
257
m/z
44
74
m/z
102
207
m/z
74
m/z
77
m/z
59
81
124
m/z
59
m/z
147
228
m/z
117
215
int.
212
24
61
180
int.
1000
35
int.
151
159
int.
311
int.
141
int.
19
42
33
int.
U
int.
226
149
int.
329
623
m/z
120
226
250
m/z
45
m/z
189
m/z
75
m/z
109
m/z
63
96
m/z
72
m/z
159
242
m/z
119
217
int.
296
47
32
int.
19
int.
388
int.
623
int.
1000
int.
23
16
int.
89
int.
132
153
int.
320
186
-------
947
m/z
41
56
73
593
m/z
51
128
170
594
m/z
50
104
595
m/z
53
91
119
596
m/z
118
237
597
m/z
42
78
598
m/z
45
65
95
599
m/z
45
82
900
m/z
113
134
266
901
m/z
77
195
267
902
m/z
50
87
189
hexanoic acid
int.
627
90
412
m/z
42
57
74
int.
535
102
56
m/z
43
60
87
int.
214
1000
98
m/z
45
61
int.
186
66
m/z
46
69
int.
19
21
m/z
55
70
int.
128
20
2-isopropylnaphthalene
int.
100
216
368
m/z
63
152
int.
111
133
m/z
76
153
int.
157
184
m/z
77
154
int.
129
114
m/z
115
155
int.
147
1000
m/z
127
156
int.
131
139
isosafrole
int.
110
441
m/z
51
131
int.
222
371
m/z
63
132
int.
127
107
m/z
77
135
int.
277
129
m/z
78
161
int.
208
250
m/z
103
162
int.
355
1000
long if dene
int.
438
1000
394
m/z
55
93
133
int.
719
611
338
m/z
65
94
161
int.
346
546
568
m/z
67
95
204
int.
453
404
172
m/z
77
105
int.
566
614
m/z
69
107
int.
713
475
malachite green
int.
113
158
m/z
126
253
int.
313
1000
m/z
165
254
int.
369
160
m/z
208
329
int.
135
189
m/z
209
330
int.
233
775
m/z
210
331
int.
181
170
methapyriline
int.
72
54
methyl
int.
178
285
137
m/z
45
79
int.
47
48
m/z
53
97
int.
40
516
m/z
58
190
int.
1000
40
m/z
71
191
int.
188
67
m/z
72
int.
225
methanesulfonate
m/z
56
78
109
int.
15
27
59
m/z
48
79
110
int.
108
821
60
m/z
50
80
int.
26
1000
m/z
63
81
int.
35
44
m/z
64
82
int.
48
33
2-methylbenzothiozole
int.
152
204
m/z
50
108
int.
133
392
m/z
58
109
int.
153
102
m/z
62
148
int.
106
279
m/z
63
149
int.
309
1000
m/z
69
150
int.
513
110
3-methylcholanthrene
int.
58
160
50
m/z
119
250
267
int.
55
56
192
m/z
125
252
268
int.
83
322
1000
m/z
126
253
269
int.
305
271
185
m/z
132
263
int.
99
59
m/z
133
265
int.
122
106
4,4'-methylenebis(2-chloroaniline)
int.
190
352
144
m/z
84
229
268
int.
107
228
358
m/z
98
231
int.
299
1000
m/z
104
233
int.
133
227
m/z
115
265
int.
226
171
m/z
140
266
int.
316
631
4,5-methylenephenanthrene
int.
50
60
900
m/z
62
94
190
int.
55
255
1000
m/z
63
95
int.
95
659
m/z
74
163
int.
69
80
m/z
81
187
int.
145
213
m/z
86
188
int.
53
137
75
-------
903
m/z
50
76
139
166
181
904
m/z
50
65
76
114
141
905
m/z
51
96
193
906
m/z
45
136
907
m/z
51
130
908
m/z
50
76
158
909
m/z
50
65
115
910
m/z
51
94
911
m/z
41
63
92
91 2
m/z
41
65
108
1 -methyl f luorene
int. m/z fnt.
66 51 87
196 83 135
54 151 73
136 176 96
99
2-methy (.naphthalene
int. m/z int.
29 51 39
19 69 56
14 77 15
13 115 303
748 142 1000
1 -methylphenanthrene
int. m/z int.
54 63 86
132 163 55
152
m/z
62
87
152
177
m/z
57
70
86
116
143
m/z
70
165
int.
57
53
124
52
int.
28
25
13
25
105
int.
62
217
m/z
63
88
163
178
m/z
58
71
87
126
m/z
74
189
int.
137
78
57
202
int.
47
126
18
13
int.
51
165
m/z
74
89
164
179
m/z
62
74
89
139
m/z
81
191
int.
64
203
58
182
int.
26
25
42
98
int.
52
532
m/z
75
90
165
180
m/z
63
75
113
140
m/z
83
192
int.
85
58
1000
686
int.
65
23
19
24
int.
164
1000
2-(methylthio)benzothiazole
int. m/z int.
790 50 212
239 148 938
m/z
63
180
int.
383
250
m/z
69
181
int.
578
1000
m/z
82
int.
233
m/z
108
int.
627
1 ,5-naphthalenecliamine
int. m/z int.
48 65 83
262 131 40
1 ,4-naphthoquinon*
int. m/z Int.
445 51 62
590 101 51
1000 159 100
alpha-naphthylamine
int. m/z int.
25 51 31
27 71 58
401 116 212
5-nitro-o-toluidine
int. m/z fnt.
194 52 159
168 104 120
2-nitroaniline
int. m/z int.
64 50 51
181 64 155
566 108 170
3-nitPoanfline
int. m/z int.
101 52 120
1000 66 114
87 138 717
m/z
77
141
m/z
52
102
m/z
57
72
142
m/z
53
106
m/z
51
65
138
m/z
53
80
139
int.
75
43
int.
52
613
int.
36
104
53
int.
121
691
int.
89
960
1000
int.
59
169
51
m/z
79
157
m/z
66
103
m/z
59
89
143
m/z
77
152
m/z
52
66
139
m/z
62
91
int.
111
89
int.
69
52
int.
46
62
1000
fnt.
766
1000
int.
207
96
63
int.
58
62
m/z
103
158
m/z
74
104
m/z
62
113
144
m/z
78
m/z
53
80
m/z
63
92
int.
86
1000
int.
189
550
int.
28
22
101
int.
176
int.
74
212
int.
143
764
m/z
118
159
m/z
75
130
m/z
63
114
m/z
79
m/z
62
91
m/z
64
93
int.
52
117
int.
205
433
int.
59
34
int.
619
int.
58
86
int.
121
62
76
-------
913
m/z
52
66
914
m/z
51
152
915
m/z
41
57
158
916
m/z
41
56
102
917
m/z
40
57
918
m/z
50
79
919
m/z
41
56
920
m/z
41
54
83
921
m/z
73
217
922
m/z
47
95
165
923
m/z
51
91
147
924
m/z
74
126
252
4-nitroaniline
int. m/z int.
228 53 160
124 80 266
4-nitrobiphenyl
int. m/z int.
131 63 104
902 153 284
m/z
62
92
m/z
76
169
int.
110
300
int.
179
374
m/z
63
108
m/z
115
199
int.
216
636
int.
134
1000
m/z
64
138
m/z
141
200
int.
164
520
int.
277
125
m/z
65
m/z
151
int.
1000
int.
259
N-nitroso-di-n-butylamine
int. m/z int.
1000 42 536
994 84 985
161
N-nitrosodiethytamine
int. m/z int.
170 42 079
525 57 492
807 103 35
m/z
43
86
m/z
43
70
int.
570
103
int.
69
24
m/z
44
99
m/z
44
71
int.
313
197
int.
1000
28
m/z
55
115
m/z
45
85
int.
129
158
int.
20
25
m/z
56
116
m/z
54
87
int.
167
237
int.
18
31
N-nitrosomethylethylamine
int. m/z int.
117 42 1000
99 59 13
m/z
43
71
int.
667
60
m/z
44
73
int.
26
57
m/z
54
88
int.
17
772
m/z
56
89
int.
189
20
N-nitrosomethylphenylaffline
int. m/z int.
181 51 434
331 104 147
N - ni t rosomorpho I i ne
int. m/z int.
181 42 192
1000 57 49
N-nitrosopiperidine
int. m/z int.
320 42 1000
58 55 444
28 84 47
pentach I orobenzene
int. m/z Int.
160 108 239
106 248 648
pentach I oroethane
int. m/z int.
203 60 398
165 117 1000
716 167 901
pent amethy I benzene
int. m/z int.
126 53 84
218 105 128
60 148 420
perytene
int. m/z int.
33 111 43
243 224 49
1000 253 219
m/z
52
106
m/z
43
85
m/z
43
56
114
m/z
125
250
m/z
62
119
169
m/z
63
115
m/z
112
248
int.
104
673
int.
52
13
int.
43
224
491
int.
102
1000
int.
119
979
422
Int.
61
120
int.
70
75
m/z
63
107
m/z
44
86
m/z
51
57
115
m/z
178
252
m/z
83
121
m/z
65
117
m/z
113
249
int.
110
220
int.
17
333
int.
14
17
26
int.
102
642
int.
378
306
int.
99
91
int.
111
52
m/z
77
212
m/z
54
87
m/z
52
67
m/z
213
254
m/z
85
130
m/z
77
133
m/z
124
250
int.
1000
137
int.
85
14
int.
12
21
int.
179
199
int.
218
293
int.
145
1000
int.
132
284
m/z
78
m/z
55
116
m/z
53
82
m/z
215
m/z
94
132
m/z
79
134
m/z
125
251
int.
194
int.
95
337
int.
32
26
int.
218
int.
114
272
int.
64
105
int.
251
86
77
-------
925
IH/Z
43
65
110
926
m/z
50
166
927
m/z
50
87
200
928
m/z
51
102
929
m/z
41
145
256
930
m/Z
40
53
78
931
m/z
50
104
163
932
m/z
53
79
109
933
m/z
47
84
181
948
m/z
61
97
196
934
m/z
45
69
135
phenacetin
int. m/z
443 51
47 79
50 137
phenothiazine
int. m/z
145 51
240 167
int.
33
31
461
int.
120
607
m/z
52
80
138
m/z
63
198
int.
112
179
40
int.
134
186
m/z
53
31
179
m/z
69
199
int.
164
154
672
int.
190
1000
m/z
63
108
180
m/z
100
200
int.
39
1000
64
int.
128
143
m/z
64
109
m/z
154
int.
30
196
int.
149
1 -phenylnaphthalene
int. m/z
132 51
101 88
144 201
int.
156
183
136
m/z
63
89
202
int.
148
162
643
m/z
74
100
203
int.
124
155
1000
m/z
75
101
204
int.
142
527
999
m/z
76
102
205
int.
136
111
159
2 -phenylnaphthalene
int. m/z
108 63
188 202
prooam[de
int. . m/z
270 66
334 147
102 257
pyridine
int. m/z
45 48
112 54
151 79
safrole
int. m/z
132 51
477 105
109
squalene
int. m/z
62 55
43 81
47 121
int.
101
398
int.
109
198
122
int.
11
12
1000
int.
369
130
int.
94
465
46
m/z
76
203
m/z
74
173
m/z
49
55
80
m/z
63
131
m/z
67
82
137
int.
136
270
int.
112
1000
int.
62
16
101
int.
108
437
int.
105
52
41
m/z
88
204
m/z
75
175
m/z
50
75
81
m/z
77
132
m/z
68
93
int.
133
1000
int.
137
615
int.
324
21
58
int.
391
166
int.
119
70
m/z
89
205
m/z
84
254
m/z
51
76
m/z
78
161
m/z
69
95
int.
158
157
int.
194
133
int.
414
19
int.
228
298
int.
1000
104
m/z
101
m/z
109
255
m/z
52
77
m/z
103
162
m/z
70
107
int.
333
int.
186
211
int.
879
22
int.
348
1000
int.
57
43
1 , 2 , 4 , 5 - tet rach 1 orobenzene
int. m/z
125 49
197 108
224 214
int.
176
284
791
m/z
61
109
216
int.
127
231
1000
m/z
72
143
218
int.
183
194
482
m/z
73
145
220
int.
332
117
101
m/z
74
179
int.
448
237
2,3,4,6-tetrachlorophenot
int. m/z
234 65
107 131
164 230
thianaphthene
int. m/z
80 50
139 74
104 136
int.
167
463
793
int.
91
55
52
m/z
66
133
232
m/z
51
89
int.
105
270
1000
int.
65
191
m/z
83
166
234
m/z
62
90
int.
134
298
471
int.
82
136
m/z
84
168
m/z
63
108
int.
178
273
int.
162
82
m/z
96
194
m/z
67
134
int.
202
168
int.
78
1000
78
-------
935
m/z
40
59
936
m/z
50
92
185
937
m/z
40
52
65
78
104
938
m/z
50
67
107
939
m/z
41
79
120
940
m/z
74
114
227
941
m/z
45
59
103
942
m/z
46
73
thio«cet*iide
int.
225
165
M/Z
42
60
int.
485
437
m/Z
43
75
int.
44
1000
m/z
46
76
int.
18
25
m/z
57
77
int.
36
43
m/z
58
int.
93
thiox«nthone
int.
262
188
137
m/z
63
108
212
int.
180
129
1000
m/z
69
139
213
int.
320
385
145
m/z
74
152
int.
116
227
m/z
69
183
int.
176
112
m/z
82
184
int.
12V
951
o-toluidine
int.
51
164
59
113
45
m/z
41
53
66
79
106
int.
38
192
24
243
1000
m/z
42
53
74
80
107
int.
35
86
19
80
90
m/z
49
62
65
89
int.
10
26
14
107
m/z
50
63
76
90
int.
88
68
21
76
m/z
51
64
77
91
int.
169
30
313
52
1 ,2,3-trimethoxybenzene
int.
257
114
190
m/z
51
77
108
int.
459
246
144
m/z
52
79
110
int.
139
132
898
m/z
53
82
125
int.
276
117
578
m/z
63
93
153
int.
112
483
759
m/z
65
95
168
int.
341
801
1000
2,4,5-trimethylaniline
int.
80
62
1000
m/z
52
91
121
int.
58
167
87
m/z
51
93
134
int.
63
51
670
m/z
53
117
135
int.
66
54
978
m/z
65
118
136
int.
150
65
99
m/z
67
119
int.
74
93
triphenylene
int.
52
181
132
m/z
87
200
228
tripropytene
int.
492
1000
57
m/z
46
60
117
int.
55
67
1000
glycol
int.
15
34
92
m/z
100
202
229
methyl
m/z
47
71
161
int.
107
56
184
ether
int.
19
16
21
m/z
101
224
m/z
55
72
int.
108
84
int.
17
44
m/z
112
225
m/z
57
73
int.
131
56
int.
68
363
m/z
113
226
m/z
58
74
int.
244
313
int.
43
232
1.3,5-trithi«ne
int.
1000
102
m/z
47
91
int.
150
92
m/z
48
92
int.
98
111
m/z
59
110
int.
93
58
m/z
60
138
int.
76
259
m/z
64
int.
136
79
-------
EPA METHOD 1618
THE CONSOLIDATED GC METHOD FOR THE
DETERMINATION OF ITD/RCRA PESTICIDES
USING SELECTIVE GC DETECTORS
-------
Introduction
Method 1618 was developed by EPA's Office of Water Regulations
and Standards to provide improved precision and accuracy of
analysis of pollutants in aqueous and solid matrices.
Method 1618 is an automated, wide-bore capillary column gas
chromatography method for analysis of organo-halide and
organo-phosphorus pesticides and phenoxy-acid herbicides and
herbicide esters and other compounds amenable to extraction
and analysis by wide-bore capillary column gas chromatography
with halogen specific and organo-phosphorus detectors.
Questions concerning the Methods or their application should
be addressed to:
W. A. telliard
USEPA
Office of Water Regulations and Standards
401 M Street SW
Washington, DC 20460
202-382-7131
OR
USEPA OURS
Sample Control Center
P.O. Box 1407
Alexandria, Virginia 22313
703-557-5040
Publication date: May 1988
-------
Method 1618, 12 February 1988 Draft
Organo-halide and Organo-phosphorus Pesticides and Phenoxy-acid
Herbicides by Capillary Column Gas Chromatography
1 SCOPE AND APPLICATION
1.1 This method is designed to meet the survey
requirements of the Environmental
Protection Agency (EPA). It is used to
determine the organo-halide and organo-
phosphorus pesticides, and the phenoxy-
acid herbicides and herbicide esters
associated with the Clean Water Act; the
Resource Conservation and Recovery Act;
the Comprehensive Environmental Response,
Compensation and Liability Act; and other
compounds amenable to extraction and
analysis by automated, wide-bore capillary
column gas Chromatography (GO with
halogen specific and organo-phosphorus
detectors.
1.2 The chemical compounds listed in tables 1
through 3 may be determined in waters,
soils, sediments, and sludges by this
method. The method is a consolidation of
EPA Methods 608, 608.1, 614, 615, 617,
622, and 701. For waters, the sample
extraction and concentration steps are
essentially the same as in these methods.
However, the extraction and concentration
steps have been extended to other sample
matrices. The method should be applicable
to other pesticides and herbicides. The
quality assurance/quality control
requirements in this method give the steps
necessary to determine this applicability.
1.3 When this method is applied to analysis of
unfamiliar samples, compound identity
shall 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. Gas
chromatography-mass spectrometry (GCMS)
can be used to confirm compounds in
extracts produced by this method when
analyte levels are sufficient.
Table 1
CHLORINATED PESTICIDES DETERMINED BY
LARGE-BORE, FUSED-SILICA, CAPILLARY COLUMN
GAS CHROMATOGRAPHY WITH HALIDE SPECIFIC
DETECTOR
EPA
EGD
089
102
103
105
104
434
433
441
091
431
094
093
092
432
478
090
095
096
097
098
099
435
100
101
437
439
430
438
436
112
108
109
106
110
107
111
440
113
442
Compound
Aldrin
alpha-BHC
beta-BHC
delta-BHC
gamma-BHC (Lindane)
Captafol
Captan
Carbophenothion
Chlordane
Chlorobenzilate
4,4'-DDD
4,4'-DDE
4, 4' -DDT
Dial late
Dichlone
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Heptachlor
Heptachlor epoxide
Isodrin
Kepone
Methoxychlor
Mi rex
Nitrofen (TOO
PCB-1016
PCB-1221
PCB-1232
PCS -1242
PCS -1248
PCS- 1254
PCB-1260
PCNB (pentachloro-
nitrobenzene)
Toxaphene
Trif luralin
CAS Registry
309-00-2
319-84-6
319-85-7
319-86-8
58-89-9
2425-06-1
133-06-2
786-19-6
57-74-9
510-15-6
72-54-8
72-55-9
50-29-3
2303-16-4
117-80-6
60-57-1
959-98-8
33213-65-9
1031-07-8
72-20-8
7421-93-4
53494-70-5
76-44-8
1024-57-3
465-73-6
143-50-0
72-43-5
2385-85-5
1836-75-5
12674-11-2
11104-28-2
11141-16-5
53469-21-9
12672-29-6
11097-69-1
11096-82-5
82-68-8
8001-35-2
1582-09-8
82
-------
Table 1 (continued)
Non-ITD organo-halide compounds
Compound
CAS Registry
Chloroneb
Chloropropylate
OBCP
Dicofol
Etridiazole
Perthane (Ethylan)
Propachlor
Strobane
2675-77-6
5836-10-2
96-12-8
115-32-2
2593-15-9
72-56-0
1918-16-7
8001-50-1
Table 2
PHOSPHORUS PESTICIDES DETERMINED BY LARGE-
BORE, FUSED-SILICA, CAPILLARY COLUMN GAS
CHROMATOGRAPHY WITH FLAME PHOTOMETRIC
DETECTOR
EPA
EGD
Compound
CAS Registry
468 Azinphos ethyl 2642-71-9
453 Azinphos methyl 86-50-0
461 Chlorfevinphos 470-90-6
469 Chlorpyrifos 2921-88-2
443 Coumaphos 56-72-4
479 Crotoxyphos 7700-17-6
471 Dane ton 8065-48-3
460 Diazinon 333-41-5
450 Dichlorvos 62-73-7
455 Dicrotophos 141-66-2
449 Dimethoate 60-51-5
452 Dioxathion 78-34-2
458 Disulfoton 298-04-4
467 EPN 2104-64-5
463 Ethion 563-12-2
446 Famphur 52-85-7
454 Fensulfothion 115-90-2
447 Fenthion 55-38-9
464 Hexamethylphosphoramide 680-31-9
474 Leptophos 21609-90-5
475 Malathion 121-75-5
456 Methyl parathion 298-00-0
444 Mevinphos 7786-34-7
470 Monocrotophos 6923-22-4
459 Naled 300-76-5
448 Parathion 56-38-2
457 Phorate 298-02-2
465 Phosmet
473 Phosphamidon
477 Sulfotepp
476 TEPP
472 Terbufos
466 Tetrachloryinphos
445 Trichlorofon
451 Tricresylphosphate
462 Trimethylphosphate
Non-ITD thiophosphate compounds
732-11-6
13171-21-6
3689-24-5
107-40-3
13071-79-9
961-11-5
42-68-6
78-30-8
512-56-1
Compound CAS Registry
Bolstar
Dichlorofenthion
Ethoprop
Merphos
Methyl chlorpyrifos
Methyl trithion
Ronnel
Tokuthton
Trichloronate
35400-43-2
97-17-6
13194-48-4
150-50-5
5598-13-0
299-84-3
34643-46-4
327-98-0
Table 3
PHENOXYACID HERBICIDES DETERMINED BY
LARGE-BORE, FUSED-SILICA, CAPILLARY
COLUMN GAS CHROMATOGRAPHY WITH
ELECTRONEGATIVE DETECTOR
EPA
EGD
481
480
482
483
Compound
2,4-D
D i noseb
2.4.5-T
2,4,5-TP
CAS Registry
94-75-7
88-85-7
93-76-5
93-72-1
Non-ITD phenoxyacid herbicides
Compound CAS Registry
Dalapon
2,4-DB
-------
1.4 The detection limit of this method is
usually dependent on the level of
interferences rather than instrumental
limitations. The limits in tables 4-5
typify the minimum quantity that can be
detected with no interferences present.
1.5 This method is for use by or under the
supervision of analysts experienced in the
use of a gas chromatograph and in the
interpretation of gas chromatograph ic
data. Each laboratory that uses this
method must demonstrate the ability to
generate acceptable results using the
procedure in section 8.2.
adsorption chromatography and concentrated
to one mL.
2.3 Gas chromatography--a one uL aliquot of
the extract is injected into the gas
chromatograph (GO. The compounds are
separated on a wide-bore, fused silica
capillary column. The organo-halide com
pounds, including the derivatized phenoxy-
acid herbicides, are detected by an
electron capture, microcoulometric, or
electrolytic conductivity detector. The
phosphorus containing compounds are
detected using a flame photometric
detector.
2 SUMMARY OF METHOD
2.1 Extract ion--the percent solids content of
a sample is determined. If the solids
content is less than one percent, a one
liter sample is extracted with methylene
chloride using continuous extraction
techniques. If the solids content is 1 -
30 percent, the sample is diluted to one
percent solids with reagent water,
homogenized ultrasonically, and extracted
with methylene chloride using continuous
extraction techniques. If the solids
content is greater than 30 percent, the
sample is extracted with methylene
chloride:acetone using ultrasonic
techniques. Samples in which phenoxy-acid
herbicides are to be determined are
acidified prior to extraction.
2.2 Concentration and cleanup--for samples in
which pesticides are to be determined,
each extract is dried over sodium sulfate,
concentrated using a Kuderna-Oanish
evaporator, cleaned up (if necessary)
using gel permeation chromatography (GPC)
and/or adsorption chromatography, and re-
concentrated to one mL. Sulfur is removed
from the extract, if required. For
samples in which the herbicides are to be
determined, each extract is processed to
remove the acids and esters. The esters
are hydrolyzed, combined with the acids,
and derivatized to form the methyl esters.
The solution containing the methyl esters
is cleaned up (if necessary) using
2.4 Identification of a pollutant (qualitative
analysis) is performed by comparing the GC
retention times of the compound on two
dissimilar columns with the respective
retention times of an authentic standard.
Compound identity is confirmed when the
retention times agree within their
respective windows.
2.5 Quantitative analysis is performed by
using an authenic standard to produce a
calibration factor or calibration curve,
and using the calibration data to
determine the concentration of a pollutant
in the extract. The concentration in the
sample is calculated using the sample
weight or volume and the extract volume.
2.6 Quality is assured through reproducible
calibration and testing of the extraction
and GC systems.
3 CONTAMINATION AND INTERFERENCES
3.1 Solvents, reagents, glassware, and other
sample processing hardware may yield
artifacts and/or elevated baselines
causing misinterpretation of
chromatograms. All materials used in the
analysis shall be demonstrated to be free
from interferences under the conditions of
analysis by running method blanks as
described in section 8.5.
84
-------
Table 4
GAS CHROMATOGRAPHY OF ORGANO-HALIDE COMPOUNDS
EPA
EGD
089
102
103
105
104
434
433
441
091
431
481
094
093
092
432
478
090
480
095
096
097
098
099
435
100
101
437
439
430
438
436
112
108
109
106
110
107
111
440
482
483
113
442
Comoound
Aldrin
aLpha-BHC
beta-BHC
delta-BHC
gamma-BHC (Lindane)
Captafol
Captan
Carbophenothion
Chlordane
Chlorobenzilate
2,4-0
4,4'-DDD
4,4'-DDE
4,4'-DDT
Diattate
Oichlone
Dieldrin
Oinoseb
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Heptachlor
Heptachlor epoxide
Isodrin
Kepone
Methoxychlor
Hi rex
Nitrofen (TDK)
PCB-1016
PCB-1221
PCS- 1232
PCB-1242
PCB-1248
PCS- 1254
PCS -1260
PCNB
2,4,5-T
2,4,5-TP
Toxaphene
Trif luralin
DB-5 Column
Concentration (Cone) and Retention Time (RT)
Cone 1 RT 1 Cone 2 RT 2 Cone 3 RT 3 HDL (1)
(ug/mL) (min) (ug/mL) (mini (uq/mL) (min) (ug/L) (ug/kg)
19.77
13.77
14.74
15.93
15.01
31.26
22.03
28.44
26.49
20.84
26.99
24.70
29.01
13.57
24.88
30.28
23.54
26.49
28.77
26.02
27.48
31.25
18.14
21.69
21.19
28.04
32.17
34.49
25.99
15.24
26.95
25.78
12.95
85
-------
Table 4 (continued)
GAS CHROMATOGRAPHY OF ORGANO-HALIDE COMPOUNDS
EPA
EGD
089
102
103
105
104
434
433
441
091
431
481
094
093
092
432
478
090
480
095
096
097
098
099
435
100
101
437
439
430
438
436
112
108
109
106
110
107
111
440
482
483
113
442
Compound
Atdrin
alpha-BHC
beta-BHC
delta-BHC
gamma -BHC (Lindane)
Captafol
Captan
Carbophenoth i on
Chlordane
Chlorobenzi late
2,4-0
4,4'-DDO
4,4'-OOE
4,4'-OOT
Diallate
Dichlone
Dieldrin
Oinoseb
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Heptachlor
Heptachlor epoxide
Isodrin
Kepone
Methoxychlor
Mi rex
Nitrofen (TDK)
PCB-1016
PCB-1221
PCB-1232
PCB-1242
PCS -1248
PCS -1254
PCB-1260
PCNB
2.4,5-T
2,4,5-TP
Toxaphene
Trif luralin
SPB-608 Column
Concentration (Cone) and Retention Time (RT)
Cone 1 RT 1 Cone 2 RT 2 Cone 3 RT 3 HDL (1)
(ufl/mL) (min) (uq/mL) (min) (uq/mL) (min) (ug/L) (ug/kg)
18.33
13.70
15.04
17.15
15.22
26.83
24.24
28.69
26.03
22.91
26.79
24.16
28.75
12.89
24.35
26.25
22.81
27.15
29.41
26.11
28.82
33.27
16.87
21.01
20.33
26.28
33.37
33.59
26.35
14.78
29.13
29.83
11.01
86
-------
Table 4 (continued)
GAS CHROHATOGRAPHY OF ORGANO-HALIDE COMPOUNDS
EPA
EGO
089
102
103
105
104
434
433
441
091
431
481
094
093
092
432
478
090
480
095
096
097
098
099
435
100
101
437
439
430
438
436
112
108
109
106
110
107
111
440
482
483
113
442
Compound
Aldrin
alpha-BHC
beta-BHC
delta-BHC
gamma-BHC (Lindane)
Captafol
Captan
Carbophenothion
Chlordane
Chlorobenzi late
2.4-0
4,4'-DDD
4,4'-OOE
4,4'-ODT
Diallate
Dichlone
Dieldrin
Dinoseb
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Heptachlor
Heptachlor epoxide
Isodrin
Kepone
Methoxychlor
Hi rex
Nitrofen (TOK)
PCB-1016
PCB-1221
PCB-1232
PCB-1242
PCS -1248
PCB-1254
PCB-1260
PCNB
2,4,5-T
2,4,5-TP
Toxaphene
Trifluralin
DB-608 Column
Concentration (Cone) and Retention Time (RT)
Cone 1 RT 1 Cone 2 RT 2 Cone 3 RT 3 HDL (1)
(ug/mL) (min) (ug/mL) (min) (ug/mL) (min) (ug/L) (ug/kg)
18.33
13.70
15.04
17.15
15.22
26.83
24.24
28.69
26.03
22.91
26.79
24.16
28.75
12.89
24.35
26.25
22.81
27.15
29.41
26.11
28.82
33.27
16.87
21.01
20.33
26.28
33.37
33.59
26.35
14.78
29.13
29.83
11.01
Column: 30 +/- 2 m x 0.50 +/- 0.05 mn i.d.
Temperature program: 1 min at 50 oC; 50 - 280 at 5 oC per min; 5 minute
hold at 250 oC
Gas velocity: 30 +/- 5 cm/sec at 30 oC
87
-------
Table 5
CALIBRATION OF THIOPHOSPHATE, COMPOUNDS ON DB-5 COLUMN
Concentration (Cone) and Retention Time (RT)
EPA
EGO
Cone 1
Compound (nq/mL)
RT 1 Cone 2
(mini (ng/mL)
RT 2 Cone 3
(min) (ng/mL)
RT 3 MOL (1)
(min) (ug/L) fug/kg)
Calibration Group #1
450
470
449
458
447
453
Dichlorvos
Monocrotophos
Oimethoate
Disulfoton
Methyl chtorpyrifos
Fenthion
Merphos
Tokuthion
Bolstar (Sulprofos)
Azinphos methyl
50
500
50
50
50
50
50
50
50
100
9.91
21.61
23.73
38.38
33.01
34.98
36.04
37.52
39.80
45.67
100
1000
100
100
100
100
100
100
100
200
9.90
21.66
23.71
28.34
32.99
34.97
36.03
37.51
39.79
45.64
1000
2000
1000
1000
1000
1000
1000
1000
1000
1000
9.91
21.54
23.69
28.38
33.01
34.98
36.03
37.52
39.80
45.66
Calibration Group #2
445
455
471
469
461
446
474
Trichlorofon
Dicrot optics
Demeton
Dichlorofenthion
Ronnel
Chlorpyrifos
Chlorfevinphos
Methyl trithion
(Carbofenthion-methyl)
Famphur
Leptophos
100
200
100
50
50
50
50
50
50
50
9.94
21.23
23.70
32.50
33.80
35.08
36.20
38.77
40.15
45.91
200
400
400
100
100
100
100
100
100
100
9.93
21.26
23.70
32.49
33.80
35.07
36.20
38.77
40.14
45.91
1000
1000
1000
•1000
1000
iooo
1000
1000
1000
1000
9.93
21.19
23.70
32.51
33.81
35.08
36.21
38.78
40.15
45.92
Calibration Group #3
444
459
477
472
473
452
448
479
454
465
468
Mevinphos
Naled
Sulfotepp
Terbufos
Phosphamidon
Dioxathion
Parathion
Crotoxyphos
Fensulfothion
Phosmet
Azinphos ethyl
50
100
50
50
200
400
100
100
100
100
100
14.23
20.71
21.78
26.35
32.43
34.03
35.05
36.37
38.90
43.08
48.32
100
200
100
100
600
800
200
200
200
200
200
14.21
20.67
21.74
26.30
32.40
34.00
35.03
36.35
38.88
43.05
48.27
1000
1000
1000
1000
1000
1000
1000
1000
1000
1000
1000
14.23
20.72
21.79
26.36
32.44
34.03
35.06
36.37
38.90
43.08
48.32
Calibration Group #4
457
460
456
475
466
463
467
443
Ethoprop
Phorate
Diazinon
Methyl parathion
Ma lath ion
Trichloronate
Tetrachlorvinphos
Ethion
EPN
Coumaphos
50
50
50
50
50
50
50
50
50
100
19.85
22.24
28.01
32.97
34.73
35.43
36.93
39.32
43.33
52.16
100
100
100
100
100
100
100
100
100
200
19.84
22.22
27.99
32.96
34.73
35.43
36.93
39.31
43.32
52.14
1000
1000
1000
1000
1000
1000
1000
1000
1000
1000
19.86
22.25
28.03
32.98
34.75
35.45
36.94
39.33
43.35
52.18
88
-------
3.2 Glassware and, where possible, reagents
are cleaned by solvent rinse and baking at
450 °C for one hour minimum in a muffle
furnace or kiln. Some thermally stable
materials, such as PCBs, may not be
eliminated by this treatment and thorough
rinsing with acetone and pesticide quality
hexane may be required.
3.3 Specific selection of reagents and
purification of solvents by distillation
in all-glass systems may be required.
3.4 Interference by phthalate esters can pose
a major problem in pesticide analysis when
using the electron capture detector.
Phthalates usually appear in the
chromatogram as large, late eluting peaks.
Phthalates may be leached from common
flexible plastic tubing and other plastic
materials during the extraction and clean-
up processes. Cross-contamination of
clean glassware routinely occurs when
plastics are handled during extraction,
especially when solvent wetted surfaces
are handled. Interferences from
phthalates can best be minimized by
avoiding the use of plastics in the
laboratory, or by using a microcoulometric
or electrolytic conductivity detector.
3.5 The acid forms of the herbicides are
strong acids that react readily with
alkaline substances and can be lost during
analysis. Glassware and glass wool must
be acid rinsed with dilute hydrochloric
acid and the sodium sulfate must be
acidified with sulfuric acid prior to use.
3.6 Organic acids and phenols cause the most
direct interference with the herbicides.
Alkaline hydrolysis and subsequent
extraction of the basic solution can
remove many hydrocarbons and esters that
may interfere with the herbicide analysis.
3.7 Interferences coextracted from samples
will vary considerably from source to
source, depending on the diversity of the
site being sampled. The cleanup
procedures given in this Method can be
used to overcome many of these
interferences, but unique samples may
require additional cleanup to achieve the
minimum levels given in tables 4-5.
4 SAFETY
4.1 The toxicity or carcinogenicity of each
compound or reagent used in this method
has not been precisely determined;
however, each chemical compound should be
treated as a potential health hazard.
Exposure to these compounds should be
reduced to the lowest possible level. 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 handling sheets
should also be made available to all
personnel involved in these analyses.
Additional information on laboratory
safety can be found in references 1-3.
4.2 The following compounds covered by this
method have been tentatively classified as
known or suspected human or mammalian
carcinogens: 4,4'-DDD, 4,4'-ODT, the BHCs
and the PCBs. Primary standards of these
compounds shall be prepared in a hood, and
a NIOSH/MESA approved toxic gas respirator
should be worn when high concentrations
are handled.
4.3 Diazomethane is a toxic carcinogen which
can decompose or explode under certain
conditions. Solutions decompose rapidly
in the presence of solid materials such as
copper powder, calcium chloride, and
boiling chips. The following operations
may cause explosion: heating above 90 °C;
use of grinding surfaces such as ground
glass joints, sleeve bearings, and glass
stirrers; and storage near alkali metals.
Diazomethane shall be used only behind a
safety screen in a well ventilated hood
and should be pipetted with mechanical
devices only.
4.4 Mercury vapor is highly toxic. If mercury
is used for sulfur removal, all operations
involving mercury shall be performed in a
hood.
89
-------
4.5 Unknown samples may contain high
concentrations of volatile toxic
compounds. Sample containers should be
opened in a hood and handled with gloves
that will prevent exposure. The oven used
for sample drying to determine percent
moisture should be located in a hood so
that vapors from samples do not create a
health hazard in the laboratory.
5 APPARATUS AND MATERIALS
5.1 Sampling equipment for discrete or
composite sampling.
5.1.1 Sample bottles and caps
5.1.1.1 Liquid samples (waters, sludges and
similar materials that contain less than
five percent solids)--sample bottle, amber
glass, 1 liter or 1 quart, with screw cap.
5.1.1.2 Solid samples (soils, sediments, sludges,
filter cake, com post, and similar
materials that contain more than five
percent solids)--sample bottle, wide
mouth, amber glass, 500 ml minimum.
5.1.1.3 If amber bottles are not available,
samples shall be protected from light.
5.1.1.4 Bottle caps--threaded to fit sample
bottles. Caps shall be lined with Teflon.
5.1.1.5 Cleaning
5.1.1.5.1 Bottles are detergent water washed, then
solvent rinsed or baked at 450 °C for one
hour minimum before use.
5.1.1.5.2 Liners are detergent water washed, then
reagent water and solvent rinsed, and
baked at approx 200 °C for one hour
minimum prior to use.
5.1.2 Compositing equipment—automatic or manual
compositing system incorporating glass
containers cleaned per bottle cleaning
procedure above. Sample containers are
kept at 0 - 4 °C during sampling. Glass
or Teflon tubing only shall be used. If
the sampler uses a peristaltic pump, a
minimum length of compressible si Iicone
rubber tubing may be used in the pump
only. Before use, the tubing shall be
thoroughly rinsed with methanol, followed
by repeated rinsings with reagent water to
minimize sample contamination. An
integrating flow meter is used to collect
proportional composite samples.
5.2 Equipment for determining percent moisture
5.2.1 Oven, capable of being temperature
controlled at 110 t 5 °C.
5.2.2 Oessicator.
5.2.3 Crucibles, porcelain.
5.2.4 Weighing pans, aluminum.
5.3 Extraction equipment.
5.3.1 Equipment for ultrasonic extraction.
5.3.1.1 Sonic disrupter--375 watt with pulsing
capability and 1/2 or 3/4 in. disrupter
horn (Ultrasonics, Inc, Model 375C, or
equivalent).
5.3.1.2 Sonabox (or equivalent), for use with
disrupter.
5.3.2 Equipment for liquid-liquid extraction
5.3.2.1 Continuous liquid-liquid extractor—Teflon
or glass connecting joints and stopcocks
without lubrication, 1.5 - 2 liter
capacity (Hershberg-Uolf Extractor, Cal-
Glass, Costa Hesa, California, 1000 or
2000 mL continuous extractor, or
equivalent).
5.3.2.2 Round-bottom flask, 500 mL, with heating
mantle.
5.3.2.3 Condenser, Graham, to fit extractor.
5.3.2.4 pH meter, with combination glass
electrode.
5.3.2.5 pH paper, wide range (Hydrion Papers, or
equivalent).
90
-------
5.2.3 Separatory funnels--250, 500, and 1000 nl,
with Teflon stop cocks.
5.3.4 Filtration apparatus
5.3.4.1 Glass powder funnels--125 - 250 mL
5.3.4.2 Filter paper for above (Whatman 41, or
equivalent)
5.3.5 Beakers
5.3.5.1 1.5 - 2 liter, calibrated to one liter
5.3.5.2 400 - 500 mL
5.3.6 Spatulas—stainless steel or Teflon
5.3.7 Drying column--400 mm x 15 to 20 mm i.d.
Pyrex chromatographic column equipped with
coarse glass frit or glass wool plug.
5.3.7.1 Pyrex glass wool—solvent extracted or
baked at 450 °C for one hour minimum.
5.4 Evaporation/concentration apparatus
5.4.1 Kuderna-Danish (K-D) apparatus
5.4.1.1 Evaporation flask--500 ml (Kontes K-
570001-0500, or equivalent), attached to
concentrator tube with springs (Kontes K-
662750-0012).
5.4.1.2 Concentrator tube--10 ml, graduated
(Kontes K-570050-1025, or equivalent) with
calibration verified. Ground glass
stopper (size 19/22 joint) is used to
prevent evaporation of extracts.
5.4.1.3 Snyder column—three ball macro (Kontes K-
503000-0232, or equivalent).
5.4.1.4 Snyder column—two ball micro (Kontes K-
469002-0219, or equivalent).
5.4.1.5 Boiling chips
5.4.1.5.1 Glass or silicon carbide--approx 10/40
mesh, extracted with methylene chloride
and baked at 450 °C for one hr minimum.
5.4.1.5.2 Teflon (optional)--extracted with
methylene chloride.
5.4.2 Water bath—heated, with concentric ring
cover, capable of temperature control (± 2
°C), installed in a fume hood.
5.4.3 Nitrogen evaporation device—equipped with
heated bath that can be maintained at 35 -
40 °C (N-Evap, Organomation Associates,
Inc., or equivalent).
5.4.4 Sample vials--amber glass, 1 - 5 mL with
Teflon-lined screw or crimp cap, to fit GC
autosampler.
5.5 Balances
5.5.1 Analytical—capable of weighing 0.1 mg.
5.5.2 Top loading—capable of weighing 10 mg.
5.6 Apparatus for sample cleanup.
5.6.1 Automated gel permeation chroma tograph
(Analytical Biochemical Labs, Inc,
Columbia, MO, Model GPC Autoprep 1002, or
equivalent).
5.6.1.1 Column--600 - 700 mm x 25 mm i.d., packed
with 70 g of SX-3 Bio-beads (Bio-Rad
Laboratories, Richmond, CA, or
equivalent).
5.6.1.2 Syringe, 10 mL, with Luer fitting.
5.6.1.3 Syringe filter holder, stainless steel,
and glass fiber or Teflon filters (Gelman
4310, or equivalent).
5.6.1.4 UV detectors--254-mu, preparative or semi-
prep flow cell: (Isco, Inc., Type 6;
Schmadzu, 5 mm path length; Beckman-Altex
152W, 8 uL micro-prep flow cell, 2 nro
path; Pharmacia UV-1, 3 mm flow cell; LDC
'Milton-Roy UV-3, monitor #1203; or
equivalent).
5.6.2 Vacuum system for eluting cleanup
cartridges.
91
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5.6.2.1 Vacuum system--capable of achieving 0.1
bar (house vacuum, vacuum pump, or water
aspirator), with vacuum gauge.
5.6.2.2 VacElute Manifold (Anatytichem
International, or equivalent).
5.6.2.3 Vacuum trap—made from 500 mL sidearm
flask fitted with single hole rubber
stopper and glass tubing.
5.6.2.4 Rack for holding 10 ml volumetric flasks
in the manifold.
5.6.3 Chromatographic column--400 mm x 22 mm
i.d., with Teflon stop cock and coarse
frit (Kontes K-42054, or equivalent).
5.6.4 Sulfur removal tubes--40 - 50 ml bottle or
test tube with Teflon lined screw cap.
5.7 Centrifuge apparatus
5.7.1 Centrifuge—capable of rotating 500 ml
centrifuge bottles or 15 mL centrifuge
tubes at 5,000 rpm minimum.
