United States
Environmental Protection
Agency
Office Of Water
(WH-585)
September 1990
£EPA
Analytical Methods For
The National Sewage Sludge
Survey
-------
Analytical Methods for the
National Sewage Sludge Survey
Prepared for:
William A. Telliard, Chief
Analytical Methods Staff
USEPA Office of Water Regulations and Standards
401 M Street, S.W.
Washington, DC 20460
Under EPA Contract No. 68-C9-0019
Publication Date: September 1990
Printed on Recycled Paper
-------
INTRODUCTION
This document is a compilation of the analytical methods that the
USEPA Office of Water Regulations and Standards (OWRS) used in the
National Sewage Sludge Survey, as well as methods proposed for
monitoring under 40 CFR Parts 257 and 503, Standards for the Disposal of
Sewage Sludge.
These methods are taken from OWRS Industrial Technology Division
(1TD) methods and from "Methods for Chemical Analysis of Water and
Wastes" (MCAWW), USEPA EMSL, Cincinnati, Ohio 45268, EPA-
600/4-79-020 (Revised March 1983). MCAWW is available from the
National Technical Information Service, Springfield, Virginia 22161,
PB84-128677.
Questions concerning this document should be addressed to:
William A. Telliaixl
USEPA OWRS
Sample Control Center
P. O. Box 1407
Alexandria, VA 22313
703/557-5040
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li
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ANALYTICAL METHODS FOR
THE NATIONAL SEWAGE SLUDGE SURVEY
TABLE OF CONTENTS
CATEGORY METHOD ANALYTE
ORGANIC 1624C Volatiles (VGA)
1625C Semivolatiles (ABN)
1618 Pesticides/Herbicides
1613 Dioxins/Furans
METALS 1620 25 elements
Antimony
Arsenic
Selenium
Thallium
Mercury
42 elements
160.3 Residue
335.2 Cyanide
340.2 Fluoride
351.3 TKN
353.2 Nitrate-Nitrite
365.2 Phosphorous
TECHNIQUE
PAGE
CLASSICALS
GCMS 1
GCMS 33
GC 81
GCMS 121
ICP 165
GFAA
GFAA
GFAA
GFAA
CVAA
ICP/Semiquantitative screen
Gravimetric 209
Spectrophotometric 213
Electrode 225
Potentiometric 235
CdReduction 243
Ascorbic Acid Reduction 253
III
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IV
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EPA METHOD 1624C
VOLATILE COMPOUNDS
BY DILUTION
EPA METHOD
SEMIVOLATILE ORGANIC COMPOUNDS
BY DILUTION
-------
-------
Introduction
Methods 1624 and 1625 were developed by the Industrial
Technology Division (ITD) within the United States
Environmental Protection Agency's (USEPA) Office of Water
Regulations and Standards (OURS) 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. Telliard
USEPA
Office of Water Regulations and Standards
401 A Street SW
Washington, DC 20460
202/382-7131
OR
USEPA OWRS
Sample Control Center
P.O. Box 1407
Alexandria, Virginia
703/557-5040
Publication date: June 1989
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4
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Method 1624 Revision C June 1989
Volatile Organic Compounds by isotope Dilution GCMS
1 SCOPE AND APPLICATION
1.1 This method is designed to meet the survey
requirements of the USEPA ITD. The method
is used to determine the volatile toxic
organic pollutants associated with the
Clean Water Act (as amended 1987); the
Resource Conservation and Recovery Act (as
amended 1986); the Comprehensive Environ-
mental Response, Compensation and
Liability Act (as amended 1986); and other
compounds amenable to purge and trap gas
chromatography-mass spectrometry (GCMS).
1.2 The chemical compounds listed in Tables 1
and 2 may be determined in waters, soils,
and municipal sludges by the method.
1.3 The detection limits of the method' are
usually dependent on the level of
interferences rather than instrumental
limitations. The levels in Table 3 typify
the minimum quantities that can be
detected with no interferences present.
1.4 The GCMS portions of the method are for
use only by analysts experienced with GCMS
VOLATILE ORGANIC COMPOUNDS DETERMINED
Table 1
BY GCMS USING ISOTOPE
Pollutant
DILUTION AND INTERNAL
STANDARD TECHNIQUES
Labeled Compound
Compound
acetone
acrolein
acrylonitrile
benzene
bromod i ch I oromethane
bromoform
bromomethane
carbon tetrachloride
chlorobenzene
ch loroethane
2-chloroethylvinyl ether
chloroform
chloromethane
di bromoch I oromethane
1 , 1 -dich loroethane
1 , 2-di ch I oroethane
1,1 -dich loroethene
trans- 1 , 2-di ch lorethene
1,2-dichloropropane
trans-1,3-dichloropropene
diethyl ether
p-dioxane
ethylbenzene
methylene chloride
methyl ethyl ketone
1,1, 2, 2- tetrach loroethane
tetrach loroethene
toluene
1 , 1 , 1 -trich loroethane
1 , 1 ,2- trich loroethane
trichloroethene
vinyl chloride
Storet
81552
34210
34215
34030
32101
32104
34413
32102
34301
34311
34576
32106
34418
32105
34496
32103
34501
34546
34541
34699
81576
81582
34371
34423
81595
34516
34475
34010
34506
34511
39180
39175
CAS Registry
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
EPA-EGD
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
Analog
d6
d4
"3
d
13c6
13c
13d3
C
d5
d5
13C
13d3
c
dj
d4
d2
«3
d6
d4
d10
d8
ddio
J
13c22
da
13*
C2
13C_
d.
CAS Registry
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
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or under the close supervision of such
qualified persons. Laboratories unfamil-
iar with analysis of environmental samples
by GCMS should run the performance tests
in Reference 1 before beginning.
2 SUMMARY OF METHOD
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. (See Table 2). 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 labeted
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
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
maximum is compared with spectra in the
EPA/NIH Mass Spectral File (Reference 4).
Tentative identification is established
when the spectrum agrees (see Section 12).
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 Table 1- and other
compounds for which standards and labeled
analogs are available, the GCMS system is
Table 2
VOLATILE ORGANIC COMPOUNDS TO BE DETERMINED BY REVERSE SEARCH AND QUANT I TAT ION USING KNOWN RETENTION TIMES,
RESPONSE FACTORS, REFERENCE COMPOUNDS, AND MASS SPECTRA
EGD
No.