5.7.2 Centrifuge bottles--500 ml, with screw
caps, to fit centrifuge.
5.7.3 Centrifuge tubes--12-15 mL, with screw
caps, to fit centrifuge.
5.7.3 Funnel, Buchner, 15 cm.
5.7.3.1 Flask, filter, for use with Buchner
funnel.
5.7.3.2 Filter paper, 15 cm (Whatman #41, or
equivalent).
5.8 Derivatization apparatus—Diazald kit with
clear seal joints for generation of
diazomethane (Aldrich Chemical Co.
Z10,025-0, or equivalent).
5.9 Miscellaneous glassware
5.9.1 Pipettes, glass, volumetric, 1.00, 5.00,
and 10.0 mL
5.9.2 Syringes, glass, with Luerlok tip, 0.1,
1.0 and 5.0 mL. Needles for syringes, two
inch, 22 gauge.
5.9.3 Volumetric flasks, 10.0, 25.0, and 50.0 mL
5.9.4
5.10
5.10.1
5.10.1.1
5.10.1.2
5.10.2
Scintillation vials, glass, 20 - 50 mL,
with Teflon-lined screw caps.
Gas chromatographs--two GCs shall be
employed. Both shall have split less or
on-column simultaneous automated injection
into separate capillary columns with a
halide specific detector or flame
photometric detector at the end of each
column, temperature program with
isothermal holds, data system capable of
recording simultaneous signals from the
two detectors, and shall meet all of the
performance specifications in section 12.
GC columns--bonded
capiIlary
phase fused silica
Primary--60 *5 m x 0.5 ± 0.05 mm i.d. 5%
phenyl, 94X methyl, 1X vinyl silicone (J &
U DR-5 Megabore, Supelco SP-5, or
equivalent).
Confirmatory--J&W DB-608, Supelco SPB-608,
or equivalent, with same dimensions as
primary column.
Data system--shall collect and record GC
data, store GC runs on magnetic disk or
tape, process GC data, compute peak areas,
store calibration data including retention
times and calibration factors, identify GC
peaks through retention times, compute
concentrations, and generate reports.
5.10.2.1 Data acquisition--GC data shall be
collected continuously throughout the
analysis and stored on a mass storage
device.
5.10.2.2 Calibration factors and calibration
curves--the data system shall be used to
record and maintain lists of calibration
factors, and multi-point calibration
curves (section 7). Computations of
relative standard deviation (coefficient
92
-------
of variation) are used for testing
calibration linearity. Statistics on
initial (section 8.2) and on-going
(section 12.7) performance shall be
computed and maintained.
5.10.2.3 Data process ing--the data system shall be
used to search, locate, identify, and
quantify the compounds of interest in each
GC analysis. Software routines shall be
employed to compute and record retention
times and peak areas. Displays of
chromatograms and library comparisons are
required to verify results.
5.10.3 Detectors
5.10.3.1 Halide specific--electron capture or
electrolytic conductivity (Micoulometric,
Hall, or O.I.), capable of detecting TBD
pg of aldrin under the analysis conditions
given in table 2.
5.10.3.2 Flame photometric--capable of detecting
TBD pg of TBD under the analysis
conditions given in table 2.
5.10.4 Chromatographs may be configured in one of
two ways: (1) Two halide specific
detectors (HSDs) in one GC; two flame
photometric detectors (FPDs) in the other.
With this configuration, the primary and
confirmatory columns and detectors are in
the same GC. (2) One HSD and one FPD in
each GC. With this configuration, the
primary columns and detectors are in one
GC, the confirmatory columns and detectors
are in the other.
6 REAGENTS AND STANDARDS
6.1 Sample preservation--sodium thiosulfate
(ACS), granular.
6.2 pH adjustment
6.2.1 Sodium hydroxide--reagent grade
6.2.1.1 Concentrated solution (10N)--dissolve 40 g
NaOH in 100 ml reagent water.
6.2.1.2 Dilute solution (O.IM)--dissolve 4 g NaOH
in 1 liter of reagent water.
6.2.2 Sulfuric acid (1 + 1)--reagent grade, 6N
in reagent water. Slowly add 50 mL H2S04
(specific gravity 1.84) to 50 mL reagent
water.
6.2.3 Potassium hydroxide--37 w/v percent.
Dissolve 37 g KOH in 100 ml reagent water.
6.3 Solution drying
6.3.1 Sodium sulfate, reagent grade, granular
anhydrous (Baker 3375, or equivalent),
rinsed with methylene chloride (20 mL/g),
baked at 450 °C for one hour minimum,
cooled in a dessicator, and stored in a
pre-cleaned glass bottle with screw cap
which prevents moisture from entering.
6.3.2 Acidified sodium sulfate--add 0.5 mL H2S04
and 30 mL ethyl ether to 100 g sodium
sulfate. Mix thoroughly. Allow the ether
to evaporate completely. Transfer the
mixture to a clean container and store at
110 t 5 °C.
6.4 Solvents--methylene chloride, hexane,
ethyl ether, acetone, isooctane, and
methanol; pesticide quality; lot certified
to be free of interferences.
6.4.1 Ethyl ether must be shown to be free of
peroxides before it is used, as indicated
by EH Laboratories Quant Test Strips
(Scientific Products P1126-8, or
equivalent). Procedures recommended for
removal of peroxides are provided with the
test strips. After cleanup, 20 mL of
ethyl alcohol is added to each liter of
ether as a preservative.
6.4.2 Acetone:hexane (1:10)--prepare by adding
10 mL acetone to 90 mL hexane
6.5 GPC calibration solution—solution
containing 300 mg/mL corn oil, 15 mg/mL
bis(2-ethylhexyl) phthalate, 1.4 mg/mL
pentachlorophenol, 0.1 mg/mL perylene, and
0.5 mg/mL sulfur
93
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6.6 Sample cleanup
6.6.1 Florisil--PR grade, 60/100 mesh, activated
at 650 - 700 °C, stored in the dark in
glass container with Teflon-lined screw
cap. Activate at 130 o for 16 h minimum
immediately prior to use. Alternatively,
500 mg cartridges (J.T. Baker, or
equivalent) may be used.
6.6.2 Diol cartridges--diol bonded silica, 1 g
cartridges with stainless steel frits
(Analytichem, Harbor City, CA, or
equivalent).
6.6.2.1 Diol cartridge calibration solution--
2,4,6-trichlorophenol, 0.1 ug/mL in
acetone.
6.6.3 Silicic acid, 100 mesh
6.6.3
6.7
removal--mercury (triple
copper powder (bright, non-
Sulfur
distilled),
oxidized), or TBA sodium
mercury is used, observe
precautions in section 4.
sulfite. If
the handling
Derivatization—diazald reagent [N-methyl-
(N-nitroso-p-toluene sulfanamide)], fresh
and high purity (Aldrich Chemical Co.)
6.8 Reference matrices
6.8.1 Reagent water—water in which the
compounds of interest and interfering
compounds are not detected by this method.
6.8.2 High solids reference matrix--playground
sand or similar material in which the
compounds of interest and interfering
compounds are not detected by this method.
Hay be prepared by extraction with
methylene chloride and/or baking at 450 °C
for 4 hours minimum.
6.9 Standard solutions--purchased as solutions
or mixtures with certification to their
purity, concentration, and authenticity,
or prepared from materials of known purity
and composition. If compound purity is 96
percent or greater, the weight may be used
without correction to compute the
6.10
6.10.1
6.10.2
6.10.3
6.11
concentration of the standard. When not
being used, standards are stored in the
dark at -20 to -10 °C in screw-capped
vials with Teflon-lined lids. A mark is
placed on the vial at the level of the
solution so that solvent evaporation loss
can be detected. The vials are brought to
room temperature prior to use. Any
precipitate is redissolved and solvent is
added if solvent loss has occurred.
Preparation of stock solutions--prepare in
isooctane per the steps below. Observe
the safety precautions in section 4.
Dissolve an appropriate amount of assayed
reference material in solvent. For
example, weigh 10 mg aldrin in a 10 ml
ground glass stoppered volumetric flask
and fill to the mark with isooctane.
After the aldrin is completely dissolved,
transfer the solution to a 15 ml vial with
Teflon-lined cap.
Stock standard solutions should be checked
for signs of degradation prior to the
preparation of calibration or performance
test standards. Quality control check
samples that can be used to determine the
accuracy of calibration standards are
available from the US Environmental
Protection Agency, Environmental
Monitoring and Support Laboratory,
Cincinnati, Ohio 45268.
Stock standard solutions shall be replaced
after six months, or sooner if comparison
with quality control check standards
indicates a change in concentration.
stock
prepare
Calibration solutions—using
solutions (section 6.9),
calibration solutions of the mixtures
shown in table 5 at the levels specified.
6.12 Surrogate spiking solutions
6.12.1 Chlorinated pesticides—prepare di butyl
chlorendate and TBD at a concentration of
10 ng/mL in acetone.
-------
6.12.2 Phosphorus containing pesticides--prepare 7.2.1
TBD and T6D at a concentration of TBD
ng/mL in TBD.
6.12.3 Phenoxyacid herbicides—prepare TBD and
TBO at a concentration of TBD ng/mL in
TBD.
6.13 DDT and endrin decomposition solution--
prepare a solution containing endrin and
dieldrin each at a concentration of 25
ug/mL and DDT at a concentration of 50
ug/mL.
6.14 Combined OC standards--used for
calibration verification (sections 7.5 and 7.2.2
14.5) and for determination of initial
(section 8.2) and on-going (section 14.6)
precision and recovery. Prepare these
solutions at the levels specified in table
5.
6.15 Stability of solutions—all standard
solutions (sections 6.9 - 6.13) shall be
analyzed within 48 hours of preparation
and on a monthly basis thereafter for
signs of degradation. Standards will
remain acceptable if the peak area remains
within t 15 percent of the area obtained
in the initial analysis of the standard.
7 SETUP AND CALIBRATION
The GC systems can be calibrated using the
external standard technique in section 7.3
or the internal standard technique in 7.2.3
section 7.4.
7.1 Configure the GC systems in one of the two
ways given in section 5.10.4 and establish
the operating conditions in table 4.
7.2 Attainment of minimum levels, retention
time reproducibility, and DDT/Endrin
decomposition—determine that each
column/detector system meets minimum level
and retent i on reproduc i biIi ty
requirements, and that the organohalide
systems meet the DDT and Endrin
decomposition test, as follows:
Analyze 1 uL each of the low level
calibration mixtures in tables 4 and 5 per
the procedure in section 13 to demonstrate
that each column/detector system meets the
minimum levels in tables 4 and 5, and that
each compound elutes within one minute of
its retention time as specified in tables
4 and 5. Note: Failure to meet the
minimum levels indicates a problem with
the column/detector system under test.
Poor GC system sensitivity is usually
traceable to a dirty detector, carrier gas
leaks, or improper detector and data
system sensitivity settings.
On each column/detector system, analyze
three replicates of Calibration Group N in
table 4 for halogenated pesticides, and of
Calibration Group 1 in table 5 for
phosphorus containing pesticides. Using
the GC data system, measure and record the
retention time at the GC peak maximum for
each of the compounds in these mixtures.
The variation between the minimum and
maximum retention time for every compound
in the mixtures shall not exceed three
seconds. Note: Failure to meet these
retention time specifications indicates a
problem with the column/detector system
under test. Poor retention time
reproducibility is usually traceable to
poor GC column temperature control (often
caused by room temperature or line voltage
fluctuations), or carrier gas teaks.
DDT and endrin decomposition—inject one
uL of the decomposition test solution
(section 6.13), and compute the areas of
the dieldrin, DDT, and endrin peaks. The
areas of the DDT and endrin peaks shall be
greater than nn and mm percent,
respectively of the area of the dieldrin
peak. Note: The decomposition of DDT
and/or endrin are usually accompanied by
the appearance of the decomposition
products of these compounds.
Decomposition of DDT and endrin can be
eliminated by a thorough cleaning and
deactivation of the GC injection port
and/or by removal of a section from the
front end of the GC column. GC column
replacement may be necessary.
95
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7.3 External standard calibration
7.3.1 Inject 1.0 uL of the mixtures in tables 4
and 5 into the GC column/detector pairs
appropriate for the mixture, beginning
with the lowest level mixture and
proceeding to the highest. For each
compound, compute and store, as a function
of the concentration injected, the
retention time and peak area on both
column/detector systems (primary and
confirmatory). For the mu Iticomponent
analytes (PCBs, chlordane, toxaphene),
store the retention time and peak area for
the five largest peaks in the
chromatogram.
7.3.2 Retention time--the polar nature of some
analytes causes the retention time to
decrease as the quantity injected
increases. To compensate this effect, the
retention time for compound identification
is correlated with the analyte level.
7.3.2.1 If the difference between the maximum and
minimum retention times for any compound
is less than five seconds over the
calibration range, the retention time for
that compound can be considered constant
and an average retention time may be used
for compound identification.
7.3.2.2 Retention time calibration curve
(retention time vs amount)-- If the
retention time for a compound in the
lowest level standard is more than five
seconds greater than the retention time
for the compound in the highest level
standard, a retention time calibration
curve shall be used for identification of
that compound.
7.3.3 Calibration factor (ratio of area to
amount injected)
7.3.3.1 Compute the coefficient of variation
(relative standard deviation) of the
calibration factor over the three point
range for each compound on each
column/detector system.
7.3.3.2 Linearity-if the calibration factor for
any compound is constant (less than the
limits specified in tables 4 and 5) over
the three point calibration range, an
average calibration factor may be used for
that compound; otherwise, the complete
calibration curve (area vs amount) for
that compound shall be used.
7.4 Internal standard caIi brat ion--The
internal standard approach may be used
when more precise and accurate results are
required than can be obtained with the
external standard method. However, this
improved precision and accuracy can be
attained only if there is no interference
with the internal standard by the
compounds of interest and compounds found
in each sample matrix. Because of this
limitation, no internal standard can be
suggested that is applicable to all
samples. Suggested internal standards are
2,2'-di fluorobiphenyl, TBD, and TBD for
halogenated compounds; and deca
fluorotriphenylphosphine (DFTPP), TBO and
TBD for the phosphorus containing
compounds.
7.4.1 Add a constant amount of internal standard
to each of the calibration solutions in
tables 4 and 5.
7.4.2 Inject 1.0 uL of the solutions in tables 4
and $ into the GC column/detector system
appropriate for the mixture, beginning
with the lowest level mixture and
proceeding to the highest. For each
compound, compute and store, as a function
of the concentration injected, the
retention time and peak area on both
column/detector systems (primary and
confirmatory). For the mutticomponent
analytes (PCBs, chlordane, toxaphene),
store the retention time and peak area for
the five largest peaks in the
chromatogram.
7.4.3 Relative retention time--Using the GC data
system, compute the relative retention
times for each compound in each of the
mixtures:
96
-------
Relative retention time = 7.4.4.2
retention time of compound
retention time of internal standard
If multiple internal standards are used,
the nearest eluted internal standard shall
be used for reference. Note: The polar
nature of some compounds causes the
retention time to decrease as the quantity 7.5
injected increases. To compensate this
effect, the relative retention time for
compound identification shall be
correlated with the level of the compound.
7.4.3.1 If the retention time difference between
the compound and its internal standard are
invariant (less than three seconds) over
the three point calibration range, the
average relative retention time may be
used for identification of that compound.
7.4.3.2 Relative retention time calibration curve
(relative retention time vs amount)--if 7.5.1
the retention time difference between the
compound and its internal standard in the
lowest level standard is more than three 7.5.2
seconds greater than this difference in
the highest level standard, a relative
retention time calibration curve shall be
used for identification of that compound.
7.4.4 Response factors--call" brat ion requires the
determination of response factors (RF)
which are defined by the following
equation:
Linearity-if the response factor (RF) for
any compound is constant (less than 15
percent coefficient of variation) over the
three point calibration range, the average
response factor may be used for that
compound; otherwise, the complete
calibration curve for that compound shall
be used.
Combined QC standards—to preclude
periodic analysis of all of the
calibration solutions listed in tables 4
and 5, the GC systems are calibrated with
the combined OC standards (section 6.14)
as a final step. Not all of the compounds
in these standards will be separated by
the GC columns used in this method.
Retention times and calibration or
response factors are verified for the
compounds that are resolved, and
calibration or response factors are
obtained for the unresolved peaks.
Analyze the combined QC standards on their
respective column/detector pairs.
External standard calibration—for those
compounds that exhibit a single, distinct
GC peak, the retention time shall be
within ± five seconds of the retention
time of the peak in the medium level
calibration standard (section 7.3.1), and
the calibration factor using the primary
column shall be within ± 20 percent of the
calibration factor in the medium level
standard (7.3.1).
RF = (Ag x Cis)/(A.s x Cg), where; Ag is
the area for the compound, A. is the area
for the internal standard, C. is the
concentration of the internal standard
(ug/mL), and C is the concentration of
the compound (ug/mL).
7.4.4.1 The response factor is determined for the
three concentrations given in tables 4 and
5. The amount of internal standard added
to each extract is the same so that C-
remains constant. The RF is plotted vs
concentration for each compound in the
standard (C ) to produce a calibration
curve.
7.5.3 Internal standard caIibrat ion--for those
compounds that exhibit a single, distinct
GC peak, the retention time difference
between the peak and its internal standard
shall be within ± three seconds of this
difference in the medium level calibration
standard (section 7.4.2), and the response
factors on both column/detector systems
shall be within ± 10 percent of the
response factor in the medium level
standard (section 7.4.2).
7.5.4 If all compounds meet the criteria in
section 7.5.2 or 7.5.3, analysis of
precision and recovery standards (section
-------
8) may begin. If, however, any compound
fails, the measurement system is not per
forming properly for that compound. In
this event, correct the problem and repeat
the test, or recalibrate the system
(section 7.3 or 7.4).
7.5.5 For the peaks containing two or more
compounds, compute and store the retention
times or relative retention times at the
peak maxima on both columns (primary and
confirmatory), and also compute and store
the calibration factors or response
factors on both columns. These results
will be used for calibration verification
(section 14.2 and 14.5) and for precision
and recovery studies (sections 8.2 and
14.6).
7.6 Florisil calibration--the cleanup proce-
dure in section 11 utilizes Florisil col-
umn chromatography. Florisil from differ-
ent batches or sources may vary in adsorp-
tive capacity. To standardize the amount
of Florisil that is used, the use of the
lauric acid value (reference 4) is sug-
gested. The referenced procedure deter-
mines the adsorption of lauric acid (in
mg/g of Florisil) from hexane solution.
The amount of Florisil to be used for each
column is calculated by dividing 110 by
this ratio and multiplying by 20 g.
8 QUALITY ASSURANCE/QUALITY CONTROL
8.1 Each laboratory that uses this method is
required to operate a formal quality
assurance program (reference 5). The
minimum requirements of this program
consist of an initial demonstration of
laboratory capability, an ongoing analysis
of standards and blanks as tests of
continued performance, and analysis of
spiked samples to assess accuracy.
Laboratory performance is compared to
established performance criteria to
determine if the results of analyses meet
the performance characteristics of the
method. If the method is to be applied
routinely to samples containing high
solids with very little moisture (e.g.,
soils, compost), the high solids reference
matrix (section 6.8.2) is substituted for
the reagent water (section 6.8.1) in all
performance tests, and the high solids
method (section 10) is used for these
tests.
8.1.1 The analyst shall make an initial
demonstration of the ability to generate
acceptable accuracy and precision with
this method. This ability is established
as described in section 8.2.
8.1.2 The analyst is permitted to modify this
method to improve sepa rations or lower
the costs of measurements, provided all
performance requirements are met. Each
time a modification is made to the method
or a cleanup procedure is added, the
analyst is required to repeat the
procedure in section 8.2 to demonstrate
method performance.
8.1.3 Analyses of blanks are required to
demonstrate freedom from contamination.
The procedures and criteria for analysis
of a blank are described in section 8.5.
8.1.4 The laboratory shall spike all samples
with at least one surrogate compound to
monitor method performance. This test is
described in section 8.3. When results of
these spikes indicate atypical method
performance for samples, the samples are
diluted to bring method performance within
acceptable limits (section 17).
8.1.5 The laboratory shall, on an on-going
basis, demonstrate through calibration
verification and the analysis of the
combined QC standard (section 6.14) that
the analysis system is in control. These
procedures are described in sections 14.1,
14.5, and 14.6.
8.1.6 The laboratory shall maintain records to
define the quality of data that is
generated. Development of accuracy
statements is described in section 8.4.
8.1.7 Other analytes may be determined by this
method. The procedure for establishing a
98
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preliminary quality control limit for a
new analyte is given in section 8.6.
8.2 Initial precision and accuracy--to
establish the ability to generate
acceptable precision and accuracy, the
analyst shall perform the following
operations:
8.2.1 For low solids (aqueous samples), extract,
concentrate, and analyze one set of four
one-liter aliquots of the combined QC
standards (section 6.14) according to the
procedure in section 10. For high solids
samples, one set of four 30 gram aliquots
of the high solids reference matrix are
used.
8.2.2 Using results of the set of four analyses,
compute the average recovery (X) in ug/mL
in the extract and the standard deviation
of the recovery (s) in ug/mL for each
compound, by the external standard
(section 7.3) or internal standard
(section 7.4) method.
8.2.3 For each compound, compare s and X with
the corresponding limits for initial
precision and accuracy in tables 6-8.
If s and X for all compounds meet the
acceptance criteria, system performance is
acceptable and analysis of blanks and
samples may begin. If, how ever, any
individual s exceeds the precision limit
or any individual X falls outside the
range for accuracy, system performance is
unacceptable for that compound.
8.3 The laboratory shall spike all samples
with at least one surrogate compound to
assess method performance on the sample
matrix.
8.3.1 Analyze each sample according to the
method beginning in section 10.
8.3.2 Compute the percent recovery (P) of the
surrogate compound(s) using the external
or internal standard method (section 7.3
or 7.4).
Table 6
PRECISION AND RECOVERY OF ORGANO-HAUDE
COMPOUNDS
EPA
EGD
089
102
103
105
104
434
433
441
091
431
094
093
092
432
478
090
095
096
097
098
099
435
100
101
437
439
430
438
436
112
108
109
106
110
107
111
440
113
442
Compound
Aldrin
alpha-BHC
beta-BHC
delta-BHC
gamma- BHC (Undane)
Captafol
Captan
Carbophenothion
Chlordane
Chlorobenzi late
4,4'-DDD
4,4'-ODE
4,4'-ODT
Diallate
Oichlone
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Heptachlor
Heptachlor epoxide
Isodrin
Kepone
Methoxychlor
Mi rex
Nitrofen (TDK)
PCS- 1016
PCB-1221
PCB-1232
PCS -1242
PCS -1248
PCB-1254
PCS -1260
PCNB
Toxaphene
Trif luralin
Recovery
(percent)
82.2
105.9
94.2
30.4
109.9
78.
37.
185.2
113.2
117.2
82.1
97.3
62.
42.
93.7
81.6
63.7
38.3
97.2
22.2
14.1
59.1
468.3
54.9
76.
104.9
90.5
90.3
97.5
111.3
RSD
5.1
7.9
9.1
48.
3.8
10.
8.
12.
8.5
32.
18.
12.
10.
16.
16.
34.
22.
23.
28.
21.
30.
12.
9.8
11.
14.
5.5
99
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Table 7
PRECISION AND RECOVERY OF THIOPHOSPHATE
COMPOUNDS
Table 8
PRECISION AND RECOVERY OF PHENOXYACID
HERBICIDES
EPA
EGD
468
461
469
443
479
471
460
450
455
449
452
458
467
463
446
454
447
453
464
474
475
456
444
470
459
448
457
465
473
477
476
472
466
445
451
462
Compound
Azinphos ethyl
Chlorfevinphos
Chlorpyrifos
Coumaphos
Crotoxyphos
Demeton
Diazinon
Dichlorvos
Dicrotophos
Ditnethoate
Oioxathion
Disulfoton
EPN
Ethion
Famphur
Fensulfothion
Fenthion
Gut hi on
Hexamethyl-
phosphoramide 120.
Leptophos
Ma lath ion
Methyl parathion
Mevinphos
Monocrotophos
Naled
Parathion
Phorate
Phosmet
Phosphamidon
Sulfotepp
TEPP
Terbufos
Tetrachlorvinphos
Trichlorofon
Tricresylphosphate
Trimethy I phosphate
Recovery
(percent)
77.0
98.
84.1
73.0
23.
86.9
80.6
95.8
42.5
79.6
69.
81.8
82.0
62.8
67.
32.
77.0
4.
77.2
89.8
82.0
85.
9.
74.
82.6
97.0
79.
61.
101.
82.0
87.3
88.3
40.4
82.
25.6
RSD
BX
17.
4.8
9.3
3.
4.6
4.5
5.8
31.4
7.2
5.
6.0
5.4
14.5
26.
2.
8.9
8.9
5.9
5.6
10.
7.4
5.0
6.
16.
5.
18.2
4.5
11.0
27.9
8.
15.1
EPA Recovery
EGD Compound (percent) RSD
481 2,4-D 92. 5.0
480 Dinoseb 58. 9.8
482 2,4,5-T 69. 9.1
483 2,4,5-TP 60. 7.6
2,4-DB 86. 9.5
8.3.3 The recovery of the surrogate compound
shall be within the limits of 20 to 200
percent. If the recovery of any compound
falls outside of these limits, method
performance is unacceptable for that
compound in that sample, and the sample is
complex. Water samples are diluted, and
smaller amounts of soils, sludges, and
sediments are reanalyzed per section 17.
8.4 Method accuracy—the laboratory shall
spike at least ten percent of the samples
from a given site type (e.g., influent to
treatment, treated effluent, produced
water, river sediment). If only one
sample from a given site type is analyzed,
a spiked analysis on that sample shall be
performed.
8.4.1 The concentration of the spike in the
sample shall be determined as follows:
8.4.1.1 If, as in compliance monitoring, the
concentration of a specific analyte in the
sample is being checked against a
regulatory concentration limit, the spike
shall be at that limit or at one to five
times higher than the background
concentration determined in section 8.4.2,
whichever concentration is larger.
8.4.1.2 If the concentration of an analyte in the
sample is not being checked against a
limit specific to that analyte, the spike
shall be at the concentration of the
combined QC standard (section 6.14) or at
one to five times higher than the
100
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background concentration, whichever
concentration is larger.
8.4.1.3 If it is impractical to determine the
background concentration before spiking
(e.g., maximum holding times will be
exceeded), the spike concentration shall
be (1) the regulatory concentration limit,
if any; otherwise, the larger of either
five times the expected background
concentration or at the concentration of
the combined QC standard (section 6.14).
8.4.2 Analyze one sample aliquot to determine
the background concentration (B) of each
analyte. If necessary, prepare a standard
solution appropriate to produce a level in
the sample one to five times the
background concentration. Spike a second
sample aliquot with the standard solution
and analyze it to determine the
concentration after spiking (A) of each
analyte. Calculate the percent recovery
(P) of each analyte:
P = 100 (A - B) / T, where
T is the true value of the spike.
8.4.3 Compare the percent recovery for each
analyte with the corresponding QC
acceptance criteria in tables 6-8. If
any analyte fails the acceptance criteria
for recovery, the sample is complex and
must be diluted and reanalyzed per section
17.
8.4.4 As part of the QA program for the
laboratory, method accuracy for samples
shall be assessed and records shall be
maintained. After the analysis of five
spiked samples of a given matrix type
(water, soil, sludge, sediment) in which
the analytes pass the tests in section
8.4, compute the average percent recovery
(P) and the standard deviation of the
percent recovery (sp) for each compound
(or co-eluting compound group). Express
the accuracy assessment as a percent
recovery interval from P - 2sp to P + 2sp
for each matrix. For example, if P = 90X
and sp = 10X for five analyses of compost,
the accuracy interval is expressed as 70 -
11 OX. Update the accuracy assessment for
each compound in each matrix on a regular
basis (e.g. after each 5-10 new accuracy
measurements).
8.5 Blanks--reagent water and high solids
reference matrix blanks are analyzed to
demonstrate freedom from contamination.
8.5.1 Extract and concentrate a one liter
reagent water blank or a high solids
reference matrix blank with each sample
lot (samples started through the
extraction process on the same 8 hr shift,
to a maximum of 20 samples). Analyze the
blank immediately after analysis of the
combined QC standard (section 14.6) to
demonstrate freedom from contamination.
8.5.2 If any of the compounds of interest
(tables 1 thru 3) or any potentially
interfering compound is found in an
aqueous blank at greater than one ug/L, or
in a high solids reference matrix blank at
greater than 10 ug/kg (assuming the same
calibration factor as aldrin and diazinon
or a response factor of 1 relative to the
nearest eluted internal standard, for
compounds not listed in tables 1 thru 3),
analysis of samples is halted until the
source of contamination is eliminated and
a blank shows no evidence of contamination
at this level.
8.6 Other analytes may be determined by this
method. To establish a quality control
limit for an analyte, determine the
precision and accuracy by analyzing four
replicates of the analyte along with the
combined QC standard per the procedure in
section 8.2. Compute the average percent
recovery (A) and the standard deviation of
percent recovery (sn) for the analyte, and
measure the recovery and standard
deviation of recovery for the other
analytes. The data for the new analyte is
assumed to be valid if the precision and
recovery specifications for the other
analytes are met. Establish a preliminary
quality control limit of A ± 2sn for the
101
-------
new analyte and add the limit to table 6,
7. or 8.
8.7 The specifications contained in this
method can be met if the apparatus used is
calibrated properly, then maintained in a
calibrated state. The standards used for
calibration (section 7), calibration
verification (section 14.5), and for
initial (section 8.2) and on-going
(section 14.6) precision and recovery
should be identical, so that the most
precise results will be obtained. The GC
instruments will provide the most
reproducible results if dedicated to the
settings and conditions required for the
analyses of the analytes given in this
method.
8.8 Depending on specific program
requirements, field replicates and field
spikes of the analytes of interest into
samples may be required to assess the
precision and accuracy of the sampling and
sample transporting techniques.
9 SAMPLE COLLECTION, PRESERVATION, AND
HANDLING
9.1 Collect samples in glass containers
following conventional sampling practices
(reference 6), except that the bottle
shall not be prerinsed with sample before
collection. Aqueous samples which flow
freely are collected in refrigerated
bottles using automatic sampling
equipment. Solid samples are collected as
grab samples using wide mouth jars.
9.2 Maintain samples at 0 - 4 °C from the time
of collection until extraction. If the
samples will not be extracted within 72
hours of collection, adjust the sample to
a pH of 5.0 to 9.0 using sodium hydroxide
or sulfuric acid solution. Record the
volume of acid or base used. If residual
chlorine is present in aqueous samples,
add 80 mg sodium thiosulfate per liter of
water. EPA methods 330.4 and 330.5 may be
used to measure residual chlorine
(reference 7).
9.3 Begin sample extraction within seven days
of collection, and analyze all extracts
within 40 days of extraction.
10 SAMPLE EXTRACTION AND CONCENTRATION
Samples containing one percent solids or
less are extracted directly using
continuous liquid/ liquid extraction
techniques (section 10.2.1 and figure 3).
Samples containing one to 30 percent
solids are diluted to the one percent
level with reagent water (section 10.2.2)
and extracted using continuous
liquid/liquid extraction techniques.
Samples containing greater than 30 percent
solids are extracted using ultrasonic
techniques (section 10.2.5) For
determination of the phenoxy-acid
herbicides, a separate sample aliquot is
extracted, derivatized, and cleaned up.
The derivatized extract is then combined
with the organo-chlorine extract.
10.1 Determination of percent solids
10.1.1 Weigh 5 - 10 g of sample into a tared
beaker. Record the weight to three
figures.
10.1.2 Dry overnight (12 hours minimum) at 110
+/- 5 °C, and cool in a dessicator.
10.1.3 Determine percent solids as follows:
X solids = weight of dry sample x 100
weight of wet sample
10.2 Preparation of samples for extraction
10.2.1 Samples containing one percent solids or
less—extract the sample directly using
continuous liquid/liquid extraction
techniques.
10.2.1.1 Measure 1.00 +/- 0.01 liter of sample into
a clean 1.5 - 2.0 liter beaker. For the
phenoxy-acid herbicides, measure a
separate one liter aliquot.
102
-------
10.2.1.2 Spike 0.5 ml of the surrogate spiking
solution (section 6.8) into the sample
aliquot. For the phenoxy-acid herbicides,
spike 0.5 ml of the herbicide surrogate
spiking solution into the herbicide
aliquot. Proceed to preparation of the QC
aliquots for low solids samples (section
10.2.3).
10.2.2 Samples containing one to 30 percent
solids
10.2.2.1 Mix sample thoroughly.
10.2.2.2
10.2.2.3
10.2.2.5
Using the percent solids found in 10.1.3,
determine the weight of sample required to
produce one liter of solution containing
one percent solids as follows:
sample weight
1000 grams
X solids
Place the weight determined in 10.2.2.2 in
a clean 1.5 - 2.0 liter' beaker. For the
phenoxy-acid herbicides, place a separate
aliquot in a clean beaker. Discard all
sticks, rocks, leaves and other foreign
material prior to weighing.
Bring the sample aliquot(s) to 100 - 200
ml volume with reagent water.
10.2.2.6 Spike 0.5 ml of the appropriate surrogate
spiking solution (section 6.12) into each
sample aliquot.
10.2.2.7 Using a clean metal spatula, break any
solid portions of the sample into small
pieces.
10.2.2.8 Place the 3/4 in. horn on the ultrasonic
probe approx 1/2 in below the surface of
each sample aliquot and pulse at 50
percent for three minutes at full power.
If necessary, remove the probe from the
solution and break any large pieces using
the metal spatula or a stirring rod and
repeat the sonication. Clean the probe
with methylene chloridetacetone (1:1)
between samples to preclude cross-
contamination.
10.2.2.9 Bring the sample volume to 1.0 +/- 0.1
liter with reagent water.
10.2.3 Preparation of QC aliquots for samples
containing low solids (<30 percent),
10.2.3.1 For each sample or sample lot (to a
maximum of 20) to be extracted at the same
time, place two 1.0 +/- 0.01 liter
aliquots of reagent water in clean 1.5 -
2.0 liter beakers. For the phenoxy-acid
herbicides, place two additional one liter
aliquots in clean beakers.
10.2.3.2 To serve as a blank, spike 0.5 ml of the
pesticide surrogate spiking solution
(section 6.12.1 and 6.12.2) into one
reagent water aliquot, and 0.5 mL of the
herbicide surrogate spiking solution
(section 6.12.3) into a second reagent
water aliquot.
10.2.3.3 Spike the combined QC standard (section
6.14) into a reagent water aliquot. For
the herbicides', spike the herbicide
standard into the remaining reagent water
aliquot.
10.2.4 Stir and equilibrate all sample and QC
solutions for 1-2 hours. Extract the
samples and QC aliquots per section 10.3.
10.2.5 Samples containing 30 percent solids or
greater
10.2.5.1 Mix the sample thoroughly
10.2.5.2 Weigh 30 +/- 0.3 grams into a clean 400 -
500 ml beaker. For the herbicides, weigh
an additional two 30 gram aliquots into
clean beakers. Discard all sticks, rocks,
leaves and other foreign material prior to
weighing.
10.2.5.3 Herbicide acidification-add 50 at. of
reagent water to one of the herbicide
sample aliquots and stir on a stirring
plate for one hour minimum. Using a pH
meter, determine and record the sample pH
while stirring. Slowly add H2S04 while
stirring and determine and record the
103
-------
amount of acid required to acidify the
sample to pH <2 Discard this aliquot.
10.2.5.4 Spike 0.5 ml of the appropriate surrogate
spiking solution (section 6.12) into the
pesticide and herbicide aliquots.
10.2.5.5 QC aliquots--for each sample or sample lot
(to a maximum of 20) to be extracted at
the same time, place two 30 +/- 0.3 gram
aliquots of the high solids reference
matrix in clean 400 - 500 ml beakers. For
the herbicides, place three additional
aliquots in clean beakers and use one of
these to determine the amount of acid
required for acidification per step
10.2.5.3. Discard this aliquot.
10.2.5.6 To serve as a blank, spike 0.5 ml of the
pesticide surrogate spiking solution
(section 6.12.1 and 6.12.2) into one
aliquot of the high solids reference
matrix, and O.S mL of the herbicide
surrogate spiking solution (section
6.12.3) into a second aliquot of the high
solids reference matrix.
10.2.5.7 Spike 1.0 mL of the combined QC standard
(section 6.14) into a high solids
reference matrix aliquot. For the
herbicides, spike the herbicide standard
into the remaining high solids reference
matrix aliquot. Extract the high solids
samples per section 10.4.
10.3 Continuous extraction of low solids
(aqueous) samples--place 100 - 150 ml
methylene chloride in each continuous
extractor and 200 - 300 ml in each
distilling flask.
10.3.1 Pour the sample(s), blank, and standard
aliquots into the extractors. Rinse the
glass containers with 50 - 100 ml
methylene chloride and add to the
respective extractors. Include all solids
in the extraction process.
10.3.2 Extraction—for the pesticides, adjust the
pH of the waters in the extractors to 5 -
9 with NaOH or H2S04 while monitoring with
a pH meter. For the herbicides, adjust
the pH to two or less Caution: some
samples require acidification in a hood
because of the potential for generating
hydrogen sulfide.
10.3.3 Begin the extraction by heating the flask
until the methylene chloride is boiling.
When properly adjusted, 1 - 2 drops of
methylene chloride per second will fall
from the condenser tip into the water.
Test and adjust the pH of the waters
during the first 1 - 2 hours of
extraction. Extract for 18 - 24 hours.
10.3.4 Remove the distilling flask, estimate and
record the volume of extract (to the
nearest 100 mL), and pour the contents
through a prerinsed drying column
containing 7 to 10 cm of anhydrous sodium
sulfate. Rinse the distilling flask with
30 - 50 mL of methylene chloride and pour
through the drying column. For pesticide
extracts and for herbicide extracts to be
cleaned up using GPC, collect the solution
in a 500 mL K-D evaporator flask equipped
with a 10 mL concentrator tube. Seal,
label the pesticide and herbicide
fractions, and concentrate per sections
10.5 to 10.6. For herbicide extracts not
to be cleaned up by GPC, collect the
solution in a 500 - 1000 mL separatory
funnel and proceed to section 12 for
hydrolysis and esterification of the
herbicides.
10.4 Ultrasonic extraction of high solids
samples
10.4.1 For the herbicide aliquots, add the amount
of acid determined in section 10.2.5.3 to
the sample aliquot and the amount
determined in section 10.2.5.5 to the QC
aliquots and mix thoroughly.
10.4.2 Add 60 - 70 grams of sodium sulfate to the
pesticide aliquots and an equal amount of
acidified sodium sulfate to the herbicide
aliquots and mix each aliquot thoroughly.
Some wet sludge samples may require more
than 70 grams for complete removal of
water. All water must be removed prior to
104
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addition of organic solvent so that the
extraction process is efficient.
10.4.3 Add 100 +/- 10 mL of acetone:methylene
chloride (1:1) to each of the aliquots and
mix thoroughly.
10.4.4 Place the 3/4 in. horn on the ultrasonic
probe approx 1/2 in below the surface of
the solvent but above the solids layer and
pulse at 50 percent for three minutes at
full power. If necessary, remove the
probe from the solution and break any
large pieces using a metal spatula or a
stirring rod and repeat the sonication.
Clean the horn with five percent aqueous
sodium bicarbonate immediately after
sonicating any of the herbicide aliquots
to prevent acid damage to the horn.
10.4.5 Decant the pesticide extracts through a
prerinsed drying column containing 7 to 10
cm anhydrous sodium sulfate into 500 -
1000 mL graduated cylinders. Decant the
herbicide extracts similarly using
acidified sodium sulfate.
10.4.6 Repeat the extraction steps (10.4.2 -
10.4.4) twice more for each sample and OC
aliquot. On the final extraction, swirl
the sample or QC aliquot, pour into its
respective drying column, and rinse with
acetone:methylene chloride. Record the
total extract volume. If necessary,
transfer the extract to a centrifuge tube
and centrifuge for 10 minutes to settle
fine particles.
10.4.7 For all pesticide extracts and for
herbicide extracts to be cleaned up using
GPC, filter these extracts through Whatman
#41 paper into a 500 mL K-D evaporator
flask equipped with a 10 mL concentrator
tube. Rinse the graduated cylinder or
centrifuge tube with 30 - 50 mL of
methylene chloride and pour through filter
to complete the transfer. Seal and label
the K-Ds as the pesticide and herbicide
fractions. Concentrate these fractions
per sections 10.5 through 10.8. For
herbicide extracts not to be cleaned up by
GPC, filter the solution through Whatman
#41 paper into a 500 - 1000 mL separatory
funnel and proceed to section 12 for
hydrolysis and esterificat ion of the
herbicides.
10.5 Macro concentration
10.5.1 Concentrate the extracts in separate 500
mL K-D flasks equipped with 10 mL
concentrator tubes. Add 1 to 2 clean
boiling chips to the flask and attach a
three-ball macro Snyder column. Prewet
the column by adding approx one mL of
methylene chloride through the top. Place
the K-D apparatus in a hot water bath so
that the entire lower rounded surface of
the flask is bathed with steam Adjust the
vertical position of the apparatus and the
water temperature as required to complete
the concentration in 15 to 20 minutes. At
the proper rate of distillation, the balls
of the column will actively chatter but
the chambers will not flood.
10.5.2 When the liquid has reached an apparent
volume of one mL, remove the K-D apparatus
from the bath and allow the solvent to
drain and cool for at least 10 minutes.
10.5.3 If the extract is to be cleaned up using
GPC, remove the Snyder column and rinse
the flask and its lower joint into the
concentrator tube with 1 - 2 mL of
methylene chloride. A 5 mL syringe is
recommended for this operation. Adjust
the final volume to 10 mL and proceed to
GPC cleanup in section 11.
10.6 Hexane exchange--extracts to be subjected
to diol or Florist I cleanup and extracts
that have been cleaned up are exchanged
into hexane.
10.6.1 Remove the Snyder column, add
approximately 50 mL of hexane and a clean
boiling chip, and reattach the Snyder
column. Concentrate the extract as in
section 10.5 except use hexane to prewet
the column. The elapsed time of the
concentration should be 5 - 10 minutes.
105
-------
10.6.2 Remove the Snyder col urn and rinse the
flask and its lower joint into the
concentrator tube with 1 - 2 mL of hexane.
Adjust the final volume of extracts that
have not been cleaned up by GPC to 10 mL
and those that have been cleaned up by GPC
to 5 mL (the difference accounts for the
50 percent loss in the GPC cleanup) Clean
up the extracts using the diol, Florisil,
and/or sulfur removal procedures in
section 11.
11 CLEANUP AND SEPARATION
11.1 Cleanup procedures may not be necessary
for relatively clean samples (treated
effluents, groundwater, drinking water).
If particular circumstances require the
use of a cleanup procedure, the analyst
may use any or all of the procedures below
or any other appropriate procedure.
However, the analyst first shall
demonstrate that the requirements of
section 8.2 can be met using the cleanup
procedure(s) as an integral part of the
method.
11.1.1 Gel permeation chromatography (section
11.2) removes many high molecular weight
interferents that cause GC column
performance to degrade. It is used for
all soil and sediment extracts and may be
used for water extracts that are expected
to contain high molecular weight organic
compounds (e.g., polymeric materials,
humic acids).