Compound
CAS Registry
532 allyl alcohol* 107-18-6
533 carbon disulfide 75-15-0
534 2-chloro-1,3-butadiene
(chloroprene) 126-99-8
535 chloroacetonitrile* 107-14-2
536 3-chloropropene 107-05-1
537 crotonaldehyde* 123-73-9
538 1,2-dibromoethane (EDS) 106-93-4
539 dibromomethane 74-95-3
540 trans-1,4-
dichloro-2-butene 110-57-6
541 1,3-dichloropropane 142-28-9
542 cis-1,3-dichloropropene 10061-01-5
543 ethyl cyanide* 107-12-0
EGD
No. Compound
CAS Registry
544 ethyl methacrylate 97-63-2
545 2-hexanone 591-78-6
546 iodomethane 74-88-4
547 isobutyl alcohol* 78-83-1
548 methacrylonitrile 126-98-7
549 methyl methacrylate 78-83-1
550 4-methyl-2-pentanone 108-10-1
551 1,1,1,2-tetrachloroethane 630-20-6
552 trichlorofluoromethane 75-69-4
553 1,2,3-trichloropropane 96-18-4
554 vinyl acetate 108-05-4
951 m-xylene 108-38-3
952 o- + p-xylene
* determined at a purge temperature of 75 - 85 "C
6
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2.4
3.1
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
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.
The quality of the analysis 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 condi.tions of the
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 may serve 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.
Table 3
GAS CHROMATOGRAPHY OF PURGEABLE ORGANIC COMPOUNDS
EGD
NO.
(1)
245
345
246
346
288
388
216
316
244
344
546
616
716
202
Retention time
Compound
chloromethane-dj
chloromethane
bromomethane- d.
bromomethane
vinyl chloride-d_
vinyl chloride
chloroethane-dg
chloroethane
methylene chlorlde-d-
methylene chloride
iodomethane
acetone-d^
acetone
acrolein-d,
4
Mean
(sec)
147
148
243
246
301
304
378
386
512
517
498
554
565
564
EGD
Ref
181
245
181
246
181
288
181
216
181
244
181
181
616
181
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 -
0.641 -
0.270
1.210
0.423
1.195
0.501
1.023
0.620
1.060
0.813
1.017
0.889
1.019
0.903
Mini-
mum
Level
(3)
(uq/mL)
50
50
50
50
50
10
50
50
10
10
50
50
(5)
Method Detection
Limit (4l
ION
solids
(ug/kg)
207*
148*
190*
789*
566*
3561*
50
high
solids
(ug/kg)
13
11
11
24
280*
322*
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Table 3 (continued)
GAS CHROHATOGRAPHY OF PURGEABLE ORGANIC COMPOUNDS
EGD
No.
(1)
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
Retention time
ComDound
acrolein
acrylonitrile-dj
acrylonitrile
carbon disulfide
trichlorof luoromethane
ethyl cyanide
1,1-dichloroethene-dp
1,1-dichloroethene
3-chloropropene
ally I alcohol
bromochloromethane (I.S.)
1,1-dichloroethane-d,
1 , 1 -dichloroethane
diethyl ether-d1Q
diethyl ether
trans-1,2-dichloroethene-d_
trans- 1 , 2-dich loroethene
methyl ethyl ketone-d.
methyl ethyl ketone
chloroform- C.
chloroform
chloroacetonitrile
1, 2-dich loroethane-d.
1,2-dichloroethane
dibromomethane
methacrylonitrile
isobutyl alcohol
1,1,1-trichloroethane- C2
1,1,1-trichloroethane
p-dioxane-dg
p-dioxane •
carbon tetrachloride- C.
carbon tetrachloride
vinyl acetate
bromodichloromethane- C.
bromodichloromethane
2-chloro-1,3-butadiene
crotonaldehyde
1,2-dichloropropane-d,
1, 2-dich I oropropane
cis-1,3-dichloropropene
trichloroethene- C2
trich loroethene
1 , 3-di ch I oropropane
benzene-d.
benzene
chlorodibromomethane- C-
chlorodibromomethane
1,1,2-trichloroethane- C2
1,1,2-trichloroethane
Mean
(sec)
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
EGD
Ref
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
Relative (2)
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 -
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
Mini-
mum
Level
(3)
(ug/mL)
50
50
50
10
10
10
10
10
50
50
10
10
50
50
10
10
10
10
10
10
50
50
10
10
10
10
10
10
10
10
10
10
10
10
10
10
Method Detection
Limit (4)
low
solids
(ug/kg)
377*
360*
31
16
63
41
241*
21
23
16
--
87
28
29
41
23
15
26
high
solids
(ug/kg)
18
9
5
1
12
3
80*
2
3
4
140*
9
3
5
2
8
2
1
8
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Table 3 (continued)
GAS CHROMATOGRAPHY Of PURGEABLE ORGANIC COMPOUNDS
EGO
No.
(1)
233
333
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
Retention time
CofflDound
trans-1 , 3-dichloropropene-d,
trans-1 ,3-dichloropropene
2-chloroethylvinyl ether
1,2-dibromoethane
2-bromo-1-chloropropane (I.S.
methyl methacrylate
bromoform- C^
bromoform
1,1,1,2-tetrachloroethane
4 -methy I -2-pentanone
1 , 2 , 3 - 1 r i ch I oropropane
1 , 1 ,2,2-tetrachloroethane-d_
1 ,1 ,2,2-tetrachloroethane
2-hexanone
tetrachloroethene- C^
tetrachloroethene
trans-1 ,4-dichloro-2-butene
1,4-dichlorobutane (int std)
ethyl methacrylate
toluene-dg
toluene
chlorobenzene-d^
chlorobenzene
ethylbenzene-d.Q
ethylbenzene
bromof I uorobenzene
m-xylene
o- + p-xylene
Mean
(sec)
1226
1226
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
EGD
Ref
182
233
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
Relative
0.922 - 0
0.993 - 1
0.983 - 1
0.98
1.000 - 1
1.06
1.048 - 1
0.992 - 1
1.08
0.92
0.98
0.969 - 0
0.890 - 1
0.98
0.966 - 0
0.997 - 1
1.00
1.000 - 1
1.03
1.016 - 1
1.001 - 1
1.066 - 1
0.914 - 1
1.144 - 1
0.981 - 1
1.255 - 1
1.51
1.57
(2)
.959
.016
.026
.000
.087
.003
.996
.016
.996
.003
.000
.054
.019
.135
.019
.293
.018
.290
Mini-
mum
Level
(3)
(ua/mL)
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
(ua/kg) (ua/ka)
(6)* (6)*
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
solids).