11.1.2 The diol cartridge (section 11.3) removes
polar organic compounds such as phenols.
It is used for all extracts.
11.1.3 The Florisil column (section 11.4) allows
for selected fractionation of the
compounds of interest and will also
eliminate polar interferences. Its use is
optional Note: Some organophosphorus
pesticides may be irreversibly absorbed by
Florisil.
11.1.4 Alumina column cleanup (section 11.5) is
used to remove polar interferences. Its
use is optional.
11.1.5 Elemental sulfur, which interferes with
the electron capture gas chromatography of
some of the pesticides and herbicides, is
removed using mercury or activated copper.
Sulfur removal (section 11.6) is required
when sulfur is known or suspected to be
present.
11.2 Gel permeation chromatography (GPC)
11.2.1 Column packing
11.2.1.1 Place 70 - 75 g of SX-3 Bio-beads in a 400
- 500 ml beaker.
11.2.1.2 Cover the beads with methylene chloride
and allow to swell overnight (12 hours
minimum).
11.2.1.3 Transfer the swelled beads to the column
and pump solvent through the column, from
bottom to top, at 4.5 - 5.5 mL/min prior
to connecting the column to the detector.
11.2.1.4 After purging the column with solvent for
1 - 2 hours, adjust the column head
pressure to 7 - 10 psig, and purge for 4 •
5 hours to remove air. Maintain a head
pressure of 7 • 10 psig. Connect the
column to the detector.
11.2.2 Column calibration
11.2.2.1 Load 5 mL of the calibration solution
(section 6.5) into the sample loop.
11.2.2.2 Inject the calibration solution and record
the signal from the detector. The elution
pattern will be corn oil, bis(2-ethyl
hexyl) phthalate, pentachlorophenol,
perylene, and sulfur.
11.2.2.3 Set the "dump time" to allow >85 percent
removal of the corn oil and >85 percent
collection of the phthalate.
11.2.2.4 Set the "collect time" to the peak minimum
between perylene and sulfur.
11.2.2.5 Verify the calibration with the
calibration solution after every 20
extracts. Calibration is verified if the
106
-------
recovery of the pentachlorophenol is
greater than 85 percent. If calibration
is not verified, the system shall be
recalibrated using the calibration
solution, and the previous 20 samples
shall be re-extracted and cleaned up using
the calibrated GPC system.
11.2.3 Extract cleanup--GPC requires that the
column not be over loaded. The column
specified in this method is designed to
handle a maximum of 0.5 gram of high
molecular weight material in a 5 ml
extract. If the extract is known or
expected to contain more than 0.5 gram,
the extract is split into fractions for
GPC and the fractions are combined after
elution from the column. The solids
content of the extract may be obtained
gravimetrically by evaporating the solvent
from a 50 uL aliquot.
11.2.3.1 Filter the extract or load through the
filter holder to remove particulates.
Load the 5.0 ml extract onto the column.
11.2.3.2 Elute the extract using the calibration
data determined in 11.2.2. Collect the
eluate in a clean 400 • 500 ml beaker.
11.2.3.3 Rinse the sample loading tube thoroughly
with methylene chloride between extracts
to prepare for the next sample.
11.2.3.4 If a particularly dirty extract is
encountered, a 5.0 mL methylene chloride
blank shall be run through the system to
check for carry-over.
11.2.3.5 Concentrate the pesticide extract and
exchange into hexane per sections 10.5 and
10.6. Proceed to section 12 with the
herbicide extract.
11.3 Diol cartridge
11.3.1 Setup
11.3.1.1 Attach the Vac-elute manifold to a water
aspirator or vacuum pump with the trap and
gauge installed between the manifold and
vacuum source.
11.3.1.2 Place the diol cartridges in the manifold,
turn on the vacuum source, and adjust the
vacuum to 5 - 10 psia.
11.3.2 Cartridge washing--pre-elute each
cartridge prior to use with 5 ml of
hexane:acetone (9:1) to remove potential
interferences.
11.3.3 Cartridge certification--each cartridge
lot must be certified to ensure recovery
of the compounds of interest and removal
of 2,4,6-trichlorophenol.
11.3.3.1 To make the diol test mixture, add 1.0 ml
of the trichlorophenol solution (section
6.6.2.1) to 1.0 ml of the combined
calibration standard (section 6.14).
Elute the mixture using the procedure in
11.3.4.
11.3.3.2 Concentrate the eluant to 1.0 ml using the
nitrogen blowdown apparatus (section
5.4.3) and inject 1.0 uL of the
concentrated eluant into the GC using the
procedure in section 13. The recovery of
all organo-halide and organo-phosphorus
analytes (including the unresolved GC
peaks) shall be in the range of 75 • 125
percent, and the peak for trichlorophenol
shall not be detectable; otherwise the
diol cartridge is not performing properly
and the cartridge lot shall to rejected.
11.3.4 Extract cleanup
11.3.4.1 After cartridge washing (section 11.3.2),
release the vacuum and place the rack
containing the 10 ml volumetric flasks
(section 5.6.2.4) in the vacuum manifold.
Reestablish the vacuum at 5 • 10 psia.
11.3.4.2 Using a pi pet or a one ml syringe,
transfer 1.0 mL of extract to a diol
cartridge.
11.3.4.3 Elute each cartridge into its volumetric
flask with 9 mL of hexane/acetone.
11.3.4.4 Release the vacuum and remove the 10 mL
volumetric flasks Quantitatively transfer
each eluted extract from its 10 mL flask
107
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into a clean centrifuge tube or sample
vial. Rinse the volumetric flask with two
1-ml aliquots of hexane to ensure
quantitative transfer.
11.3.4.5 Concentrate the eluted extracts to 1.0 mL
using the nitrogen blow-down apparatus.
If sulfur crystals are evident in the
eluted extract, or if sulfur is suspected
to be present, proceed to section 11.5 for
sulfur removal. If sulfur is not known or
expected to be present, adjust the final
volume to 5 or 10 mL (per section 10.6),
depending on whether or not the extract
was subjected to GPC cleanup, and proceed
to section 13 for GC analysis.
11.4.5 Concentrate the fractions as in section
10.6, except use hexane to prewet the
column. Readjust the final volume to 5 or
10 mL as in section 10.6, depending on
whether the extract was subjected to GPC
cleanup, and analyze by gas chromatography
per the procedure in section 13.
11.5 Alumina column
11.5.1 Reduce the volume of the extract to 0.5 mL
and bring to 1.0 mL with acetone.
11.5.2 Add 3 g of activity III neutral alumina to
a 10 mL chromatographic column. Tap the
column to settle the alumna.
11.4 Florisil column
11.4.1 Place a weight of Florisil (nominally 20
g) predetermined by calibration (section
7.6) in a chromatographic column. Tap the
column to settle the Florisil and add 1 -
2 cm of anhydrous sodium sulfate to the
top.
11.4.2 Add 60 mL of hexane to wet and rinse the
sodium sulfate and Florisil. Just prior
to exposure of the sodium sulfate layer to
the air, stop the elution of the hexane by
closing the stopcock on the
chromatographic column. Discard the
eluate.
11.4.3 Transfer the concentrated extract (section
10.6.2) onto the column. Complete the
transfer with two 1-mL hexane rinses.
11.4.4 Place a clean 500 ml K-D flask and
concentrator tube under the column. Drain
the column into the flask until the sodium
sulfate layer is nearly exposed. Elute
fraction 1 with 200 mL of six per cent
ethyl ether in hexane (v/v) at a rate of
approx 5 mL/min Remove the K-D flask.
Elute fraction 2 with 200 mL of 15 percent
ethyl ether in hexane (v/v) into a second
K-D flask. Elute fraction 3 with 200 mL
of 50 percent ethyl ether in hexane (v/v).
The elution patterns for the organo-halide
pesticides and PCBs are shown in table 9.
11.5.3 Transfer the extract to the top of the
column and collect the eluate in a clean
10 mL concentrator tube. Rinse the
extract container with 1 - 2 mL portions
of hexane (to a total volume of 9 mL) and
add to the alumina column. Do not allow
the column to go dry.
11.5.4 Concentrate the extract to 1.0 mL if
sulfur is to be removed, or adjust the
final volume to 5 or 10 mL as in section
10.6, depending on whether the extract was
subjected to GPC cleanup, and analyze by
gas chromatography per section 13.
11.6 Sulfur removal--elemental sulfur will
usually elute entirely in fraction 1 of
the Florisil column cleanup.
11.6.1 Transfer the concentrated extract into a
clean concentrator tube or Teflon-sealed
vial. Add 1 - 2 drops of mercury or 100
mg of activated copper powder and seal
(reference 9). If TBA sulfite is used,
add 1 mL of the TBA sulfite reagent and 2
mL of isopropanol.
11.6.2 Agitate the contents of the vial for 1 - 2
hours on a reciprocal shaker. If the
mercury or copper appears shiny, or if
precipitated sodinn sulfite crystals from
the TBA sulfite reagent are present, and
if the color remains unchanged, all sulfur
has been removed; if not, repeat the
addition and shaking.
108
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11.6.3.1 If mercury or copper is used, centrifuge
and filter the extract to remove all
residual mercury or copper. Dispose of
the mercury waste properly. Bring .the
final volume to 1.0 mL and analyze by gas
chromatography per the procedure in
section 13.
11.6.3.2 If TBA sulfite is used, add 5 mL of
reagent water and shake for 1 - 2 minutes.
Centrifuge and filter the extract to
remove all precipitate. Transfer the
hexane (top) layer to a sample vial and
adjust the final volume to 5 or 10 mL as
in section 10.6, depending on whether the
extract was subjected to GPC cleanup, and
analyze by gas chromatography per section
13.
12 HYDROLYSIS AND ESTERIFICATIOM OF PHENOXY-
ACID HERBICIDES - Sample extracts that
have been cleaned up by GPC are diluted to
100 - 200 mL in a 500 - 1000 mL separatory
funnel prior to separation of the acids
from the esters.
12.1 Separation of phenoxy-acids and phenoxy-
acid esters
12.2 Ester hydrolysis
12.2.1 Transfer the organic layer to a K-D flask
and concentrate to 20 - 30 mL per section
10.5.1.
12.2.2 After the flask has cooled, remove the
Snyder column and add 5 mL of 37 percent
aqueous KOH, 30 mL of reagent water, and
40 mL of methanol.
12.2.3 Add one or two boiling chips to the flask,
install a condenser, and return the
apparatus to the water bath. Reflux the
mixture for 2-3 hours. Remove the flask
from the water bath and allow to drain and
cool for at least 10 minutes.
12.2.4 Transfer the hydrolysate to a 100 - 500 mL
separatory funnel Add 50 mL of methylene
chloride to the funnel and extract the
hydrolysate by shaking the funnel for two
minutes with periodic venting to release
excess pressure. Allow the layers to
separate for a minimum of ten minutes.
Discard the organic phase. Repeat the
extraction twice more. The aqueous phase
contains the free acids.
12.1.1 Add 100 - 200 mL of 0.1 N aqueous sodium
hydroxide solution to the separatory
funnel containing the methylene chloride
extract (section 10.3.4), the methylene
chloride/acetone extract (section 10.4.7),
or the GPC cleaned up sample extract
(10.6.2).
12.1.2 Insert the stopper into the funnel and
shake for two minutes with periodic
venting to release excess pressure. Allow
the organic layer to separate from the
aqueous layer for a minimum of ten
minutes.
12.3 Extraction/concentration of the free acids
12.3.1 Combine the aqueous phases from the
separation (12.1.3) and hydrolysis
(12.2.4) steps in the separatory funnel.
12.3.2 Adjust the pH of the solution to <2 with
H2S04 and extract three times with 100 mL
portions of methylene chloride. Combine
the organic extracts and pour through a
prerinsed drying column containing 7 to 10
cm of acidified anhydrous sodium sulfate.
Collect in a K-D flask fitted with a 10 mL
condenser.
12.1.3 Drain the organic and aqueous layers into
separate clean beakers. Return the
organic phase to the extractor and repeat
the extraction twice more. The aqueous
layer contains the free acids; the organic
layer contains the herbicide esters that
must be hydroIyzed.
12.3.3 Concentrate the extract to approximately 5
mL per section 10.5 and further
concentrate the extract to near dryness
using the nitrogen blowdown apparatus.
Bring the volume to 5 mL with isooctane.
If desired, the extract may be transferred
to a 10 mL sample vial and stored at -20
to -10 °C.
109
-------
12.4 Esterification--observe the safety
precautions regarding diazomethane in
section 4.
12.4.1 Set up the diazomethane generation
apparatus as given in the instructions in
the Oiazald kit.
12.4.2 Transfer one mL of the isooctane solution
(section 12.3.3) to a clean vial and add
0.5 mL of methanol and 3 mL of ether. For
extracts that have been cleaned up by GPC,
use 2 mL.
12.4.2 Add two mL of diazomethane solution and
let the sample stand for 10 minutes with
occasional swirling. The yellow color of
diazomethane should persist throughout
this period. If the yellow color
disappears, add two mL of diazomethane
solution and allow to stand, with
occasional swirling, for another 10
minutes. Colored or complex samples will
require at least 4 ml of diazomethane to
ensure complete reaction of the
herbicides. Continue adding diazomethane
in 2 mL increments until the yellow color
persists for the entire 10 minute period
or until 10 mL of diazomethane solution
has been added.
12.4.3 Rinse the inside wall of the container
with 0.2 - 0.5 mL of diethyl ether and add
10 - 20 mg of silicic acid to react excess
diazomethane. Filter through Whatman #41
paper into a clean sample vial. If the
solution is colored or cloudy, evaporate
to near dryness using the nitrogen
blowdown apparatus, bring to 10 mL with
hexane, and proceed to section 11.1 for
diol cleanup. If the solution is clear
and colorless, evaporate to near dryness,
bring to 1.0 mL with hexane and proceed to
section 13 for GC analysis.
13 GAS CHROMATOGRAPHY - tables 4 - 5
summarize the recommended operating
conditions for the gas ctiromatographs.
Included in these tables are the retention
times and estimated detection limits that
can be achieved under these conditions.
Examples of the separations achieved by
the primary and secondary columns are
shown in figures 1 through 10.
13.1 Calibrate the system as described in
section 7.
13.2 Combination of pesticide and herbicide
extracts
13.2.1 Pesticide extracts cleaned up by diol
cartridge—combine the 1.0 mL final
pesticide extract (section 11.3.4.5 or
11.5.3) with the 1.0 mL final herbicide
extract (section 11.3.4.5 or 11.5.3 if the
herbicide extract required cleanup;
section 12.4.3 if it did not).
13.2.1 Pesticide extracts cleaned up by Florisil-
• combine 1.0 mL of the 5.0 mL or 10.0 mL
pesticide extract (section 11.4.5) with
the 1.0 mL final herbicide extract
(section 11.3.4.5 or 11.5.3 if the
herbicide extract required cleanup;
section 12.4.3 if it did not).
13.3 Addition of internal standard--if the
internal standard calibration procedure is
being used, add the internal standard
solution to the extract immediately prior
to loading the extract into the auto
sampler to minimize the possibility of
loss by evaporation, adsorption, or
reaction. Mix thoroughly.
13.4 Set the injection volume on the
autosampler to inject 1.0 uL of all
standards and extracts of blanks and
samples.
13.5 Set the data system or GC control to start
the temperature program upon sample
injection, and begin data collection after
the solvent peak elutes. Set the data
system to stop data collection at the end
of the temperature program and to return
the column to the initial temperature.
14 SYSTEM AND LABORATORY PERFORMANCE
14.1 At the beginning of each eight hour shift
during which analyses are performed, GC
system performance and calibration are
110
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verified for all pollutants and surrogates
on all column/detector systems For these
tests, analysis of the combined QC
standard (tables 4 and 5) shall be used to
verify all performance criteria.
Adjustment and/or recalibrat ion (per
section 7) shall be performed until all
performance criteria are met. Only after
all performance criteria are met may
samples, blanks, and precision and
recovery standards be analyzed.
14.2 Retention times
14.2.1 External standard--the absolute retention
times of the peak maxima shall be within
+/- 10 seconds of the retention times in
the initial calibration (section 7.3.1 and
7.5.5).
14.2.1 Internal standard--the absolute retention
times of the peak maxima shall be within
+/- 30 seconds of the retention times in
the initial calibration (section 7.4.2)
and the retention time difference between
a compound of interest and its internal
standard shall be within +/- 5 seconds of
this retention time in the initial
calibration (section 7.4.2 and 7.5.5).
14.3 GC resolution--resolution is acceptable if
the valley height between two peaks (as
measured from the baseline) is less than
50 percent of the taller of the two peaks.
14.3.1 OTgano-halide compounds
14.3.1.1 Primary column (DB-S)--TBD and TBD.
14.3.1.2 Confirmatory column (SPB-608)--TBO and
TBD.
14.3.2 Thiophosphorus compounds
14.3.2.1 Primary column (DB-S)--TBD and TBD.
14.3.2.2 Confirmatory column (SPB-608)--TBD and
TBD.
14.4 Decomposition of DDT and endrin--TBD
14.5 Calibration verification--calibration is
verified for the combined QC standard
only. If verification requirements are
met, the calibration is assumed to be
valid for the multicomponent analytes
(PCBs, chlordane, toxaphene).
14.5.1 External standard--compute the area of
each peak in the combined calibration
standard. This area shall be within +/-
50 percent of the area in the initial
analysis of this standard (section 7.5.5).
14.5.1 Internal standard--compute the response
factor of each peak in the combined QC
standard. The response factor shall be
within +/- 25 percent of the response
factor in the initial analysis of this
standard (section 7.5.5).
14.6 On-going precision and accuracy
14.6.1 Analyze the extract of the precision ami
recovery standard extracted with each
sample lot.
14.6.2 Compute the concentration of each analyte
by the internal or external standard
method.
14.6.3 For each analyte, compare the
concentration with the initial recovery
determined in the initial test (section
8.2). The concentration of each analyte
shall be within +/- two standard
deviations of the average concentration
determined in the initial test of the
method (section 8.2). If all analytes
pass, the extraction, concentration, and
cleanup processes are in control and
analysis of blanks and samples may
proceed. If, however, any of the analytes
fail, these processes are not in control.
In this event, correct the problem, re-
extract the sample lot, and repeat the on-
going precision and recovery test.
14.6.4 Add results which pass the specifications
in 12.6.3 to initial and previous on-going
data. Update QC charts to form a graphic
representation of continued laboratory
performance. Develop a statement of
111
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laboratory data quality for each analyte
by calculating the average percent
recovery (R) and the standard deviation of
percent recovery sr. Express the accuracy
as a recovery interval from R - 2sr to R +
2sr. For example, if R * 95X and sr = 5X,
the accuracy is 85 - 10SX.
15 QUALITATIVE DETERMINATION
15.1 Qualitative determination is accomplished
by comparison of data from analysis of a
sample or blank with data from analysis of
the shift standard (section 13.1), and
with data stored in the retention time and
calibration libraries (section 7.2.3 and
7.4.3) Identification is confirmed when
retention time and amounts agree per the
criteria below.
15.2 External standard--for each compound on
each column/detector system, establish a
retention time window +/• 20 seconds on
either side of the retention time in the
calibration data (section 7.3) For
compounds that have a retention time curve
(section 7.3.2.2), establish this window
as the minimum -20 seconds and maximum +20
seconds. For the multi-component
analytes, use the retention times of the
five largest peaks in the chromatogram
from the calibration data (section 7.3.1).
15.2.1 Compounds not requiring a retention time
calibration curve--if a peak from the
analysis of a sample or blank is within a
window (as defined in section 15.2) on the
primary column/detector system, it is
considered tentatively identified. A
tentatively identified compound is
confirmed when (1) the retention time for
the compound on the confirmatory
column/detector system is within the
retention time window on that system, and
(2) the computed amounts (section 16) on
each system (primary and confirmatory)
agree within a factor of three.
15.2.2 Compounds requiring a retention time
calibration curve--if a peak from the
analysis of a sample or blank is within a
window (as defined in section 15.2) on the
primary column/detector system, it is
considered tentatively identified. A
tentatively identified compound is
confirmed when (1) the retention times on
both systems (primary and confirmatory)
are within +/- 30 seconds of the retention
times for the computed amounts (section
16), as determined by the retention time
calibration curve (section 7.3.2.2), and
(2) the computed amounts (section 16) on
each system (primary and confirmatory)
agree within a factor of three.
15.3 Internal standard—for each compound on
each column/detector system, establish a
relative retention time window equivalent
to +/- 10 seconds on either side of the
relative retention time in the calibration
data (section 7.4). For compounds that
have a retention time curve (section
7.4.3.2), establish this window as the
minimum -20 seconds and the maximum +20
seconds. For the multi-component
analytes, use the relative retention times
of the five largest peaks in the
chromatogram from the calibration data
(section 7.4.3).
15.3.1 Compounds not requiring a relative
retention time calibration curve--if a
peak from the analysis of a sample or
blank is within a window (as defined in
section 15.3) on the primary
coI urn/detector system, it is considered
tentatively identified. A tentatively
identified compound is confirmed when (1)
the relative retention time for the
compound on the confirmatory
column/detector system is within the
relative retention time window on that
system, and (2) the computed amounts
(section 16) on each system (primary and
confirmatory) agree within a factor of
three.
15.3.2 Compounds requiring a relative retention
time calibration curve--if a peak from the
analysis of a sample or blank is within a
window (as defined in section 15.3) on the
primary column/detector system, it is
considered tentatively identified. A
tentatively identified compound is
-------
confirmed when (1) the relative retention
times on both systems (primary and
confirmatory) are within the relative
retention time equivalent of +/-20 seconds
of the relative retention times for the
computed amounts (section 16), as
determined by the relative retention time
calibration curve (section 7.4.3.2), and
(2) the computed amounts (section 16) on
each system (primary and confirmatory)
agree within a factor of three.
16 QUANTITATIVE DETERMINATION
16.1 External standard
16.1.1 Using the GC data system, compute the
concentration of the analyte detected in
the extract (in ug/mL) using the
calibration factor or calibration curve
(section 7.3.3.2).
16.2 Internal standard
16.2.1 Using the GC data system, compute the
concentration of the analyte detected in
the extract (in ug/mL) using the response
fac tor or calibration curve (section
7.4.4.2) using the following equation:
ex
where C is the concentration of the
analyte in the extract, and the other
terms are as defined in section 7.4.4
16.2.2 Liquid samples--compute the concentration
in the sample using the following
equation:
10
-------
which the concentration is in the
calibration range.
17 ANALYSIS OF COMPLEX SAMPLES
17.1 Some samples may contain high levels
(>1000 ug/L) of the com pounds of
interest, interfering compounds, and/or
polymeric materials. Some samples may not
concentrate to 10 mL (section 10.6);
others may overload the GC column and/or
detector.
17.2 The analyst shall attempt to clean up all
samples using GPC (section 10.2), Florisil
(section 10.4), diol cartridge (section
10.3), and sulfur removal (section 10.5).
If these techniques do not remove the
interfering compounds, the extract is
di luted by a fac tor of 10 and reanalyzed
(section 16.2).
17.3 Recovery of surrogates--in most samples,
surrogate recoveries will be similar to
those from reagent water or from the high
solids reference matrix. If the surrogate
recovery is outside the range of 20 - 200
percent, the sample shall be reextracted
and reanalyzed. If the surrogate recovery
is still outside this range, the method
does not work on the sample being analyzed
and the result may not be reported for
regulatory compliance purposes.
18 METHOD PERFORMANCE
18.1 Development of this method is detailed in
reference 10.
References
1. "Working with Carcinogens," DHEU, PHS,
CDC, NIOSH, Publication 77-206, (Aug
1977).
2. "OSHA Safety and Health Standards, General
Industry" OSHA 2206, 29 CFR 1910 (Jan
1976).
3. "Safety in Academic Chemistry
Laboratories," ACS Committee on Chemical
Safety (1979).
4. Mills, P. A., "Variation of Florisil
Activity: Simple Method for Measuring
Adsorbent Capacity and Its Use in
Standardizing Florisil Columns," J. Assoc.
Off. Analytical Chemists, 51, 29 (1968).
5. "Handbook of Analytical Quality Control in
Water and Uastewater Laboratories," USERA,
EMSL, Cincinnati, OH 45268, EPA-600/4-79-
019 (March 1979).
6. "Standard Practice for Sampling Water,"
ASTM Annual Book of Standards, ASTM,
Philadelphia, PA, 76 (1980).
7. "Methods 330.4 and 330.5 for Total
Residual Chlorine," USEPA, EMSL,
Cincinnati, OH 45268, EPA 600/4-70-020
(March 1979).
8. "Determination of Pesticides and PCBs in
Industrial and Municipal Wastewaters,"
EPA-600/4-82-023, US Environmental
Protection Agency, Environmental
Monitoring and Support Laboratory,
Cincinnati, Ohio, 45268, June 1982.
9. Goerlitz, D.F., and Law, L.M. "Bulletin
for Environmental Contamination and
Toxicology," 6, 9 (1971).
10. "Consolidated GC Method for the
Determination of ITD/RCRA Pesticides using
Selective GC Detectors," Report Reference
32145-01, Document R70, S-CUBED, A
Division of Maxwell Laboratories, Inc, PO
Box 1620, La Jot la, CA, 92038-1620
(September 1986)
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EPA METHOD 8290
ANALYSIS OF PCDD/PCDF'S
115
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DRAFT
24 May 1987
METHOD 8290
ANALYTICAL PROCEDURES AND QUALITY ASSURANCE
FOR MULTIMEDIA ANALYSIS
OF
POLYCHLORINATED DIBENZO-p-DIOXINS
AND
POLYCHLORINATED DIBENZOFURANS
BY
HIGH-RESOLUTION GAS CHROMATOGRAPHY/HIGH-RESOLUTION MASS
SPECTROMETRY
(Exhibits D and E)
by
Yves Tondeur
June 1987
Project Officer
Werner F. Beckert
Environmental Monitoring Systems Laboratory
Las Vegas, Nevada 89193-3478
116
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Draft
24 May 1987
NOTICE
This document is a preliminary draft. It has not been formally released
by the University of Nevada Environmental Research Center or the U.S. Environ-
mental Protection Agency, and it should not at this stage be construed to
represent University or Agency policy. It is circulated for comments on its
technical merit and policy implications.
-------
Draft
24 May 1987
FOREWORD
In January 1986, the Environmental Protection Agency published an analy-
tical protocol, Protocol for the Analysis of 2,3,7,8-Tetrachlorodibenzo-p-
Dioxin (TCDD) by High-Resolution Gas Chroraatography/High-Resolution Mass
Spectrometry (HRGC/HRMS) (EPA 600/4-86-004), aimed at the determination of part-
per-trillion and sub-part-per-trillion levels of 2,3,7,8-TCDD and of total TCDD
in soil, sediment and aqueous samples. The January 1986 document was intended
to be a stepping stone for the realization of a more comprehensive method that
would include all the polychlorinated dibenzodioxin (PCDD) and polychlorinated
dibenzofuran (PCDF) congeners present in a broader spectrum of environmentally
significant matrices.
The present report constitutes a draft addressing the analytical proce-
dures (Exhibit D) and quality assurance (Exhibit E, quality assessment and
control) requirements sections of the future analytical protocol for the
analysis of PCDDs and PCDFs by HRGC/HRMS; i.e., Method 8290. At times, refer-
ence to other exhibits (e.g., Exhibit C) are made, even though these sections
have not been prepared. The format used for this report is similar to the
format used for other EPA TCDD protocols. Figures and tables are, however,
grouped at the end of Exhibit D. A final version of Method 8290 is expected
following peer review of this draft report and the completion of the single-
laboratory evaluation. Elements included in this Method 8290 have been taken
from a variety of sources, such as the EPA Region VII low-resolution mass
spectrometry (LRMS) TCDD protocol, the aforementioned high-resolution mass
ii
118
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Draft
24 May 1987
spectrometry TCDD protocol, the RCRA Method 8280 (LRMS) protocol, the method
evaluation study final report by the Midwest Research Institute on "Analysis
for Polychlorinated Dibenzo-p-Dioxins and Dibenzofurans in Human Adipose Tissue"
(EPA-560/5-86-020), the National Dioxin Study Analytical Procedures and Quality
Assurance Plan for the Analysis of 2,3,7,8-TCDD in Tier 3-7 Samples (EPA/600/3-
85/019), and the analytical protocol for the analysis for PCDDs and PCDFs by
HRGC/HRMS submitted recently for review by Region VII. Also, we wish to
acknowledge the contributions from experts in the analysis of PCDDs and PCDFs
in environmental samples. The cooperation of P. W. Albro (National Institute
of Environmental Health Sciences, Research Triangle Park, NC), L. Alexander
(Center for Disease Control, Atlanta, GA), J. R. Hass and D. J. Harvan (Triangle
Laboratories, Inc., Research Triangle Park, NC), R. Harless (US EPA, Research
Triangle Park, NC), R. D. Kleopfer (US EPA, Region VII, Kansas City, MO), D. W.
Kuehl (US EPA, Duluth, MN), M. J. Miille (California Analytical Laboratories,
Sacramento, CA), R. W. Noble (Monsanto Company, Dayton, OH), T. M. Sack
and J. S. Stanley (Midwest Research Institute, Kansas City, MO), and T. S.
Viswanathan (Ecology and Environment, Inc., Kansas City, MO) is particularly
appreciated.
iii
119
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Draft
24 May 1987
TABLE OF CONTENTS
Foreword ii
Abbreviations and Symbols v
Analytical Methods (Exhibit D)
1. Scope and Application D-l
2. Summary of the Method D-2
3. Definitions D-5
4. Interferences D-10
5. Safety D-ll
6. Apparatus and Equipment D-l6
7. Reagents and Standard Solutions D-23
8. System Performance Criteria. . . .' D-27
9. Calibration D-32
10. Quality Assessment/Quality Control Procedures D-AO
11. Sample Preservation D-41
12. Extraction and Cleanup Procedures D-44
13. Analytical Procedures D-59
14. Calculations D-63
APPENDIX A: PROCEDURE FOR THE COLLECTION, HANDLING, ANALYSIS, AND
REPORTING REQUIREMENTS OF WIPE TESTS PERFORMED WITHIN THE
LABORATORY D-71
APPENDIX B: STANDARDS TRACEABILITY PROCEDURE D-75
APPENDIX C: SIGNAL-TO-NOISE DETERMINATION METHOD D-81
Figures D-84
Tables D-93
Quality Assurance Requirements (Exhibit E)
1. Summary of QA/QC Analyses E-l
2. Quality Assessment/Quality Control E-2
3. Laboratory Evaluation Procedures E-12
iv
120
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Draft
24 May 1987
LIST OF ABBREVIATIONS AND SYMBOLS
A
ADC
AX-21
C
CDC
CDWG
8 C
cm
DB-5
DS
EDL
EMPC
EMSL-LV
EPA
g
GC
GC/MS
HEPA
HpCDD
HpCDF
HRGC/HRMS
HxCDD
HxCDF
IFB
IS
KD
L
MB
MCL
mL
mm
M/AM
MS
MSD
OCDD
OCDF
OSHA
PCB
PCDD
PCDPE
PCDF
PE
Integrated Ion abundance
Analogue-to-digltal- conversion
Type of carbon adsorbent
Concentration
Center for Disease Control
Chlorinated Dloxlns Workgroup
Degree centigrade
Carbon-13 labeled
Centimeter
Type of fused-sllica capillary column
Data system
Estimated detection limit
Estimated maximum possible concentration
Environmental Monitoring System Laboratory, Las Vegas
Environmental Protection Agency
Gram
Gas chroraatography or gas chromatograph
Gas chromatography/mass spectrometry
High-efficiency particulate absorbent
Heptachlorodibenzodioxln
Heptachlorodibenzofuran
High-resolution gas chromatography /high-resolution
mass spectrometry
Hexachlorodibenzodioxin
Hexachlorodibenzofuran
Invitation for Bid
Internal Standard
Kuderna-Danish
Liter
Method blank
Method calibration limit
Milliliter
Millimeter
Mass spectrometer resolving power
Matrix spike
matrix spike duplicate
Octachlorodibenzodloxin
Octachlorodibenzofuran
Occupational Safety and Health Administration
Polychlorinated biphenyl
Polychlorinated dibenzodloxin
Polychlorinated dlphenyl ether
Polychlorinated dibenzofuran
Performance evaluation
121
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Draft
24 May 1987
PEM
PeCDD
PeCDF
PFK
Pg
ppm
ppt
Q
QA
QA/QC
rpm
RPD
RRF
RRF
RRT
RS
S
SAS
SES
SICP
SIM
SMO
S/N
SOP
SP-2330
Still-
bottom
TCDD
TEF
V
v/v
W
WTE
uL
Performance evaluation material
Pentachlorodlbenzodloxln
Pentachlorodlbenzofuran
Perfluorokerosene
Plcogram
Part per million
Part per trillion
Amount of substance
Quality Assurance or Quality Assessment
Quality Assessment/Quality Control
Revolutions per minute
Relative percent difference
Relative response factor
Mean relative response factor
Relative retention time
Recovery standard
EPA reference standard solution
Special Analytical Service
Site evaluation sheet
Selected ion current profile
Selected ion monitoring
Sample Management Office
Signal-to-noise ratio
Standard Operating Procedure
Type of fused-silica capillary column
Name of a matrix that is used as a noun
Tetrachlorodibenzodioxin
Toxicity Equivalency Factor
Volume
Volume/volume
Weight or laboratory working standard
Wipe test experiment
Microliter
vi
122
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Draft
24 May 1987
ANALYTICAL METHODS
(EXHIBIT D)
123
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Draft
24 May 1987
EXHIBIT D
1. Scope and Application
1.1 This method provides procedures for the detection and quantitative measure-
ment of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD), 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) concentrations. 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 calibra-
tion limits (MCLs) and other germane information. Analysis of a one-tenth
aliquot of the sample permits measurement of concentrations up to 10 times
the upper MCL (Table 1). Samples containing concentrations of specific
congeneric analytes (PCDDs and PCDFs) considered within the scope of this
method that are greater than the upper MCL must be analyzed by a protocol
designed for such concentration levels. An optional method for reporting
the analytical results using a 2,3,7,8-TCDD toxicity equivalency factor
(TEF) is described.
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1.2 The sensitivity of this method is dependent upon the level of interferences
within a given matrix. Actual limits of detection and quantification will
be provided based on the single- or multi-laboratory evaluation of this
protocol, and on examining the data gathered by the Sample Management
Office (SMO) from Special Analytical Services (SAS) performed over the
past few years.
1.3 This method is designed for use by analysts who are experienced with
residue analysis and skilled in high-resolution gas chromatography/high-
resolution mass spectrometry (HRGC/HRMS).
1.4 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.
2. Summary of the Method
2.1 This procedure uses matrix-specific extraction, analyte-specific cleanup,
and high-resolution capillary column gas chromatography/high-resolution
mass spectrometry (HRGC/HRMS) 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 shown in Figure 1.
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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^)
labeled PCDDs/PCDFs listed in Column 1 of Table 2. The sample is then
extracted according to a matrix-specific extraction procedure. The extrac-
tion procedures are: a) toluene (or benzene) Soxhlet extraction for soil,
sediment and fly ash samples; b) methylene chloride liquid-liquid extrac-
tion for water samples; c) toluene (or benzene) Dean-Stark extraction for
fuel oils and aqueous sludges; d) toluene (or benzene) extraction for
still-bottoms; e) hexane/methylene chloride Soxhlet extraction for fish
tissue and paper pulp; and f) raethylene chloride extraction for human
adipose tissue. 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 residue is cleaned up by column
chromatography on neutral alumina and carbon on Celite 545®. The extract
from adipose tissue is treated with silica gel impregnated with sulfuric
acid before chromatography on acidic silica gel, neutral alumina, and
carbon on Celite 545®. Fish tissue and paper pulp are subjected to an
acid wash treatment only prior to chroraatography or neutral alumina and
carbon/Celite. The preparation of the final extract for HRGC/HRMS
analysis is accomplished by adding, to the concentrated carbon column
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eluate, 10 to 50 uL uL (depending on the matrix type) of a tridecane
solution containing 50 pg/uL of each of the two recovery standards
13C12-1,2,3,4-TCDD and 13C12-1,2,3,7,8,9-HxCDD (Table 2). The former is
used to determine the percent recoveries of tetra- and pentachlorinated
PCDD/PCDF congeners while the latter is used for the determination of
hexa-, hepta- and octa-chlorinated PCDD/PCDF congeners percent recoveries.
2.5 One to two uL 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).
2.6 The identification of OCDD and nine of the fifteen 2,3,7,8-substituted
congeners (Table 3), for which a !3C-labeled standard is available in the
sample fortification and recovery standard solutions (Table 2), is based
on their elution at their exact retention time (-1 to +3 seconds from the
respective internal or recovery standard signal) 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 estab-
lished by using a GC column performance evaluation solution, 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
D-4
127
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1 O
relative to A Cj2~OCDD and the simultaneous detection of the two most
abundant ions in the molecular ion region. Confirmation is based on a
comparison of the ratio of the integrated ion abundance of the molecular
ion species to their theoretical abundance ratio.
2.7 Quantification of the individual congeners, total PCDDs and total PCDFs is
achieved in conjunction with the establishment of a multipoint (seven
points) calibration curve for each homologue, during which each cali-
bration solution is analyzed once.
3. Definitions
3.1 Polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans
(PCDFs): Compounds (Figure 2) that contain from one to eight chlorine
atoms. The fifteen 2,3,7,8-substituted PCDDs (totaling 75) and PCDFs
(totaling 135) are shown in Table 3. The number of isomers at different
chlorination levels is shown in Table 4.
3.2 Homologous series: Defined as a group of chlorinated dibenzodioxins or
dibenzofurans having a specific number of chlorine atoms.
3.3 Isomer: Defined by the arrangement of chlorine atoms within an
homologous series. For example, 2,3,7,8-TCDD is a TCDD isomer.
3.4 Congener: Any isomer of any homologous series.
D-5
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3.5 Internal Standard: An Internal standard Is a C,o~labeled analogue of a
congener chosen from the compounds listed in Table 3 and of OCDD. Internal
standards are added to all samples including method blanks and quality con-
trol samples before extraction, and they are used to measure the concentra-
tion of the analytes. Nine internal standards are used in this method.
There is one for each of the dioxin and furan homologues (except for OCDF)
with the degree of chlorination ranging from four to eight.
3.6 Recovery Standard: Recovery standards (two) are used to determine the
percent recoveries for PCDDs and PCDFs. The 13C12~1»2,3,4-TCDD is used to
measure the percent recoveries of the tetra- and pentachlorinated dioxins
i "3
and furans while iJCj2~l|2,3,7,8,9-HxCDD permits the recovery determination
of the hexa-, hepta- and octachlorinated homologues. They are added to
the final sample extract before HRGC/HRMS analysis. Furthermore, ^C,--
1,2,3,7,8,9-HxCDD is used for the identification of the unlabeled analogue
present in sample extracts (this exhibit, Section 2.6).
3.7 High-Resolution Concentration Calibration Solutions (Table 5): Solutions
(tridecane) containing known amounts of 17 selected PCDDs and PCDFs, nine
internal standards ( C^-labeled PCDDs/PCDFs), and two carbon-labeled
recovery standards (this exhibit, Section 3.6); the set of seven solutions
is used to determine the instrument response of the unlabeled analytes
relative to the internal standards and of the internal standards relative
to the recovery standards.
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3.8 Sample Fortification Solution (Table 2): A solution (isooctane) containing
the nine internal standards, which is used to spike all samples before
extraction and cleanup.
3.9 Recovery Standard Solution (Table 2): A tridecane solution containing the
two recovery standards, which is added to the final sample extract before
HRGC/HRMS analysis.
3.10 Field Blank: A portion of a sample representative of the matrix under
consideration, which is free of any PCDDs/PCDFs.
3.11 Laboratory Method Blank: A blank prepared in the laboratory and carried
through all analytical procedure steps except the addition of a sample
aliquot to the extraction vessel.
3.12 Rinsate: A portion of solvent used to rinse sampling equipment. The
rinsate is analyzed to demonstrate that samples were not contaminated
during sampling.
3.13 GC Column Performance Check Mixture: A tridecane solution containing a
mixture of selected PCDD/PCDF standards including the first and last
eluters for each homologous series, which is used to demonstrate continued
acceptable performance of the capillary column (i.e., £ 25 percent valley
separation of 2,3,7,8-TCDD from all the other 21 TCDD isomers) and to
define the homologous PCDD/PCDF retention time windows.
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This Page Was Intentionally Left Blank
131
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3.14 Performance Evaluation Materials: Representative sample portions
containing known amounts of certain unlabeled PCDD/PCDF congeners (in
particular the ones having a 2,3,7,8-substitution pattern). Representa-
tive interferences may be present. PEMs are obtained from the EPA EMSL-LV
and submitted to potential contract laboratories, who must analyze these
and obtain acceptable results before being awarded a contract for sample
analyses (see IFB Pre-Award Bid Confirmations). PEMSs are also included
as unspecified ("blind") quality control (QC) samples in any sample batch
submitted to a laboratory for analysis.
3.15 Relative Response Factor: Response of the mass spectrometer to a known
amount of an analyte relative to a known amount of an internal standard.
3.16 Estimated Level of Method Blank Contamination: The response from a signal
occurring in the homologous PCDD/PCDF retention time windows, at any of
the masses monitored, is used to calculate the level of contamination
in the method blank, as described in Section 14 (this exhibit). The
results from such calculations must be reported along with the data
obtained on the samples belonging to the batch associated with the method
blank.
Reporting a method blank contamination level for any of the 2,3,7,8-
substituted congeners except OCDD and OCDF that exceeds 10 percent
of the desired detection limit would invalidate the results and require
automatic sample reruns (Exhibit C) for all positive samples found in
that batch of samples. A positive sample is defined as a sample found to
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contain at least one 2,3,7,8-substltuted PCDD/PCDF congener (except OCDD
and OCDF). A valid method blank run is an analysis during which all
internal standard signals are characterized by S/N of at least 10:1.
3.17 Sample Rerun: Extraction of another portion of the sample followed by
extract cleanup and extract analysis.
3.18 Extract Reanalysis: Analysis by HRGC/HRMS of another aliquot of the
final extract.
3.19 Mass Resolution Check: Standard method used to demonstrate a static
resolving power of 10,000 minimum (10 percent valley definition).
3.20 Method Calibration Limits (MCLs): For a given sample size, a final
extract volume, and the lowest and highest concentration calibration
solutions, the lower and upper MCLs delineate the region of quantification
for which the HRGC/HRMS system was calibrated with standard solutions.
3.21 HRGC/HRMS Method Blank (MB): This additional QC check analysis corresponds
to a 2-uL injection of the method blank extract into the GC column and a
complete (tetra- through octachlorinated congeners) HRGC/HRMS analysis.
Such a QC check is required following a calibration run and before the
daily analysis of the first sample extract. Acceptable HRGC/HRMS method
blanks (see this exhibit, Section 3.16, for guidelines) must be obtained
before sample extracts can be analyzed.
D-9
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3.22 Matrix Spike (MS): A sample which is spiked with a known amount of the
matrix spike fortification solution (this exhibit, Section 3.24) prior
to the extraction step. The recoveries of the matrix spike compounds are
determined; they are used to estimate the effect of the sample matrix
upon the analytical methodology.
3.23 Matrix Spike Duplicate (MSD): A second portion of the same sample as
used in the matrix spike analysis and which is treated like the matrix
spike sample.
3.24 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. The solution also contains all
internal standards used in the sample fortification solution at concen-
trations as shown in Table 2.
4. Interferences
4.1 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 at the
end of this Section). All of these materials must be demonstrated to
be free from interferents under the conditions of analysis by running
laboratory method blanks. Analysts should avoid using PVC gloves.
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4.2 The use of high-purity reagents and solvents helps minimize interference
problems. Purification of solvents by distillation in all-glass systems
may be necessary.
4.3 Interferents co-extracted 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 xanthenes that may be found at con-
centrations 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 8.1.3 (this exhibit). While certain clean-
up techniques are provided as part of this method, unique samples may
require additional cleanup steps to achieve lower detection limits.
4.4 A high-resolution capillary column (60 m DB-5) is used to resolve as many
PCDD and PCDF isomers as possible; however, no single column is known to
resolve all isomers. The use of several capillary columns will, in fact,
be necessary during the determination of the toxicity equivalency factors
(TEFs) (this exhibit, Section 14.7).
References:
1. "Control of Interferences in the Analysis of Human Adipose Tissue
for 2,3,7,8-Tetrachlorodibenzo-p-dioxin". D. G. Patterson et al.,
Environ. Toxicol. Chem. 5, 355-360 (1986).
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2. "Protocol for the Analysis of 2,3,7,8-TCDD by HRGC/HRMS".
J. S. Stanley and• T. M. Sack, EPA 600/4-86-004.
5. Safety
5.1 The following safety practices are exerpted directly from EPA Method 613,
Section 4 (July 1982 version) and amended for use In conjunction with
this method.
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 inhala-
tion and ingestion. It is recommended that such samples be processed in
a confined environment, such as a hood or a glove bo*. Laboratory
personnel handling these types of samples should also wear masks fitted
with charcoal filter absorbent media to prevent inhalation of dust.
5.2 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 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.
D-12
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Additional references to laboratory safety are given in references 1-3
(see end of Section 5, this exhibit). Benzene and 2,3,7,8-TCDD have been
identified as suspected human or mammalian carcinogens.
5.3 Each laboratory must develop a strict safety program for the handling of
2,3,7,8-TCDD. The laboratory practices listed below are recommended.
5.3.1 Contamination of the laboratory will be minimized by conducting most of
the manipulations in a hood.
5.3.2 The effluents of sample splitters for the gas chromatograph and roughing
pumps on the HRGC/HRMS system should pass through either a column of ac-
tivated charcoal or be bubbled through a trap containing oil or high-
boiling alcohols.
5.3.3 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 liquid wastes and dispose of the solutions when 2,3,7,8-TCDD can no
longer be detected.
5.4 Some of 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.
5.4.1 The following statements on safe handling are as complete as possible on
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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. Techniques used in handling radioactive and
infectious materials are applicable to 2,3,7,8-TCDD.
5.4.1.1 Protective Equipment: Throw-away plastic gloves, apron or lab coat,
safety glasses and laboratory hood adequate for radioactive work.
5.4.1.2 Training: Workers must be trained in the proper method of removing
contaminated gloves and clothing without contacting the exterior
surfaces.
5.4.1.3 Personal Hygiene: Thorough washing of hands and forearms after each
manipulation and before breaks (coffee, lunch, and shift).
5.4.1.4 Confinement: Isolated work area, posted with signs, segregated glass-
ware and tools, plastic-backed absorbent paper on benchtops.
5.4.1.5 Waste: Good technique includes minimizing contaminated waste.
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Plastic bag liners should be used in waste cans.
5.4.1.6 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.
5.4.1.7 Decontamination: Personnel - any mild soap with plenty of scrubbing
action. Glassware, tools and surfaces - Chlorothene NU Solvent (Trade-
mark 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 any detergent and water. Dish
water may be disposed to the sewer after percolation through a char-
coal bed filter. It is prudent to minimize solvent wastes because
they require special disposal through commercial sources that are
expensive.
5.4.1.8 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.
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5.4.1.9 Wipe Tests: A useful method of 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: Appendix A describes a procedure for the collection, handling,
analysis, and reporting requirements of wipe tests performed within
the laboratory. The results and decision making processes are based
on the presence of 2,3,7,8-substituted PCDD/PCDFs.
5.4.1.10 Inhalation: Any procedure that may produce airborne contamination
must be carried out with good ventilation. Gross losses to a venti-
lation 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.
5.4.1.11 Accidents: Remove contaminated clothing immediately, taking precau-
tions not to contaminate skin or other articles. Wash exposed skin
vigorously and repeatedly until medical attention is obtained.
References:
1. "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.
2. "OSHA Safety and Health Standards, General Industry", (29 CFR 1910),
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Occupational Safety and Health Administration, OSHA 2206 (revised
January 1976).
3. "Safety in Academic Chemistry Laboratories", American Chemical Society
Publication, Committee on Chemical Safety (3rd Edition, 1979.)
6. Apparatus and Equipment
6.1 High-Resolution Gas Chromatograph/High-Resolution Mass Spectrometer/Data
System (HRGC/HRMS/DS).
6.1.1 The GC must be equipped for temperature programming, and all required
accessories must be available, such as syringes, gases, and capillary
columns. The GC injection port must be designed for capillary
columns. The use of splitless injection techniques is recommended.
On-column 1-ul 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-uL injection volume is used
consistently (i.e., the injection volumes for all extracts, blanks,
calibration solutions and the performance check samples are 2 uL).
One-uL injections are allowed; however, laboratories are encouraged to
remain consistent throughout the analyses by using the same injection
volume at all times.
6.1.2 Gas Chromatograph/Mass Spectrometer (GC/MS) Interface—The GC/MS interface
components should withstand 350° C. The interface must be designed so
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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. Vespel1" or equivalent ferrules are recommended.
6.1.3 Mass Spectrometer—The static resolving power of the instrument must be
maintained at a minimum of 10,000 (10 percent valley). The mass spec-
trometer 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 (this exhibit, Section 9.1.4.1). 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 the
descriptors contain 10 ions. The selection (Table 6) of the molecular
ions M and M+2 for 13c-HxCDF and 13<>HpCDF rather than M+2 and M+4 (for
consistency) is to eliminate, even under high-resolution mass spectrometric
conditions, interferences occuring 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. The recommended mass spectrometer tuning conditions (this
exhibit, Section 8.2.3) are based on the groups of monitored ions shown
in Table 6.
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6.1.4 Data System—A dedicated data system is employed to control the rapid
multiple ion monitoring process and to acquire the data. Quantification
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.
Quantifications 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
(this exhibit, Section 8.2.4) and provide hard copies of peak profiles
to demonstrate the required resolving power. The data system should
also permit the measurement of noise on the base line.
NOTE: The detector ADC zero setting must allow peak-to-peak measurement
of the noise on the base line of every monitored channel and allow for
good estimation of the instrument resolving power. In Figure 3, the
effect of different zero settings on the measured resolving power is shown.
6.2 GC Column
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
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HRGC/HRMS analysis, the 60-m DB-5 fused-silica capillary column is recom-
mended. Minimum acceptance criteria must be demonstrated and documented
(this exhibit, Section 8.1). At the beginning of each 12-hour period
'(after mass 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 Table 7.
6.3 Miscellaneous Equipment and Materials
The following list of items does not necessarily constitute an exhaustive
compendium of the equipment needed for this analytical method.
6.3.1 Nitrogen evaporation apparatus with variable flow rate.
6.3.2 Balances capable of accurately weighing to 0.01 g and 0.0001 g.
6.3.3 Centrifuge.
6.3.4 Water bath, equipped with concentric ring covers and capable of being
temperature-controlled within + 2° C.
6.3.5 Stainless steel or glass container large enough to hold contents of
one-pint sample containers.
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6.3.6 Glove box.
6.3.7 Drying oven.
6.3.8 Stainless steel spoons and spatulas.
6.3.9 Laboratory hoods.
6.3.10 Pipets, disposable, Pasteur, 150 mm long x 5 mm ID.
6.3.11 Pipets, disposable, serological, 10 mL, for the preparation of the
carbon column specified in Section 7.1.2.
6.3.12 Reacti-vial, 2 mL, silanized amber glass.
6.3.13 Stainless steel meatgrinder with a 3- to 5-mm hole size inner plate.
6.3.14 Separatory funnels, 125 mL.
6.3.15 Kuderna-Danish concentrator, 500 mL, fitted with 10-mL concentrator
tube and three-ball Snyder column.
6.3.16 Teflon™ boiling chips (or equivalent), washed with hexane before use.
6.3.17 Chromatographic column, glass, 300 mm x 10.5 mm, fitted with Teflon
stopcock.
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6.3.13 Adaptors for concentrator tubes.
6.3.19 Glass fiber filters.
6.3.20 Dean-Stark trap, 5 or 10 mL, with T-joints, condenser and 125-mL flask.
6.3.21 Continuous liquid-liquid extractor.
6.3.22 All-glass Soxhlet apparatus, 500-mL flask.
6.3.23 Glass funnels, sized to hold 170 mL of liquid.
6.3.24 Desiccator.
6.3.25 Solvent reservoir (125 mL), Kontes; 12.35 cm diameter (special order
item), compatible with gravity carbon column.
6.3.26 Rotary evaporator with a temperature-controlled water bath.
6.3.27 High-speed tissue homogenizer, equipped with an EN-8 probe or
equivalent.
6.3.28 Glass wool, extracted with methylene chloride, dried and stored in a
clean glass jar.
NOTE: Reuse of glassware should be minimized to avoid the risk of
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contamination. All glassware that is reused must be scrupulously
cleaned as soon as possible after use, applying the following procedure:
Rinse glassware with the last solvent used in it, then with high-purity
acetone and hexane. Wash with hot detergent water. Rinse with copious
amounts of tap water and several portions of distilled water. Drain, dry
and heat in a muffle furnace at 400° C for 15 to 30 minutes. Volumetric
glassware must not be heated in a muffle furnace. Some thermally stable
materials (such as PCBs) may not be removed by heating in a muffle
furnace. In these cases, rinsing with high-purity acetone and
hexane may be substituted for muffle-furnace heating. After the
glassware is dry and cool, rinse it with hexane and store it inverted
or capped with solvent-rinsed aluminum foil in a clean environment.
7. Reagents and Standard Solutions
7.1 Column Chromatography Reagents
7.1.1 Alumina, neutral, Super 1, Woelm®, 80/200 mesh. Store in a sealed
container at room temperature in a desiccator over self-indicating
silica gel.
7.1.2 Carbopak C (80 to 100 mesh, Supelco 1-1025) and Celite 545® (Supelco).
Preparation of the Carbopak C/Celite 545® column: Thoroughly mix
3.6 g Carbopak C (80 to 100 mesh) and 16.4 g Celite 545® in a 40-mL
vial. Activate the mixture at 130° C for 6 hours, then store it in a
desiccator. Cut off both ends of a 10-mL disposable serological pipet
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DRAFT
to give a 4-lnch long column. Fine-polish both ends and flare, if
desired. Insert a glass-wool plug at one end, then pack the column with
0.64 g of the activated Carbopak C/Celite 545® mixture to form a 2-cm
long absorbant bed. Cap the packing with another glass-wool plug.
7.2 Reagents
7.2.1 Sulfuric acid, concentrated, ACS grade, specific gravity 1.84.
7.2.2 Potassium hydroxide, ACS grade, 20 percent (w/v) in distilled water.
7.2.3 Sodium chloride, analytical reagent, 5 percent (w/v) in distilled
water.
7.2.4 Potassium carbonate, anyhdrous, analytical reagent.
7.3 Desiccating Agent
7.3.1 Sodium sulfate, granular, anhydrous; use as such.
7.4 Solvents
7.4.1 High-purity, distilled-in-glass or highest available purity: methylene
chloride, hexane, benzene, methanol, tridecane, isooctane, toluene,
cyclohexane, and acetone.
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7.5 Calibration Solutions
/.b.l Hign-K.esolution Concentration Calibration Solutions Arable 5) — Seven
triaecane solutions containing uniabeied (totaling 17) and carbon-labeled
(.totaling j.i; rcL>i>s and fuuts at Known concentrations used to calibrate
tne instrument. Tne concentration ranges are nomoiogue dependent, witn
the lowest values associated with the tetra- and pentachiorinated
dioxins and turans (2.5 pg/uL,; and the highest for the octachlorinated
congeners (.1000 pg/uL;.
1.1.2 These high-resolution concentration calibration solutions may be obtained
from the Quality Assurance Division, US EPA, Las Vegas, Nevada. However,
additional secondary standards must be obtained from commercial sources,
and solutions must be prepared in the contractor laboratory. Trace-
ability (Appendix B) of standards must be verified against EPA-supplied
standard solutions. Such procedures will be documented by laboratory
standard operating procedures (SOP) as required in IFB Preaward Bid
Confirmations, part 2.f.(4). 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. A recommended traceability procedure for PCDD/PCDF standards
is described in Appendix B.
7.5.3 Store the concentration calibration solutions in 1-mL minivials at
room temperature in the dark.
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7.6 GC Column Performance Check Solution
This solution contains the firstand last-eluting isomers for each homolo-
gous series from tetra- through hepta-chlorinated congeners. The solution
also contains a series of other TCDD isomers for the purpose of documenting
the chromatographic resolution. The ^g -2,3,7,8-TCDD is also present.
The laboratory is required to use tridecane as the solvent and adjust the
volume so that the final concentration does not exceed 100 pg/uL per
congener. Table 8 summarized the qualitative composition (minimum
requirement) of this performance evaluation solution.
NOTE: The use of a PCDD/PCDF-containing fly-ash extract is allowed but
the qualitative equivalency of the fly-ash extract to the EPA solution
should be demonstrated for each fly-ash extract.
7.7 Sample Fortification Solution
This isooctane 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 13C-12-OCDF
is not present in the solution.)
7.8 Recovery Standard Solution
This tridecane solution contains two recovery standards (^c.2-1,2,3,4-
TCDD and ^c^-l,2,3,7,8,9-HxCDD) at a nominal concentration of 50 pg/uL
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per compound. Ten to titty UL of this solution will be spliced into each
sample extract betore the final concentration step and tiKUu/HKMb analysis.
a. System Performance Criteria
System performance criteria are presented oeiow. me laboratory may use the
recommended UC column described in Section 6.2 (.this exhibit;. It must be
documented tnat ail applicable system performance criteria specified in
Section 8.1 (this exhibit; were met before analysis of any sample is per-
formed. Table / provides recommended uu conditions tnat can be used to
satisiy tne required criteria, figure 4 provides a typical i^-nour analysis
sequence wnereoy tne response factors and mass spectrometer resolving
power checks must be performed at the beginning and the end of each .u-hour
period of operation. A uc 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 (this exhibit, Section 3.21) is required .between
a calibration run and the first sample run. The same method blank extract
may thus be analyzed more than once if the number of samples within a batch
requires more than 12 hours of analyses.
8.1 GC Column Performance
8.1.1 Inject 2 uL (this exhibit, Section 6.1.1) of the column performance
check solution (this exhibit, Section 7.6) and acquire selected ion
monitoring (SIM) data as described in Section 6.1.3 (this exhibit) within
a total cycle time of £ 1 second (this exhibit, Section 9.1.4.1).
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8.1.2 The chromatographic separation between 2,3,7,8-TCDD and the peaks repre-
senting any other TCDD isomers must be resolved with a valley of < 25
percent (Figure 5), where
Valley Percent = (x/y) (100)
x = measured as in Figure 5 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 (this
exhibit, Section 13.4.1) and quantitative purposes. All peaks (that
Includes 13C12-2,3,7,8-TCDD) must be labeled and identified on the
chromatograms. Furthermore, all first eluters of a homologous series
must be labeled with the letter F, and all last eluters of a homologous
series must be labeled with the letter L (Figure 5 shows an example of
peak labeling for TCDD isomers). Any individual selected ion current
profile (SICP) (for the tetras, this would be the S1CP for ra/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)
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constitutes an acceptable form of data presentation. An SICP for
the labeled compounds (e.g. , m/z 334 for labeled TCDD) is also required.
8.1.3 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 toler-
ance 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 excercised
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 indicate in the case narrative of its report
which congener (only one is permitted) was missed.
8.2 Mass Spectrometer Performance
8.2.1 The mass spectrometer must be operated in the electron ionization mode.
A static resolving power of at least 10,000 (10 percent valley defini-
tion) must be demonstrated at appropriate masses before any analysis is
performed (this exhibit, Section 13). 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 visual check (i.e.,
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documentation is not required) of the static resolution be made by using
the peak matching unit before and after each analysis. Corrective
actions must be implemented whenever the resolving power does not meet
the requirement.
8.2.2 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 the instrument performances.
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
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downtime for source cleaning.
8.2.3 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 the peak matching unit and the aforementioned PFK reference
peak, verify that the exact mass of m/z 380.9760 (PFK) is within 5 ppm
of the required value. Note that the selection of the low- and high-mass
ions must be such that they provide the largest voltage jump performed
in any of the five mass descriptors (Table 6).
8.2.4 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 6) must allow manual determina-
tion of the resolution, i.e., the horizontal axis must be a calibrated
mass scale (amu or ppm per division). The result of the peak width
measurement (performed at 5 percent of the maximum, which corresponds to
the 10-percent valley definition) must appear on the hard copy and
cannot exceed 100 ppm at m/z 380.9760 (or 0.038 amu at that particular
mass).
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9. Calibration
9.1 Initial Calibration
Initial calibration is required before any samples are analyzed for PCDDs
and PCDFs. Initial calibration is also required if any routine calibration
(this exhibit, Section 9.3) does not meet the required criteria listed in
Section 9.4 (this exhibit).
9.1.1 All seven high-resolution concentration calibration solutions listed in
Table 5 must be used for the initial calibration.
9.1.2 Tune the instrument with PFK as described in Section 8.2.3 (this exhibit).
9.1.3 Inject 2 uL of the GC column performance check solution (this exhibit,
Section 7.6) and acquire SIM mass spectral data as described earlier in
Section 8.1 (this exhibit). The total cycle time must be £ 1 second.
The laboratory must not perform any further analysis until it is demon-
strated and documented that the criterion listed in Section 8.1.2 (this
exhibit) was met.
9.1.4 By using the same GC (this exhibit, Section 6.2) and mass spectrometer
(this exhibit, Section 6.1.3) conditions that produced acceptable results
with the column performance check solution, analyze a 2-uL portion of
each of the seven concentration calibration solutions once with the
following mass spectrometer operating parameters.
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9.1.4.1 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.
9.1.4.2 Acquire SIM data for all the ions listed in the five descriptors of
Table 6.
9.1.4.3 The ratio of integrated ion current for the ions appearing in Table 9
(homologous series quantification ions) must be within the indicated
control limits (set for each homologous series).
9.1.4.4 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 9.
NOTE: Sections 9.1.4.3 and 9.1.4.4 (this exhibit) require that 17 ion
ratios from Section 9.1.4.3 and 11 ion ratios from Section 9.1.4.4 be
within the specified control limits simultaneously in one run. It is
the laboratory's responsibility to take corrective action if the ion
abundance ratios are outside the limits.
9.1.4.5 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. Appendix C describes the
procedure to be followed for the measurement of the S/N from con-
spicuously weak signals. This measurement is required for any GC
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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.
9.1.4.6 Referring to Table 10, 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 13C12 internal standards (RRF(ra); m - 18 to 26)] relative
to the two recovery standards according to the following formulae:
RRF(n)
Ax ' Qis
QX ' Ais
Ais
RRF(m) - —
Ars
where
Ax - sum of the integrated ion abundances of the quantification
ions (Tables 6 and 9) for unlabeled PCDDs/PCDFs,
" 8um °f the integrated ion abundances of the quantification
ions (Tables 6 and 9) for the labeled internal standards,
Ars • sum of the integrated ion abundances of the quantification
ions (Tables 6 and 9). for the labeled recovery standards,
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Q!S " quantity of the internal standard injected (pg),
Qrs « quantity of the recovery standard injected (pg), and
Qx - quantity of the unlabeled PCDD/PCDF analyte injected (pg).
The RRF(n) and RRF(m) are dimensionless quantities; the
units used to express QIS, Qrs and Qx must be the same.
9.1.4.7 Calculate the RRF(n)s and their respective percent relative standard
deviations (%RSD) for the seven calibration solutions:
7
RRF(n) - 1/7 I RRF4(n) ,
where n represents a particular PCDD/PCDF (2,3,7,8-substituted) con-
gener (n - 1 to 17; Table 10), and j Is the injection number (or
calibration solution number; j - 1 to 7).
9.1.4.8 The relative response factors to be used for the determination of the
concentration of total isomers in a homologous aeries (Table 10) are
calculated as follows:
9.1.4.8.1 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
isotner (Table 4; TCDD, PeCDD, HpCDD, and TCDF), the mean RRF used
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will be the same as the mean RRF determined in Section 9.1.4.7 (this
exhibit).
NOTE: The calibration solutions do not contain Cj2~0CDF 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 13C12-OCDF with [M]+ of OCDD).
Therefore, the RRF for OCDF is calculated relative to 13C,2-OCDD.
9.1.4.8.2 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:
_
RRF(k)
1
t n=l
RRF
n »
where
27 to 30 (Table 10), 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 each homologous
series (e.g., two for PeCDF, four for HxCDF, three for
HxCDD, two for HpCDF).
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NOTE: Presumably, the HRGC/HRMS response factors of different isomers
within a homologous series are different. However, this analytical
protocol will make the assumption that the HRGC/HRMS responses of all
isomers in a homologous series that do not have the 2,3,7,8-substitution
pattern are the same as the responses of one or more of the 2,3,7,8-
substituted isomer(s) In that homologous series.
9.1.4.9 Relative response factors [RRF(m)] to be used for the determination
of the percent recoveries for the nine internal standards are calcu-
lated as follows:
RRF(m)
Aism *
Ars
1 7
RRF(m) - - Z RRFj(m),
7 J-l
where:
m = 18 to 26 (congener type) and j = 1 to 7 (injection number),
j m = sum of the integrated ion abundances of the quantification ions
(Tables 6 and 9) for a given internal standard (m = 18 to 26),
Ars - sum of the Integrated ion abundances of the quantification Ions
(Tables 6 and 9) for the appropriate recovery standard (see Table 5,
footnotes),
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Q and Q. m = quantities of, respectively, the recovery standard (rs)
and a particular internal standard (is « m) injected
(pg),
RRF(m) ** relative response factor of a particular internal
standard (m) relative to an appropriate recovery
standard, as determined from one injection, and
RRF(ffl) » calculated mean relative response factor of a particular
internal standard (m) relative to an appropriate recovery
standard, as determined from the seven initial calibra-
tion injections (j).
9.2 Criteria for Acceptable Calibration
The criteria listed below for acceptable calibration must be met before
the analysis is performed.
9.2.1 The percent relative standard deviations for the mean response factors
[RRF(n) and RRF(m)] from each of the 26 determinations (17 for the
unlabeled standards and 9 for the labeled reference compounds) must be
less than 20 percent.
9.2.2 The S/N for the GC signals present in every S1CP (including the
ones for the labeled standards) must be >^ 2.5.
9.2.3 The isotopic ratios (Table 9) must be within the specified control limits.
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NOTE: If the criterion for acceptable calibration listed in Section
9.2.1 (this exhibit) is met, the analyte-specific RRF can then be con-
sidered independent of the analyte quantity for the calibration concen-
tration range. The mean RRFs will be used for all calculations until
the routine calibration criteria (this exhibit, Section 9.4) are no
longer met. At such time, new mean RRFs will be calculated from a new
set of injections of the calibration solutions.
9.3 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.
9.3.1 Inject 2 uL of the concentration calibration solution HRCC-3 containing
10 pg/uL of tetra- and pentachlorinated congeners, 25 pg/uL of hexa-
and heptachlorinated congeners, 50 pg/uL of octachlorinated congeners,
and the respective internal and recovery standards (Table 5). By using
the same HRGC/HRMS conditions as used in Sections 6.1.3 and 6.2 (this
exhibit), determine and document an acceptable calibration as provided in
Section 9.4 (this exhibit).
9.4 Criteria for Acceptable Routine Calibration
The following criteria must be met before further analysis is performed.
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If these criteria are not met, corrective action must be taken.
9.4.1 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 (this exhibit, Section
9.1.4.7).
9.4.2 The measured RRFs [RRF(m) for the labeled standards] obtained during
the routine calibration runs must be within 20 percent of the mean
values established during the initial calibration (this exhibit, Section
9.1.4.9).
9.4.3 The ion-abundance ratios (Table 9) must be within the allowed control
limits.
9.4.4 If either one of the above criteria (this exhibit, Sections 9.4.1 and
9.4.2) is not satisfied, the entire initial calibration process (this
exhibit, Section 9.1) must be repeated. If the ion-abundance ratio
criterion (this exhibit, Section 9.4.3) is not satisfied, refer to the
note in Section 9.1.4.4 (this exhibit) 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.
10. Quality Assessment/Quality Control Procedures
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See Exhibit E for QA/QC requirements.
11. Sample Preservation
11.1 The sample collection, shipping, handling, and chain-of-custody procedures
are not described in this document. Sample collection personnel will, 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 judg-
ment, based on the appearance of the sample, regarding the necessity for
additional mixing. If the sample is clearly inhomogeneous, 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.
11.2 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.
11.3 Grinding or Blending of Fish Samples.
If not otherwise specified by the EPA, the whole fish (frozen) should be
blended or ground to provide a homogeneous sample. The use of a stain-
less steel meatgrinder with a 3- to 5-mm hole size inner plate is recom-
mended. In some circumstances, analysis of fillet or specific organs of
fish may be requested by the EPA. If so requested by the EPA, the above
whole fish requirement is superseded.
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11.4 With the exception of the fish and adipose tissues, which must be stored
at -20° C, all samples must be stored at 4° C, extracted within 30 days
and completely analyzed within 45 days of collection.
11.5 Phase Separation - This is a guideline for phase separation on very wet
(>25 percent water) soil and sediment samples. Place a 50-g portion in a
suitable centrifuge bottle and centrifuge for 30 minutes at 2,000 rpm.
Remove the bottle and mark the interface level on the bottle. Estimate
the relative volume of each phase. With a disposable pipet, transfer the
liquid layer into a clean bottle. Mix the solid with a stainless steel
spatula and remove a portion to be weighed and analyzed (percent moisture
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 and sediment method. Take note of and report the estimated
volume of liquid before disposing of the liquid as a liquid waste.
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).
11.6 Soil, Sediment or Paper Sludge (Pulp) Percent Moisture Determination.
The percent moisture of soil or sediment samples showing detectable
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levels (see note below) of at least one 2,3,7,8-substituted PCDD/PCDF
congener is determined according to the following recommended procedure.
Weigh a 9.5- to 10.5-g portion of the soil or sediment sample (+ 0.5 g)
to three significant figures. Dry it to constant weight at 100° 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 moisture on Form (to be determined). Do not use this solid
portion of the sample for extraction, but instead dispose of it as
hazardous waste. The pulp sample (10 g) should be dried overnight in a
fume hood.
NOTE: Until detection limits are determined (Section 1.2, this exhibit),
the lower MCLs (Table 1) may be used to estimate the minimum detectable
levels.
Weight of wet soil - Weight of dry soil
Percent moisture - —————————————— x
Weight of wet soil
11.7 Fish Tissue Lipid Content Determination
The percent lipid of fish samples showing detectable levels (see Section
11.6 note; this exhibit) of at least one 2,3,7,8-substituted PCDD/PCDF
congener is determined as follows:
Use a separate portion (2 g) of the ground frozen fish sample. Blend it
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with 6 g anhydrous sodium sulfate, pour the mixture in a 1-cm i.d.
glass column and extract the lipids by passing two 25-mL portions of
methylene chloride through the column and collecting the extract in a
tared 100-mL round-bottom flask. Concentrate the extract on a rotary
evaporator until constant weight is attained. The percent lipid is
calculated using the following expression:
Weight of residue from extraction (in g)
Percent lipid * x
Weight of fish tissue portion (in g)
Dispose of the lipid residue as a hazardous waste if the results of the
analysis indicate the presence of PCDDs or PCDFs.
1.8 Adipose Tissue Lipid Content Determination
Details for the determination of the adipose tissue lipid content are
provided in Section 12.11.3 (this exhibit).
12. Extraction and Cleanup Procedures
12.1 Internal standard addition. Use a portion of 1 g to 1000 g (typical sam-
ple size requirements for each type of matrix are given in Section 12.2
of this exhibit and in Table 1) of the sample to be analyzed. Transfer
the sample portion to a tared flask and determine its weight. Except for
adipose tissue, add an appropriate quantity of the sample fortification
mixture (this exhibit, Section 3.8) to the sample. All samples should be
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spiked with 100 uL of the sample fortification mixture to give internal
standard concentrations as indicated in Table 1. As an example, for
13C12-2,3,7,8-TCDD, a 10-g soil sample requires the addition of 1000 pg of
1^C12-2,3,7,8-TCDD to give the requisite 100 ppt fortification level. For
the fortification of soil, sediment, fly ash, water and fish tissue
samples, mix the 100 uL sample fortification solution with 1.5 mL ace-
tone. Do not dilute the isooctane solution for the other matrices. The
fortification of adipose tissue is carried out at the time of horaogeniza-
tion (this exhibit, Section 12.11.2.3).
12.2 Extraction
The extraction and purification procedures for biological tissue samples
are described in Sections 12.10 (fish tissue) and 12.11 (adipose tissue)
of this exhibit.
12.2.1 Sludge/Fuel Oil. Extract aqueous sludge samples by refluxing a sample
(e.g., 2 g) with 50 mL toluene (or benzene) in a 125-mL flask fitted
with a Dean-Stark water separator. Continue refluxing the sample
until all the water is removed. Cool the sample, filter the toluene
(or benzene) extract through a glass-fiber filter, or equivalent, into
a 100-mL round-bottom flask. Rinse the filter with 10 mL toluene (or
benzene), and combine the extract and rinsate. Concentrate the combined
solutions to near dryness on a rotary evaporator at 50° C (toluene) or
a Kuderna-Danish (KD) apparatus (benzene). Use of an inert gas to
concentrate the extract is also permitted. Proceed with Section 12.2.4
below.
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NOTE: If the labeled sludge sample dissolves in toluene, treat it
according to the instructions in Section 12.2.2 below. If the labeled
sludge sample originates from pulp (paper mills), treat it according
to the instructions starting in Section 12.10.1 but without the addition
of sodium sulfate.
12.2.2 Still-Bottom. Extract still-bottom samples by mixing a sample portion
(e.g., 1.0 g) with 10 mL toluene (or benzene) in a small beaker and
filtering the solution through a glass-fiber filter (or equivalent)
into a 50-mL round-bottom flask. Rinse the beaker and filter with 10
mL toluene (or benzene). Concentrate the combined toluene (or benzene)
solutions to near dryness on a rotary evaporator at 50° C. A KD appa-
ratus can be used if benzene is the extraction solvent. Proceed with
Section 12.2.4 below.
12.2.3 Fly Ash. Extract fly ash samples by placing a sample portion (e.g., 10
g) and an equivalent amount of anhydrous sodium sulfate in a Soxhlet
extraction apparatus charged with 100 mL toluene (or benzene), and
extract for 16 hours using a three cycle/hour schedule. Cool and
filter the toluene (or benzene) extract through a glass-fiber filter
into a 500-tnL round-bottom flask. Rinse the filter with 5 mL toluene
(or benzene). Concentrate the combined toluene (or benzene) solutions
to near dryness on a rotary evaporator (toluene) at 50° C or a KD
apparatus (benzene). Proceed with Section 12.2.4 below.
12.2.4 Transfer the residue to a 125-mL separatory funnel using 15 mL hexane.
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Rinse the flask with two 5-mL portions of hexane and add the rinses to
the funnel. Shake two minutes with 50 mL of 5 percent sodium chloride
solution, discard the aqueous layer and proceed with Section 12.3
(this exhibit).
12.2.5 Soil. Add 10 g anhydrous sodium sulfate to the soil 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). Add 200 to 250 mL benzene (or toluene) to the
Soxhlet apparatus and reflux for 24 hours. The solvent must cycle
completely through the system at least three times per hour.
12.2.5.1 Transfer the extract from Section 12.2.5 to a KD apparatus mounted
with a three-ball Snyder column (or to a 500-mL round-bottom flask
for evaporating the toluene on a rotary evaporator).
12.2.5.2 Add a Teflon1" or an equivalent boiling chip. Concentrate in a 70° C
water bath to an apparent volume of 10 mL. Remove the apparatus from
the water bath and allow it to cool for 5 minutes.
12.2.5.3 Add 50 mL hexane and a new boiling chip to the KD flask. Concen-
trate 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.
12.2.5.4 Remove and invert the Snyder column, and rinse it down into the KD
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apparatus with two 1-raL 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 rinsates to the funnel. Proceed with Section
12.3 (this exhibit).
12.2.6 Aqueous Samples. Mark the water meniscus on the side of the 1-L sample
bottle for later determination of the exact sample volume. Pour the
entire sample (approximately 1-L) into a 2-L separatory funnel. Proceed
with Section 12.2.6.1 (this exhibit).
NOTE: A continuous liquid-liquid extractor may be used in place of a
separatory funnel when experience with a sample from a given source
indicates that a serious emulsion problem will result or an emulsion is
encountered when using a separatory funnel. Add 60 mL methylene chloride
to the sample bottle, seal, and shake for 30 seconds to rinse the inner
surface. Transfer the solvent to the extractor. Repeat the sample bot-
tle rinse with an additional 50- to 100-mL portion of methylene chloride
and add the rinsate to the extractor. Add 200 to 500 mL methylene
chloride to the distilling flask, add sufficient reagent water 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 12.2.6.1 and 12.2.6.2 (this exhibit). Proceed with Section
12.2.6.3 (this exhibit).
12.2.6.1 Add 60 mL methylene chloride to the sample bottle, seal, and shake for
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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. Allow the organic layer to sepa-
rate from the water phase for a minimum of 10 minutes. If the emul-
sion interface between layers is more than one-third the volume of
the solvent layer, the analyst must employ mechanical techniques to
complete the phase separation. Collect the methylene chloride 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. Repeat the extraction twice with
fresh 60-ml portions of methylene chloride. After the third extrac-
tion, rinse the sodium sulfate with an additional 30 mL methylene
chloride to ensure quantitative transfer. Combine all extracts and
the rinsate in the KD apparatus.
12.2.6.2 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.
Remove the Snyder column, add 50 mL hexane, re-attach the Snyder
column and concentrate to approximately 5 mL. Add a new boiling chip
to the KD apparatus before proceeding with the second concentration
step. Rinse the flask and the lower joint with two 5-mL portions
of hexane and combine the rinsates with the extract to give a final
volume of about 15 mL.
12.2.6.3 Determine the original sample volume by filling the sample bottle to
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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 12.3 (this exhibit).
12.3 Partition the extract (15 mL hexane) 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.
12.4 Partition the extract (15 mL hexane) against 40 mL of 5 percent (w/v)
aqueous sodium chloride. Shake for two minutes. Remove and discard the
aqueous layer (bottom).
12,5 Partition the extract against 40 mL 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).
12.6 Partition the extract against 40 mL of five percent (w/v) sodium chloride.
Shake for two minutes. Remove and discard the aqueous layer (bottom).
Dry the extract by pouring it through a funnel containing anhydrous
sodium sulfate and collect it in a 50-tnL round-bottom flask. Rinse the
sodium sulfate with two 15-mL portions of hexane, add the rinsates to the
50-mL flask, and concentrate the hexane solution to near dryness on a
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rotary evaporator (35° C water bath), making sure all traces of toluene
(when applicable) are removed. (Use of blow-down with an inert gas to
concentrate the extract is also permitted.)
12.7 Pack a gravity column (glass, 300 mm x 10.5 mm), fitted with a Teflon*
stopcock, in the following manner: Insert a glass-wool plug into the
bottom of the column. Add a 4-g layer of sodium sulfate. Add a 4-g
layer of Woelm® Super 1 neutral alumina. Tap the top of the column
gently. Woe1m® 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 present, discard the column. Do not tap a wetted column.
12.8 Dissolve the residue from Section 12.6 (this exhibit) in 2 mL hexane and
apply the hexane solution to the top of the column. Elute with enough
hexane (3-4 mL) to complete the transfer of the sample cleanly to the
surface of the alumina. Discard the eluate.
12.8.1 Elute with 10 mL of 8 percent (v/v) methylene chloride in hexane.
12.8.2 Elute the PCDDs and PCDFs from the column with 15 mL of 60 percent
(v/v) methylene chloride in hexane and collect this fraction in a
conical shaped (15 mL) concentrator tube.
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12.9 Carbon Column Cleanup
Prepare a Carbopak C/Cellte 545® column as described in Section 7.1.2
(this exhibit).
12.9.1 With a carefully regulated stream of nitrogen, concentrate the
60-percent fraction (this exhibit, Section 12.8.2) to about 2 mL.
Rinse the Carbopak C/Celite 545® with 5 mL toluene followed by 2 mL of
75:20:5 methylene chloride/methanol/benzene, 1 mL of 1:1 cyclohexane/
methylene chloride, and 5 mL hexane. The flow rate should be less than
0.5 mL/min. Discard the rinsates. While the column is still wet with
hexane, add the sample concentrate to the top of the column. Rinse the
concentrator tube which contained the sample concentrate twice with
1 mL hexane and add the rinsates to the top of the column. Elute the
column sequentiallly with two 2-mL portions of hexane, 2 mL cyclohexane/
methylene chloride (50:50, v/v), and 2 mL methylene chloride/methanol/
benzene (75:20:5, v/v). Combine these eluates; this combined fraction
may be used as a check on column efficiency. Now 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.
12.9.2 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 uL using a stream of nitrogen and a sand bath.
Rinse the rotary evaporator flask three times with 300 uL of a solution
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of 1 percent toluene in methylene chloride. Add 10 uL for soil, sedi-
ment, and water, or 50 uL for sludge, still-bottom and fly ash of the
tridecane recovery standard solution. Store the sample at room tempera-
ture in the dark.
12.10 Extraction and Purification Procedures for Fish and Paper Pulp Samples
12.10.1 Add 30 g anhydrous sodium sulfate to a 10-g portion of a homogeneous
fish sample (this exhibit, Section 11.3)'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 glass-
wool plug. Add 200 mL hexane/methylene chloride (1:1) to the Soxhlet
apparatus and reflux for 12 hours. The solvent must cycle completely
through the system at least three times per hour. Follow the same
procedure for the dried (this exhibit, Section 11.6) paper pulp samples.
12.10.2 Transfer the fish or paper pulp extract from Section 12.10.1 to a KD
apparatus equipped with a Snyder column.
12.10.3 Add a Teflon1" or an 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.
12.10.4 Add 50 mL isooctane 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.
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NOTE: The methylene chloride must have been completely removed before
proceeding with the next step.
12.10.5 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 rinsates to the funnel. Proceed with the cleanup according to
the instructions starting in Section 12.5 (this exhibit).
12.11 Extraction and Purification Procedures for Human Adipose Tissue
12.11.1 Human adipose tissue samples must be stored at -20° C from the time of
collection until the time of analysis. The use of chlorinated mate*
rials during the collection of the sample must be avoided. Samples
are handled with stainless steel forceps, spatulas, or scissors. All
sample bottles (glass) are cleaned as specified in the note appearing
in Section 6.3 (this exhibit). Teflonm-lined caps should be used.
12.11.2 Adipose Tissue Extraction Procedure
12.11.2.1 Weigh to the nearest 0.01 g a 10-g portion of a frozen adipose
tissue sample into a culture tube (2.2 x 15 cm).
NOTE: The sample size may be smaller, depending on availability.
In such a situation, the analyst is required to adjust the volume of
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the Internal standard solution added to the sample to meet the for-
tification level stipulated in Table 1.
12.11.2.2 Allow the adipose tissue specimen to reach room temperature (up to 2
hours).
12.11.2.3 Add 10 mL methylene chloride and 100 uL of the sample fortification
solution. Homogenize the mixture for approximately 1 minute with a
tissue homogenizer.
12.11.2.4 Allow the mixture to separate, and 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.
12.11.2.5 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 (this exhibit, Section 12.11.2.A).
12.11.2.6 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.
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12.11.2.7 Adjust the volume to the 100-mL,mark with methylene chloride.
12.11.3 Adipose Tissue Lipid Content Determination
12.11.3.1 Preweigh a clean 1-dram glass vial to the nearest 0.0001 g on an
analytical balance tared to zero.
12.11.3.2 Accurately transfer 1.0 mL of the final extract (100 mL) from Section
12.11.2.6 (this exhibit) to the 1-dram 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 blow-down
is continued until a constant weight is achieved.
12.11.3.3 Accurately weigh the 1-dram 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.
12.11.3.A Calculate the percent lipid content of the original sample to the
nearest 0.1 percent as shown below:
wlr x vext
Lipid Content, LC (%) = ———^^— x 100
Wat * Val
where
wlr = weight of the lipid residue to the nearest 0.0001 g
calculated from Section 12.11.3.3 (this exhibit),
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Vext = total volume (100 mL) of the extract In mL from
Section 12.11.2.6 (this exhibit)
weight of the original adipose tissue sample to the
nearest 0.01 g from Section 12.11.2.1 (this exhibit),
and
Vai = volume of the aliquot of the final extract in mL
used for the quantitative measure of the lipid residue
(1.0 mL).
12.11.3.5 Record the lipid residue measured in Section 12.11.3.3 (this exhibit)
and the percent lipid content from Section 12.11.3.4 (this exhibit).
12.11.4 Adipose Tissue Extract Concentration
12.11.4.1 Quantitatively transfer the remaining extract volume (99.0 mL) to a
500-mL round-bottom flask. Rinse the volumetric flask with 20 to 30
mL of additional methylene chloride to ensure quantitative transfer.
12.11.4.2 Concentrate the extract on a rotary evaporator and a water
bath at 40°C until an oily residue remains.
12.11.5 Adipose Tissue Extract Cleanup Procedures
12.11.5.1 Add 200 mL hexane to the lipid residue in the 500-mL Erlenmeyer
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flask and swirl the flask to dissolve the residue.
•12.11.5.2 Slowly add, with stirring, 100 g of AO-percent w/w sulfuric-acid-
impregnated silica gel. Stir with a magnetic stirrer for two hours
at room temperature.
12.11.5.3 Allow the solid phase to settle and decant the liquid through a
powder funnel containing 20 g anhydrous sodium sulfate into another
500-mL Erlenmeyer flask.
12.11.5.4 Rinse the solid phase with two 50-mL portions of hexane. Stir each
rinse for 15 minutes, decant, and dry as described under Section
12.11.5.3. Combine the hexane extracts from Section 12.11.5.3
(this exhibit) with the rinses.
12.11.5.5 Rinse the sodium sulfate in the powder funnel with an additional
25 mL hexane and combine this rinse with the hexane extracts from
Section 12.11.5.4 (this exhibit).
12.11.5.6 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 4 g silica gel and allow to settle, then add 16 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.
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12.11.5.7 Quantitatively transfer the hexane extract from the Erlenraeyer flask
(this exhibit, Sections 12.11.5.3 through 12.11.5.5) 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.
12.11.5.8 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 inL. Use nitrogen blow-down 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
12.11.5.1 (this exhibit).
12.11.5.9 The extract Is ready for the alumina and carbon cleanups described
In Sections 12.7 through 12.9.2 (this exhibit).