(5) Specification derived from related compound.
(6) An unknown interference in the particular sludge studied precluded measurement of the Method Detection
Limit (MDL) for this compound.
* 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 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.
-------
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 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 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 NIOSH/MESA
approved toxic gas respirator should be
worn when high concentrations are handled.
5 APPARATUS AND MATERIALS
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.
EXIT 1/4 IN. O.D.
INLET 1'4 IN. O.D.
EXIT 1'4 IN. O.D.
10 MM GLASS FRIT
MEDIUM POROSITY
SAMPLE INLET
2-WAY SYRINGE VALVE
17 CM 20 GAUGE SYRINGE NEEDLE
MM O.D. RUBBER SEPTUM
INLET W4 IN. O.O.
1/t6 IN. O.D.
/~ STAINLESS STEEL
MOLECULAR SIEVE
PURGE GAS FILTER
PURGE GAS
I FLOW CONTROL
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
FIGURE 1 Purging Device for 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.
10
-------
PURGE INLET FITTING
SAMPLE OUTLET FITTING
3" X 6 MM QD. GLASS TUBING
PACKING DETAIL
•- 5 MM GLASS WOOL
1
77 CM SILICA GEL
15 CM TENAX GC
»- 1 CM 3S OV-1
GLASS WOOL
II *- 5 MM (
\_TflAP INLI
CONSTRUCTION DETAIL
COMPRESSION
FITTING NUT
AND FERRULES
14 FT. 70. /FOOT
RESISTANCE WIRE
WRAPPED SOLID
FIGURES Trap Construction and Packings
FIGURE 2 Purging Device for Soils or Waters
5.2.2 Trap--25 to 30 cm x 2.5 mm i.d. minimum,
containing the following:
5.2.4 The purge and trap device may be a
separate unit, or coupled to a GC as shown
in Figures 4 and 5.
5.2.2.1 Methyl silicone packing—one ± 0.2 cm, 3
percent OV-1 on 60/80 mesh Chromosorb W,
or equivalent.
5.2.2.2 Porous polymer--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.
5.2.3 Desorber--shall heat the trap to 175 t 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.
CARRIER GAS
FLOW CONTROL
LIQUID INJECTION PORTS
COLUMN OVEN
CONFIRMATORY COLUMN
TO DETECTOR
ANALYTICAL COLUMN
PURGE G«S
FLOW CONTROL
1'JX MOLECULAR
SIEVE FILTER
NOTE.
ALL LINES BETWEEN TRAP
AND GC SHOULD BE HEATED
TOBO-C
FIGURE 4 Schematic of Purge and Trap
Device-Purge Mode
11
-------
CARRIER GAS
FLOW CON'
—v.
ITROL NX
V*
LIQUID INJECTION PORTS
COLUMN OVEN
r
OPTIONAL 4-PORT COLUMN
SELECTION VALVE
CONFIRMATORY COLUMN
TO DETECTOR
ANALYTICAL COLUMN
PURGE GA!
FLOW COI
1M MOLECULAR
SIEVE FILTER
| PURGING
1 DEVICE
NOTE:
ALL LINES BETWEEN TRAP
AND GO SHOULD BE HEATED
TO 80'C.
FIGURE 5 Schematic of Purge and Trap
Device-Desorb Mode
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
(Section 7), quality assurance (Section
8), and performance tests (Section 11) of
this method.
5.3.1 Column--2.8 ± O.b m x 2 ± 0.5 m 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.
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
5.5.2
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 174
176 95 to 101 percent of m/z 174
177 5 to 9 percent of m/z 176
Data acquisition—mass spectra shall be
collected continuously throughout the
analysis and stored on a mass storage
device.
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).
Data process ing- -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.
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.
5.5.3
5.5.4
12
-------
5.8 Syringe valves--2-nay, with Luer ends
(Teflon or Kel-F).
5.9 Syringe--5 mL, gas-tight, with shut-off
valve.
5.10 Bottles--15 mL, screw-cap with Teflon
liner.
5.11 Balances
5.11.1 Analytical, capable of weighing 0.1 mg.
5.11.2 Top loading, capable of weighing 10 rag.
5.12 Equipment for determining percent moisture
5.12.1 Oven, capable of being temperature
controlled at 110 ± 5 °C.
5.12.2 Dessicator.
5.12.3 Beakers--50 - 100 mL.
6 REAGENTS AND STANDARDS
6.1 Reagent water—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 (Hillipore 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
per the steps below. Observe the safety
precautions given in Section 4.
6.5.1 Place approximately 9.8 ml of methanol in
a 10 mL ground glass stoppered volumetric
flask. Allow the flask to stand unstop-
pared for approximately 10 minutes or un-
til all methanol wetted surfaces have
dried.
In each case, weigh the flask, immediately
add the compoundc 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.
6.5.1.2 Gases (chloromethane, bromomethane,
chloroethane, vinyl chloride)—fil I 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.
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
13
-------
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 soIution--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, p-dioxane, 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.
6.7 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.
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
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 tig/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. Examine the spectrum to
determine that only a single compound is
present. Fragments not attributable to
the compound under study indicate the
presence of an interfering compound.
Adjust the analytical conditions and scan
14
-------
rate (for this test only) to produce an
undistorted spectrum at the GC peak
maximum. An undistorted spectrum Mill
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.
7.2.2 The authentic reference spectrum is
obtained under BFB tuning conditions
(Section 7.1 and Table 4) to normalize it
to spectra from other instruments.
7.2.3 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.
7.2.4 For the compounds in Table 2 and 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 (Table 3); the response
factor and the quantisation m/z (Table 5);
and spectrum (Appendix A) to the reverse
search library. Edit the spectrum per
Section 7.2.3, if necessary.