13. Analytical Procedures.
13.1 Remove the sample extract or blank from storage. With a stream of dry,
purified nitrogen, reduce the extract volume to 10 uL or 50 uL (the
volume of the tridecane recovery standard solution) as stipulated above
(this exhibit, Section 12.9.2).
13.2 Inject a 2-uL aliquot of the extract into the GC, operated under the
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conditions previously used (this exhibit, Section 6.2) to produce accept-
able results with the performance check solution.
13.3 Acquire SIM data according to Section 6.1.3 (this exhibit). Use the same
acquisition and mass spectrometer operating conditions previously used to
determine the relative response factors (this exhibit, Sections 9.1.A.6
through 9.1.4.9). Ions characteristic for polychlorinated diphenyl
ethers are included in the descriptors listed in Table 6. Their presence
is to monitor their interference during the characterization of PCDFs.
NOTE: The acquisition period must at least encompass the PCDD/PCDF
overall retention time window previously determined (Section 8.1, this
exhibit). Selected ion current profiles (SICP) for the lock-mass ions
(one per mass descriptor) must also be recorded and included in the data
package as deliverables. These SICPs must be true representations of the
evolution of the lock-mass ions amplitudes during the HRGC/HRMS run.
(See this exhibit, Section 8.2.2 for the proper level of reference compound
to be metered into the ion chamber.) 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
[Y. Tondeur et al., Anal. Chem. 56, 1344 (1984)]. Report any discrepancies
in the case narrative.
13.4 Identification Criteria
For a gas chromatographic peak to be identified as a PCDD or PCDF, it
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must meet all of the following criteria:
13.4.1 Relative Retention Times.
13.4.1.1 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 relative retention time (RRT; at maximum peak height) of the
sample components (i.e., the two ions used for quantification purposes
listed in Table 6) must be within -1 and +3 seconds of the retention
time of the peak for the isotopically labeled internal or recovery
standard at m/z corresponding to the first characteristic ion (of the
set of two; Table 6) to obtain a positive identification of these
nine 2,3,7,8-substituted PCDDs/PCDFs and OCDD.
13.4.1.2 For 2,3,7,8-substituted compounds, that do not have an isotopically
labeled internal standard present in the sample extract (this repre-
sents a total of six congeners; Table 3), the relative retention time
must fall within the established homologous retention time windows by
analyzing the column performance check solution (this exhibit, Section
8.1.3). Identification of OCDF is based on its retention time rela-
i1
tive to 1JCj2~OCDD as determined from the daily routine calibration
results.
13.4.1.3 For non-2,3,7,8-substituted compounds (tetra through octa; totaling
119 congeners), the retention time must be within the corresponding
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homologous retention time windows established by analyzing the column
performance check solution (this exhibit, Section 8.1.3).
13.4.1.4 The ion current responses for both ions used for quantitative pur-
poses (e.g., for TCDDs: m/z 319.8465 and 321.8936) must reach maximum
simultaneously (+ 2 seconds).
13.4.1.5 The ion current responses for both ions used for the labeled stan-
dards (e.g., for 13C12-TCDD: m/z 331.9368 and m/z 333.9339) must
reach maximum simultaneously (+ 2 seconds).
NOTE: The analyst is required to verify the presence of 1,2,8,9-TCDD
and 1,3,4,6,8-PeCDF (this exhibit, Section 8.1.3) in the SICPs of the
daily performance checks. Should either one compound be missing, the
analyst is required to report that observation with the results
associated with the sample batch as it may indicate a potential
problem with the ability to detect all the PCDDs/PCDFs.
13.4.2 Ion Abundance Ratios
13.4.2.1 The integrated ion current for the two ions used for quantification
purposes must have a ratio between the lower and upper limits
established for the homologous series to which the peak is assigned.
See Sections 9.1.4.3 and 9.1.4.4 (this exhibit) and Table 9 for
details.
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13.4.3 Signal-to-Noise Ratio
13.4.3.1 All ion current intensities must be ^> 2.5 times noise level for posi-
tive identification of a PCDD/PCDF compound or a group of coeluting
isomers. Appendix C describes the procedure to be followed for the
determination of the S/N.
13.4.4 Polychlorinated Diphenyl Ether Interferences
13.4.4.1 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 PCDPE
channel.
14. Calculations
14.1 For gas chromatographic peaks that have met the criteria outlined in
Sections 13.4.1.1 through 13.4.3.1 (this exhibit), calculate the concen-
tration of the PCDD or PCDF compounds using the formula:
Ax x Qls
C
x
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,
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Ax - sum of the integrated ion abundances of the quantification
ions (Table 6) for unlabeled PCDDs/PCDFs,
sum °f ^6 integrated ion abundances of the quantification ions
(Table 6) for the labeled internal standards,
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(n) = calculated mean relative response factor for the analyte
[RRF(n) with n = 1 to 17; Section 9.1.4.7, this exhibit].
If the analyte 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
9.1.4.7 (this exhibit). 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 9.1.4.8.2 (this exhibit). [RRF(k) with k = 27 to 30.]
14.2 Calculate the percent recovery of the nine internal standards measured in
the sample extract, using the formula:
x Qrs
Internal standard percent recoverr = — x 100
x Ars x
where
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Ais = sum °f tne Integrated ion abundances of the quantification
ions (Table 6) for the labeled internal standard,
Ars = sum of the integrated ion abundances of the quantification
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 9.1.4.9 (this exhibit) [RRF(m) with
m » 18 to 26].
NOTE: For human adipose tissue, adjust the percent
recoveries by adding 1 percent to the calculated value.
14.3 If the concentration in the 10-uL or 50-uL final extract of any of the
fifteen 2,3,7,8-substituted PCDD/PCDF compounds (Table 3) exceeds the
upper method calibration limits (MCL) listed in Table 1 (e.g., 200 pg/uL.
D-65
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for TCDD in soil), the linear range of response versus concentration may
have been exceeded, and, after contacting EPA/SMO, a reanalysis of the
sample (using 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 (this exhibit, Sections 12.1 to
12.9.3). For the other congeners (including OCDD), however, report the
measured concentration and indicate that the value exceeds the MCL.
14.4 The total concentration for each homologous series of PCDD and PCDF is
calculated by summing up the concentrations of all positively identified
isomers of each homologous series. Therefore, the total should also
include the 2,3,7,8-substituted congeners. The total number of GC
signals included in the homologous total concentration value must be
specified in the report.
14.5 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.
14.5.1 Samples giving a response for both quantification ions (Tables 6 and 9)
that is less than 2.5 times the background level.
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14.5.1,1 Use the expression for EDL (specific 2,3,7,8-substituted PCDD/PCDF)
below to calculate an EDL for eiach 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 quanti-
fication 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 product
height.
Use the formula:
2.5 x Ax x Qls
EDL (specific 2,3,7,8 subst.-PCDD/PCDF) «
Ais x W x RRF(n)
where
EDL = estimated detection limit for homologous 2,3,7,8-substituted
PCDDs/PCDFs.
Ax, Ajs, W, RRF(n), and Qjg retain the same meanings as defined
in Section 14.1.
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14.5.2 Samples characterized by a response above the background level with a
S/N of at least 2.5 for at least one of the quantification ions
(Tables 6 and 9).
14.5.2.1 When the response of a signal having the same retention time as a
2,3,7,8-substituted congener has a S/N in excess of 2.5 and does not
meet any of the other qualitative identification criteria listed in
Section 13.4, calculate the "Estimated Maximum Possible Concentration'
(EMPC) according to the expression shown in Section 14.1.
14.6 The relative percent difference (RPD) is calculated as follows:
I Si - S2 |
RPD = x 100
( S1 + S2 ) / 2
Sj and 82 represent sample and duplicate sample results.
14.7 The 2,3,7,8-TCDD toxic equivalents (TE) of PCDDs and PCDFs present in the
sample are calculated, only at the data user's request, 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 the non-2,3,7,8-substituted
compounds as shown in Table 11. 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
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listed in Table 11. The exclusion of other homologous series such as
mono-, di-, tri- and octachlorinated dibenzodioxins and dibenzofurans
does not mean that they are non-toxic. Their toxicity, as known at this
time, is much less than the toxicity of the compounds listed in Table 11.
The above procedure for calculating the 2,3,7,8-TCDD toxic 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 14.1 and 14.4.
14.7.1 Two-GC Column TEF Determination
Isomer specificity for all 2,3,7,8-substituted PCDDs/PCDFs cannot be
achieved on the 60-m DB-5 GC column alone. In order to determine the
proper concentrations of the individual 2,3,7,8-substituted congeners,
the sample extract must be reanalyzed on a 60-m SP-2330 (or SP-2331) GC
column.
14.7.1.1 The concentrations of 2,3,7,8-TCDD (see note below), 2,3,4,7,8-PeCDF,
l,2,3,4,6,7,8HpCDD, 1,2,3,4,6,7,8-HpCDF, and 1,2,3,4,7,8,9-HpCDF are
calculated from the analysis of the sample extract on the 60-m DB-5
fused-silica column. The experimental conditions remain the same as the
conditions described previously in Section 13 (this exhibit), and the
calculations are performed as outlined in Section 14 (this exhibit).
D-69
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14.7.1.2 The concentrations of 2,3,7,8-TCDF, 1,2,3,7,8-PeCDD and -PeCDF,
1,2,3,4,7,8-HxCDD and -HxCDF, 1,2,3,6,7,8-HxCDD and -HxCDF,
1,.2,3,7,8,9-HxCDD and -HxCDF, and 2,3,4,6,7,8-HxCDF are obtained from
the analysis of the sample extract on the second fused-silica capil-
lary column (confirmation GC column: 60 m SP-2330). However, the
GC/MS conditions must be altered so that: (1) only the first three
descriptors (i.e., tetra-, penta-, and hexachlorinated congeners)
of Table 6 are used; and (2) the switching time between descriptor 2
(pentachlorinated congeners) and descriptor 3 (hexachlorinated
congeners) takes place following the elution of 1^C12-1,2,3,7,8-PeCDD.
The concentration calculations are performed as outlined in Section
14 (this exhibit).
NOTE: The confirmation and quantification of 2,3,7,8-TCDD (this
exhibit, Section 14.7.1.1) may be accomplished on the SP-2330 GC
column instead of the DB-5 column, provided the criteria listed in
Section 8.1.2 (this exhibit) are met and the requirements described
in Section 2.2 (Exhibit E) are followed.
14.7.1.3 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 13.4.2 and 13.4.3
(this exhibit), respectively. In addition, the retention time
identification criterion described in Section 13.4.1.1 (this exhibit)
applies here for congeners for which a carbon-labeled analogue is
available in the sample extract. However, the relative retention
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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"12 and the results from the
routine calibration run on the SP-2330 column.
D-71
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APPENDICES
196
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APPENDIX A
Procedure for the Collection, Handling, Analysis, and Reporting
Requirements of Wipe Tests Performed within the Laboratory
D-72
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This procedure is designed for the periodic evaluation of potential con-
tamination by 2,3,7,8-substituted PCDD/PCDF congeners of the working areas •
inside the laboratory. •
PERFORMING WIPE TEST
Perform the wipe tests on surface areas of two inches by one foot with
laboratory wipers 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.
COMPOSITE SAMPLE PREPARATION
Close the jar containing the wipers and 200 mL 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. Add 100 uL of the
sample fortification solution to the concentrator tube (Section 3.8, this
exhibit), 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.
D-73
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Prepare a neutral alumina column as described in Section 12.7 (this
exhibit) and follow the steps outlined in Sections 12.8 thru 12.8.2 (this
exhibit).
Add 10 uL of the recovery standard solution as described in Section
12.9.2 (this exhibit).
EXTRACT ANALYSIS
Concentrate the contents of the vial to a final volume of 10 uL (either in
a minivial or in a capillary tube). Inject two 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 Section 13 (this exhibit). Perform
calculations according to Section 14 (this exhibit).
REPORTING FORMAT
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 outlined in
this analytical protocol, a lower limit of calibration of 25 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 8 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 25 x 5 - 125 pg/WTE and the positive response for the blank
D-74
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would be 8 x 5 = 40 pg). Also, report the recoveries of the internal standards
during the simplified cleanup procedure.
FREQUENCY OF WIPE TESTS
At a minimum, wipe tests should be performed when there is evidence of
contamination in the method blanks.
CORRECTIVE ACTION
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 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 absorbant (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.
D-75
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APPENDIX B
Standards Traceability Procedure
NOTE: The content of this appendix is based on the assumption that EPA
will have within its repository a mixture (named S2) containing known
concentrations (e.g., 100 pg/uL) of the eight 13C-labeled 2,3,7,8-substi-
tuted PCDD/PCDF congeners marked with an asterisk in Table 3 of this
exhibit, and a second solution (named SI, with the same concentration as
used for S2) containing the eight corresponding unlabeled analogues.
D-76
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All laboratories are expected to maintain traceability of their standard
solutions by verifying that all standard solutions used for direct quantifica-
tion of samples agree in chemical identity and concentration with the EPA
primary standard solutions. The specific procedures are described below:
Each time a new laboratory working standard solution (W) is prepared, the
identities and concentrations of the components of this solution must be veri-
fied. Verifications of the identities of the compounds are to be carried out
by HRGC/HRMS. The EPA reference standard (S) and the laboratory working stan-
dard (W) are to be analyzed under the instrumental conditions described in this
exhibit, which are appropriate for the analysis of PCDDs and PCDFs. Two
criteria must be satisfied to verify the identifications:
o Elution of the component(s) of the laboratory working standard must
be at the same retention time(s) as those of the component(s) of the
EPA reference standard solution.
o Concentration^) of the laboratory working standard component(s) must
be equal to or less than 20 percent different from the EPA reference
standard component(s).
Qualitative Characterization
Due to the complexity brought by the large number of possible PCDD and
PCDF congeners, the requirement for qualitative verification by comparison of
the retention times applies only to the eight 2,3,7,8-substituted PCDD/PCDF
D-77
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congeners marked with an asterisk in Table 3 and for which a carbon-labeled
analogue is available. Two situations need to be considered:
a) The laboratory is required to trace back its unlabeled PCDD/PCDF standards
to EPA standards. This is accomplished by adding an appropriate aliquot
of the EPA 13c_iabe;Le(j standard solution (S2) to an aliquot of the labora-
tory working solution (Wl) so that the concentrations are comparable; the
new mixture is then analyzed by HRGC/HRMS. The retention times of the
eight unlabeled PCDDs/PCDFs discussed above must fall within -1 to +3
seconds of the EPA ^Q_iabeled analogues.
b) In addition to a), the laboratory is required to trace back its
labeled standards to EPA standards. Proceed as follows: Add an aliquot
of the laboratory working standard solution (W2) containing the carbon-
labeled compounds to an aliquot of the EPA standard solution (SI) containing
the eight unlabeled 2,3,7,8-substituted PCDD/PCDF congeners discussed
above, and analyze by HRGC/HRMS. The concentrations must be comparable.
The retention times for the eight carbon-labeled compounds must fall
within -3 to +1 seconds of the EPA unlabeled analogues.
Quantitative Characterization
To establish that the concentration of the laboratory working standard is
correct with respect to the EPA reference standard, the relative response
factors (RRFs) for the eight 2,3,7,8-substituted PCDD/PCDF congeners (marked
with asterisks in Table 3) must be determined as described in this exhibit.
D-78
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The concentrations of the EPA reference and laboratory working standards should
be approximately the same (e.g., 50 pg/uL/congener). Proceed as follows:
1) Mix equal portions of the two EPA standard solutions (SI and S2) and
analyze by HRGC/HRMS. Calculate two RRFs for each of the eight analytes
as shown below:
Response factor of unlabeled congener (i) relative to carbon-labeled
analogue (j):
RRF (Sl,i)
Qi
Response factor of carbon-labeled congener (j) relative to unlabeled
analogue (i):
AJ x Qt
RRF (S2,j) =
Qj x At
where Aj and AJ represent the integrated ion abundances of, respectively,
the unlabeled congener and carbon-labeled congener, and Q^ and QJ the
quantities of, respectively, the unlabeled congener and carbon-labeled
congener, with i = 1 to 8, j - 1 to 8.
D-79
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2) Add an appropriate aliquot of the laboratory working solution Wl (or W2)
to an aliquot of the EPA solution S2 (or SI). Analyze the mixture by
HRGC/HRMS and calculate the corresponding response factors as indicated
below:
RRF (Wl,i)
Ql x A-j
or
Aj x Qt
RRF (W2,j) = -
Qj x Ai
A and Q have the same meanings as in (1).
3) When the percent difference between each congener relative response factor
— RRF (Sl,i) and RRF (Wl.i), and RRF (S2,j) and RRF (W2,j) — does not
exceed 20 percent, the concentration of the laboratory working standard is
correct. (RPD * relative percent difference.)
| RRF (Sl,i) - RRF (Wl,i) |
RPD - ——————————• x 100
RRF (SI, i)
and
| RRF (S2,j) - RRF(W2,j) |
RPD - ————————— x 100
RRF (S2,j)
D-80
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Traceability Requirements
If any or all of the above conditions for qualitative and quantitative
verifications for the laboratory working standard are not met, the standard is
not traceable to the EPA reference standard and can therefore not be used for
the analysis of samples.
NOTE: The procedure outlined above is required for laboratories which use
different batches of analytical standard compounds in the preparation of
the sample fortification and recovery standard solutions and in the prepara-
tion of the HRCC solutions. Laboratories which use the same batch of
analytical standards during the preparation of the sample fortification
and recovery, standard solutions and the HRCC solutions are exempt from
following the above procedure, provided proper traceability documentation
is available.
In addition, the records pertaining to the above qualitative and
quantitative requirements, records of all verifications, documentation of the
preparation, and all inventory must be kept for all contract laboratory pri-
mary, secondary, and working standards that are generated for the purpose of
analyzing samples for EPA. These records should include the signed and dated
logbooks containing the information pertaining to the preparation of the
laboratory standards (weight of compound(s), volume and nature of the solvent,
laboratory code name, EPA reference standard lot number) and of any modification
made to the EPA reference standard. All standards should be used on a first
in, first out basis. The raw data, quantification reports and calculations
must be kept on file.
D-81
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APPENDIX C
Slgnal-to-Noise Ratio Determination
D-82
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SIGNAL-TO-NOISE RATIO DETERMINATION
MANUAL DETERMINATION
This method describes a manual determination of the signal-to-noise ratio
(S/N) from a GC/MS signal, based on the measurement of its peak height relative
to the baseline noise. The procedure is composed of four steps as outlined
below. (Refer to Figure 7 for the following discussion.)
1. Estimate the peak-to-peak noise (N) by tracing the two lines (El and E2)
defining the noise envelope. The lines should pass through the estimated
statistical mean of the positive and the negative peak excursions as shown
on Figure 7. In addition, the signal offset (0) should be set high enough
such that negative-going noise (except for spurious negative spikes) is
recorded.
2. Draw the line (C) corresponding to the mean noise between the segments
defining the noise envelope.
3. Measure the height of the GC/MS signal (S) at the apex of the peak relative
to the mean noise C. For noisy GC/MS signals, the average peak height
should be measured from the estimated mean apex signal D between E3 and
EA.
D-83
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4. Compute the S/N.
This method of S/N measurement is a conventional, accepted method of noise
measurement in analytical chemistry.
D-84
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FIGURES CAPTIONS
1. Method flow chart for sample extraction and cleanup as used for the
analysis of PCDDs and PCDFs in complex waste and biological samples.
2. General structures of dibenzodioxin and dibenzofuran.
3. 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. (Not aesthetic.)
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.
4. Typical 12-hour analysis sequence of events.
5. 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 Table 7.
D-85
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6. 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/AM of 10,500 (10 percent
valley definition).
7. 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 such that negative-going baseline noise is recorded.
D-86
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Complex
Waste
Sample
Soil/
Sediment
ois*tur
Fish and
Adipose
Tissues
1) Internal
Standards
2)Extraction
Sample Extract
1) Acid-Base Cleanup
2)Chromatographic Cleanup
3) Recovery Standards
HRGC/HRMS
Figure 1
D-87
212
-------
8
7
0
0
Dibenzodioxin
8
6 ' 0 ' 4
Dibenzofuran
Figure 2
D-88
213
-------
M/AM
5,600
B
5,600
8,550
Figure 3
D-89
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D-93
218
-------
Table 1. Types of Matrices, Sample Sizes and 2,3,7,8-TCDD-Based
Method Calibration Limits (Parts per Trillion)
Lower MCL^3'
Upper MCL^a)
Weight (g)
IS Spiking
Levels (ppt)
Final Extr.
Vol. (uL)
Soil
Sediment
2.5
200
10
100
10
Fly
Ash
2.5
200
10
100
50
Sludges
Water Fuel Oil
0.025 12.5
2 1000
1000 2
1 500
10 50
Still-
Bottom
25
2000
1
1000
50
Fish
Tissue
Paper
Pulp
2.5
200
10
100
10
Human
Adipose
Tissue
2.5
200
10
100
10
(fl)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.
NOTE: Chemical reactor residues are treated as still-bottoms if
their appearances suggest so.
D-94
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Table 2. Composition of the Sample Fortification
and Recovery Standard Solutions
Analyte
Sample Fortification
Solution
Concentration
(pg/uL; Solvents
Isooctane)
Recovery Standard
Solution
Concentration
(pg/uL; Solvent:
Tridecane)
J3c12-2,3,7,8-TCDD
}3c12-2,3,7,8-TCDF
1JC12-1,2,3,4-TCDD
"c12-l,2,3,7,8-PeCDD
1JC12-l,2,3,7,8-PeCDF
13C,2-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,2,3,4,6f7,8-HpCDD
13C12-l,2,3,4,6,7,8-HpCDF
10
10
™
10
10
25
25
—
25
25
—
50
—
—
50
—
—
13C,o-OCDD
50
D-95
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Table 3. The Fifteen 2,3,7,8-Substttuted PCDD and PCDF Congeners
PCDD PCDF
2,3,7,8-TCDD<*) 2,3,7,8-TCDF(*>
l,2,3,7,8-PeCDD(*) 1,2,3,7,8-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
1,2,3,7,8,9-HxCDD<+> 1,2,3,7,8,9-HxCDF
l,2,3,4,6,7,8-HpCDD(*> 1,2,3,4,7,8-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-iabeled analogue is used as an Internal standard.
13c_iabe].e(i analogue is used as a recovery standard.
D-96
221
-------
Table 4. Isomers of Chlorinated Dioxins and Furans as a
Function of the Number of Chlorine Atoms
Number of
Chlorine
Atoms
Numbejr of
Dioxin
Isomers
Number of
2,3,7,8
Isomers
Number of
Fur an
Isomers
Number of
2,3,7,8
Isomers
1
2
3
4
5
6
7
8
2
10
14
22
14
10
2
1
1
1
3
1
1
4
16
28
38
28
16
4
1
1
2
4
2
1
Total
75
135
10
D-97
222
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Table 5. High-Resolution Concentration Calibration Solutions
Concentration (pg/uL)
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
7 77777 j* w~- »
OCDD 1 ,
OCDF 1 ,
Internal Standards
}3c12-2,3,7,8-TCDD
J3C12-2,3,7,8-TCDF
1JC19-l,2,3,7,8-PeCDD
1 1 li ' '
|3C12-l,2,3,7,8-PeCDF
;3C12-l,2,3,6,7,8-HxCDD
|3C12-l,2,3,4,7,8-HxCDF
J3C12-l,2,3,4,6,7,8-HpCDD
|3C12-l,2,3,4,6,7,8-HpCDF
1-3C12-OCDD
Recovery Standards
13C10-l,2,3,4-TCDD(a)
n±2 ' ' '
C -1 7 ? 7 R Q-
12 1'/zJ-3'/'°>y
HxCDD
-------
Table 6. Ions Monitored for HRGC/HRMS analysis of PCDD/PCDFs
( S * Internal/recovery standard)
Descriptor Accurate(a)
Mass
1 303.9016
305.8987
315.9419
317.9389
319.8965
321.8936
331.9368
333.9339
375.8364
[354.9792]
2 339.8597
341.8567
351.9000
353.8970
355.8546
357.8516
367.8949
369.8919
409.7974
[354.9792]
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
Elemental
Composition
C12H435C140
C12H435C1337C10
13C12H435C140
13C12H435C1337C10
C12H435C14°2
C12H435C1337C102
13C12H435C1402
13C12H435C1337C102
C12H435C160
C9F13
C12H335C1437C10
C12H335C1337C120
13C12H335C1437C10
13C12H335C1337C120
C12H335C1437C102
C12H335C1337C1202
13C12H335C1437C102
13C12H335C1337C1202
C12H335C170
C9F13
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
(Continued)
D-99
224
-------
Table 6. Continued
Descriptor Accurate
Mass
3 373.
375.
383.
385.
389.
391.
401.
403.
445.
[354.
4 407.
409.
417.
419.
423.
425.
435.
437.
479.
[430.
8208
8178
8642
8610
8156
8127
8559
8529
7555
9792]
7818
7789
8253
8220
7766
7737
8169
8140
7165
9728]
Ion
ID
M+2
M+4
M
M+2
M+2
M+4
M+2
M+4
M+4
LOCK
M+2
M+4
M
M+2.
M+2
M+4
M+2
M+4
M+4
LOCK
Elemental
Composition
/-" U -JJf*-\
L12H2 U.5
C H 35C1
13C12H235C16
13C H
35ci5
C12H2
C12H
1 J r> u
012H
1 j r> TJ
C12H
2
2
2
C12H2
35
35
35
35
35
Cl
Cl
Cl
Cl
Cl
Analyte
37C10
37ci2o
0
37C10
5
4
5
4
6
37
37
37
37
37
CIO 2
ci2o2
C102
ci2o2
ci2o
C9F13
Cii
1 *)
35ci637cio
C12H35C15
13C
c12
13r
c12
C12
c12
13C
C12
13r
c12
C12
CgF
H
H
H
H
H
H
H
1
35
35
35
35
35
35
35
7
37ci2o
ci7o
Cl
Cl
Cl
Cl
Cl
Cl
6
6
5
6
5
7
37
37
37
37
37
37
CIO
C102
ci2o2
cio2
ci2o2
ci2o
HxCDF
HxCDF
HxCDF
HxCDF
HxCDD
HxCDD
HxCDD
HxCDD
OCDPE
PFK
HpCDF
HpCDF
HpCDF
HpCDF
HpCDD
HpCDD
HpCDD
HpCDD
NCDPE
PFK
(S)
(S)
(S)
(S)
(S)
(S)
(S)
(S)
(Continued)
D-100
225
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Table 6. Continued
Descriptor Accurate Ion
Mass ID
Elemental
Composition
Analyte
5 441
443
457
459
469
471
513
[430.
.7428
.7399
.7377
.7348
.7779
.7750
.6775
9728 ]
M+2
M+4
M+2
M+4
M+2
M+4
M+4
LOCK
C
C
C
C
13C
^3C
C
12
12
12
12
12
12
12
35
35
Cl
Cl
35C1
35
35
35
35
Cl
Cl
Cl
Cl
7
6
7
6
7
6
8
37
37
37
37
37
37
37
CIO
ci2o
cio2
ci2o2
cio2
ci2o2
ci2o
C9F17
OCDF
OCDF
OCDD
OCDD
OCDD
OCDD
DCDPE
PFK
(S)
(S)
(a)lhe following nuclidic masses were used:
H = 1.007825 0 = 15.994915
C = 12.000000 35ci » 34.968853
13C = 13.003355 37C1 = 36.965903
D-101
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Table 7. Recommended GC Operating Conditions
Column coating DB-5
Film thickness 0.25 urn
Column dimension 60 m x 0.32 mm
Injector temperature 270° C
Splitless valve time 45 s
Interface temperature Function of the final temperature
Temperature program
Stage Init. Temp. Init. Hold. Temp. Fin. Temp. Fin.
(° C) Time (rain) Ramp (° C) Hoi.
(° C/min) Time
1 200 25 220 16
2 5 235 7
3 5 330 5
Total time: 60 min
D-102
227
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Table 8. 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(a)
5
PCDD-Positional Isomer
Early Late
Eluter Eluter
1,3,6,8 1,2,8,9
1,2,4,6,87 1,2,3,8,9
PCDF-Positional
Early
Eluter
1,3,6,8
1,3,4,6,8
Isomer
Late
Eluter
1,2,8
1,2,3,8
,9
,9
6
7
8
1,2,4,7,9
1,2,3,4,6,8 1,2,3,4,6,7
1,2,3,4,6,7,8 1,2,3,4,6,7,9
1,2,3,4,6,7,8,9
1,2,3,4,6,8 1,2,3,4,8,9
1,2,3,4,6,7,8 1,2,3,4,6,7,9
1,2,3,4,6,7,8,9
(a)ln addition to these two PCDD isomers, the 1,2,3,4-, 1,2,3,7-,
1,2,3,8-, 2,3,7,8-, 13C12-2,3,7,8-, and 1,2,3,9-TCDD isomers
must also be present;
D-103
228
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Table 9. Theoretical Ion Abundance Ratios and Their
Control Limits for PCDDs and PCDFs
Number of
Chlorine
Atoms
4
5
6
6(a)
7
7
8
Ion Theoretical
Type Ratio
M
0.77
M+2
M+2
1.55
M+4
M+2
1.24
M+4
M
0.51
M+2
M
0.44
M+2
M+2
1.04
M+4
M+2
0.89
M+4
Control Limits
lower upper
0.65 0.89
1.24 1.86
1.05 1.43
0.43 0.59
0.37 0.51
0.88 1.20
0.76 0.89
(a)Used only for 13C-HxCDF (IS).
-------
Table 10. 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 OCDF
18 l3C12-2,3,7,8-TCDD
19 13C,2-2,3,7,8-TCDF
20 13C12-l,2,3,7,8-PeCDD
21 13C12-l,2,3,7,8-PeCDF
22 13C12-l,2,3,6,7,8-HxCDD
23 13C12-l,2,3,4,7,8-HxCDF
24 13C,2-l,2,3,4,6,7,8-HpCDD
25 J3C12-1,2,3,4,6,7,8-HPCDF
26 1:iC12-OCDD
27 Total PeCDFs
28 Total HxCDFs
29 Total HxCDDs
30 Total HpCDFs
D-105
230
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TABLE 11. 2,3,7,8-TCDD Equivalent Factors (TEFs) for the
Polychlorinated Dibenzodioxins and Dibenzofurans
Number
Compound(s)
*Excludlng the 2,3,7,8-substituted congeners.
TEF
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
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
* Total - TCDD
* Total - PeCDD
* Total - HxCDD
* Total - HpCDD
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-HpCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
* Total - TCDF
* Total - PeCDF
* Total - HxCDF
* Total - HpCDF
1.00
0.50
0.04
0.04
0.04
0.001
0.01
0.005
0.0004
0.00001
0.10
0.10
0.10
0.01
0.01
0.01
0.01
0.001
0.001
0.001
0.001
0.0001
0.00001
D-106
231
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Table 12. Toxiclty Equivalency Factor: 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 13C12-1,2,3,4,7,8-HxCDF
1,2,3,7,8,9-HxCDF 13C12-1,2,3,4,7,8-HxCDF
2,3,4,6,7,8-HxCDF 13C1 ?-l,2, 3,4, 7,8-HxCDF
The retention time of 2,3,4,7,8-PeCDF on the DB-5 column is
relative to 13C12-l,3,7,8-PeCDF and the retention
time of 1,2,3,4,7,8,9-HpCDF relative to 13C12-1,2,3,4,6,7,8-
measured
time c
HpCDF.
D-107
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QUALITY ASSURANCE REQUIREMENTS
(Quality Assessment and Quality Control)
(Exhibit E)
233
-------
1. SUMMARY OF QA/QC ANALYSES
° Initial and periodic calibration and instrument performance checks.
o HRGC/HRMS method blank analysis.
0 Field blank analyses (Section 2.4.2, this exhibit); a minimum of one
fortified field blank shall be analyzed with each sample batch; an
additional fortified field blank must be analyzed when a new lot of
absorbent or solvent is used. A matrix spike may be used in place of
a fortified field blank.
o Analysis of a batch of samples with accompanying QA/QC analyses:
Sample Batch — £ 24 samples, including field blank and rinsate
sample(s).
Additional QA/QC analyses per batch:
Fortified field blank or matrix spike (MS) 1
Method blank (MB) 1
Duplicate sample or matrix spike duplicate (MSD) 1
Total 3
o "Blind" QC samples (soil, sediment, water) may be submitted to the
laboratory as ordinary samples included in the sample batch.
E-l
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Blind samples include:
Uncontarainated soil, sediment, or water samples
Split samples,
Unidentified duplicates, and
Performance evaluation samples.
2. QUALITY ASSESSMENT/QUALITY CONTROL
2.1 Performance Evaluation Samples — Included among the samples in all
batches will be samples (blind or double blind) containing known amounts
of unlabeled 2,3,7,8-substituted PCDDs/PCDFs or other PCDD/PCDF congeners.
2.2 Performance Check Solutions
2.2.1 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)
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).
These procedures are described in Section 8 of Exhibit D. If the
required criteria are not met, remedial action must be taken before any
E-2
235
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samples are analyzed.
2.2.2 To validate positive sample data, the routine or continuing calibration
(HRCC-3) 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.
2.2.2.1 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.
2.2.2.2 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.
2.2.3 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.
2.2.4 Deviations from criteria specified for the GC performance check or for
E-3
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the mass resolution check (Section 8, Exhibit D) invalidate all positive
sample data collected between analyses of the performance check solu-
tion, and the extracts from those positive samples shall be reanalyzed
(Exhibit C).
If the routine calibration run fails at the beginning of a 12-hour shift,
the instructions in Exhibit D, Section 9.4.4 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, 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. 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.
2.3 The GC column performance check mixture, high-resolution concentration
calibration solutions, and the sample fortification solutions may be
obtained from the EMSL-LV. However, if not available from the EMSL-LV,
standards can be obtained from other sources, and solutions can be pre-
pared in the laboratory. Concentrations of all solutions containing
2,3,7,8-substituted PCDDs/PCDFs, which are not obtained from the EMSL-
LV, must be verified by comparison with the EPA standard solutions that
E-4
237
-------
are available from the EMSL-LV. (Refer to Appendix B, Exhibit D, for
details on the recommended standards traceability procedure.)
2.4 Blanks
2.4.1 Method Blank
One method blank is required per batch of samples. To that effect,
perform all steps detailed in the analytical procedure (Section 12,
Exhibit D) using all reagents, standards, equipment, apparatus, glass-
ware and solvents that would be used for a sample analysis, but omit
addition of the soil, aqueous or any other matrix sample portion.
2.4.1.1 The method blank must contain the same amount of Cj2~labeled
internal standards that is added to samples before extraction.
2.4.1.2 An acceptable method blank exhibits no positive response as stated in
Section 3.16, Exhibit D. If the method blank, which was extracted
along with a batch of samples, is contaminated, all positive samples
must be rerun (Exhibit C).
2.4.1.2.1 If the above criterion is not met, check solvents, reagents, forti-
fication solutions, apparatus and glassware to locate and eliminate
the source of contamination before any further samples are extracted
and analyzed.
2.4.1.2.2 If new batches of reagents or solvents contain interfering
E-5
238
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contaminants, purify or discard them.
2.4.2 Field Blanks
Each batch of samples contains a field blank sample of uncontaminated
soil, sediment or water that is to be fortified before analysis accord-
ing to Section 2.4.2.1 (this exhibit). In addition to this field blank,
a batch of samples may include a rinsate, which is a portion of the sol-
vent (usually trichloroethylene) that was used to rinse sampling equip-
ment. The rinsate is analyzed to assure that the samples were not
contaminated by the sampling equipment.
2.4.2.1 Fortified Field Blank
2.4.2.1.1 Weigh a 10-g portion or use 1 L (for aqueous samples) of the speci-
fied field blank sample and add 100 uL of the solution containing
the nine internal standards (Table 2, Exhibit D) diluted with 1.5 mL
acetone (Section 12.1, Exhibit D).
2.4.2.1.2 Extract by using the procedures beginning in Sections 12.2.5 or
12.2.6 of Exhibit D, as applicable, add 10 uL of the recovery stan-
dard solution (Section 12.9.2, Exhibit D) and analyze a 2-uL aliquot
of the concentrated extract.
2.4.2.1.3 Calculate the concentration (Section 14.1, Exhibit D) of 2,3,7,8-
substituted PCDDs/PCDFs and the percent recovery of the internal
standards (Section 14.2, Exhibit D). If the percent recovery at the
E-6
239
-------
measured concentration of any 2,3,7,8-substituted PCDD/PCDF congener
is <40 percent or >120 percent, report the results to SMO before
proceeding with the samples.
2.4.2.1.4 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.
2.4.2.2 Rinsate Sample
2.4.2.2.1 The rinsate sample must be fortified like a regular sample.
2.4.2.2.2 Take a 100-mL (+ 0.5 mL) portion of the sampling equipment rinse
solvent (rinsate sample), filter, if necessary, and add 100 uL of the
solution containing the nine internal standards (Table 2, Exhibit D).
2.4.2.2.3 Using a Kuderna-Danish appparatus, concentrate to approximately
5 mL.
2.4.2.2.4 Transfer the 5-mL concentrate from the K-D concentrator tube in 1-mL
portions to a 1-mL minivial, reducing the volume in the minivial as
necessary with a gentle stream of dry nitrogen.
2.4.2.2.5 Rinse the K-D 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.
E-7
-------
2.4.2.2.6 Just before analysis, add 10 uL trldecane recovery standard solution
(Table 2, Exhibit D), and reduce the volume to a final volume of 10
uL, or 50 uL, as necessary (Section 12.9.2, Exhibit D). No column
chromatography is required.
2.4.2.2.7 Analyze an aliquot following the same procedures used to analyze
samples (Section 13, Exhibit D).
2.4.2.2.8 Report percent recovery of the internal standard and the presence
of any PCDD/PCDF compounds on Form (to be determined) in pg/mL of
rinsate solvent.
2.5 Duplicate Analyses
2.5.1 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.
2.5.1.1 The results of the laboratory duplicates (percent recovery and concen-
trations of 2,3,7,8-substituted PCDD/PCDF compounds) must agree within
25 percent relative difference (difference expressed as percentage of
the mean). If the relative difference is >25 percent for any one of
the fifteen 2,3,7,8-substituted PCDDs/PCDFs, the laboratory shall
immediately contact the Sample Management Office f,or resolution of the
problem. Report all results.
E-8
241
-------
2.5.1.2 Recommended actions to help locate problems:
2.5.1.2.1 Verify satisfactory instrument performance (Section 8, Exhibit D).
2.5.1.2.2 If possible, verify that no error was made while weighing the sample
portions.
2.5.1.2.3 Review the analytical procedures with the performing laboratory
personnel.
2.6 Matrix Spike and Matrix Spike Duplicate
2.6.1 Locate the sample for the MS and MSD analyses (the sample may be labeled
"double volume").
2.6.2 Add on appropriate volume of the matrix spike fortification solution
(Exhibit D, Section 3.24), adjusting the fortification level as specified
in Exhibit D, Table 1, under IS Spiking Levels.
2.6.3 Analyze the MS and MSD samples as described in Exhibit D, Section 12.
2.6.A The results obtained from the MS and MSD samples (percent recovery and
concentrations of 2,3,7,8-substituted PCDDs/PCDFs) must agree within 20
percent relative difference.
2.7 Percent Recovery of the Internal Standards
E-9
242
-------
For each sample, method blank and rinsate, calculate the percent recovery
(Section 14.2, Exhibit D). It is recommended that the percent recovery be
>40 percent and <120 percent for all 2,3,7,8-substituted internal standards.
NOTE: A low or high percent recovery for a blank does not require dis-
carding the analytical data but it may indicate a potential problem with
future analytical data.
2.8 Identification Criteria
2.8.1 If either one of the identification criteria appearing in Sections
13.4.1.1 through 13.4.1.4, Exhibit D, 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 14.5, Exhibit D).
2.8.2 If the first initial identification criteria (Sections 13.4.1.1 through
13.4.1.4) are met, but the criteria appearing in Sections 13.4.1.5 and
13.4.2.1, Exhibit D, are not met, that sample is presumed to contain
interfering contaminants. This must be noted on the analytical report
form, and the sample must be rerun or the extract reanalyzed. Detailed
sample rerun and extract reanalysis requirements are presented in
Exhibit C.
2.9 Blind QA/QC Samples
Included among soil, sediment and aqueous samples may be QA/QC samples
E-10
243
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that are not specified as such to the performing laboratory. Types that
may be included are:
2.9.1 Uncontaminated soil, sediment, or water.
2.9.1.1 If a false positive is reported for such a sample, the laboratory
shall be required to rerun the entire associated batch of samples
(Section to be determined, Exhibit C).
2.9.2 Split samples — composited sample portions sent to more than one
laboratory.
2.9.3 Unlabeled field duplicates — two portions of a composited sample.
2.9.4 Performance evaluation samples — soil/sediment or water samples con-
taining a known amount of unlabeled 2,3,7,8-substituted PCDDs/PCDFs
and/or other PCDD/PCDF compounds.
2.9.4.1 If the performance evaluation sample result falls outside the accept-
ance windows established by the EPA, the laboratory shall be required
to rerun the entire associated batch of samples (Exhibit C).
NOTE: EPA acceptance windows are based on previously generated data.
2.10 Quality Control Charts
The performance of the entire measurement system (i.e., from the extraction
E-ll
244
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of the sample to the mass spectrometrlc determination) must be documented
by using germane control charts. The selection and design of a specific
measurement control chart must be accomplished in a rational manner so
that the measurement process can be adequately surveyed. By using the
standard deviations obtained from control samples or control runs, the
laboratory must delineate control limits, i.e., statistically congruous
extreme values, which should warn the operator of possible problems. It
is recommended to consider the values corresponding to two standard devi-
ations as warning limits and the values from three standard deviations as
control limits (i.e., corrective actions are required). For some par-
ticular applications, however, the control limits must not exceed the
limits set forth by the EPA (e.g., ion-abundance ratios). [Specific and
required QC charts, such as mass and GC resolutions, ion abundance ratios,
RRF values, etc., will be described in the final version of this protocol.]
2.11 Standard Operating Procedures (SOPs)
As part of the quality assurance program, the laboratory must use in-house
SOPs describing how the basic operations executed within the laboratory
are done.
2.12 Internal Audits
Internal audits of records, instrumentation performances and calibration
data are highly encouraged in order to identify defects that could
compromise the quality of the results.
E-12
245
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2.13 Records
At each laboratory, records must be maintained on site for six months
after contract completion to document the quality of all data generated
during the contract period. Before any records are disposed, written
concurrence from the Contracting Officer must be obtained.
2.14 Unused portions of samples and sample extracts must be preserved for six
months after sample receipt; appropriate samples may be selected by EPA
personnel for further analyses.
2.15 Reuse of glassware is to be minimized to avoid the risk of contamination.
3. Laboratory Evaluation Procedures
3.1 On a quarterly basis, the EPA Project Officer or his/her designated repre-
sentatives may conduct an evaluation of the laboratory to ascertain that
the' laboratory is meeting contract requirements. This section outlines
the procedures which may be used by the Project Officer or his/her author-
ized representative in order to conduct a successful evaluation of
laboratories conducting dioxin analyses according to this protocol. The
evaluation process consists of the following steps: 1) analysis of a
performance evaluation (PE) sample, and 2) on-site evaluation of the
laboratory to verify continuity of personnel, instrumentation, and quality
assurance/quality control functions. The following is a description of
these two steps.
E-13
246
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3.2 Performance Evaluation (PE) Sample Analysis
3.2.1 The PE sample set will be sent to a participating laboratory to verify
the laboratory's continuing ability to produce acceptable analytical
results. The PE sample will be representative of the types of samples
that will be analyzed under this contract.
3.2.2 When the PE sample results are received, they are scored using the PE
Sample Score Sheet shown in Figure (to be determined). If a false
positive (e.g., a PE sample not containing 2,3,7,8-TCDD or other PCDD/
PCDF but reported by the laboratory to contain it or them) is reported,
the laboratory has failed the PE analysis requirement. The Project
Officer will notify the laboratory immediately if such an event occurs.
3.2,3 As a general rule, a laboratory should achieve 75 percent or more of the
total possible points for all three categories listed on the PE Sample
Score Sheet, and 75 percent or more of the maximum possible points in
each category, to be considered acceptable for this program. However,
the Government reserves the right to accept scores of less than 75
percent.
3.2.4 If unanticipated difficulties with the PE samples are encountered, the
total points may be adjusted by the Government evaluator in an impartial
and equitable manner for all participating laboratories.
3.3 On-site Laboratory Evaluation
E-14
247
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3.3.1 An on-slte laboratory evaluation is performed to verify that (1) the
laboratory is maintaining the necessary minimum level in instrumentation
and levels of experience in personnel committed to the contract and (2)
that the necessary quality assurance activities are being carried out.
It also serves as a mechanism for discussing laboratory weaknesses
identified through routine data audits, PE sample analyses results, and
prior on-site evaluations. Photographs may be taken during the on-site
laboratory evaluation tour.
3.3.2 The sequence of events for the on-site evaluations is shown in Figure
(to be determined). A Site Evaluation Sheet (SES) is used to document
the results of the evaluation.
E-15
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EPA METHOD 3050
ACID DIGESTION OF SEDIMENTS, SLUDGES AND SOILS
Modifications: a) Sb digestion not to exceed 95° C
b) HC1 reflux for ICP fraction
249
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METHOD 3050
ACID DIGESTION OF SEDIMENTS. SLUDGES, AND SOILS
1.0 SCOPE AND APPLICATION
1.1 This method Is an acid digestion procedure used to prepare sedi-
ments, sludges, and soil samples for analysis by flame or furnace atomic
absorption spectroscopy (FLAA and GFAA, respectively) or by inductively
coupled argon plasma spectroscopy (ICP). Samples prepared by this method may
be analyzed by ICP for all the listed metals, or by FLAA or GFAA as indicated
below (see also Paragraph 2.1):
Aluminum
Bari urn
Beryllium
Cadmi urn
Calcium
Chromium
Cobalt
Copper
Iron
Lead
FLAA
Magnesium
Manganese
Molybdenum
Nickel
Potassium
Sodiurn
Thallium
Vanadium
Zinc
GFAA
Arsenic
Beryl 1i urn
Cadmi urn
Chromium
Cobalt
Iron
Molybdenum
Selenium
Thallium
Vanadium
2.0 SUMMARY OF METHOD
2.1 A representative
acid and hydrogen peroxide.
acid or hydrochloric acid.
reflux acid for (1) the ICP
analysis of Al, Ba, Be, Ca,
Zn. Dilute nitric acid is
AA analysis of As, Be, Cd,
shall be dried for a total
1- to 2-g (wet weight) sample is digested in nitric
The digestate is then refluxed with either'nitric
Dilute hydrochloric acid is used as the final
analysis of As and Se, and (2) the flame AA or ICP
Cd, Cr, Co, Cu, Fe, Mo, Pb, Ni, K, Na, Tl, V, and
employed as the final dilution acid for the furnace
Cr, Co, Pb, Mo, Se, Tl, and V. A separate sample
solids determination.
3.0 INTERFERENCES
3.1 Sludge samples can contain diverse matrix types, each of which may
present its own analytical challenge. Spiked samples and any relevant
standard reference material should be processed to aid in determining whether
Method 3050 is applicable to a given waste.
Revision Q
Date September 1986
50
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4.0 APPARATUS AND MATERIALS
4.1 Conical Phillips beakers; 250-mL.
4.2 Watch glasses.
4.3 Drying ovens: That can be maintained at 30*C.
4.4 ThermometerT That covers range of 0 to 200* C.
4.5 Whatman NoT 41 filter paper (or equivalent).
4.6 Centrifuge and centrifuge tubes.
5.0 REAGENTS
5.1 ASTM Type II water (ASTM D1193): Water should be monitored for
impurities.
5.2 Concentrated nitric add, reagent grade (HNOs) : Add should be
analyzed to determine level of Impurities. If method blank 1s
-------
Using a ribbed watch glass, allow the solution to evaporate to 5 ml without
boiling, while maintaining a covering of solution over the bottom of the
beaker.
7.3 After Step 7.2 has been completed and the sample has cooled, add 2
ml of Type II water and 3 ml of 30% H20?. Cover the beaker with a watch glass
and return the covered beaker to the not plate for warming and to start the
peroxide reaction. Care must be taken to ensure that losses do not occur due
to excessively vigorous effervescence. Heat until effervescence subsides and
cool the beaker.
7.4 Continue to add 30% HpC^ 1n 1-mL allquots with warming until the
effervescence 1s minimal or until the general sample appearance 1s unchanged.
NOTE: Do not add more than a total of 10 ml 30% H202-
7.5 If the sample 1s being prepared for (a) the ICP analysis of As and
Se, or (b) the flame AA or ICP analysis of Al, Ba, Be, Ca, Cd, Cr, Co, Cu, Fe,
Pb, Mg, Mn, Mo, N1, K, Na, Tl, V, and Zn, then add 5 ml of concentrated HC1
and 10 ml of Type II water, return the covered beaker to the hot plate, and
reflux for an additional 15 m1n without boiling. After cooling, dilute to
100 mL with Type II water. Particulates in the digestate that may clog the
nebulizer should be removed by filtration, by centrifugation, or by allowing
the sample to settle.
7.5.1 Filtration: Filter through Whatman No. 41 filter paper (or
equivalent) and dilute to 100 ml with Type II water.
7.5.2 Centrifugation: Centrifugation at 2,000-3,000 rpm for 10 m1n
1s usually sufficient to clear the supernatant.
7.5.3 The diluted sample has an approximate acid concentration of
5.0% (v/v) HC1 and 5.0% (v/v) HN03. The sample 1s now ready for
analysis.
7.6 If the sample 1s being prepared for the furnace analysis of As, Be,
Cd, Cr, Co, Pb, Mo, Se, Tl, and V, cover the sample with a ribbed watch glass
and continue heating the acid-peroxide digestate until the volume has been
reduced to approximately 5 ml. After cooling, dilute to 100 ml with Type II
water. Participates in the digestate should then be removed by filtration, by
centrifugation, or by allowing the sample to settle.
7.6.1 Filtration: Filter through Whatman No. 41 filter paper (or
equivalent) and dilute to 100 ml with Type II water.
7.6.2 Centrifugation: Centrifugation at 2,000-3,000 for 10 m1n 1s
usually sufficient to clear the supernatant.
7.6.3 The diluted digestate solution contains approximately 5%
(v/v) HN03. For analysis, withdraw allquots of appropriate volume and
add any required reagent or matrix modifier. The sample 1s now ready for
analysis.
Revision 0
Date September 1986
252
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7.7 Calculations:
7.7.1 The concentrations determined are to be reported on the basis
of the actual weight of the sample. If a dry weight analysis 1s desired,
then the percent sol Ids of the sample must also be provided.
7.7.2 If percent solids 1s desired, a separate determination of
percent solids must be performed on a homogeneous aliquot of the sample.
8.0 QUALITY CONTROL
8.1 For each group of samples processed, preparation blanks (Type II
water and reagents) should be carried throughout the entire sample preparation
and analytical process. These blanks will be useful 1n determining 1f samples
are being contaminated.
8.2 Duplicate samples should be processed on a routine basis. Duplicate
samples will be used to determine precision. The sample load will dictate the
frequency, but 20% 1s recommended.
8.3 Spiked samples or standard reference materials must be employed to
determine accuracy. A spiked sample should be Included with each group of
samples processed and whenever a new sample matrix is being analyzed.
8.4 The concentration of all calibration standards should be verified
against a quality control check sample obtained from an outside source.
9.0 METHOD PERFORMANCE
9.1 No data provided.
10.0 REFERENCES
10.1 None required.
Revision 0
Date September 1986
253
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METHOD 3O3O
ACID DIGESTION OF SEDIMENTS. SUUOGES. AND SOILS
C
7. 1
' Mix
• ample, take
l-Z g portion
for eacn
alged H^Oj.;
• for
t react.
7.4 j
Add M101
•nd warm until
•f f ervemc«nc«
1« •inlin«l
Q
Revision Q
Date September 1986
254
-------
METHOD 3OSO
AGIO DIGESTION OF SEDIMENTS. SLUDGES. AND SOILS
(Continued)
Furnace analysts of
A«. Be. Ca. Cr. Co. Pb.
Mo. Se. Tl. and V
7.6
ICP analysts at As ana Se
or flame AA or ICP
analysis of Al.Ba.8e.
Be. Ca. Ca. Cr. Cp. Cu.
. PO. Mg. Mn. Mo. HI.
K. Na. Tl. V. and Zn
Continue
heating to
reduce volume
7.6
7.5 I
Add
concentrated
HCL ana Type II
water: re lux
Dilute with
Type II water
7.6
7.5
Cool:
dilute
with Type II
water: filter
oartlculates In
the dlgestate
Filter
Oartlculates
In dlgestate
7.7.1|Determine
I percent
sol Ids on
homogeneous
sample aliquot
for calculation
7.7.21
I Oat era me
concentration*:
report percent
aollda of
•ample
f Stop j
Revision 0
Date September 1986
255
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3.3 METHODS FOR DETERMINATION OF METALS
This manual contains six analytical techniques for trace metal
determinations: Inductively coupled argon plasma emission spectrometry (ICP),
direct-aspiration or flame atomic absorption spectrometry (FAA), graphite-
furnace atomic absorption spectrometry (GFAA), hydride-generation atomic
absorption spectrometry (HGAA), cold-vapor atomic absorption spectrometry
(CVAA), and several procedures for hexavalent chromium analysis. Each of
these 1s briefly discussed below 1n terms of advantages, disadvantages, and
cautions for analysis of wastes.
ICP's primary advantage 1s that 1t allows simultaneous or rapid
sequential determination of many elements 1n a short time. The primary
disadvantage of ICP 1s background radiation from other elements and the plasma
gases. Although all ICP Instruments utilize high-resolution optics and back-
ground correction to minimize these Interferences, analysis for traces of
metals 1n the presence of a large excess of a single metal is difficult.
Examples would be traces of metals 1n an alloy or traces of metals in a limed
(high calcium) waste. ICP and Flame AA have comparable detection limits
(within a factor of 4) except that ICP exhibits greater sensitivity for
refractories (Al, Ba, etc.). Furnace AA, in general, will exhibit lower
detection limits than either ICP or FLAA.
Flame AAS (FLAA) determinations, as opposed to ICP, are normally
completed as single element analyses and are relatively free of interelement
spectral Interferences. Either a nitrous-oxide/acetylene or air/acetylene
flame is used as an energy source for dissociating the aspirated sample into
the free atomic state making analyte atoms available for absorption of light.
In the analysis of some elements the temperature or type of flame used 1s
critical. If the proper flame and analytical conditions are not used,
chemical and 1on1zat1on Interferences can occur.
Graphite Furnace AAS (GFAA) replaces the flame with an electrically
heated graphite furnace.The furnace allows for gradual heating of the sample
aliquot 1n several stages. Thus, the processes of desolvatlon, drying,
decomposition of organic and Inorganic molecules and salts, and formation of
atoms which must occur In a flame or ICP 1n a few milliseconds may be allowed
to occur over, a much longer time period and at controlled temperatures in the
furnace. This allows an experienced analyst to remove unwanted matrix
components by using temperature programming and/or matrix modifiers. The
major advantage of this technique is that it affords extremely low detection
limits. It 1s the easiest to perform on relatively clean samples. Because
this technique 1s so sensitive, Interferences can be a real problem; finding
the optimum combination of digestion, heating times and temperatures, and
matrix modifiers can be a challenge for complex matrices.
Revision 0
Date September 1986
256
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Hydride AA utilizes a chemical reduction to reduce and separate arsenic
or selenium selectively from a sample dlgestate. The technique therefore has
the advantage of being able to Isolate these two elements from complex samples
which may cause Interferences for other analytical procedures. Significant
Interferences have been reported when any of the following 1s present: 1)
easily reduced metals (Cu, Ag, Hg); 2) high concentrations of transition
metals (>200 mg/L); 3) oxidizing agents (oxides of nitrogen) remaining
following sample digestion.
Cold-Vapor AA uses a chemical reduction to reduce mercury selectively.
The procedure 1s extremely sensitive but 1s subject to Interferences from some
volatile organlcs, chlorine, and sulfur compounds.
Revision 0
Date September 1986
-------
EPA METHOD 204.2
ANTIMONY
ATOMIC ABSORPTION, FURNACE TECHNIQUE
259
-------
ANTIMONY
Method 204.2 (Atomic Absorption, furnace technique)
STORET NO. Total 01097
Dissolved 01095
Suspended 01096
Optimum Concentration Range: 20-300 ug/1
Detection Limit: 3 ug/1
Preparation of Standard Solution
1. Stock solution: Prepare as described under "direct aspiration method".
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. These solutions are also to be used for "standard additions".
3. The calibration standard should be diluted to contain 0.2% (v/v) HNOj.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual. .
Sample Preparation
1. The procedures for preparation of the sample as given in parts 4.1.1 thru 4.1.3 of the
Atomic Absorption Methods section of this manual should be followed including the
addition of sufficient 1:1 HC1 to dissolve the digested residue for the analysis of
suspended or total antimony. The sample solutions used for analysis should contain 2%
(v/v) HNO3.
Instrument Parameters (General)
1. Drying Time and Temp: 30sec-125°C.
2. Ashing Time and Temp: 30 sec-800'C.
3. Atomizing Time and Temp: lOsec-2700'C.
4. Purge Gas Atmosphere: Argon
5. Wavelength: 217.6 nm
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation, see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Approved for NPDES
Issued 1978
260
-------
Notes
1. 1 he above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injection, continuous flow purge gas and non-pyrotytic
graphite. Smaller size furnace devices or those employing faster rates of atomization can
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. The use of background correction is recommended.
3. Nitrogen may also be used as the purge gas.
4. If chloride concentration presents a matrix problem or causes a loss previous to
atomization, add an excess of 5 mg of ammonium nitrate to the furnace and ash using a
ramp accessory or with incremental steps until the recommended ashing temperature is
reached.
5. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
6. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
7. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. Precision and accuracy data are not available at this time.
261
-------
EPA METHOD 206.2
ARSENIC
ATOMIC ABSORPTION, FURNACE TECHNIQUE
263
-------
ARSENIC
Method 206.2 (Atomic Absorption, furnace technique)
STORET NO. Total 01002
Dissolved 01000
Suspended 01001
Optimum Concentration Range: 5-100 ug/1
Detection Limit: 1 ug/1
Preparation of Standard Solution
1. Stock solution: Dissolve 1.320 g of arsenic trioxide, As203 (analytical reagent grade) in
100 ml of deionized distilled water containing 4 g NaOH. Acidify the solution with 20 ml
cone. HNO3 and dilute to 1 liter. 1 ml = 1 mg As(lOOOmg/l).
2. Nickel Nitrate Solution, 5%: Dissolve 24.780 g of ACS reagent grade Ni(NO3)2-6H:O in
deionized distilled water and make up to 100ml.
3. Nickel Nitrate Solution, 1%: Dilute 20 ml of the 5% nickel nitrate to 100 ml with
deionized distilled water.
4. Working Arsenic Solution: Prepare dilutions of the stock solution to be used as
calibration standards at the time of analysis. Withdraw appropriate aliquots of the stock
solution, add 1 ml of cone. HNO3, 2ml of 30% H,O2 and 2ml of the 5% nickel nitrate
solution. Dilute to 100 ml with deionized distilled water.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. Transfer 100 ml of well-mixed sample to a 250 ml Griffin beaker, add 2 ml of 30% H:O,
and sufficient cone. HNO3 to result in an acid concentration of l%(v/v). Heat for 1 hour
at 95°C or until the volume is slightly less than 50 ml.
2. Cool and bring back to 50 ml with deionized distilled water.
3. Pipet 5 ml of this digested solution into a 10-ml volumetric flask, add 1 ml of the 1%
nickel nitrate solution and dilute to 10 ml with deionized distilled water. The sample is
now ready for injection into the furnace.
Approved for NPDES and SDWA
Issued 1978
264
-------
NOTE: If solubilization or digestion is not required, adjust the HNO3 concentration of
the sample to 1% (v/v) and add 2 ml of 30%H2Oj and 2 ml of 5% nickel nitrate to each
100 ml of sample. The volume of the calibration standard should be adjusted with
deionized distilled water to match the volume change of the sample.
Instrument Parameters (General)
1. Drying Time and Temp: 30 sec-125'C.
2. Ashing Time and Temp: 30 sec-1100°C.
3. Atomizing Time and Temp: 10860-2700*0.
4. Purge Gas Atmosphere: Argon
5. Wavelength: 193.7 nm
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation, see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injection, purge gas interrupt and non-pyrolytic
graphite. Smaller size furnace devices or those employing faster rates of atomtzation can
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. The use of background correction is recommended.
3. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
4. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
5. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
6. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. In a single laboratory (EMSL), using a mixed industrial-domestic waste effluent
containing 15 ug/1 and spiked with concentrations of 2, 10 and 25 t/g/l, recoveries of
85%, 90% and 88% were obtained respectively. The relative standard deviation at these
concentrations levels were ±8.8%, ±fc.2%, ±5.4% and ±8.7%, respectively.
2. In a single laboratory (EMSL), using Cincinnati, Ohio tap water spiked at concentrations
of 20, 50 and 100 ug As/1, the standard deviations were ±0.7, ±1.1 and ±1.6
respectively. Recoveries at these levels were 105%, 106% and 101%, respectively.
265
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EPA METHOD 270.2
SELENIUM
ATOMIC ABSORPTION, FURNACE TECHNIQUE
267
-------
SELENIUM
Method 270.2 (Atomic Absorption, furnace technique)
STORET NO. Total 01147
Dissolved 01145
Suspended 01146
Optimum Concentration Range: 5-100 ug/l
Detection Limit: 2 ug/l
Preparation of Standard Solution
1. Stock Selenium Solution: Dissolve 0.3453 g of selenous acid (actual assay 94.6% H2SeO3)
in deionized distilled water and make up to 200 ml. 1 ml == 1 mgSe(1000mg/l).
2. Nickel Nitrate Solution, 5%: Dissolve 24.780 g of ACS reagent grade Ni(NO;):'6H,O in
deionized distilled water and make up to 100 ml.
3. Nickel Nitrate Solution, 1%: Dilute 20 ml of the 5% nickel nitrate to 100 ml with
deionized distilled water.
4. Working Selenium Solution: Prepare dilutions of the stock solution to be used as
calibration standards at the time of analysis. Withdraw appropriate aliquots of the stock
solution, add 1 ml of cone. HNO3, 2 ml of 30% H2O2 and 2 ml of the 5% nickel nitrate
solution. Dilute to 100 ml with deionized distilled water.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. Transfer 100 ml of well-mixed sample to a 250 ml Griffin beaker, add 2 ml of 30% H:O,
and sufficient cone. HNO3 to result in an acid concentration of 1 %(v/v). Heat for 1 hour
at 95°C or until the volume is slightly less than 50 ml.
2. Cool and bring back to 50 ml with deionized distilled water.
3. Pipet 5 ml of this digested solution into a 10-ml volumetric flask, add 1 ml of the 1%
nickel nitrate solution and dilute to 10 ml with deionized distilled water. The sample is
now ready for injection into the furnace. NOTE: If solubilization or digestion is not
required adjust the HNO3 concentration of the sample to 1% (v/v) and add 2 ml of 30%
H,O, and 2 ml of 5% nickel nitrate to each 100 ml of sample. The volume of the
calibration standard should be adjusted with deionized distilled water to match the
volume change of the sample.
Approved for NPDES and SOW A
Issued 1978
268
-------
Instrument Parameters
1. Drying time and temperature: 30 sec © 125*C
2. Charring time and temperature: 30 sec @ 1200°C
3. Atomizing time and temperature: 10 sec © 2700°C
4. Purge Gas Atmosphere: Argon
5. Wavelength: 196.0 nm.
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100. based on the use of a 20 ul injection, puige gas interrupt and non-pyiolyik
graphite. Smaller size furnace devices or those employing faster rates of atomization can
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. The use of background correction is recommended;
3. Selenium analysis suffers interference from chlorides (> 800 mg/1) and sulfate (> 200
mg/1). For the analysis of industrial effluents and samples with concentrations of sulfate
from 200 to 2000 mg/1, both samples and standards should be prepared to contain 1%
nickel.
4. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
5. For quality control requirements and optional recommendations for use in drinking
water analyses, see part 10 of the Atomic Absorption Methods section of this manual.
6. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
7. Data to entered into STORET must be reported as ug/1.
Precision and Accuracy
1. Using a sewage treatment plant effluent containing <2 ug/1 and spiked with a
concentration of 20 ug/1, a recovery of 99% was obtained.
2. Using a series of industrial waste effluents spiked at a 50 ug/1 level, recoveries ranged
from 94 to 112%.
3. Using a 0.1% nickel nitrate solution as a synthetic matrix with selenium concentrations
of 5, 10, 20, 40, 50, and 100 ug/1, relative standard deviations of 14.2, 11.6, 9.3, 7.2, 6.4
and 4.1 %, respectively, were obtained at the 95% confidence level.
269
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4. In a single laboratory (EMSL), using Cincinnati, Ohio tap water spiked at concentrations
of 5, 10, and 20 t/g Se/1, the standard deviations were ±0.6, ±0.4, and ±0.5,
respectively. Recoveries at these levels were 92%, 98%, and 100%, respectively.
Reference:
"Determining Selenium in Water, Wastewater, Sediment and Sludge By Flameless Atomic
Absorption Spectroscopy", Martin, T. D., Kopp, J. F. and Ediger, R. D. Atomic Absorption
Newsletter 14,109(1975).
270
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EPA METHOD 279.2
THALLIUM
ATOMIC ABSORPTION, FURNACE TECHNIQUE
271
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THALLIUM
Method 279.2 (Atomic Absorption, furnace technique)
STORET NO. Total 01059
Dissolved 01057
Suspended 01058
Optimum Concentration Range: 5-100 ug/1
Detection Limit: 1 ug/1
Preparation of Standard Solution
1. Stock solution: Prepare as described under "direct aspiration method".
2. Prepare dilutions of the stock solution to be used as calibration standards at the time of
analysis. These solutions are also to be used for "standard additions".
3. The calibration standard should be diluted to contain 0.5% (v/v) HNO3.
Sample Preservation
1. For sample handling and preservation, see part 4.1 of the Atomic Absorption Methods
section of this manual.
Sample Preparation
1. Prepare as described under "direct aspiration method". Sample solutions for analysis
should contain 0.5% (v/v) HNO3.
Instrument Parameters (General)
1. Drying Time and Temp: 30 sec @ 125'C
2. Ashing Time and Temp: 30 sec @ 400°C
3. Atomizing Time and Temp: 10 sec @ 2400°C
4. Purge Gas Atmosphere: Argon
5. Wavelength: 276.8 nm
6. Other operating parameters should be set as specified by the particular instrument
manufacturer.
Analysis Procedure
1. For the analysis procedure and the calculation, see "Furnace Procedure" part 9.3 of the
Atomic Absorption Methods section of this manual.
Approved for NPDES
Issued 1978
172
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Notes
1. The above concentration values and instrument conditions are for a Perkin-Elmer HGA-
2100, based on the use of a 20 ul injection, continuous flow purge gas and non-pyrolytic
graphite. Smaller size furnace devices or those employing faster rates of atomization can
be operated using lower atomization temperatures for shorter time periods than the
above recommended settings.
2. The use of background correction is recommended.
3. Nitrogen may also be used as the purge gas.
4. For every sample matrix analyzed, verification is necessary to determine that method of
standard addition is not required (see part 5.2.1 of the Atomic Absorption Methods
section of this manual).
5. If method of standard addition is required, follow the procedure given earlier in part 8.5
of the Atomic Absorption Methods section of this manual.
6. Data to be entered into STORET must be reported as ug/1.
Precision and Accuracy
1. Precision and accuracy data are not available at this time.
273
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EPA 245.5
MERCURY IN SEDIMENT
MANUAL COLD VAPOR TECHNIQUE
275
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MERCURY IN SEDIMENT
Method 245.5 (Manual Cold Vapor Technique)
1. Scope and Application
1.1 This procedure'" measures total mercury (organic f inorganic) in soils, sediments,
bottom deposits and sludge type materials.
1.2 The range of the method is 0.2 to 5 ug/g. The range may be extended above or below the
normal range by increasing or decreasing sample size or through instrument and
recorder control.
2. Summary of Method
2.1 A weighed portion of the sample is digested in aqua regia for 2 minutes at 95"C, followed
by oxidation with potassium permanganate. Mercury in the digested sample is then
measured by the conventional cold vapor technique.
2.2 An alternate digestion'2' involving the use of an autoclave is described in (8.2).
3. Sample Handling and Preservation
3.1 Because of the extreme sensitivity of the analytical procedure and the omnipresence of
mercury, care must be taken to avoid extraneous contamination. Sampling devices and
sample containers should be ascertained to be free of mercury; the sample should not be
exposed to any condition in the laboratory that may result in contact or air-borne
mercury contamination.
3.2 While the sample may be analyzed without drying, it has been found to be more
convenient to analyze a dry sample. Moisture may be driven off in a drying oven at a
temperature of 60°C. No mercury losses have been observed by using this drying step.
The dry sample should be pulverized and thoroughly mixed before the aliquot is
weighed.
4. Interferences
4.1 The same types of interferences that may occur in water samples are also possible with
sediments, i.e., sulfides, high copper, high chlorides, etc.
4.2 Volatile materials which absorb at 253.7 nm will cause a positive interference. In order to
remove any interfering volatile materials, the dead air space in the BOD bottle should be
purged before the addition of stannous sulfate.
5. Apparatus
5.1 Atomic Absorption Spectrophotometer (See Note 1): Any atomic absorption unit
having an open sample presentation area in which to mount the absorption cell is
suitable. Instrument settings recommended by the particular manufacturer should be
followed.
NOTE 1: Instruments designed specifically for the measurement of mercury using the
cold vapor technique are commercially available and may be substituted for the atomic
absorption Spectrophotometer.
Issued 1974
276
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5.2 Mercury Hollow Cathode Lamp: Westinghouse WL-22847, argon filled, or equivalent.
5.3 Recorder: Any multi-range variable speed recorder that is compatible with the UV
detection system is suitable.
5.4 Absorption Cell: Standard spectrophotometer cells 10 cm long^ having quartz end
windows may be used. Suitable cells may be constructed from plexiglass tubing, 1" O.D.
X 4-1/2". The ends are ground perpendicular to the longitudinal axis and quartz
windows (1" diameter X 1/16" thickness) are cemented in place. Gas inlet and outlet
ports (also of plexiglass but 1/4" O.D.) are attached approximately 1/2" from each end.
The cell is strapped to a burner for support and aligned in the light beam to give the
maximum transmittance,
NOTE 2: Two 2" X 2" cards with one inch diameter holes may be placed over each end
of the cell to assist in positioning the cell for maximum transmittance.
5.5 Air Pump: Any peristaltic pump capable of delivering 1 liter of air per minute may be
used. A Masterflex pump with electronic speed control has been found to be satisfactory.
(Regulated compressed air can be used in an open one-pass system.)
5.6 Flowmeter: Capable of measuring an air flow of 1 liter per minute.
5.7 Aeration Tubing: Tygon tubing is used for passage of the mercury vapor from the sample
bottle to the absorption cell and return. Straight glass tubing terminating in a coarse
porous frit is used for sparging air into the sample.
5.8 Drying Tube: 6" X 3/4" diameter tube containing 20 g of magnesium perchlorate (See
Note 3). The apparatus is assembled as shown in the accompanying diagram.
NOTE 3: In place of the magnesium perchlorate drying tube, a small reading lamp with
60W bulb may be used to prevent condensation of moisture inside the cell. The lamp is
positioned to shine on the absorption cell maintaining the air temperature in the cell
about 10°C above ambient.
6. Reagents
6.1 Aqua Regia: Prepare immediately before use by carefully adding three volumes of cone.
HC1 to one volume of cone. HNO3.
6.2 Sulfuric Acid, 0.5 N: Dilute 14.0 ml of cone, sulfuric acid to 1 liter.
6.3 Stannous Sulfate: Add 25 g stannous sulfate to 250 ml of 0.5 N sulfuric acid (6.2). This
mixture is a suspension and should be stirred continuously during use.
6.4 Sodium Chloride-Hydroxylamine Sulfate Solution: Dissolve 12 g of sodium chloride
and 12 g of hydroxylamine sulfate in distilled water and dilute to 100ml.
NOTE 4: A 10% solution of stannous chloride may be substituted for (6.3) and
hydroxylamine hydrochloride may be used in place of hydroxylamine sulfate in (6.4)
6.5 Potassium Permanganate: 5% solution, w/v. Dissolve 5 g of potassium permanganate in
100 ml of distilled water.
6.6 Stock Mercury Solution: Dissolve 0.1354 g of mercuric chloride in 75 ml of distilled
water. Add 10 ml of cone, nitric acid and adjust the volume to 100.0 ml. 1.0 ml = 1.0
mgHg.
6.7 Working Mercury Solution: Make successive dilutions of the stock mercury solution
(6.6) to obtain a working standard containing 0.1 ug/ml. This working standard and the
dilution of the stock mercury solutions should be prepared fresh daily. Acidity of the
277
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working standard should be maintained at 0.15% nitric acid. This acid should be added
to the flask as needed before the addition of the aliquot.
7. Calibration
7.1 Transfer 0, 0.5, 1.0, 2.0, 5.0 and 10 ml aliquots of the working mercury solution (6.7)
containing 0 to 1.0 ug of mercury to a series of 300 ml BOD bottles. Add enough distilled
water to each bottle to make a total volume of 10 ml. Add 5 ml of aqua regia (6.1) and
heat 2 minutes in a water bath at 95°C. Allow the sample to cool and add 50 ml distilled
water and 15 ml of KMnO4 solution (6.5) to each bottle and return to the water bath for
30 minutes. Cool and add 6 ml of sodium chloride-hydroxylamine sulfate solution (6.4)
to reduce the excess permanganate. Add 50 ml of distilled water. Treating each bottle
individually, add 5 ml of stannpus sulfate solution (6.3) and immediately attach the
bottle to the aeration apparatus. At this point, the sample is allowed to stand quietly
without manual agitation. The circulating pump, which has previously been adjusted to
rate of 1 liter per minute, is allowed to run continuously. The absorbance, as exhibited
either on the spectrophotometer or the recorder, will increase and reach maximum
within 30 seconds. As soon as the recorder pen levels off, approximately 1 minute, open
the bypass value and continue the aeration until the absorbance returns to its minimum
value (See Note 5). Close the bypass value, remove the fritted tubing from the BOD
bottle and continue the aeration. Proceed with the standards and construct a standard
curve by plotting peak height versus micrograms of mercury.
NOTE 5: Because of the toxic nature of mercury vapor precaution must be taken to avoid
its inhalation. Therefore, a bypass has been included in the system to either vent the
mercury vapor into an exhaust hood or pass the vapor through some absorbing media,
such as:
a) equal volumes of 0.1 N KMnO4 and 10% H2SO4
b) 0.25% iodine in a 3% KI solution.
A specially treated charcoal that will absorb mercury vapor is also available from
Barnebey and Cheney, E. 8th Ave., and North Cassidy St., Columbus, Ohio 43219,
Cat. #580-13 or #580-22.
8. Procedure
8.1 Weigh triplicate 0.2 g portions of dry sample and place in bottom of a BOD bottle. Add 5
ml of distilled water and 5 ml of aqua regia (6.1). Heat 2 minutes in a water bath at 95°C.
Cool, add 50 ml distilled water and 15 ml potassium permanganate solution (6.5) to each
sample bottle. Mix thoroughly and place in the water bath for 30 minutes at 95CC. Cool
and add 6 ml of sodium chloride-hydroxylamine sulfate (6.4) to reduce the excess
permanganate. Add 55 ml of distilled water. Treating each bottle individually, add 5 ml
of stannous sulfate (6.3) and immediately attach the bottle to the aeration apparatus.
Continue as described under (7.1).
8.2 An alternate digestion procedure employing an autoclave may also be used. In this
method 5 ml of cone. H2SO4 and 2 ml of cone. HNO, are added to the 0.2 g of sample. 5
ml of saturated KMnO4 solution is added and the bottle covered with a piece of
aluminum foil. The samples are autoclaved at 121°C and 15 Ibs. for 15 minutes. Cool,
make up to a volume of 100 ml with distilled water and add 6 ml of sodium chloride-
-------
hydroxylamine sulfate solution (6.4) to reduce the excess permanganate. Purge the dead
air space and continue as described under (7. 1).
9. Calculation
9. 1 Measure the peak height of the unknown from the chart and read the mercury value from
the standard curve.
9.2 Calculate the mercury concentration in the sample by the formula:
usHg/B = in the aliquot
"6 66 wt of the aliquot in gms
9.3 Report mercury concentrations as follows: Below 0.1 ug/gm, <0.1; between 0.1 and 1
ug/gm, to the nearest 0.01 ug; between 1 and 10 ug/gm, to nearest 0.1 ug; above 10
ug/gm, to nearest ug.
10. Precision and Accuracy
10.1 The following standard deviations on replicate sediment samples were recorded at the
indicated levels; 0.29 ug/g ±0.02 and 0.82 ug/g ±0.03. Recovery of mercury at these
levels, added as methyl mercuric chloride, was 97% and 94%, respectively.
Bibliography
1. Bishop, J. N., "Mercury in Sediments", Ontario Water Resources Comm., Toronto, Ontario,
Canada, 1971.
2. Salma, M., private communication, EPA Cal/Nev Basin Office, Almeda, California.
279
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EPA METHOD 200.7
INDUCTIVELY COUPLED PLASMA - ATOMIC EMISSION
SPECTROMETRIC METHOD FOR TRACE ELEMENT ANALYSIS
OF WATER AND WASTES METHOD
Modification: + 42 Element Screen
281
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United States
Environmental Protection
Agency
Environmental Monitoring and
Support Laboratory
Cincinnati OH 45268
Research and Development
EPA
Test Method
Inductively Coupled Plasma—
Atomic Emission Spectrometric
Method for Trace Element
Analysis of Water and
Wastes—Method 200.7
1. Scope and Application
1.1 This method may be used for
the determination of dissolved,
suspended, or total elements in
drinking water, surface water,
domestic and industrial wastewaters
1.2 Dissolved elements are
determined in filtered and acidified
samples. Appropriate steps must be
taken in all analyses to ensure that
potential interference are taken into
account This is especially true when
dissolved solids exceed 1500 mg/ L
(See 5.)
1.3 Total elements are determined
after appropriate digestion procedures
are performed Since digestion
techniques increase the dissolved
solids content of the samples,
appropriate steps must be taken to
correct for potential interference
effects (See 5 )
1.4 Table! lists elements for which
this method applies along with
recommended wavelengths and
typical estimated instrumental
detection limits using conventional
pneumatic nebulization Actual
working detection limits are sample
dependent and as the sample matrix
varies, these concentrations may also
vary In time, other elements may be
Dec 1982
added as more information becomes
available and as required
1.5 Because of the differences
between various makes and models of
satisfactory instruments, no detailed
instrumental operating instructions
can be provided Instead, the analyst
is referred to the instructions provided
by the manufacturer of the particular
instrument.
2. Summary of Method
2.1 The method describes a
technique for the simultaneous or
sequential multielement
determination of trace elements in
solution. The basis of the method is
the measurement of atomic emission
by an optical spectroscopic technique
Samples are nebulized and the
aerosol that is produced is transported
to the plasma torch where excitation
occurs Characteristic atomic-line
emission spectra are produced by a
radio-frequency inductively coupled
plasma (ICP) The spectra are
dispersed by a grating spectrometer
and the intensities of the lines are
monitored by photomultiplier tubes
The photocurrents from the
photomultiplier tubes are processed
and controlled by a computer system
A background correction technique is
required to compensate for variable
background contribution to the
282
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determination of trace elements
Background must be measured
adjacent to analyte lines on samples
during analysis. The position selected
for the background intensity
measurement, on either or both sides
of the analytical line, will be
determined by the complexity of the
spectrum adjacent to the analyte line
The position used must be free of
spectral interference and reflect the
same change in background
intensity as occurs at the analyte
wavelength measured Background
correction is not required in cases of
line broadening where a background
correction measurement would
actually degrade the analytical result.
The possibility of additional
interferences named in 5 1 (and tests
for their presence as described in 5 2)
should also be recognized and
appropriate corrections made
3. Definitions
3.1 Dissolved — Those elements
which will pass through a 0 45 fjrr\
membrane filter.
3.2 Suspended — Those elements
which are retained by a 0 45 /ym
membrane filter
3.3 Total — The concentration
determined on an unfiltered sample
following vigorous digestion (9 3), or
the sum of the dissolved plus
suspended concentrations (9 1 plus
92.)
3.4 Total recoverable — The
concentration determined on an
unfiltered sample following treatment
with hot, dilute mineral acid (9 4)
3.5 Instrumental detection limit —
The concentration equivalent to a
signal, due to the analyte. which is
equal to three times the standard
deviation of a series of ten replicate
measurements of a reagent blank
signal at the same wavelength
3.6 Sensitivity — The slope of the
analytical curve, i e functional
relationship between emission
intensity and concentration
3.7 Instrument check standard — A
multielement standard of known
concentrations prepared by the
analyst to monitor and verify
instrument performance on a daily
basis (See 761)
3.8 Interference check sample — A
solution containing both interfering
and analyte elements of known
concentration that can be used to
verify background and interelement
correction factors. (See 762)
3.9 Quality control sample — A
solution obtained from an outside
source having known, concentration
values to be used to verify the
calibration standards. (See 7.6.3)
3.10 Calibration standards — a
series of know standard solutions
used by the analyst for calibration of
the instrument (i e . preparation of the
analytical curve). (See 7 4)
3.11 Linear dynamic range — The
concentration range over which the
analytical curve remains linear
3.12 Reagent blank — A volume of
deionized. distilled water containing
the same acid matrix as the
calibration standards carried through
the entire analytical scheme (See
752)
3.13 Calibration blank — A volume
of deionized, distilled water acidified
with HN03 and HCI (See 751)
3.14 Method of standard addition —
The standard addition technique
involves the use of the unknown and
the unknown plus a known amount of
standard (See 1061)
4. Safety
4.1 The toxicity or carcmogenicity 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 analysis Additional
references to laboratory safety are
available and have been identified
(14 7, 14 8 and 14 9) for the
information of the analyst
5. Interferences
5.1 Several types of interference
effects may contribute to inaccuracies
in the determination of trace
elements They can be summarized as
follows
511 Spectral interfere/tees can be
categorized as 1) overlap of a spectral
line from another element. 2)
Dec 1982
unresolved overlap of molecular band
spectra. 3) background contribution
from continuous or recombination
phenomena, and 4) background
contribution from stray light from the
line emission of high concentration
elements. The first of these effects
can be compensated by utilizing a
computer correction of the raw data.
requiring the monitoring and
measurement of the interfering
element The second effect may
require selection of an alternate
wavelength. The third and fourth
effects can usually be compensated by
a background correction adjacent to
the analyte line. In addition, users of
simultaneous multielement
instrumentation must assume the
responsibility of verifying the absence
of spectral interference from an
element that could occur in a sample
but for which there is no channel in
the instrument array Listed in Table 2
are some interference effects for the
recommended wavelengths given in
Table 1 The data in Table 2 are
intended for use only as a
rudimentary guide for the indication of
potential spectral interferences For
this purpose, linear relations between
concentration and intensity for the
analytes and the interferents can be
assumed
The interference information, which
was collected at the Ames Laboratory,1
is expressed at analyte concentration
eqivalems (i e. false analyte concen-
trations) arising from 100 mg L of the
interferent element The suggested use
of this information is as follows
Assume that arsenic (at 193 696 nm)
is to be determined in a sample
containing approximately 10 mg L of
aluminum According to Table 2. 100
mg L of aluminum would yield a false
signal for arsenic equivalent to
approximately 1 3 mg L Therefore.
10 mg/ L of aluminum would result in
a false signal for arsenic equivalent to
approximately 0 1 3 mg L The reader
is cautioned that other analytical
systems may exhibit somewhat
different levels of interference than
those shown in Table 2. and that the
interference effects must be evaluated
for each individual system
Only those interferents listed were
investigated and the blank spaces in
Table 2 indicate that measurable inter-
ferences were not observed for the
interferent concentrations listed in
Table 3 Generally, interferences were
discernible if they produced peaks or
background shifts corresponding to
2 5"'o of the peaks generated by the
Ami", l.ilior.Uiw USDOE low., Sli • ,Jnn.-r,iu
Am.-s low.i 'jOOl 1
28J
-------
analyte concentrations also listed in
Table 3.
At present, information on the listed
silver and potassium wavelengths are
not available but it has been reported
that second order energy from the
magnesium 383.231 nm wavelength
interferes with the listed potassium line
at 766491 nm.
5.1.2 Physical interferences are
generally considered to be effects
associated with the sample nebuliza-
tion and transport processes Such
properties as change in viscosity and
surface tension can cause significant
inaccuracies especially in samples
which may contain high dissolved
solids and/or acid concentrations. The
use of a peristaltic pump may lessen
these interferences. If these types of
interferences are operative, they must
be reduced by dilution of the sample
and/or utilization of standard addition
techniques. Another problem which
can occur from high dissolved solids
is salt buildup at the tip of the
nebulizer. This affects aersol flow-rate
causing instrumental drift Wetting
the argon prior to nebulization, the
use of a tip washer, or sample dilution
have been used to control this
problem. Also, it has been reported
that better control of the argon flow
rate improves instrument
performance. This is accomplished
with the use of mass flow controllers.
5.1.3 Chemical Interferences are
characterized by molecufar compound
formation, ionization effects and
solute vaporization effects. Normally
these effects are not pronounced with
the ICP technique, however, if
observed they can be minimized by
careful selection of operating
conditions (that is, incident power,
observation position, and so forth), by
buffering of the sample, by matrix
matching, and by standard addition
procedures These types of
interferences can be highly dependent
on matrix type and the specific
analyte element
5.2 It is recommended that
whenever a new or unusual sample
matrix is encountered, a series of
tests be performed prior to reporting
concentration data for analyte
elements. These tests, as outlined in
5.2.1 through 5 2.4, will ensure the
analyst that neither positive nor
negative interference effects are
operative on any of the analyte el-
ements thereby distorting the
accuracy of the reported values
5.2.1 Serial dilution—tf the analyte
concentration is sufficiently high (min-
imally a factor of 10 above the instru-
mental detection limit after dilution),
an analysis of a dilution should agree
within 5 % of the original determina-
tion (or within some acceptable con-
trol limit (14.3) that has been estab-
lished for that matrix) If not, a
chemical or physical interference ef-
fect should be sgspected.