7.3 Assemble the purge and trap device. Pack
the trap as shown in Figure 3 and
condition overnight at 170 - 180 °C by
backflushing 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.
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
methanolic 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 GCHS system. The
test is repeated only if the purge and
Table 5
VOLATILE ORGANIC COMPOUND CHARACTERISTIC M/Z'S
Compound
acetone
acrolein
acrylonitrile
allyl alcohol
benzene
2-bromo-1-chloropropane (4)
bromochloromethane (4)
bromodi ch loromethane
bromoform
bromomethane
carbon disulfide
carbon tetrachloride
2-chloro-1,3-butadiene
chloroacetonitrile
chlorobenzene
chloroethane
2-chloroethylvinyl ether
Labeled
Analog
d6
d4
d6
C
C
d.
13C
ds
d5
d7
Primary
m/z (1)
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
Reference
compound
(2)
181
181
182
181
Response factor at
purge temp, ofs
20 °C 80 °C
(3) 0.20
1.93 2.02
0.29 0.50
(3) 1.12
-------
Table 5 (continued)
VOLATILE ORGANIC COMPOUND CHARACTERISTIC M/Z'S
Compound
chloroform
chloromethane
3-chloropropsne
crotonaldehyde
dibromochloromethane
1,2-dibromoethane
dibromomethane
1,4-dichlorobutane (4)
trans-1,4-dichloro-2-butene
1,1-dich I oroethane
1,2-dichloroethane
1,1-dichloroethene
trans-1,2-dichlorethene
1 , 2-di ch I oropropane
1 , 3 - d i ch I oropropane
cis-1,3-dichloropropene
trans-1,3-dichloropropene
diethyl ether
p-dioxane
ethyl cyanide
ethyl methacrylate
ethylbenzene
2-hexanone
iodome thane
isobutyl alcohol
methylene chloride
methyl ethyl ketone
methyl methacrylate
4-methyl-2-pentanone
methacryloni tri le
1,1,1 , 2- tetrach loroethane
1 , 1 , 2 , 2 - tet rach I oroethane
tetrach I oroethene
toluene
1,1, 1-triehl oroethane
1 , 1,2- tri ch I oroethane
t rich I oroethene
tpichlorof luoromethane
1 ,2, 3-trichloropropane
vinyl acetate
vinyl chloride
m-xylene
o- + p-xylene
Labeled
Analog
13C
•fs
•t-9
13C
*s
?
d2
d2
d6
d4
d10
da
d10
d2
4
13d2
C2
d8
13*3
M
C2
"3
Primary
m/z (1)
85/86
50/53
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
72/80
69
58
67
131
83/84
164/172
92/100
97/102
83/84
95/136
101
75
86
62/65
106
106
Reference
compound
(2)
181
182
182
181
183
182
182
181
183
183
181
181
182
183
181
182
181
183
182
183
183
Response factor at
purge temp, of
20 °C 80 °C
0.43
<3>
0.86
1.35
0.093
0.89
0.29
(3)
0.69
0.076
4.55
(3)
0.23
0.15
0.25
0.20
2.31
0.89
0.054
1.69
3.33
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
0.79
0.25
2.19
0.72
0.19
-
-
(1) native/labeled
(2) 181 = bromoehloromethane 182 = 2-bromo-1-chloropropane 183 = 1,4-dichlorobutane
(3) not detected at a purge temperature of 20 °C
(4) internal standard
NOTE: Because the composition and purity of commercially-supplied isotopically labeled standards may vary, the
primary m/z of the labeled analogs given in this table should be used as guidance. The appropriate m/z of the
labeled analogs should be determined prior to use for sample analysis. Deviations from the m/z's listed here
must be documented by the laboratory and submitted with the data.
16
-------
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-dg) produces an area at m/z 91 (or
99) approximately 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.
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
calibration curve for toluene using
toluene-dg is given in Figure 6.
10-
tn.
1
1.0-
5
UJ
oc
0.1-
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
Also shown are the ± 10 percent error
limits (dotted lines). Relative response
is determined according to the procedures
described below. A minimum of five data
points are required for calibration
(Section 7.4.4).
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 = the isotope ratio measured in the
pure pollutant (Figure 7A).
R = the isotope ratio of pure labeled
compound (Figure 7B).
R = the isotope ratio measured in the an-
alytical mixture of the pollutant and la-
beled compounds (Figure 7C).
(A)
AREA=168920
• M/Z 100
• M/Z 92
(B)
AREA=60960
• M/Z 100
• M/Z 92
(C)
M/Z 92 . 96666
M/Z 100" 82508
• M/Z 100
• M/Z 92
FIGURE? Extracted Ion Current Profiles for (A)
Toluene, (B) Toluene-ds, and (C) a Mixture of
Toluene and Toluene-da
The correct way to calculate RR is:
RR *
If Rffl is not between 2R and 0.5Rx/ the
method does not apply and the sample is
17
-------
analyzed by the internal standard method
(Section 7.5).
1
60960
0.00001640
7.4.2
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:
R = (area at m/z)
R = 96868
m 5i508
1.174
(area at m./z)
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 n^/z = 0, then
R = 50721 = 50720
1
The data from these analyses are reported
to three significant figures (see Section
13.6). In order to prevent rounding
errors from affecting the values to be
reported, all calculations performed prior
to the final determination of
concentrations should be carried out using
at least four significant figures.
Therefore, the calculation of R above is
rounded to four significant figures.
The m/z's are always selected such that R
> V
tent 10
[ion times (RT) between the pollutant
and labeled compounds, special precautions
are required to determine the isotope ra-
tios.
RX, R , and R are defined as follows:
Rv = [area m,/z (at RT.)]
x 1 1
Ry =
[area m-/z (at RT-)]
[area m-/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-dg. For these
data:
R = 168920 = 168900
The RR for the above data is then calcu-
lated using the equation given in Section
7.4.1. For the example, rounded to four
significant figures, RR = 1.174. Not all
labeled compounds elute before their
pollutant analogs.
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 bromochloromethane, 2-bromo-
1-chloropropane, and 1,4-dichlorobutane.
Concentrations of the labeled compounds
and pollutants without labeled analogs are
computed relative to the nearest eluting
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 =
iA x C. 2,
where
18
-------
A is the EICP area at the characteristic
m/z for the compound in the daily stan-
dard.