5.2.2 Spike addition—The recovery
of a spike addition added at a
minimum level of 10X the in-
strumental detection limit (maximum
100X) to the original determination
should be recovered to within 90 to
110 percent or within the established
control limit for that matrix. If not, a
matrix effect should be suspected. The
use of a standard addition analysis
procedure can usually compensate for
this effect Caution: The standard ad-
dition technique does not detect coin-
cident spectral overlap. If suspected,
use of computerized compensation, an
alternate wavelength, or comparison
with an alternate method is recom-
mended (See 5 2.3)
5.2.5 Comparison with alternate
method of analysis—When investi-
gating a new sample matrix, compari-
son tests may be performed with other
analytical techniques such as atomic
absorption spectrometry, or other
approved methodology
5.2.4 Wavelength scanning of
analyte line region—If the appropriate
equipment is available, wavelength
scanning can be performed to detect
potential spectral interferences
6. Apparatus
6.1 Inductively Coupled Plasma-
Atomic Emission Spectrometer
6.1.1 Computer controlled atomic
emission spectrometer with background
correction
6.1.2 Radiofrequency generator
6.1.3 Argon gas supply, welding
grade or better.
6.2 Operating conditions — Because
of the differences between various
makes and models of satisfactory
instruments, no detailed operating
instructions can be provided Instead,
the analyst should follow the
instructions provided by the
manufacturer of the particular
instrument Sensitivity, instrumental
detection limit, precision, linear dy-
namic range, and interference effects
must be investigated and established
for each individual analyte line on that
particular instrument It is the
Dec 1982
responsibility of the analyst to verify
that the instrument configuration and
operating conditions used satisfy the
analytical requirements and to
maintain quality control data
confirming instrument performance
and analytical results.
7. Reagents and standards
7.1 Acids used in the preparation
of-standards and for sample processing
must be ultra-high purity grade or
equivalent Redistilled acids are
acceptable
7.1.1 Acetic acid, cone, (sp gr 1.06).
7.1.2 Hydrochloric acid. cone, (sp gr
1.19)
7.1.3 Hydrochloric acid. (1 +1). Add
500 mL cone HCl (sp gr 1.19) to 400
mL deiomzed, distrilled water and
dilute to 1 liter.
7.1.4 Nitric acid, cone (spgr1.41).
7.1.5 Nitric acid,C\ + "\): Add 500 mL
cone HNO3(sp gr 1.41) to 400 ml
deiomzed, distilled water and dilute to
1 liter.
7.2 Diomzed. distilled water: Prepare
by passing distilled water through a
mixed bed of cation and anion ex-
change resins Use deiomzed, distilled
water for the preparation of all
reagents, calibration standards and as
dilution water The purity of this water
must be equivalent to ASTM Type II
reagent water of Specification D 1193
(14.6).
7.3 Standard stock solutions may be
purchased or prepared from ultra high
purity grade chemicals or metals All
salts must be dried for 1 h at 105°C
unless otherwise specified
(CAUTION Many metal salts are ex-
tremely toxic and may be fatal if swal-
lowed Wash hands thoroughly after
handling ) Typical stock solution pre-
paration procedures follow
7.3.1 Aluminum solution, stock. 1
mL = 100 fig Al Dissolve 0.100 g of
aluminum metal in an acid mixture of 4
mL of (1+1) HCl and 1 mL of cone. HN03
m a beaker Warm gently to effect
solution. When solution is complete,
transfer quantitatively to a liter flask,
add an additional 10 mL of (1 + 1) HCl
and dilute to 1,000 mL with deiomzed,
distilled water
7.3.2 Antimony solution stock. 1 mL
= 100 fjg Sb Dissolve 0.2669 g K(SbO)
C4HjOe in deiomzed distilled water,
add 10 mL (1+1) HCl and dilute
to 1000 ml with deiomzed. distilled
water
284
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7.3.3 Arsenic solution, stock. 1 mL r
100 ug As. Dissolve 0.1320 g of As203
in 100 mL of deionized, distilled water
containing 0.4 g NaOH. Acidify the
solution with 2 mL cone. HMOs and
dilute to 1,000 ml with deionized,
distilled water.
7.3.4 Barium solution, stock. 1 mL
= 100 Mg Ba: Dissolve 0.1516 g BaCI2
(dried at 250°C for 2 hrs) in 10 mL
deionized, distilled water with 1 mL
(1+DHC1 Add 10.0 mL(1+1) HCI
and dilute to 1,000 mL with deionized.
distilled water
7.3.5 Beryllium solution, stock. 1
mL = 100 ug Be: Do not dry. Dis-
solve 1.966 g BeS04 • 4' 4H20, in
deionized, distilled water, add 10.0 mL
cone. HNCb and dilute to 1,000 mL
with deionized, distilled water
7.3.6 Boron solution, stock. 1 mL
= 100 ug B. Do not dry. Dissolve
0.5716 g anhydrous H3B03 m deionized
distilled water dilute to 1.000 mL.
Use a reagent meeting ACS specifica-
tions, keep the bottle tightly stoppered
and store in a desiccator to prevent
the entrance of atmospheric moisture
7.3.7 Cadmium solution, stock. 1
mL = 100 fug Cd: Dissolve 0 1142 g
CdO in a minimum amount of (1*1)
HN03 Heat to increase rate of dis-
solution. Add 10.0 mLconc HN03
and dilute to 1,000 mLVvith deionized,
distilled water.
7.3.5 Calcium solution, stock, 1 mL
= 100 ng Ca: Suspend 0.2498 g
CaC03 dried at 180°C for 1 h before
weighing m deionized, distilled water
and dissolve cautiously with a min-
imum amount of (1 + 1) HNC>3 Add
100 mLconc HN03 and dilute to
1,000 mL with deionized, distilled
water
7.3.5 Chromium solution, stock. 1
mL - 100 fjg Cr Dissolve 0 1923
g of CrO3 m deionized, distilled
water When solution is complete.
acidify with 10 mL cone HN03 and
dilute to 1.000 mL with deionized.
distilled water
7.3.10 Cobalt solution, stock. 1
mL - 100 ug Co Dissolve 0 1000 g
of cobalt metal in a minimum amount
of (1*1) HN03 Add 10 0 mL < 1*11 HCI
and dilute to 1.000 mL with deionized
distilled water
7.3. / 1 Copper solution, stock, 1
mL = 100 fjg Cu Dissolve 0 1252 g
CuO m a minimum amouni of 0*1)
HNO3 Add 100 mLconc HN03 and
dilute to 1,000 mL with deionized.
distilled water
7.3.12 Iron solution, stock. 1 mL
= 100/jg Fe Dissolve 0.1430 g
FejOs in a warm mixture of 20 mL
(1 +1) HCI and 2 mL of cone. HNO3.
Cool, add an additional 5 mL of cone.
"HNO3 and dilute to 1000 mL with
deionized, distilled water.
7.3.13 Lead solution, stock. 1 mL
= 100/ug Pb: Dissolve 0.1599 g
Pb(NC>3)2 in minimum amount of
(1+1) HN03. Add 10.0 mLconc. HMOs
and dilute to 1.000 mL with deionized.
distilled water
7.3.14 Magnesium solution, stock. 1
mL = 100 ug Mg: Dissolve 0.1658 g
MgO m a minimum amount of (1 + 1)
HNO3. Add 10.0 mL cone. HN03 and
dilute to 1,000 mL with deionized,
distilled water
7.3.15 Manganese solution, stock. 1
mL = 100 ug Mn Dissolve 0 1000 g
of manganese metal in the acid mix-
ture 10 mL cone HCI and 1 mL cone
HN03, and dilute to 1.000 mL with
deionized, distilled water.
7.3.16 Molybdenum solution, stock.
1 mL = 100 vg Mo: Dissolve 0 2043 g
(NH^jMoO* m deionized, distilled
water and dilute to 1.000 mL.
7.3.17 Nickel solution, stock. 1
mL=100/;gNi Dissolve 0.1000 g
of nickel metal in 10 mL hot cone
HN03, cool and dilute to 1,000 mL
with deionized, distilled water
7.3.18 Potassium solution, stock, 1
mL = \ 00 fjg K. Dissolve 0.1907 g
KCI, dried at 110°C, m deionized,
distilled water dilute to 1.000 mL
7 3.19 Selenium solution, stock. 1
mL = 100 /jg Se Do not dry Dissolve
0 1727 g H?SeO3 (actual assay 94 6%)
in deionized, distilled water and dilute
to 1.000 mL
7.3.20 Silica solution, stock. 1 mL
= 100 fjg SiO2 Do not dry Dissolve
0 4730 g Na2SiO3 • 9H?O in deionized.
distilled water Add 100 mL cone
HN03 and dilute to 1,000 mL with
deionized, distilled water
7.3.21 Silver solution, stock. 1
mL = 100^gAg Dissolve 0 1575 g
AgN03 in 100 mL of deionized. dis-
tilled water and 10 mL cone HNO3
Dilute to 1.000 mL with deionized,
distilled water
7 3 22 Sodium solution, stock. 1
mL = 100 fjg Na Dissolve 0 2542 g
NaCI m deionized, distilled water
Add 100 mLconc HNO . and dilute
to 1 000 mL with deionized, distilled
waller
Dec 1982
7.3.23 Thallium solution, stock, 1
mL = 100 ug Tl. Dissolve 0 1303 g
TIN03 in deionized, distilled water.
Add 10.0 mL cone. HN03 and dilute
to 1,000 mL with deionized, distilled
water.
7.3.24 Vanadium solution, stock. 1
mL = 100 ug V Dissolve 0.2297
NH4V03 m a minimum amount of
cone. HNO3 Heat to increase rate
of dissolution. Add 10.0 mL cone.
HN03 and dilute to 1,000 mL with
deionized. distilled water
7.3.25 Zinc solution, stock. 1 mL
= 100 ug Zn: Dissolve 0.1245 g ZnO
in a minimum amount of dilute HN03.
Add 10 0 mL cone. HN03 and dilute
to 1,000 mL with deionized, distilled
water
7.4 Mixed calibration standard so-
lutions—Prepare mixed calibration
standard solutions by combining ap-
propriate volumes of the stock solu-
tions m volumetric flasks (See 7.4 1
thru 7 4.5) Add 2 mL of (Ul)
HCI and dilute to 100 mL with
deionized, distilled water (See Notes
1 and 6 ) Prior to preparing the mixed
standards, each stock solution should
be analyzed separately to determine
possible spectral interference or the
presence of impurities Care should
be taken when preparing the rruxed
standards that the elements are com-
patible and stable Transfer the mixed
standard solutions to a FEP fluoro-
carbon or unused polyethylene bottle
for storage Fresh mixed standards
should be prepared as needed with
(tie realization that concentration can
change on aging Calibration stand-
ards must be initially verified using
a quality control sample and moni-
tored weekly for stability (See 763)
Although not specifically required,
some typical calibration standard com-
binations follow when using those
specific wavelengths listed m Table
1
7.4.1 Mixed standard solution I —
Manganese, beryllium, cadmium, lead,
and zinc
7 4.2 Mixed standaid solution II -
Barium, copper, iron, vanadium and
cobalt
743 Mixed standard solution III -
Molybdenum silica arsenic and
selenium
744 Mixed standard solution IV -
Calcium sodium potassium alumi-
num, chroiruutn and nickel
285
-------
7 4.5 Mixea standard solution V—
Antimony boron, magnesium, silver,
and thallium
NOTE 1 If the addition of silver
to the recommended acid combination
results in an initial precipitation,
add 15 ml_ of deionized distilled
water and warm the flask until the
solution clears Cool and dilute to 100
ml with deionized, distilled water For
this acid combination the silver con-
centration should be limited to 2
mg L Silver under these conditions
is stable in a tap water matrix
for 30 days Higher concentrations
of silver require additional HCI
7.5 Two types of blanks are required
for the analysis The calibration blank
'3 13) is used in establishing the
analytical curve while the reagent
olank (3 12) is used to correct for
possible contamination resulting from
varying amounts of the acids used in
the sample processing
7.5.1 The calibration blank is pre-
pared by diluting 2 ml of (1 + 1) HNOj
and 10 ml of (1*1) HCI to 100 ml
with deionized, distilled water (See
Note 6 ) Prepare a sufficient quantity
to be used to flush the system be-
tween standards and samples
7.5.2 The reagent blank must con-
contain all the reagents and in the
same volumes as used m the pro-
cessing of the samples The reagent
blank must be carried through the
complete procedure and contain the
same acid concentration in the final
solution as the sample solution
used for analysis
7.6 In addition to the calibration
standards, an instrument check stan-
dard (3 7), an interference check
sample (3 81 and a quality control
sample (3 9) are also required for the
analyses
761 The instrument check standard
s oreoared by the analyst by com-
oining compatible elements at a con-
centration equivalent to the midpoint
nf their respective calibration curves
See 12 I 1)
762 The interference check sample
3 prepared by the analyst in the
oMowing manner Select a
eoresentative sample which contains
•Animal concentrations of the
analyses of interest by known con-
rentration of interfering elements that
will provide an adequate test of the
:orrection factors Spike the sample
.vitn the elements of interest at the
3:)oroximate concentration of either
* 00 wg/L or 5 times the estimated
detection limits given in Table 1 (For
effluent samples of expected high
concentrations, spike at an
appropriate level ) If the type of
samples analyzed are varied, a
synthetically prepared sample may be
used if the above criteria and intent
are met A limited supply of a
synthetic interference check sample
will be available from the Quality
Assurance Branch of EMSL-
Cmcinnati (See 1212)
7.6.3 The quality control sample
should be prepared in the same acid
matrix as the calibration standards
at a concentration near 1 mg/L and in
accordance with the instructions
provided by the supplier. The Quality
Assurance Branch of EMSL-Cmcinnati
will either supply a quality control
sample or information where one of
equal quality can be procured (See
121 3)
8. Sample handling an
preservation
8.1 For the determination of trace
elements, contamination and loss are
of prime concern Dust in the labora-
tory environment, impurities in
reagents and impurities on laboratory
apparatus which the sample contacts
are all sources of potential
contamination Sample containers can
introduce either positive or negative
errors in the measurement-of trace
elements by (a) contributing con-
taminants through leaching or surface
desorption and (b) by depleting
concentrations through adsorption.
Thus the collection and treatment of
the sample prior to analysis requires
particular attention Laboratory
glassware including the sample bottle
(whether polyethylene, polyproplyene
or FEP-fluorocarbon) should be
thoroughly washed with detergent
and tap water, rinsed with (1 + 1) nitric
acid, tap water. (1+1) hydrochloric
acid, tap and finally deionized, distilled
water in that order (See Notes 2 and
3).
NOTE 2 Chromic acid may be useful to
remove organic deposits from glass-
ware, however, the analyst should be
be cautioned that the glassware must
be thoroughly rinsed with water to
remove the last traces of chromium.
This is especially important if chromium
is to be included in the analytical
scheme A commercial product, NOCH-
ROMIX, available from Godax Labor-
atories, 6 Varick St., New York, NY
10013, may be used in place of
chromic acid. Gnomic acid should not
be used with plastic bottles
NOTE3 If it can be documented through
Dec 1982
an active analytical quality control
program using spiked samples and re-
agent blanks, that certain steps in the
cleaning procedure are not required for
routine samples, those steps may be
eliminated from the procedure
8.2 Before collection of the sample a
decision must be made as to the type
of data desired, that is dissolved,
suspended or total, so that the appro-
priate preservation and pretreatment
steps may be accomplished Filtration.
acid preservation, etc., are to be per-
formed at the time the sample is
collected or as soon as possible
thereafter.
8.2.7 For the determination of dis-
solved elements the sample must be
filtered through a 0.45-pm membrane
filter as soon as practical after collec-
tion. (Glass or plastic filtering appara-
tus are recommended to avoid possi-
ble contamination.) Use the first 50-
100 mL to rinse the filter flask. Dis-
card this portion and collect the
required volume of filtrate. Acidify the
filtrate with (1 +1) HNOs to a pH of 2
or less Normally, 3 mL of (1 + 1) acid
per liter should be sufficient to pre-
serve the sample
8.2.2 for the determination of sus-
pended elements a measured volume
of unpreserved sample must be fil-
tered through a 0.45-//m membrane
filter as soon as practical after
collection. The filter plus suspended
material should be transferred to a
suitable container for storage and/or
shipment. No preservative is required
8.2.3 For the determination of total
or total recoverable elements, the
sample is acidified with (1 +1) HNC"3
to pH 2 or less as soon as possible.
preferable at the time of collection
The sample is not filtered before
processing.
9. Sample Preparation
9.1 For the determinations of dis-
solved elements, the filtered,
preserved sample may often be
analyzed as received The acid matrix
and concentration of the samples and
calibration standards must be the
same. (See Note 6.) If a precipitate
formed upon acidification of the
sample or during transit or storage, it
must be redissolved before the
analysis by adding additional acid
and/or by heat as described in 9 3
9.2 For the determination of sus-
pended elements, transfer the mem-
brane filter containing the insoluble
material to a 1 50-mL Griffin beaker
and add 4 mL cone HNO3. Cover the
286
-------
beaker with a watch glass and heat
gently The warn acid will soon dis-
solve the membrane.
Increase the temperature of the
hot plate and digest the material.
When the acid has nearly evaporated,
cool the beaker and watch glass and
add another 3 mL of cone. HNOa
Cover and continue heating until the
digestion is complete, generally indi-
cated by a light colored digestate
Evaporate to near dryness (2 mL), cool.
add 10 mL HCI (U1) and 15 mL
deionized. distilled water per 100 mL
dilution and warm the beaker gently
for 15 mm to dissolve any precipi-
tated or residue material Allow to
cool, wash down the watch glass and
beaker walls with deionized distilled
water and filter the sample to remove
insoluble material that could clog the
nebulizer (See Note 4.) Adjust the
volume based on the expected con-
centrations of elements present This
volume will vary depending on the
elements to be determined (See Note
6) The sample is now ready for
analysis. Concentrations so determined
shall be reported as "suspended "
NOTE 4 In place of filtering, the
sample after diluting and mixing may
be centnfuged or allowed to settle by
gravity overnight to remove insoluble
material
9.3 For the determination of total
elements, choose a measured, volume
of the well mixed acid preserved
sample appropriate for the expected
level of elements and transfer to a
Griffin beaker (See Note 5 ) Add 3 mL
of cone HN03 Place the beaker on
a hot plate and evaporate to near dry-
ness cautiously, making certain that
the sample does not boil and that no
area of the bottom of the beaker is
allowed to go dry Cool the beaker and
add another 5 mL portion of cone
HNOa Cover the beaker with a watch
glass and return to the hot plate
Increase the temperature of the hot
plate so that a gentle reflux action
occurs Continue heating, adding addi-
tional acid as necessary, until the
digestion is complete (generally indi-
cated when the digestate is light
in color or does not change in appear-
ance with continued refluxmg ) Again,
evaporate to near dryness and cool
the beaker Add 10 mL of 1*1 HCI
and 1 5 mL of deionized, distilled
water per 100 mL of final solution
and warm the beaker gently for 15
mm to dissolve any precipitate or
residue resulting from evaporation
Allow to cool, wash down the beaker
walls and watch glass with deionized
distilled water and filter the sample to
remove insoluble material that could
clog the nebulizer (See Note 4 ) Adjust
the sample to a predetermined volume
based on the expected concentrations
of elements present. The sampie is
now ready for analysis (See Note 6).
Concentrations so determined shall be
reported as "total."
NOTE 5 If low determinations of
boron are critical, quartz glassware
should be use.
NOTE 6. If the sample analysis solution
has a different acid concentration
from that given in 9.4, but does not
introduce a physical interference or
affect the analytical result, the same
calibration standards may be used.
9.4 For the determination of total
recoverable elements, choose a mea-
sured volume of a well mixed, acid
preserved sample appropriate for the
expected level of elements and trans-
fer to a Griffin beaker (See Note 5 )
Add 2 mL of (1 -M) HNOa and 10 mL
of (1 + 1) HCI to the sample and heat
on a steam bath or hot plate until the
volume has been reduced to near 25
mL making certain the sample does
not boil After this treatment, cool
the sample and filter to remove inso-
luble material that could clog the
nebulizer. (See Note 4 ) Adjust the
volume to 100 mL and mix The sample
is now ready for analysis Concentra-
tions so determined shall be reported
as "total "
10. Procedure
10.1 Set up instrument with proper
operating parameters established in
6 2 The instrument must be allowed
to become thermally stable before be-
ginning This usually requires at least
30 mm of operation prior to calibra-
tion
10.2 Initiate appropriate operating
configuration of computer
10.3 Profile and calibrate instru-
ment according to instrument
manufacturer's recommended
procedures, using the typical mixed
calibration standard solutions
described in 7 4 Flush the system
with the calibration blank (7 5.1)
between each standard (See Note 7 )
(The use of the average intensity of
multiple exposures for both
standardization and sample analysis
has been found to reduce random
error )
NOTE 7 For boron concentrations
greater than 500 */g/L extended flush
times of 1 to 2 mm may be required
10.4 Before beginning the sample
run, reanalyze the highest mixed
calibration standard as if it were a
Cec 1982
sample Concentration values obtained
should not deviate from the actual
values by more than z. 5 percent
(or the established control limits
whichever is lower) If they do. follow
the recommendations of the instru-
ment manufacturer to correct for this
condition
10.5 Begin the sample run flushing
the system with the calibration blank
solution (7 5.1) between each sample
(See Note 7 ) Analyze the instrument
check standard (761) and the calibra-
tion blank (751) each 10 samples
10.6 If it has been found that
method of standard addition are
required, the following procedure is
recommended.
JO.6.1 The standard addition tech-
nique (14 2) involves preparing new
standards in the sample matrix by
adding known amounts of standard to
one or more aliquots of the processed
sample solution This technique com-
pensates for a sample constituent that
enhances or depresses the analyte
signal thus producing a different slope
from that of the calibration standards
It will not correct for additive inter-
ference which causes a baseline shift
The simplest version of this technique
is the single-addition method The
procedure is as follows Two identical
aliquots of the sample solution, each
of volume V,, are taken To the
first (labeled A) is added a small
volume Vs of a standard analyte
solution of concentration cs To the
second (labeled 8) is added the same
volume Vs of the solvent The analy-
tical signals of A and B are measured
and corrected for nonanalyte signals
The unknown sample concentration
c. is calculated
cx = SaVsCs
IS* - Se) Vx
where SA and SB are the analytical
signals (corrected for the blank) of
solutions A and B. respectively V$
and cs should be chosen so that SA
is roughly twice SB on the average It
is best if Vs is made much less than
Vx, and thus Cs is much greater than
cx, to avoid excess dilution of the
sample matrix If a separation or
concentration step is used, the
additions are best made first and
carried through the entire procedure
For the results from this technique to
be valid, the following limitations
must be taken into consideration
1 The analytical curve must be linear
2 The chemical form of the analyte
added must respond the same as the
analyte m the sample
287
-------
3 The interference effect must be
constant over the working range of
concern
4. The signal must be corrected for
any additive interference
11. Calculation
11.1 Reagent blanks (7 5.2) should
be subtracted from all samples. This is
particularly important for digested
samples requiring large quantities of
acids to complete the digestion.
11.2 If dilutions were performed,
the appropriate factor must be applied
to sample values.
11.3 Data should be rounded to the
thousandth place and all results
should be reported in mg/L up to
three significant figures.
12. Quality Control
(Instrumental)
12.1 Check the instrument
standardization by analyzing
appropriate quality control check
standards as follow.
12.1.1 Analyze an appropriate
instrument check standard (761)
containing the elements of interest at
a frequency of 10%. This check
standard is used to determine
instrument drift. If agreement is not
within ±5% of the expected values or
within the established control limits,
whichever is lower, the analysis is out
of control. The analysis should be
terminated, the problem corrected,
and the instrument recalibrated
Analyze the calibration blank (751)
at a frequency of 10% The result
should be within the established
control limits of two standard devia-
tions of the mean value If not, repeat
the analysis two more times and
average the three results If the
average is not within the control limit,
terminate the analysis, correct the
problem and recalibrate the
instrument
12.1.2 To verify interelement and
background correction factors analyze
the interference check sample (-7 6 2)
at the beginning, end, and at periodic
intervals throughout the sample run
Results should fall within the
established control limits of 1 5 times
the standard deviation of the mean
value. If not, terminate the analysis.
correct the problem and recalibrate
the instrument
12.1.3 A quality control sample
(7 6 3) obtained from an outside
source must first be used for the
initial verification of the calibration
standards A fresh dilution of this
sample shall be anlayzed every week
thereafter to monitor their stability If
the results are not within ±5% of the
true value listed for the control
sample, prepare a new calibration
standard and recalibrate the
instrument If this does not correct the
problem, prepare a new stock
standard and a new calibration
standard and repeat the calibration
Precision and Accuracy
13.1 In an EPA round robin phase 1
study, seven laboratories applied the
ICP technique to acid-distilled water
matrices that had been dosed with
various metal concentrates. Table 4
lists the true value, the mean reported
value and the mean % relative
standard deviation
References
1 Wmge, R K , V J Peterson, and
VA Fassel, "Inductively Coupled
Plasma-Atomic Emission
Spectroscopy Prominent Lines," EPA-
600/4-79-017
2. Winefordner. J D , "Trace
Analysis Spectroscopic Methods for
Elements," Chemical Analysis, Vol
46, pp 41-42
3 Handbook for Analytical Quality
Control in Water and Wastewater
Laboratories, EPA-600'4-79-01 9
4 Garbarmo, J R and Tavlor, H E ,
"An Inductively-Coupled Plasma
Atomic Emission Spectrometric
Method for Routine Water Quality
Testing," Applied Spectroscopy 33,
No 3(1979)
5 "Methods for Chemical Analysis of
Water and Wastes," EPA-600 4 79-
020
6 Annual Book of ASTM Standards,
Part 31
7 "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, Aug 1977
8 "OSHA Safety and Health Stan-
dards, General Industry," (29 CFR
1910). Occupational Safety and Health
Administration, OSHA 2206, (Revised,
January 1976)
9 'Safety m Academic Chemistry
Laboratories, American Chemical So
ciety Publication, Committee on
Chemical Safely, 3rd Edition, 1979
1982
288
-------
Table 1. Recommended Wavelengths ' and Estimated Instrumental
Detection Limits
Element
Aluminum
Arsenic
Antimony
Barium
Beryllium
Boron
Cadmium
Calcium
Chromium
Cobalt
Copper
Iron
Lead
Magnesium
Manganese
Molybdenum
Nickel
Potassium
Selenium
Silica (SiOzJ
Silver
Sodium
Thallium
Vanadium
Zinc
. Wavelength, nm
308.215
193.696
206.833
455.403
313.042
249 773
226.502
317.933
267.716
228.616
324.754
259.940
220.353
279.079
257.610
202.030
231 604
766.491
196.026
288. 158
328.068
588.995
190.864
292.402
213.856
Estimated detection
limit. ug/L1
45
53
32
2
0.3
5
4
to
7
7
6
7
42
30
2
a
15
see'
75
58
7
29
40
8
2
' The wavelengths listed are recommended because of their sensitivity and
overall acceptance. Other wavelengths may be substituted if they can
provide the needed sensitivity and are treated with the same corrective
techniques for spectral interference. (See 5.1 I.).
2The estimated instrumental detection limits as shown are taken from
"Inductivefy Coupled Plasma-Atomic Emission Spectroscopy-Prominent
Lines. "EPA-600/4-79-017. They are given as a guide for an instrumental
limit. The actual method detection limits are sample dependent and may vary
as the sample matrix varies.
'^Highly dependent on operating conditions and plasma position.
Dec '982
289
-------
Table 2. Ana/yte Concentration Equivalents (mg/LI Arising From tnterferents at the 100 mg/L Level
Analyte Wavelength, nm Interferent
A lummum
Antimony
Arsenic
308.215
206.833
193.696
Al
0.47
1.3
Ca
—
Cr
2.9
0.44
Cu
"^^
Fe
0.08
Mg
—
Mn
0.21
Ni
—
Ti
25
V
1 4
0.45
1.1
Barium
Beryllium
Boron
Cadmium
Calcium
Chromium
Cobalt
Copper
Iron
Lead
Magnesium
Manganese
Molybdenum
Nickel
Selenium
Silicon
Sodium
Thallium
Vanadium
Zinc
455 403
313042
249.773
226 502
317933
267716
228.616
324 754
259.940
220.353
279 079
257.610
202.030
231 604
196026
288 158
588.995
190.864
292.402
213.856
0.04 0.05
0.04 —
0.08 -
— 0.03 —
0.17 - —
— 0.02 0 11
0.005 - 0.01
005 — —
023 - -
0.32 —
0.03 -
0.01 O.O1
0.003 —
0.005 —
0.003 -
0.02 -
0.04
0.04
003
0.12 -
0.13 — 0.25
0.002 0.002 -
0.03 - -
0.09 - -
0.03 0.03
— 004
0.15 —
0.05 002
0.07 0.12
007 —
030 —
005 -
0005 -
014— —
- o.o;
005 -
0.02 -
029 -
Table 3. Interferent and Analyte Elemental Concen-
trations Used for Interference Measurements
in Table 2.
Analytes (mg/LI
Interferents
(mg/L)
Al
As
B
Ba
Be
Ca
Cd
Co
Cr
Cu
Fe
Mg
Mn
Mo
Va
V;
3b
Sb
Se
S,
TI
/
7n
10
10
10
1
1
1
10
1
1
1
1
1
1
10
10
10
10
10
10
1
10
J
10
Al
Ca
Cr
Cu
Fe
Mg
Mn
Ni
Ti
V
1000
1000
200
200
1000
WOO
200
200
200
200
Dec 1982
290
-------
Table 4.
Element
Be
Mn
V
As
Cr
Cu
Fe
Ai
Cd
Co
Ni
Pb
In
Se
ICP Precision and Accuracy Data
Sample K t
True
Value
ug/L
750
350
750
200
150
250
600
700
50
500
250
250
200
40
Mean
Reported
Value
H9/L
733
345
749
208
149
235
594
696
48
512
245
236
201
32
Mean
Percent
BSD
6.2
2.7
1.8
7.5
3.8
S.I
3.0
5.6
12
10
5.8
16
5.6
21.9
True
Value
uff/L
20
15
70
22
10
11
20
60
2.5
20
30
24
16
6
Sample tt2
Mean
Reported
Value
ug/L
20-
15
69
19
10
11
19
62
2.9
20
28
30
19
8.5
Mean
Percent
RSD
9.8
6.7
2.9
23
18
40
15
33
16
4.1
11
32
45
42
True
Value
fjg/L
180
wo
170
60
50
70
180
160
14
120
60
80
80
10
Sample tt3
Mean
Reported
Value '
uff/L
176
99
169
63
50
67
178
161
13
108
55
80
82
85
Mean
Percent
RSD
5.2
33
1 1
17
33
79
60
13
16
21
14
14
94
83
Not all elements were analyzed by all laboratories.
Dec 1982
291
-------
MODIFICATION TO EPA METHOD 200.7 FOR 42 ELEMENT ICP SCREEN
Attached is a table of recommended wavelengths and minimum levels for this
procedure. This procedure requires a sequential ICP instrument (2 channel
minimum) interfaced with a computerized data system capable of the short
sampling times and narrow survey windows necessary to perform a semi-
quantitative ICP screen.
293
-------
ICP SCREEN ELEMENTS,
Requested Element
Symbol
WAVELENGTHS, 8. LTL
Wavelength* LTL «» (Note: LTL
Minimum Level)
.-.i_...:.._T.
4ta**n»
Zirconium
Al-SS
Sb-SS
As-SS
Ba-SS
Be-SS
Bi-SS
B-SS
Cd-SS
Ca-SS
Ce-SS
Cr-SS
Co-SS
Cu-SS
Dy-SS
Ei— SS
Eu-SS
Gd-SS
Ga-SS
Ge-SS
Au-SS
Hf-SS
Ho-SS
In-SS
I-SS
Ir-SS
Fe-SS
La-SS
Pb-SS
Li-SS
Lu-SS
Mg-SS
MttwQQ
nn~"oa
Hn—
-------
EPA METHOD 160.3
RESIDUE, TOTAL
GRAVIMETRIC, DRIED AT 103-105°C
295
-------
RESIDUE, TOTAL
Method 160.3 (Gravimetric, Dried at 103-lOS'O
STORET NO. 00500
1. Scope and Application
1.1 This method is applicable to drinking, surface, and saline waters, domestic and industrial
wastes.
1.2 The practical range of the determination is from 10 mg/1 to 20,000 mg/1.
2. Summary of Method
2.1 A well mixed aliquot of the sample is quantitatively transferred to a pre-weighed
evaporating dish and evaporated to dryness at 103-105'C.
3. Definitions
3.1 Total Residue is defined as the sum of the homogenous suspended and dissolved
materials in a sample.
4. Sample Handling and Preservation
4.1 Preservation of the sample is not practical; analysis should begin as soon as possible.
Refrigeration or icing to 4*C, to minimize microbiological decomposition of solids, is
recommended.
5. Interferences
5.1 Non-representative particulates such as leaves, sticks, fish and lumps of fecal matter
should be excluded from the sample if it is determined that their inclusion is not desired
in the final result.
5.2 Floating oil and grease, if present, should be included in the sample and dispersed by a
blender device before aliquoting.
6. Apparatus
6.1 Evaporating dishes, porcelain, 90 mm, 100 ml capacity. (Vycor or platinum dishes may
be substituted and smaller size dishes may be used if required.)
7. Procedure
7.1 Heat the clean evaporating dish to 103-105*C for one hour, if Volatile Residue is to be
measured, heat at 550 ±50'C for one hour in a muffle furnace. Cool, desiccate, weigh and
store in desiccator until ready for use.
7.2 Transfer a measured aliquot of sample to the pre-weighed dish and evaporate to dryness
on a steam bath or in a drying oven.
7.2.1 Choose an aliquot of sample sufficient to contain a residue of at least 25 mg. To
obtain a weighable residue, successive aliquots of sample may be added to the same
dish.
7.2.2 If evaporation is performed in a drying oven, the temperature should be lowered to
approximately 98*C to prevent boiling and splattering of the sample.
Approved for NPDES
Issued 1971
-------
7.3 Dry- the evaporated sample for at least 1 hour at 103-105*C. Cool in a desiccator and
weigh. Repeat the cycle of drying at 103-105'C, cooling, desiccating and weighing until a
constant weight is obtained or until loss of weight is less than 4% of the previous weight,
or 0.5 mg, whichever is less.
8. Calculation
8.1 Calculate total residue as follows:
Total residue, mg/1-(A " Bjxl'°°°
where:
A = weight of sample + dish in mg
B = weight of dish in mg
C = volume of sample in mi
9. Precision and Accuracy
9.1 Precision and accuracy data are not available at this time.
Bibliography
1. Standard Methods for the Examination of Water and Wastewater, 14th Edition, p 91, Method
208A,(1975).
297
-------
EPA METHOD 335.2
CYANIDE, TOTAL
TITRIMETRIC, SPECTROPHOTOMETRIC
299
-------
CYANIDE, TOTAL
Method 335.2 (Titrimetric; Spectrophotometric)
STORET NO. 00720
1. Scope and Application
1.1 This method is applicable to the determination of cyanide in drinking, surface and saline
waters, domestic and industrial wastes.
1.2 The titration procedure using silver nitrate with p-dimethylamino-benzal-rhodanine
indicator is used for measuring concentrations of cyanide exceeding 1 mg/1 (0.25
mg/250 ml of absorbing liquid).
1.3 The colorimetric procedure is used for concentrations below 1 mg/1 of cyanide and is
sensitive to about 0.02 mg/1.
2. Summary of Method
2.1 The cyanide as hydrocyanic acid (HCN) is released from cyanide complexes by means of
a reflux-distillation operation and absorbed in a scrubber containing sodium hydroxide
solution. The cyanide ion in the absorbing solution is then determined by volumetric
titration or colorimetrically.
2.2 In the colorimetric measurement the cyanide is converted to cyanogen chloride, CNC1,
by reaction with chloramine-T at a pH less than 8 without hydrolyzing to the cyanate.
After the reaction is complete, color is formed on the addition of pyridine-pyrazolone or
pyridine-barbituric acid reagent. The absorbance is read at 620 nm when using pyridine-
pyrazolone or 578 nm for pyridine-barbituric acid. 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 titrimetric measurement uses a standard solution of silver nitrate to titrate cyanide in
the presence of a silver sensitive indicator.
3. Definitions
3.1 Cyanide is defined as cyanide ion and complex cyanides converted to hydrocyanic acid
(HCN) by reaction in a reflux system of a mineral acid in the presence of magnesium ion.
4. Sample Handling and Preservation
4.1 The sample should be collected in plastic or glass bottles of 1 liter or larger size. All
bottles must be thoroughly cleansed and thoroughly rinsed to remove soluble material
from containers.
4.2 Oxidizing agents such as chlorine decompose most of the cyanides. Test a drop of the
sample with potassium iodide-starch test paper (Kl-starch paper); a blue color indicates
the need for treatment. Add ascorbic acid, a few crystals at a time, until a drop of sample
produces no color on the indicator paper. Then add an additional 0.06 g ol .i-.«>il>ic
acid foi each litei of sample volume.
Approved for NPDES
Issued 1974
Editorial revision 1974 and 1978
Technical Revision 1980
-------
4.3 Samples must be preserved with 2 ml of 10 N sodium hydroxide per liter of sample
(pH > 12) at the time of collection.
4.4 Samples should be analyzed as rapidly as possible after collection. If storage is required,
the samples should be stored in a refrigerator or in an ice chest filled with water and ice to
maintain temperature at 4*C.
5. Interferences
5.1 Interferences are eliminated or reduced by using the distillation procedure described
in Procedure 8.1, 8.2 and 8.3.
5.2 Sulfides adversely affect the colorimetric and titration procedures. Samples that
contain hydrogen sulfide, metal sulfides or oilier compounds that may produce
hydrogen sulfide during the distillation should be distilled by the optional procedure
described in Procedure 8.2. The apparatus for this procedure is shown in Figure 3.
5.3 Fatty acids will distill and form soaps under the alkaline titration conditions, making the
end point almost impossible to detect.
5.3.1 Acidify the sample with acetic acid (1 +9) to pH 6.0 to 7.0.
Caution: This operation must be performed in the hood and the sample left there
until it can be made alkaline again after the extraction has been performed.
5.3.2 Extract with iso-octane, hexane, or chloroform (preference in order named) with a
solvent volume equal to 20% of the sample volume. One extraction is usually
adequate to reduce the fatty acids 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 NaOH solution.
5A 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 formed will decompose under
(esi conditions to generate HCN. The interference of nitrate and nitrite is eliminated
In pretreatmem with sulfamic acid.
6. Apparatus
6.1 Reflux distillation apparatus such as shown in Figure 1 or Figure 2. The boiling flask
should be of 1 liter size with inlet tube and provision for condenser. The gas absorber may
be a Fisher-Milligan scrubber.
6.2 Microburet, 5.0 ml (for titration).
6.3 Spectrophotometer suitable for measurements at 578 nm or 620 nm with a 1.0 cm cell or
larger.
().{ Reflux distil l.u ion appaiaiux (or sulfide removal ax shown in Figuic'l The boiling
I laxk same .is 6.1. I'he Mill ule s< i iibbo may be a Wheaton Bubhei «70%82 with 2«.) 1'J
joinix. xi/e 100 nil. I'lic air inlet tube should not be trilled. The ( yanidc .ibsoi piion
\ esxel should be the same as the xullide s< rubbei. The air inlet tube should be li ined.
(i.") Flow meiei. xu< li ax Lab Cresi with slain less sieel lloal (Fishei 11 -1 (i I-.">()).
7. Reagents
7.1 Sodium hydroxide solution, 1.25N: Dissolve 50 g of NaOH in distilled water, and dilute
to 1 liter with distilled water.
30
-------
7.2 Leadacetate: Dissolve 30 g of Pb(C2H3O2)«3H2O in 950 ml of distilled water. Adjust
the pH to 4.5 with acetic acid. Dilute to 1 liter.
7.5 Sulfuric acid; 18N: Slowly add 500 ml of concentrated HzSO* to 500 ml of distilled
water.
7.6 Sodium dihydrogenphosphate, 1 M: Dissolve 138 g of NaH2PO4»H2O in 1 liter of
distilled water. Refrigerate this solution.
7.7 Stock cyanide solution: Dissolve 2.51 g of KCN and 2 g KOH in 900 nil of distilled
water. Standardize with 0.0192 N AgNOa. Dilute to appropriate concentration so thai
1 ml = 1 ing CN.
7.8 Standard cyanide solution, intermediate: Dilute 100.0 ml of stock (1 ml = l mgC\)to
1000 ml with distilled water (1 ml = 100.0 ug).
7.9 Working standard cyanide solution: Prepare fresh daily by diluting 100.0 ml of
intermediate cyanide solution to 1000 ml with distilled water and store in a glass
stoppered bottle. 1 ml = 10.0 ug CN.
7.10 Standard silver nitrate solution, 0.0192 N: Prepare by crushing approximately 5 g
AgNO3 crystals and drying to constant weight at 40°C. Weigh out 3.2647 g of dried
AgNO3, dissolve in distilled water, and dilute to 1000 ml (1 ml = Img CN).
7.11 Rhodanine indicator: Dissolve 20 mg of p-dimethyl-amino-benzalrhodanine in 100 ml of
acetone.
7.12 Chloramine T solution: Dissolve 1.0 g of white, water soluble Chloramine T in 100 ml of
distilled water and refrigerate until ready to use. Prepare fresh daily.
7.13 Color Reagent — One of the following may be used:
7.13.1 Pyridine-Barbituric Acid Reagent: Place 15 g of barbituric acid in a 250 ml
volumetric flask and add just enough distilled water to wash the sides of the
flask and wet the barbituric acid. Add 75 ml of pyridine and mix. Add 15 ml
of cone. HC1, mix, and cool to room temperature. Dilute to 250 ml with
distilled water and mix. This reagent is stable for approximately six months
if stored in a cool, dark place.
7.13.2 Pyridine-pyrazolone solution:
7.13.2.1 3-MethyM-phenyl-2-pyrazolin-5-one reagent, saturated solution: Add
0.25 g of 3-methyl-l-phenyl-2-pyrazolin-5-one to 50 ml of distilled
water, heat to 60°C with stirring. Cool to room temperature.
7.13.2.2 3,3'Dimethyl-l, l'-diphenyI-(4,4'-bi-2 pyrazoline]-5,5'dion€ (bispyra-
zolone): Dissolve 0.01 g of bispyrazolone in 10 ml of pyridine.
7.13.2.3 Pour solution (7.13.2.1) through non-acid-washed filter paper. Collect
the filtrate. Through the same filter paper pour solution (7.13.2.2)
collecting the filtrate in the same container as filtrate from (7.13.2.1).
Mix until the filtrates are homogeneous. The mixed reagent develops a
pink color but this does not affect the color production with cyanide if
used within 24 hours of preparation.
7.14 Magnesium chloride solution: Weight 510 g of MgCl,»6H2O into a 1000 ml flask, dissolve
and dilute to 1 liter with distilled water.
7.1") .Sillf.iitlK ;a id.
02
-------
8. Procedure
8.1 For samples without sulfide.
8.1.1 Place 500 ml of sample, or an aliquot diluted to 500 ml in the 1 liter boiling
flask. Pipet 50 ml of sodium hydroxide (7.1) into the absorbing tube. If the
apparatus in Figure 1 is used, add distilled water until the spiral is covered.
Connect the boiling flask, condenser, absorber and trap in the train. (Figure 1
or 2)
8.1.2 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
enters the boiling flask through the air inlet tube. Proceed to 8.4.
8.2 For samples that contain sulfide.
8.2.1 Place 500 ml of sample, or an aliquot diluted to 500 ml in the 1 liter boiling
flask. Pipet 50 ml of sodium hydroxide (7.1) to the absorbing tube. Add 25 ml of
lead acetate (7.2) to the sulfide scrubber. Connect the boiling flask, condenser.
scrubber and absorber in the train. (Figure 3) The flow meter is connected to the
outlet tube of the cyanide absorber.
8.2.2 Start a stream of air entering the boiling flask by adjusting the vacuum source.
Adjust the vacuum so that approximately i.5 liters per minute enters the
boiling flask through the air inlet tube. The bubble rate may not remain
constant while heat is being applied to the flask. It may be necessary to readjust
the air rate occasionally. Proceed to 8.4.
8.3 If samples contain NO3 and or NO2 add 2 g of sulfamic acid solution (7.15) after the air
rate is set through the air inlet tube. Mix for 3 minutes prior to addition of HjSO^
8.4 Slowly add 50 ml 18N sulfuric acid (7.5) through the air inlet tube. Rinse the tube with
distilled water and allow the airflow to mix the flask contents for 3 min. Pour 20 ml of
magnesium chloride (7.14) into the air inlet and wash down with a stream of water.
8.5 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, disconnect absorber and
close off the vacuum source.
8.6 Drain the solution from the absorber into a 250 ml volumetric flask. Wash the absorber
with distilled water and add the washings to the flask. Dilute to the mark with distilled
water.
8.7 Withdraw 50 ml or less of the solution from the flask and transfer to a 100ml volumetric
flask. If less than 50 ml is taken, dilute to 50 ml with 0.25N sodium hydroxide solution
(7.4). Add 15.0 ml of sodium phosphate solution (7.6) and mix.
8.7.1 Pyridine-barbituric acid method: Add 2 ml of chloramine T (7.12) and mix.
See Note 1. After 1 to 2 minutes, add 5 ml of pyridine-barbituric acid solution
(7.13.1) and mix. Dilute to mark with distilled water and mix again. Allow 8
minutes for color development then read absorbance at 578 nm in a 1 cm cell
within 15 minutes.
8.7.2 Pyridine-pyra/olene method: Add 0.5 ml of chlorawine T (7.12) and mix. Scr
Note 1 and 2. After 1 to 2 minutes add 5 ml of pyridine-pyrazolone solution
-------
(7.13.1) and mix. Dilute to mark with distilled water and mix again. After 40
minutes read absorbance at 620 nm in a 1 cm cell.
NOTE 1: Some distillates may contain compounds that have a chlorine
demand. One minute after the addition of chloramine T, test for
residual chlorine with KI-starch paper. If the test is negative, add an
additional 0.5 ml of chlorine T. After one minute, recheck the sample.
NOTE 2: More than 05. ml of chloramine T will prevent the color from
developing with pyridine-pyrazolone.
8.8 Standard curve for samples without sulfide.
8.8.1 Prepare a series of standards by pipeting suitable volumes of standard solution
(7.9) into 250 ml volumetric flasks. To each standard add 50 ml of 1.25 N
sodium hydroxide and dilute to 250 ml with distilled water. Prepare as follows:
ML of Working Standard Solution Cone, fjg CN
(1 ml = 10/t/gCN) per 250 ml
0 BLANK
1.0 10
2.0 20
5.0 50
10.0 100
15.0 150
20.0 200
8.8.2 It is not imperative that all standards be distilled in the same manner as the
samples. It is recommended that at least two standards (a high and low) be
distilled and compared to similar values on the curve to insure that the distil-
lation 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.
8.8.3 Prepare a standard curve by plotting absorbance of standard vs. cyanide
concentrations.
8.8.4 To check the efficiency of the sample distillation, add an increment of cyanide
from either the intermediate standard (7.8) or the working standard (7.9) to
500 ml of sample to insure a level of 20 /vg/1. Proceed with the analysis as in
Procedure (8.1.1).
8.9 Standard curve for samples with sulfide.
8.9.1 It is imperative that all standards be distilled in the same manner as the samples.
Standards distilled by this method will give a linear curve, but as the concen-
tration increases, the recovery decreases. It is recommended thai at least 3
standards be distilled.
8.9.2 Prepare a standard curve by plotting absorbance of standard vs. cyanide con-
centrations.
-------
8.10 Titrimetric method.
8.10.1 If the sample contains more than 1 mg/1 of CN, transfer the distillate or a
suitable aliquot diluted to 250 ml, to a 500 ml Erlenmeyer flask. Add 10-12 drops
of the benzalrhodanine indicator.
8.10.2 Titrate with standard silver nitrate to the first change in color from yellow to
brownish-pink. Titrate a distilled water blank using the same amount of sodium
hydroxide and indicator as in the sample.
8.10.3 The analyst should familiarize himself with the end point of the titration and the
amount of indicator to be used before actually titrating the samples.
9. Calculation
9.1 If the colorimetric procedure is used, calculate the cyanide, in ug/1, in the original
sample as follows:
CN,ug/l = A x 1.000 x 50
B C
where:
A = ug CN read from standard curve
B = ml of original sample for distillation
C = ml taken for colorimetric analysis
305
-------
9.2 Using the titrimetric procedure, calculate concentration of CN as follows:
CN, mg/t = v~ ~ °"-uw 25°
ml ong. sample ml of aliquot titrated
where:
A = volume of AgNO3 for titration of sample.
B = volume of AgNO3 for titration of blank.
10. Precision and Accuracy
10.1 In a single laboratory (EMSL), using mixed industrial and domestic waste samples at
concentrations of 0.06, 0.13, 0.28 and 0.62 mg/1 CN, the standard deviations were
±0.005, iO.007, ±0.031 and ±0.094, respectively.
10.2 In a single laboratory (EMSL), using mixed industrial and domestic waste samples at
concentrations of 0.28 and 0.62 mg/1 CN, recoveries were 85% and 102%, respectively.
Bibliography
1. Bark, L. S., and Higson, H. G. "Investigation of Reagents for the Colorimetrie Determination
of Small Amounts of Cyanide", Talanta, 2:471-479 (1964).
2. Elly, C. T. "Recovery of Cyanides by Modified Serfass Distillation". Journal Water Pollution
Control Federation 40:848-856 (1968).
3. Annual Book of ASTM Standards, Part 31, "Water", Standard D2036-75, Method A, p 503
(1976).
4. Standard Methods for the Examination of Water and Wastewater, 14th Edition, p 367 and 370,
Method 413B and D (1975).
5. Egekeze, J. O., and Oehne. F. W., "Direct Potentiometric Determination of Cyanide in
Biological Materials." J. Analytical Toxicology, Vol. 3, p. 119, May/June 1979.
6. Casey, J. P., Bright, J. W., and Helms. B. D., "Nitrosation Interference in Distillation Tests
for Cyanide," Gulf Coast Waste Disposal Authority. Houston, Texas.
306
-------
ALLIHN CONDENSER
AIR INLET TUBE
— CONNECTING TUBING
ONE LITER
BOILING FLASK
SUCTION
FIGURE 1
CYANIDE DISTILLATION APPARATUS
307
-------
COOLING WATER
INLET
TO LOW VACUUM
SOURCE
* ABSORBER
^ DISTILLING FLASK
HEATER-*
FIGURE 2
CYANIDE DISTILLATION APPARATUS
308
-------
EPA METHOD 340.2
FLUORIDE
POTENTIOMETRIC, ION SELECTIVE ELECTRODE
309
-------
FLUORIDE
Method 340.2 (Potentiometric, Ion Selective Electrode)
STORE! NO: Total 00951
Dissolved 00950
1. Scope and Application
1.1 This method is applicable to the measurement of fluoride in drinking, surface and saline
waters, domestic and industrial wastes.
1.2 Concentration of fluoride from 0.1 up to 1000 rag/liter may be measured.
1.3 For Total or Total Dissolved Fluoride, the Bellack distillation is required for NPDES
monitoring but is not required for SDWA monitoring.
2. Summary of Method
2.1 The fluoride is determined potentiometrically using a fluoride electrode in conjunction
with a standard single junction sleeve-type reference electrode and a pH meter having an
expanded millivolt scale or a selective ion meter having a direct concentration scale for
fluoride.
2.2 The fluoride electrode consists of a lanthanum fluoride crystal across which a potential is
developed by fluoride ions. The cell may be represented by Ag/Ag Cl, Cl"(0.3),
FT(O.OOl) LaF/test solution/SCE/
3. Interferences
3.1 Extremes of pH interfere; sample pH should be between 5 and 9. Polyvalent cations of
Si*4, Fe*3 and A1+J interfere by forming complexes with fluoride. The degree of
interference depends upon the concentration of the complexing cations, the
concentration of fluoride and the pH of the sample. The addition of a pH 5.0 buffer
(described below) containing a strong chelating agent preferentially complexes
aluminum (the most common interference), silicon and iron and eliminates the pH
problem.
4. Sampling Handling and Preservation
4.1 No special requirements.
5. Apparatus
5.1 Electrometer (pH meter), with expanded mv scale, or a selective ion meter such as the
Orion 400 Series.
5.2 Fluoride Ion Activity Electrode, such as Orion No. 94-09'"
5.3 Reference electrode, single junction, sleeve-type, such as Orion No. 90-01, Beckman No.
40454, or Corning No. 476010.
5.4 Magnetic Mixer, Teflon-coated stirring bar.
Approved for NPDES and SDWA
Issued 1971
Editorial revision 1974
310
-------
6. Reagents
6.1 Buffer solution, pH 5.0-5.5: To approximately 500 ml of distilled water in a 1 liter beaker
add 57 ml of glacial acetic acid, 58 g of sodium chloride and 4 g of CDTA'2'. Stir to
dissolve and cool to room temperature. Adjust pH of solution to between 5.0 and 5.5 with
5 N sodium hydroxide (about 150 ml will be required). Transfer solution to a 1 liter
volumetric flask and dilute to .the mark with distilled water. For work with brines,
additional NaCl should be added to raise the chloride level to twice the highest expected
level of chloride in the sample.
6.2 Sodium fluoride, stock solution: 1.0 ml = 0.1 mg F. Dissolve 0.2210 g of sodium fluoride
in distilled water and dilute to 1 liter in a volumetric flask. Store in chemical-resistant
glass or polyethylene.
6.3 Sodium fluoride, standard solution: 1.0 ml = 0.01 mg F. Dilute 100.0 ml of sodium
fluoride stock solution (6.2) to 1000 ml with distilled water.
6.4 Sodium hydroxide, 5N: Dissolve 200 g sodium hydroxide in distilled water, cool and
dilute to 1 liter.
7. Calibration
7.1 Prepare a series of standards using the fluoride standard solution (6.3) in the range of 0 to
2.00 mg/1 by diluting appropriate volumes to 50.0 ml. The following series may be used:
Millimeters of Standard Concentration when Diluted
(1.0 ml = 0.01 mg/F) to 50 ml, mg F/liter
0.00 0.00
1.00 0.20
2.00 0.40
3.00 0.60
4.00 0.80
5.00 1.00
6.00 1.20
8.00 1.60
10.00 2.00
7.2 Calibration of Electrometer: Proceed as described in (8.1). Using semilogarithmic graph
paper, plot the concentration of fluoride in mg/liter on the log axis vs. the electrode
potential developed in the standard on the linear axis, starting with the lowest
concentration at the bottom of the scale. Calibration of a selective ion meter: Follow the
directions of the manufacturer for the operation of the instrument.
8. Procedure
8.1 Place 50.0 ml of sample or standard solution and 50.0 ml of buffer (See Note) in a 150 ml
beaker. Place on a magnetic stirrer and mix at medium speed. Immerse the electrodes in
the solution and observe the meter reading while mixing. The electrodes must remain in
the solution for at least three minutes or until the reading has stabilized. At
concentrations under 0.5 mg/liter F, it may require as long as five minutes to reach a
stable meter reading; high concentrations stabilize more quickly. If a pH meter is used,
record the potential measurement for each unknown sample and convert the potential
311
-------
reading to the fluoride ion concentration of the unknown using the standard curve. If a
selective ion meter is used, read the fluoride level in the unknown sample directly in
mg/1 on the fluoride scale.
NOTE: For industrial waste samples, this amount of buffer may not be adequate.
Analyst should check pH first. If highly basic (> 9), add 1 N HC1 to adjust pH to 8.3.
9. Precision and Accuracy
9.1 A synthetic sample prepared by the Analytical Reference Service, PHS, containing 0.85
mg/1 fluoride and no interferences was analyzed by 111 analysts; a mean of 0.84 mg/1
with a standard deviation of ±0.03 was obtained.
9.2 On the same study, a synthetic sample containing 0.75 mg/1 fluoride, 2.5 mg/1
polyphosphate and 300 mg/1 alkalinity, was analyzed by the same 111 analysts; a mean
of 0.75 mg/1 fluoride with a standard deviation of ±0.036 was obtained.
Bibliography
1. Patent No. 3,431,182 (March 4, 1969).
2. CDTA is the abbreviated designation of 1,2-cyclohexylene dinitrilo tetraacetic acid. (The
monohydrate form may also be used.) Eastman Kodak 15411, Mallinckrodt 2357, Sigma D
1383, Tridom-Fluka 32869-32870 or equivalent.
3. Standard Methods for the Examination of Water and Wastewaters, p 389, Method No. 414A,
Preliminary Distillation Step (Bellack), and p 391, Method No. 414B, Electrode Method, 14th
Edition (1975).
4. Annual Book of ASTM Standards, Part 31, "Water", Standard Dl 179-72, Method B, p 312
(1976).
312
-------
EPA METHOD 351.2
NITROGEN, KJELDAHL, TOTAL
COLORIMETRIC, SEMI-AUTOMATED BLOCK DIGESTER, AAII
313
-------
NITROGEN, KJELDAHL, TOTAL
Method 351.2 (Colorimetric, Semi-Automated Block Digester, AAII)
STORET NO. 00625
1. Scope and Application
1.1 This method covers the determination of total Kjeldahl nitrogen in drinking and surface
waters, domestic and industrial wastes. The procedure converts nitrogen components of
biological origin such as amino acids, proteins and peptides to ammonia, but may not
convert the nitrogeneous compounds of some industrial wastes such as amines, nitro
compounds, hydrazones. oximes, semicarbazones and some refractory tertiary amines.
The applicable range of this method is 0.1 to 20 mg/1 TKN. The range may be extended
with sample dilution.
2. Summary of Method
2.1 The sample is heated in the presence of sulfuric acid, K2SO4 and HgSO4 for two and one
half hours. The residue is cooled, diluted to 25 ml and placed on the AutoAnalyzer for
ammonia determination. This digested sample may also be used for phosphorus
determination.
3. Definitions
3.1 Total Kjeldahl nitrogen is defined as the sum of free-ammonia and organic nitrogen
compounds which are converted to ammonium sulfate (NH4)2SO4, under the conditions
of digestion described below.
3.2 Organic Kjeldahl nitrogen is defined as the difference obtained by subtracting the free-
ammonia value (Method 350.2, Nitrogen, Ammonia, this manual) from the total
Kjeldahl nitrogen value.
4. Sample Handling and Preservation
4.1 Samples may be preserved by addition of 2 ml of cone H2SO4 per liter and stored at 4"C.
Even when preserved in this manner, conversion of organic nitrogen to ammonia may
occur. Therefore, samples should be analyzed as soon as possible.
5. Apparatus
5.1 Block Digestor-40
5.2 Technicon Manifold for Ammonia (Figure 1)
5.3 Chemware TFE (Teflon boiling stones), Markson Science, Inc., Box 767, Delmar, CA
92014)
6. Reagents
6.1 Mercuric Sulfate: Dissolve 8 g red mercuric oxide (HgO) in 50 ml of 1:4 sulfuric acid (10
ml cone H2SO4:40 ml distilled water) and dilute to 100 ml with distilled water.
6.2 Digestion Solution: (Sulfuric acid-mercuric sulfate-potassium sulfate solution): Dissolve
133 g of K2SO4 in 700 ml of distilled water and 200 ml of cone H:SO4. Add 25 ml of
mercuric sulfate solution and dilute to 1 liter.
Pending approval for NPDES
Issued 1978
314
-------
6.3 Sulfuric Acid Solution (4%): Add 40 ml of cone, sulfuric acid to 800 ml of ammonia free
distilled water, cool and dilute to 1 liter.
6.4 Stock Sodium Hydroxide (20%): Dissolve 200 g of sodium hydroxide in 900 ml of
ammonia-free distilled water and dilute to 1 liter.
6.5 Stock Sodium Potassium Tartrate Solution (20%): Dissolve 200 g sodium potassium
tartrate in about 800 ml of ammonia-free distilled water and dilute to 1 liter.
6.6 Stock Buffer Solution: Dissolve 134.0 g of sodium phosphate, dibasic (Na2HPO4) in
about 800 ml of ammonia free water. Add 20 g of sodium hydroxide and dilute to 1 liter.
6.7 Working Buffer Solution: Combine the reagents in the stated order; add 250 ml of stock
sodium potassium tartrate solution (6.5) to 200 ml of stock buffer solution (6.6) and mix.
Add xx ml sodium hydroxide solution (6.4) and dilute to 1 liter. See concentration
ranges, Table I, for composition of working buffer.
•6.8 Sodium Salicylate/Sodium Nitroprusside Solution: Dissolve 150 g of sodium salicylate
and 0.3 g of sodium nitroprusside in about 600 ml of ammonia free water and dilute to 1
liter.
6.9 Sodium Hypochlorite Solution: Dilute 6.0 ml sodium hypochlorite solution (clorox) to
100 ml with ammonia free distilled water.
6.10 Ammonium chloride, stock solution: Dissolve 3.819 g NH4C1 in distilled water and bring
to volume in a 1 liter volumetric flask. 1 ml = 1.0 mg NH3-N.
7. Procedure
Digestion
7.1 To 20 or 25 ml of sample, add 5 ml of digestion solution (6.2) and mix (use a vortex
mixer).
7.2 Add (4-8) Teflon boiling stones (5.3). Too many boiling chips will cause the sample to
boil over.
7.3 With Block Digestor in manual mode set low and high temperature at 160°C and preheat
unit to 160°C. Place tubes in digester and switch to automatic mode. Set low temperature
timer for 1 hour. Reset high temperature to 380CC and set timer for 2 1/2 hours.
7.4 Cool sample and dilute to 25 ml with ammonia free water.
Colorimetric Analysis
7.5 Check the level of all reagent containers to ensure an adequate supply.
7.6 Excluding the salicylate line, place all reagent lines in their respective containers, connect
the sample probe to the Sampler IV and start the proportioning pump.
7.7 Flush the Sampler IV wash receptacle with about 25 ml of 4.0% sulfuric acid (6.3).
7.8 When reagents have been pumping for at least five minutes, place the salicylate line in its
respective container and allow the system to equilibrate. If a precipitate forms after the
addition of salicylate, the pH is too low. Immediately stop the proportioning pump and
flush the coils with water using a syringe. Before restarting the system, check the
concentration of the sulfuric acid solutions and/or the working buffer solution.
315
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7.9 To prevent precipitation of sodium salicylate in the waste tray, which can clog the tray
outlet, keep the nitrogen flowcell pump tube and the nitrogen Colorimeter "To Waste"
tube separate from all other lines or keep tap water flowing in the waste tray.
7.10 After a stable baseline has been obtained start the Sampler.
8. Calculations
8.1 Prepare standard curve by plotting peak heights of processed standards against
concentration values. Compute concentrations by comparing sample peak heights with
standard curve.
9. Precision and Accuracy
9.1 In a single laboratory (EMSL), using sewage samples of concentrations of 1.2, 2.6, and
1.7 mgN/1, the precision was ±0.07, ±0.03 and ±0.15, respectively.
9.2 In a single laboratory (EMSL), using sewage samples of concentrations of 4.7 and 8.74
mg N/l, the recoveries were 99 and 99%, respectively.
Bibliography
1. McDaniel, W.H., Hemphill, R.N. and Donaldson, W.T., "Automatic Determination of Total
Kjeldahl Nitrogen in Estuarine Water", Technicon Symposia, pp. 362-367, Vol. 1, 1967.
2. Gales, M.E., and Booth; R.L., "Evaluation of Organic Nitrogen Methods", EPA Office of
Research and Monitoring, June, 1972.
3. Gales, M.E. and Booth, R.L., "Simultaneous and Automated Determination of Total
Phosphorus and Total Kjeldahl Nitrogen", Methods Development and Quality Assurance
Research Laboratory, May, 1974.
4. Technicon "Total Kjeldahl Nitrogen and Total Phosphorus BD-40 Digestion Procedure for
Water", August, 1974.
5. Gales, M.E., and Booth, P...L., "Evaluation of the Block Digestion System for the
Measurement of Total Kjeldahl Nitrogen and Total Phosphorus", EPA-600/4-78-015,
Environmental Monitoring and Support Laboratory, Cinncinnati, Ohio.
317
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EPA METHOD 353.2
NITROGEN, NITRATE-NITRITE
COLORIMETRIC, AUTOMATED, CADMIUM REDUCTION
319
-------
NITROGEN, NITRATE-NITRITE
Method 353.2 (Colorimetric, Automated, Cadmium Reduction;
STORET NO. Totai 00630
1. Scope and Application
1.1 This method pertains to the determination of nitrite singly, or nitrite and nitrate
combined in surface and saline waters, and domestic and industrial wastes. The
applicable range of this method is 0.05 to 10.0 mg/1 nitrate-nitrite nitrogen. The range
may be extended with sample dilution.
2. Summary of Method
2.1 A filtered sample is passed through a column containing granulated copper-cadmium ro
reduce nitrate to nitrite. The nitrite (that originally present plus reduced nitrate) is
determined by diazotizing with sulfanilamide and coupling with N-(l-naphthyl)-
ethylenediamine dihydrochloride to form a highly colored azo dye which is measured
colorimetrically. Separate, rather than combined nitrate-nitrite, values are r^dily
obtained by carrying out the procedure first with, and then without, the Cu-Cd reduction
step.
3. Sample Handling and Preservation
3.1 Analysis should be made as soon as possible. If analysis can be made within 24 hours, the
sample should be preserved by refrigeration at 4°C. When samples must be si^reJ for
more than 24 hours, they should be preserved with sulfuric acid (2 ml cone. H .SO., per
liter) and refrigeration.
Caution: Samples for reduction column must not be preserved with mercuric chloride.
4. Interferences
4.1 Build up of suspended matter in the reduction column will restrict sample flow sine;:
nitrate-nitrogen is found in a soluble state, the sample may be pre-filtered.
4.2 Low results might be obtained for samples that contain high concentrations ot' iron.
copper or other metals. EDTA is added to the samples to eliminate this interference.
4.3 Samples that contain large concentrations of oil and grease will coat the surface of the
cadmium. This interference is eliminated by pre-extracting the sample with an organic
solvent.
5. Apparatus
5.1 Technicon AutoAnalyzer (AAI or AAII) consisting of the following components.
5.1.1 Sampler.
5.1.2 Manifold (AAI) or analytical cartridge (AAII).
5.1.3 Proportioning Pump
5.1.4 Colorimeter equipped with a 15 mm or 50 mm tubular flow cell and
5.1.5 Recorder.
Approved for NPDES and SDWA
Issued 1971
Editorial revision 1974 and 1978
320
-------
5.1.6 Digital printer for AAII (Optional).
6. Reagents
6.1 Granulated cadmium: 40-60 mesh (MCB Reagents).
6.2 Copper-cadmium: The cadmium granules (new or used) are cleaned with-dilute HC1
(6.7) and copperized with 2% solution of copper sulfate (6.8) in the following manner:
6.2.1 Wash the cadmium with HC1 (6.7) and rinse with distilled water. The color of the
cadmium so treated should be silver.
6.2.2 Swirl 10 g cadmium in 100 ml portions of 2% solution of copper sulfate (6.8) for
five minutes or until blue color partially fades, decant and repeat with fresh copper
sulfate until a brown colloidal precipitate forms.
6.2.3 Wash the cadmium-copper with distilled water (at least 10 times) to remove all the
precipitated copper. The color of the cadmium so treated should be black.
6.3 Preparation of reduction column AAI: The reduction column is an 8 by 50 mm glass tube
with the ends reduced in diameter to permit insertion into the system. Copper-cadmium
granules (6.2) are placed in the column between glass wool plugs. The packed reduction
column is placed in an up-flow 20° incline to minimize channeling. See Figure 1.
6.4 Preparation of reduction column AAII: The reduction column is a U-shaped, 35 cm
length, 2 mm I.D. glass tube (Note 1). Fill the reduction column with distilled water to
prevent entrapment of air bubbles during the filling operations. Transfer the copper-
cadmium granules (6.2) to the reduction column and place a glass wool plug in each end.
To prevent entrapment of air bubbles in the reduction column be sure that all pump tubes
are filled with reagents before putting the column into the analytical system.
NOTE 1: A 0.081 I.D. pump tube (purple) can be used in place of the 2 mm glass tube.
6.5 Distilled water: Because of possible contamination, this should be prepared by passage
through an ion exchange column comprised of a mixture of both strongly acidic-cation
and strongly basic-anion exchange resins. The regeneration of the ion exchange column
should be carried out according to the manufacturer's instructions.
6.6 Color reagent: To approximately 800 ml of distilled water, add, while stirring, 100 ml
cone, phosphoric acid, 40 g sulfanilamide, and 2 g N-1-naphthylethylenediamine
dihydrochloride. Stir until dissolved and dilute to 1 liter. Store in brown bottle and keep
in the dark when not in use. This solution is stable for several months.
6.7 Dilute hydrochloric acid, 6N: Dilute 50 ml of cone. HC1 to 100 ml with distilled water.
6.8 Copper sulfate solution, 2%: Dissolve 20 g of CuSO4«5H2O in 500 ml of distilled water
and dilute to 1 liter.
6.9 Wash solution: Use distilled water for unpreserved samples. For samples preserved with
H2SO4, use 2 ml H2SO4 per liter of wash water.
6.10 Ammonium chloride-EDTA solution: Dissolve 85 g of reagent grade ammonium
chloride and 0.1 g of disodium ethylenediamine tetracetate in 900 ml of distilled water.
Adjust the pH to 8.5 with cone, ammonium hydroxide and dilute to 1 liter. Add 1/2 ml
Brij-35 (available from Technicon Corporation).
321
-------
INDENTATIONS FOR
SUPPORTING CATALYST
Cd-TURNINGS
GLASS WOOL
TILT COLUMN TO 20° POSTION
FIGURE 1. COPPER CADMIUM REDUCTION COLUMN
(1 1/2 ACTUAL SIZE)
322
-------
6.11. Stock nitrate solution: Dissolve 7.218 g KNO3 and dilute to 1 liter in a volumetric flask
with distilled water. Preserve with 2 ml of chloroform per liter. Solution is stable for 6
months. 1 ml = 1.0mgNO3-N.
6.12 Stock nitrite solution: Dissolve 6.072 g KNO: in 500 ml of distilled water and dilute to 1
liter in a volumetric flask. Preserve with 2 ml of chloroform and keep under refrigeration.
1.0ml= 1.0mgNO:-N. .
6.13 Standard nitrate solution: Dilute 10.0 ml of stock nitrate solution (6.11) to 1000ml.
1.0 ml = 0.01 mgNO3-N. Preserve with 2 ml of chloroform per liter. Solution is stable
for 6 months.
6.14 Standard nitrite solution: Dilute 10.0 ml of stock nitrite (6.12) solution to 1000 ml.
1.0ml = 0.01 mgNO:-N. Solution is unstable; prepare as required.
6.15 Using standard nitrate solution (6.13), prepare the following standards in 100.0 ml
volumetric flasks. At least one nitrite standard should be compared to a nitrate standard
at the same concentration to verify the efficiency of the reduction column^
Cone., mgNO2-N or NO3-N/1
0.0
0.05
0.10
0.20
0.50
1.00
2.00
4.00
6.00
ml Standard Solution/100 ml
0
0.5
1.0
2.0
5.0
10.0
20.0
40.0.
60.0
NOTE 2: When the samples to be analyzed are saline waters, Substitute Ocean Water
(SOW) should be used for preparing the standards; otherwise, distilled water is used. A
tabulation of SOW composition follows:
NaCl - 24.53 g/l
Cad, - 1.16 g/l
KBr - 0.10 g/l
NaF - 0.003 g/l
MgCl2 - 5.20 g/l
KC1 - 0.70 g/!
H3BO3 - 0.03 g/l
Na,SO4 - 4.09 g/l
NaHCO3 - 0.20 g/l
SrCl, - 0.03 g/l
7. Procedure
7.1 If the pH of the sample is below 5 or above 9, adjust to between 5 and 9 with either cone.
HClorconc. NH4OH.
7.2 Set up the manifold as shown in Figure 2 (AAI) or Figure 3 (AAII). Note that reductant
column should be in 20° incline position (AAI). Care should be taken not to introduce air
into reduction column on the AAII.
7.3 Allow both colorimeter and recorder to warm up for 30 minutes. Obtain a stable baseline
with all reagents, feeding distilled water through the sample line.
NOTE 3: Condition column by running 1 mg/1 standard for 10 minutes if a new
reduction column is being used. Subsequently wash the column with reagents tor 20
minutes.
323
-------
7.4 Place appropriate nitrate and/or nitrite standards in sampler in order of decreasing
concentration of nitrogen. Complete loading of sampler tray with unknown samples.
7.5 For the AAI system, sample at a rate of 30/hr, 1:1. For the AAII, use a 40/hr, 4:1 cam
and a common wash.
7.6 Switch sample line to sampler and start analysis.
8. Calculations
8.1 Prepare appropriate standard curve or curves derived from processing NO, and/or NO3
standards through manifold. Compute concentration of samples by comparing sample
peak heights with standard curve.
9. Precision and Accuracy
9.1 Three laboratories participating in an EPA Method Study, analyzed four natural water
samples containing exact increments of inorganic nitrate, with the following results:
Increment as
Nitrate Nitrogen
mg N/liter
0.29
0.35
2.31
2.48
Precision as
Standard Deviation
mg N/liter
0.012
0.092
0.318
0.176
Accuracy as
Bias,
+ 5.75
+ 18.10
+ 4.47
- 2.69
Bias,
mg N/liter
+ 0.017
+ 0.063
-1-0.103
-0.067
Bibliography
1. Fiore, J., and O'Brien, J. E., "Automation in Sanitary Chemistry - parts 1 & 2 Determination
of Nitrates and Nitrites", Wastes Engineering 33,128 & 238 (1962).
2. Armstrong, F. A., Stearns, C. R., and Strickland, J. D., "The Measurement of Upwelling and
Subsequent Biological Processes by Means of the Technicon AutoAnalyzer and Associated
Equipment", Deep Sea Research 14, p 381-389 (1967).
3. Annual Book of ASTM Standards, Part 31, "Water", Standard D1254, p 366 (1976).
4. Chemical Analyses for Water Quality Manual, Department of the Interior, FWPCA, R. A.
Taft Sanitary Engineering Center Training Program, Cincinnati, Ohio 45226 (January, 1966).
5. Annual Book of ASTM Standards, Part 31, "Water", Standard D 1141-75, Substitute Ocean
Water, p 48 (1976).
324
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EPA METHOD 365.2
PHOSPHOROUS, ALL FORMS
COLORIMETRIC, ASCORBIC ACID, SINGLE REAGENT
327
-------
PHOSPHORUS, ALL FORMS
Method 365.2 (Colorimetric, Ascorbic Acid, Single Reagent)
STORET NO. See Section 4
1. Scope and Application
1.1 These methods cover the determination of specified forms of phosphorus in drinking,
surface and saline waters, domestic and industrial wastes.
1.2 The methods are based on reactions that are specific for the orthophosphate ion. Thus,
depending on the prescribed pre-treatment of the sample, the various forms of
phosphorus given in Figure 1 may be determined. These forms are defined in Section 4.
1.2.1 Except for in-depth and detailed studies, the most commonly measured forms are
phosphorus and dissolved phosphorus, and orthophosphate and dissolved
orthophosphate. Hydrolyzable phosphorus is normally found only in sewage-type
samples and insoluble forms of phosphorus are determined by calculation.
1.3 The methods are usable in the 0.01 to 0.5 mg P/l range.
2. Summary of Method
2.1 Ammonium molybdate and antimony potassium tartrate react in an acid medium with
dilute solutions of phosphorus to form an antimony-phospho-molybdate complex. This
complex is reduced to an intensely blue-colored complex by ascorbic acid. The color is
proportional to the phosphorus concentration.
2.2 Only orthophosphate forms a blue color in this test. Polyphosphates (and some organic
phosphorus compounds) may be converted to the orthophosphate form by suifunc acid
hydrolysis. Organic phosphorus compounds may be converted to the orthophosphate
form by persulfate digestion'21.
3. Sample Handling and Preservation
3.1 If benthic deposits are present in the area being sampled, great care should be taken not
to include these deposits.
3.2 Sample containers may be of plastic material, such as cubitainers, or of Pyrex glass.
3.3 If the analysis cannot be performed the day of collection, the sample should be preserved
by the addition of 2 ml cone. H,SO4 per liter and refrigeration at 4°C.
4. Definitions and Storet Numbers
4.1 Total Phosphorus (P) — all of the phosphorus present in the sample, regardless of form,
as measured by the persulfate digestion procedure. (00665)
4.1.1 Total Orthophosphate (P, ortho) — inorganic phosphorus [(POJ"1] in the sample
as measured by the direct colorimetric analysis procedure. (70507)
4.1.2 Total Hydrolyzable Phosphorus (P, hydro) - phosphorus in the sample as
measured by the suifunc acid hydrolysis procedure, and minus pre-deiormir.ed
onhophosphates. This hydrolyzable phosphorus includes poiyplu'spiioius.
[(P,O7)4, (P3OIU)', etc.] plus some organic phosphorus. (00669)
Approved for NPDES
Issued 1971
328
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4.1.3 Total Organic Phosphorus (P, org) — phosphorus (inorganic plus oxidizable
organic) in the sample measured by the persulfate digestion procedure, and minus
hydrolyzable phosphorus and orthophosphate. (00670)
4.2 Dissolved Phosphorus (P-D) — all of the phosphorus present in the filtrate of a sample
filtered through a phosphorus-free filter of 0.45 micron pore size and measured by the
persulfate digestion procedure. (00666)
4.2.1 Dissolved Orthophosphate (P-D, ortho) — as measured by the direct colorimetric
analysis procedure. (00671)
4.2.2 Dissolved Hydrolyzable Phosphorus (P-D, hydro) — as measured by the sulfuric
acid hydrolysis procedure and minus pre-determined dissolved orthophosphates.
(00672)
4.2.3 Dissolved Organic Phosphorus (P-D, org) — as measured by the persulfate
digestion procedure, and minus dissolved hydrolyzable phosphorus and
orthophosphate. (00673)
4.3 The following forms, when sufficient amounts of phosphorus are present in the sample to
warrant such consideration, may be calculated:
4.3.1 Insoluble Phosphorus (P-I) = (P)-(P-D). (00667)
4.3.1.1 Insoluble orthophosphate (P-I, ortho) = (P, ortho)-(P-D, ortho).
(00674)
4.3.1.2 Insoluble Hydrolyzable Phosphorus (P-I, hydro) = (P, hydro)-(P-D,
hydro). (00675) \
4.3.1.3 Insoluble Organic Phosphorus (P-I, org) = (P, org) - (P-D, org).
(00676)
4.4 All phosphorus forms shall be reported as P, mg/1, to the third place.
5. Interferences
5.1 No interference is caused by copper, iron, or silicate at concentrations many times
greater than their reported concentration in sea water. However, high iron
concentrations can cause precipitation of and subsequent loss of phosphorus.
5.2 The salt error for samples ranging from 5 to 20% salt content was found to be less than
1%.
5.3 Arsenate is determined similarly to phosphorus and should be considered when present
in concentrations higher than phosphorus. However, at concentrations found in sea
water, it does not interfere.
6. Apparatus
6.1 Photometer - A spectrophotometer or filter photometer suitable for measurements at
650 or 880 nm with a light path of 1 cm or longer.
6.2 Acid-washed glassware: All glassware used should be washed with hot 1:1 HC1 and
rinsed with distilled water. The acid-washed glassware should be filled with distilled
water and treated with all the reagents to remove the last traces of phosphorus that might
be adsorbed on the glassware. Preferably, this glassware should be used only for the
determination of phosphorus and after use it should be rinsed with distilted water and
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kept covered until needed again. If this is done, the treatment with 1:1 HC1 and reagents
is only required occasionally. Commercial detergents should never be used.
7. Reagents
7.1 Sulfuric acid solution, 5N: Dilute 70 ml of cone. H2SO4 with distilled water to 500 ml.
7.2 Antimony potassium tartrate solution: Weigh 1.3715 g K(SbO)C4H4O6«l/2HA
dissolve in 400 ml distilled water in 500 ml volumetric flask, dilute to volume. Store at
4°C in a dark, glass-stoppered bottle.
7.3 Ammonium molybdate solution: Dissolve 20 g(NH4)6Mo7O24«4H2O in 500 ml of distilled
water. Store in a plastic bottle at 4°C.
7.4 Ascorbic acid, 0. LM: Dissolve 1.76 g of ascorbic acid in 100 ml of distilled water. The
solution is stable for about a week if stored at 4°C.
7.5 Combined reagent: Mix the above reagents in the following proportions for 100 ml of the
mixed reagent: 50 ml of 5N H2SO4, (7.1), 5 ml of antimony potassium tartrate solution
(7.2), 15 ml of ammonium molybdate solution (7.3), and 30 ml of ascorbic acid solution
(7.4). Mix after addition of each reagent. All reagents must reach room temperature
before they are mixed and must be mixed in the order given. If turbidity forms in the
combined reagent, shake and let stand for a few minutes until the turbidity disappears
before proceeding. Since the stability of this solution is limited, it must be freshly
prepared for each run.
7.6 Sulfuric acid solution, 11 N: Slowly add 310 ml cone. H2SO4 to 600 ml distilled water.
When cool, dilute to 1 liter.
7.7 Ammonium persulfate.
7.8 Stock phosphorus solution: Dissolve in distilled water 0.2197 g of potassium dihydrogen
phosphate, KH2PO4, which has been dried in an oven at 105°C. Dilute the solution to
1000ml; 1.0ml = 0.05 mg P.
7.9 Standard phosphorus solution: Dilute 10.0 ml of stock phosphorus solution (7.8) to 1000
ml with distilled water; 1.0 ml = 0.5 ug P.
7.9.1 Using standard solution, prepare the following standards in 50.0 ml volumetric
flasks:
ml of Standard
Phosphorus Solution (7.9) Cone., mg/1
0 0.00
1.0 0.01
3.0 0.03
5.0 0.05
10.0 0 10
20.0 0.20
30.0 0.30
40 0 0.40
50 0 0 50
7.10 Sodium hydroxide, 1 N: Dissolve 40 g NaOH in 600 ml distilled water. Cool and dilute
to 1 liter.
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8. Procedure
8.1 Phosphorus
8.1.1 Add 1 ml of H2SOj solution (7.6) to a 50 ml sample in a 125 ml Erlenmeyer flask.
8.1.2 Add 0.4 g of ammonium persulfate.
8.1.3 Boil gently on a pre-heated hot plate for approximately 30-40 minutes or until a
final volume of about 10 ml is reached. Do not allow sample to go to dryness.
Alternatively, heat for 30 minutes in an autoclave at 121°C (15-20 psi).
8.1.4 Cool and dilute the sample to about 30 ml and adjust the pH of the sample to 7.0
±0.2 with 1 N NaOH (7.10) using a pH meter. If sample is not clear at this point,
add 2-3 drops of acid (7.6) and filter. Dilute to 50 ml.
Alternatively, if autoclaved see NOTE 1.
8.1.5 Determine phosphorus as outlined in 8.3.2 Orthophosphate.
8.2 Hydrolyzable Phosphorus
8.2.1 Add 1 ml of H:SO4 solution (7.6) to a 50 ml sample in a 125 ml Erlenmeyer flask.
8.2.2 Boil gently on a pre-heated hot plate for 30-40 minutes or until a final volume of
about 10 ml is reached. Do not allow sample to go to dryness. Alternatively, heat
for 30 minutes in an autoclave at 121 °C (15-20 psi).
8.2.3 Cool and dilute the sample to about 30 ml and adjust the pH of the sample to 7.0
±0.2 with NaOH (7.10) using a pH meter. If sample is not clear at this point, add
2-3 drops of acid (7.6) and filter. Dilute to 50 ml.
Alternatively, if autoclaved see NOTE 1.
8.2.4 The sample is now ready for determination of phosphorus as outlined in 8.3.2
Orthophosphate.
8.3 Orthophosphate
8.3.1 The pH of the sample must be adjusted to 7±0.2 using a pH meter.
8.3.2 Add 8.0 ml of combined reagent (7.5) to sample and mix thoroughly. After a
minimum of ten minutes, but no longer than thirty minutes, measure the color
absorbance of each sample at 650 or 880 nm with a spectrophotometer, using the
reagent blank as the reference solution.
NOTE 1: If the same volume of sodium hydroxide solution is not used to adjust the
pH of the standards and samples, a volume correction has to be employed.
9. Calculation
9.1 Prepare a standard curve by plotting the absorbance values of standards versus the
corresponding phosphorus concentrations.
9.1.1 Process standards and blank exactly as the samples. Run at least a blank and two
standards with each series of samples. If the standards do not agree within ±2% of
the true value, prepare a new calibration curve.
9.2 Obtain concentration value of sample directly from prepared standard curve. Report
results as P, mg/1. SEE NOTE 1.
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10. Precision and Accuracy
10.1 Thirty-three analysts in nineteen laboratories analyzed natural water samples containing
exact increments of organic phosphate, with the following results:
Increment as
Total Phosphorus
mg P/liter
0.110
0.132
0.772
0.882
Precision as
Standard Deviation
mg P/liter
0.033
0.051
0.130
0.128
Accuracy as
Bias,
+ 3.09
+ 11.99
4-2.96
-0.92
Bias
mg P/liter
+0.003
+0.016
+0.023
-0.008
(FWPCA Method Study 2, Nutrient Analyses)
10.2 Twenty-six analysts in sixteen laboratories analyzed natural water samples containing
exact increments of orthophosphate, with the following results:
Increment as
Orthophosphate
mg P/liter
0.029
0.038
0.335
0.383
Precision as
Standard Deviation
mg P/liter
0.010
0.008
0.018
0.023
Accuracy as
Bias,
-4.95
-6.00
-2.75
-1.76
Bias,
mg P/liter
-0.001
-0.002
-0.009
-0.007
(FWPCA Method Study 2, Nutrient Analyses)
Bibliography
1. Murphy, J., and Riley, J., "A modified Single Solution for the Determination of Phosphate in
Natural Waters", Anal. Chim. Acta., 27, 31 (1962).
2. Gales, M., Jr., Julian, E., and Kroner, R., "Method for Quantitative Determination of Total
Phosphorus in Water", Jour. AWWA, 58, No. 10, 1363 (1966).
3. Annual Book of ASTM Standards, Part 31, "Water", Standard D515-72, Method A, p 389
(1976).
4. Standard Methods for the Examination of Water and Wastewater, 14th Edition, p 476 and 481,
(1975).
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