A. is the EICP area at the characteristic
i s
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
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
isotopically labeled compounds and
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 ± 2 °C (Section 10).
For these samples, the analytical system
may be calibrated using a purge
temperature of 40 t 2 °C in order to more
closely approximate the behavior of the
compounds of interest in high solids
samples.
8 QUALITY ASSURANCE/QUALITY CONTROL
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
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 14.2).
8.1.5 The laboratory shall, on an ongoing 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.
19
-------
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.
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
from P • 2s to P + 2s . For example, if
P = 90% and s = ^0%, the accuracy
interval is expressed as 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 wi11 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.
20
-------
Table 6
ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS
Acceptance criteria at 20 ug/L or as noted
EGD
No.
(1)
516
002
003
004
048
047
046
006
007
016
019
023
045
051
013
010
029
030
032
033
515
527
038
044
514
015
085
086
011
014
087
088
Labeled and native
compound initial
precision and accuracy
(Sect. 8.2.3)
Compound
acetone*
acrolein*
acrylonitrile*
benzene
bromodi eh 1 oromethane
bromoform
bromomethane
carbon tetrachlon'de
chlorobenzene
chloroethane
2-ehloroethylvinyl ether
chloroform
ch 1 oromethane
dibromochloromethane ,
1,1-dichloroethane
1 ,2-dichloroethane
1,1-diehloroethene
trans- 1 , 2-dich I oroethene
1 , 2-di ch I oropropane
t rans- 1 , 3 -di ch I oropropene
diethyl ether*
p-dioxane*
ethylbenzene
methylene chloride
methyl ethyl ketone*
1 , 1 ,2,2-tetrachloroethane
tetrachloroethene
toluene
1 , 1 , 1-trichloroethane
1 , 1 ,2-tri chloroethane
trichloroethene
vinyl chloride
s (ug/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
15.0
44.0
7.2
9.6
9.7
57.0
9.6
6.6
6.3
5.9
7.1
8.9
28.0
X (ug/L)
77 -
32 -
70 -
13 -
7 -
7 -
d -
16 -
14 -
d -
d -
12 -
d -
11 -
11 -
12 -
d -
11 -
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
40
146
27
29
50
159
30
29
29
33
30
30
59
Labeled
compound
recovery
(Sect. 8.3
and 14.2)
P (%)
35 -
37 -
ns -
ns -
ns -
ns -
ns -
42 -
ns -
ns -
ns -
18 -
ns -
16 -
23 -
12 -
ns -
15 -
ns -
ns -
44 -
ns -
ns -
ns -
36 -
5 -
31 -
4 -
12 -
21 -
35 -
ns -
165
163
204
196
199
214
414
165
205
308
554
172
410
185
191
192
315
195
343
284
156
239
203
316
164
199
181
193
200
184
196
452
Labeled
and native
compound
on- go ing
accuracy
(Sect. 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
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
- 44
145
- 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.
(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.
21
-------
When a sample contains this concentration 9.2
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
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 9.3
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.
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.
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.
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.
9.1
SAMPLE COLLECTION,
HANDLING
PRESERVATION, AND
Grab samples are collected in glass
containers having a total volume greater
than 20 rnL. 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.
9.4
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.
All samples shall be analyzed within 14
days of collection.
10 PURGE, TRAP, AND GCMS 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 utilizing one of
two methods, depending on the levels of
pollutants in the sample. Samples
containing one percent solids or greater,
and low to moderate levels of pollutants
are analyzed by purging a known weight of
sample added to 5 mL of reagent water
(Section 10.5). Samples containing one
percent solids or greater, and high levels
of pollutants are extracted with methanol,
and an aliquot of the methanol extract is
added to reagent water and purged (Section
10.6).
-------
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 a 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 ± 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 ± 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 10.7.
10.5 Samples containing one percent solids or
greater, and low to moderate levels of
pollutants.
10.5.1 Mix the sample thoroughly using a clean
spatula.
10.5.2 Weigh 5 t 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, and purge the sample per Section
10.7.
10.6 Samples containing one percent solids or
greater, and high levels of pollutants, or
samples requiring dilution by a factor of
more than 100 (see Section 13.4).
10.6.1 Mix the sample thoroughly using a clean
spatula.
10.6.2 Weigh 5 ± 1 grams of sample into a
calibrated 15 - 25 mL centrifuge tube.
Record the weight of the sample to three
significant figures.
10.6.3 Add 10.0 ml of methanol to the centrifuge
tube. Cap the tube and shake it
vigorously for 15 - 20 seconds to disperse
the sample in the methanol. Allow the
sample to settle in the tube. If
necessary, centrifuge the sample to settle
suspended particles.
10.6.4 Remove approximately 0.1 percent of the
volume of the supernatant methanol using a
15 - 25 uL syringe. This volume will be
in the range of 10 - 15 uL.
10.6.5 Add this volume of the methanol extract to
5 mL reagent water in a 5 ml syringe, and
analyze per Section 10.4.1.
10.6.6 For further dilutions, dilute 1 ml of the
supernatant methanol (10.6.4) to 10 ml,
100 mL, 1000 mL, etc., in reagent water.
Remove a volume of this methanol
extract/reagent water mixture equivalent
to the volume in Step 10.6.4, add it to 5
23
-------
mL reagent water in a 5 ml syringe, and
analyze per Section 10.4.1.
10.7 Purge the sample for 11.0 t 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 i 5
°C is used.
10.8 After the 11 minute purge time, attach the
trap to the chromatosraph 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
back-flushing with carrier gas at 20 - 60
mL/rnin 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
or subambient temperatures to sharpen
these peaks.
10.9 After desorbing the sample for four
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.
10.10 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
greater than 150 °C.
11 SYSTEM PERFORMANCE
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.
11.4 CC resolution--the valley height between
toluene and toluene-d- (at m/z 91 and 99
plotted on the same graph) shall be less
than 10 percent of the taller of the two
peaks.
11.5 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 which has no labeled analog
by the internal standard method (Section
7.5). Compute the concentrations of the
labeled compounds themselves by the
internal standard method. These
concentrations are computed based on the
calibration data determined in Section 7.
11.5.1 For each pollutant and labeled compound,
compare the concentration with the
corresponding limit for ongoing accuracy
in Table 6. 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.
24
-------
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:
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 s 95% and sr = 5%, the
accuracy is 85 - 105 percent.
11.5.1.1
11.5.1.2
11.5.2
<
.s
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
system (Section 7), and repeat the entire
test (Section 11.1) for all compounds,
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).
120,000
100,000
80,000
TOLUENE-D,
e
9
1
i i i i
9
e •
9
0
-+3s
— 3s
8 9 10
ANALYSIS NUMBER
i?!
fl3,
LU 5
53
1.1U -
non -
TOLUENE
: 9 « ~» , . • "
B 9 9
- 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
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 Labeled compounds and pollutants having no
labeled analog (Tables 1 and 2):
12.1.1 The signals for all characteristic m/z's
stored in the spectral library (Section
7.2.3) shad 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 In order for the compounds for which the
system has been calibrated (Table 1) to be
identified, their relative retention times
shall be within the retention time windows
specified in Table 3.
12.1.4 The system has not been calibrated for the
compounds Listed in Table 2, however, the
relative retention times and mass spectra
of these compounds are known. Therefore,
for a compound in Table 2 to be
identified, its relative retention time
must fall within a retention time window
of ± 60 seconds or t 20 scans (whichever
is greater) of the nominal retention time
of the compound specified in Table 3.
12.2 Pollutants having a labeled analog (Table
1):
25
-------
12.2.1 The signals for all characteristic rn/z's
stored in the spectral library (Section
7.2.3) 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.
12.2.3 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 The m/z's present in the sample mass
spectrum that are not present in the
reference mass spectrum shall be accounted
for by contaminant or background ions. If
the sample 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 -- Because the pollutant
and its labeled analog exhibit the same
effects upon purging, desorption, and gas
chromatography, correction for recovery of
the pollutant can be made by adding a
known amount of a labeled compound to
every sample prior to purging. Relative
response (RR) values for sample mixtures
are used in conjunction with the
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—for the compounds for
which the system was calibrated (Table 1)
according to Section 7.5, use the response
factor determined during the calibration
to calculate the concentration from the
following equation.
Concentration = (A& x C. )
(A-s x RF)
where the terms are as defined in Section
7.5.1. For the compounds for which the
system was not calibrated (Table 2), use
the response factors in Table 5 to
calculate the concentration.
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) =
0.005 L x aqueous cone (ug/L)
0.01 x % solids (g)
where "% solids" is from Section 10.1.3.
13.4 Dilution of samples—if the EICP area at
the quantitation 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 For 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 If dilution of high solids samples by
greater than a factor of 100 is required,
then extract the sample with methanol, as
described in Section 10.6.
13.5 Dilution of samples containing high
concentrations of compounds not in Table 1
-- When the EICP area of the quantitat ion
26
-------
m/z of a compound to be identified per
Section 12.3 exceeds the linear range of
the GCMS system, or when any peak in the
mass spectrum is saturated, dilute the
sample per Sections 13.4.1-13.4.3.
13.6 Report results for all pollutants, labeled
compounds, and tentatively identified
compounds found in all standards, blanks,
and samples to three significant figures.
For samples containing less than one
percent solids, the units are ug/L, and
ug/kg for undiluted samples containing one
percent solids or greater.
13.6.1 Results for samples which have been
diluted 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), 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 the GC
column.
14.2 When the recovery of any labeled compound
is outside the range given in Table 6,
dilute 0.5 mi 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. 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.
If the recovery for the aqueous
performance standard is within the range
given in Table 6, then the method does not
apply to the sample being analyzed, and
the result may not be reported for
regulatory compliance purposes.
14.3 When a high level of the pollutant is
present, reverse search computer programs
may misinterpret the spectrum of chromato-
graphically unresolved pollutant and
labeled compound pairs with overlapping
spectra. 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.
15.2 A chromatogram of the 20 ug/L aqueous
performance standards (Sections 6.7.2 and
11.1) is shown in Figure 9.
-------
HASS CHROHATOGRAH DATA: UOAID1945 HI
B9/91/84 23195:80 CALI: UOAID1945 »1
SAMPLE: UO,S,OPR,00020,09,V,NA:NA,HAS
CONOS.: 1624B, 3. 0H, 2MM, 3845,45-24068,158249,20ML/MINS
RANGE: G 1,1200 LABEL: N 9, 4.9 QUAN: A 9, 1.9 J
SCANS
1 TO 1209
100.0-1
BASE: U 20, 3
47
251
222976.
46.514
250.575
1009
34:10
1209 SCAN
41:09 TIME
FIGURE 9 Chromatogram of Aqueous Performance Standard
28
-------
REFERENCES
"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).
Bellar, T. A. and Lichtenberg, J. J.,
"Journal American Water Works Assoc-
iation," 66, 739 (1974).
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).
National Standard Reference Data System,
"Mass Spectral Tape Format", US National
Bureau of Standards (1979 and later
attachments).
"Working with Carcinogens," DHEW, PHS,
NIOSH, Publication 77-206 (1977).
"OSHA Safety and Health Standards, General
Industry," 29 CFR 1910, OSHA 2206, (1976).
"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 Wasteuater 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).
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
Purge and Trap Volatiles Analysis", S-
CUBED Division of Maxwell Laboratories,
Inc., Prepared for W. A. Telliard,
Industrial Technology Division (UH-552),
USEPA, 401 H 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
Uasteuater 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).
29
-------
Appendix A
Mass Spectra in the Form of Mass/Intensity Lists
532 allyl alcohol
m/z int. m/z
42 30 43
56 58 57
533 carbon disulfide
m/z i nt . m/z
44 282 46
534 2-chloro-1,3-butadiene
m/z int. m/z
48 21 49
54 41 61
87 12 88
535 chloroacetonitrile
m/z int. m/z
47 135 48
74 43 75
536 3-chloropropene
m/z int. m/z
35 39 36
49 176 51
76 1000 77
537 crotonaldehyde
m/z int. m/z
35 26 40
50 40 51
69 511 70
538 1,2-dibromoethane (EDB)
m/z i nt . m/z
79 50 80
105 32 106
186 13 188
539 dibromomethane
m/z int. m/z
43 99 44
91 142 92
172 375 173
int.
39
1000
int.
10
m/z
44
58
m/z
64
int.
232
300
int.
14
m/z
45
61
m/z
76
JflSi
12
15
Mi
1000
m/z
53
m/z
77
int.
13
int.
27
m/z
55
m/z
78
int.
59
int.
82
(chloroprene)
IttL.
91
30
452
int.
1000
884
int.
40
64
74
int.
28
20
1000
int.
13
29
27
int.
101
61
14
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
int.
223
54
22
int.
88
39
int.
44
31
324
int.
339
21
43
int.
51
1000
13
int.
30
1000
719
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
int.
246
11
137
int.
294
278
int..
206
29
int.
48
31
int.
15
38
int.
184
64
12
m/z
52
64
m/z
51
m/z
47
73
m/z
44
55
m/z
93
109
m/z
80
95
176
int.
241
16
int.
12
int.
40
22
int.
335
55
int.
54
922
int.
35
875
342
m/z
53
73
m/z
73
m/z
58
75
m/z
49
68
m/z
95
110
m/z
81
160
int.
1000
21
int.
22
int..
35
138
int.
27
24
int.
42
19
int.
175
18
540 trans-1,4-dichloro-2-butene
m/z i nt . m/z
49 166 50
62 286 64
90 93 91
int.
171
91
129
m/z
51
75
124
int.
~289
1000
138
m/z
52
77
126
int.
85
323
86
m/z
53
88
128
int.
878
246
12
m/z
54
89
int.
273
415
30
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
541
m/z
40
61
77
542
m/z
37
77
543
m/z
44
55
544
m/z
42
69
96
545
m/z
42
59
546
m/z
44
142
547
m/z
34
43
59
548
m/z
38
51
65
549
m/z
42
59
98
1,3-dichloropropane
int.
15
18
46
m/z
42
62
78
int.
44
22
310
m/z
47
63
79
int.
19
131
12
m/z
48
65
int.
20
38
m/z
49
75
int.
193
47
m/z
51
76
int.
55
1000
cis-1,3-dichloropropene
int.
262
328
m/z
38
110
M..
269
254
m/z
39
112
iDL,
998
161
m/z
49
int.
596
m/z
51
Mi
189
m/z
75
int.
1000
ethyl cyanide
int.
115
193
m/z
50
int.
34
m/z
51
int.
166
m/z
52
int.
190
m/z
53
int.
127
m/z
54
int.
1000
ethyl methacrylate
int.
127
1000
17
2-hexanone
int.
61
21
i odomethane
int.
57
1000
m/z
43
70
99
int.
48
83
93
m/z
45
71
113
int.
155
25
11
m/z
55
85
114
int.
32
14
119
m/z
58
86
int.
39
169
m/z
68
87
int.
60
21
(methyl butyl ketone)
m/z
43
71
m/i
127
143
int.
1000
36
int.
328
12
m/z
44
85
m/z
128
int.
24
37
int.
17
m/z
55
100
m/z
139
int.
12
56
int.
39
m/z
57
m/z
140
int.
130
int.
34
m/z
58
m/z
141
int.
382
int.
120
isobutyl alcohol
int.
21
1000
25
m/z
35
44
73
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
methacrylonitrile
int.
24
214
55
m/i
39
52
66
int.
21
446
400
m/z
41
53
67
int.
26
19
1000
m/z
42
62
68
int.
100
24
51
m/z
49
63
int.
19
59
m/z
50
64
int.
60
136
methyl methacrylate
Int..
127
124
20
m/z
43
68
99
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
31
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
550
m/z
42
57
100
551
m/z
47
84
121
552
m/z
44
68
105
553
m/z
49
76
99
554
m/z
36
951
m/z
65
951
m/z
51
4-methyl-2-pentanone (methyl isobutyl ketone; MIBK)
int.
69
205
94
m/z
43
58
int.
1000
346
m/z
44
59
int.
54
20
m/z
53
67
int.
11
12
m/z
55
69
int.
15
10
m/z
56
85
int.
13
96
1,1,1 , 2- tetrachloroethane
int.
144
31
236
m/z
49
95
131
int.
163
416
1000
m/z
60
96
133
int.
303
152
955
m/z
61
97
135
int.
330
270
301
m/z
62
98
int.
98
84
m/z
82
117
mt^
45
804
trichlorof luoromethane
int.
95
53
102
m/z
47
82
117
int.
153
40
16
m/z
49
84
119
int.
43
28
14
m/z
51
101
int.
21
1000
m/z
52
102
int.
14
10
m/z
66
103
int.
162
671
1 ,2,3-trichloropropane
int.
285
38
103
vinyl acetate
int.
5
m-xylene
int.
62
o- + p-xylene
int.
88
m/z
51
77
110
m/z
42
m/z
77
m/z
77
int.
87
302
265
int..
103
int.
124
int.
131
m/z
61
83
111
m/z
43
m/z
91
m/z
91
int.
300
23
28
int.
1000
int.
1000
int.
1000
m/z
62
96
112
m/z
44
m/z
105
m/z
105
int.
107
29
164
int.
70
int.
245
int.
229
m/z
63
97
114
m/z
45
m/z
106
m/z
106
int.
98
166
25
int.
8
int.
580
int.
515
m/z
75
98
m/z
86
m/z
m/z
int.
1000
20
int.
57
jnt^
int.
32
-------
Method 1625 Revision C June 1989
Semivolatile Organic Compounds by Isotope Dilution GCMS
1 SCOPE AND APPLICATION
1.1 This method is designed to meet the survey
requirements of the USEPA ITD. The method
is used to determine the semivolatile
toxic organic pollutants associated with
the Clean Water Act (as amended 1987); the
Resource Conservation and Recovery Act (as
amended 1986); the Comprehensive Environ-
mental Response, Compensation and
Liability Act (as amended 1986); and other
compounds amenable to extraction and
analysis by capillary 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 the
method.
1.3 The detection limits of the method are
usually dependent on the level of
interferences rather than instrumental
limitations. The limits in Tables 5 and 6
typify the minimum quantities that can be
detected with no interferences present.
1.4 The GCMS portions of the method are for
use only by analysts experienced with GCMS
or under the close supervision of such
qualified persons. Laboratories unfamil-
iar with analysis of environmental samples
by GCHS should run the performance tests
in Reference 1 before beginning.
Table 1
BASE/NEUTRAL EXTRACTABLE COMPOUNDS DETERMINED BY GCMS USING ISOTOPE DILUTION AND INTERNAL STANDARD TECHNIQUES
Pollutant
Compound
acenaphthene
acenaphthylene
anthracene
benzidine
benzo(a)anthracene
benzo( b) f I uoranthene
benzo(k)f luoranthene
benzo(a)pyrene
benzo(ghi)perylene
biphenyl (Appendix C)
bis(2-chloroethyl) ether
bis(2-chloroethoxy)methane
bis(2-chloroisopropyl) ether
bis(2-ethylhexyl) ph thai ate
4-bromophenyl phenyl ether
butyl benzyl phthalate
n-C10 (Appendix C)
n-d2 (Appendix C)
n-CU (Appendix C)
n-d6 (Appendix C)
n-dS (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)
Storet
34205
34200
34220
39120
34526
34230
34242
34247
34521
81513
34273
34278
34283
39100
34636
34292
77427
77588
77691
77757
77804
77830
77859
77886
77901
78116
78117
CAS Registry
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
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
EPA-EGD
001 B
077 B
078 B
005 B
072 B
074 B
075 B
073 B
079 B
512 B
018 B
043 B
042 B
066 B
041 B
067 B
517 B
506 B
518 B
519 B
520 B
521 B
522 B
523 B
524 B
525 B
526 B
NPDES
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
618 B
620 B
622 B
624 B
625 B
Analog
d10
d8
d10
d8
d12
d12
d12
d\l
d10
d8
d8
d12
d4
d5
d4
d22
d26
"34
d42
d50
d62
Labeled Compound
CAS Registry
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
16416-30-1
15716-08-2
62369-67-9
16416-32-3
93952-07-9
EPA-EGD
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 B
267 B
617 B
606 B
619 B
621 B
623 B
626 B
33
-------
Table 1 (continued)
BASE/NEUTRAL EXTRACTABLE COMPOUNDS DETERMINED BY GCMS USING ISOTOPE DILUTION AND INTERNAL STANDARD TECHNIQUES
Pollutant
Labeled Compound
Compound
carbazole (4c)
2-chloronaphthalene
4-ehlorophenyl phenyl ether
ehrysene
p-cymene (Appendix C)
dibenzo(a,h)anthracene
dihenzofuran (Appendix C & 4c)
dibeniothiophene (Synfuel)
di-n-butyl phthalate
1,2-dichlorobenzene
1 ,3-dichlorobenzene
1 ,4-dichlorobenzene
3,3'-diehlorobenzidine
diethyl phthalate
2,4-dimethylphenol
dimethyl phthalate
2,4-dinitrotoluene
2,6-dini trotoluene
di-n-octyl phthalate
di phenyl ami ne (Appendix C)
diphenyl ether (Appendix C)
1 ,2-diphenylhydrazine
f luoranthene
f luorene
hexach I orobenzene
hexach lorobutadiene
hexach loroethane
hexach lorocyclopentadiene
indeno(1,2,3-cd)pyrene
isophorone
naphthalene
beta-naphthylamine (Appendix C)
nitrobenzene
N-nitrosodimethylamine
N-nitrosodi-n-proplyamine
N-nitrosodiphenylamine
phenanthrene
phenol
alpha-picoline (Synfuel)
pyrene
styrene (Appendix C)
alpha-terpineol (Appendix C)
1,2,3-trichlorobenzene (4c)
1,2,4-trichlorobenzene
Storet
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
CAS Registry
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
EPA-EGD
528 B
020 B
040 B
076 B
513 8
082 8
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 B
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 8
NPDES
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 8
040 B
041 B
042 B
043 B
044 B
010 A
045 B
046 B
Analog
d8
d?,
d5
d12
d14
d14
d8
d8
d4
V
d4
d4
d6
d4
d4
"3
d.
d4
d10
d10
d10
d10
d.
^3C
13c'
13c4
13c4
d8
d8
d7
d5
d6
d14
d6
d10
ds
d7
d10
d5
"3
fs
"3
CAS Registry
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
17029-05-9
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
EPA- EGO
628 B
220 B
240 B
276 B
613 B
282 B
605 B
604 B
268 B
225 B
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 8
254 B
255 B
602 B
256 B
261 0
263 B
262 B
281 B
265 A
603 B
284 B
610 B
609 B
629 B
208 B
34
-------
Table 2
ACID EXTRACTABLE COMPOUNDS DETERMINED BY GCMS USING ISOTOPE DILUTION AND INTERNAL STANDARD TECHNIQUES
Pollutant
Labeled Compound
Compound Storet
4-chloro-3-methylphenol 34452
2-chlorophenol 34586
2,4-dichlorophenol 34601
2,4-dinitrophenol 34616
2-methyl-4,6-dinitrophenol 34657
2-nitrophenol 34591
4-nitrophenol 34646
pentachlorophenol 39032
2, 3, 6-trichlorophenol <4c) 77688
2,4,5-trichlorophenol (4c)
2,4,6-triehlorophenol 34621
CAS Registry
59-50-7
95-57-8
120-83-2
51-28-5
534-52-1
88-75-5
100-02-7
87-86-5
933-75-5
95-95-4
88-06-2
Table
EPA-EGD NPDES Analog CAS Registry
022 A
024 A
031 A
059 A
060 A
057 A
058 A
064 A
530 A
531 A
021 A
3
BASE/NEUTRAL EXTRACTABLE COMPOUNDS TO BE DETERMINED
USING KNOWN RETENTION TIMES,
EGD
No. Compound
555 acetophenone
556 4-aminobiphenyl
557 aniline
558 o-anisidine
559 aramite
560 benzanthrone
561 1,3-benzenediol(resorcinol)
562 benzenethiol
563 2, 3-benzof luorene
564 benzyl alcohol
565 2-bromochlorobenzene
566 3-bromochlorobenzene
567 4-chloro-2-nitroaniline
568 5-chloro-o-toluidine
569 4-chloroaniline
570 3-ch loroni trobenzene
571 o-eresol
572 erotoxyphos
573 2,6-di-tert-butyl-p-benzoquihone
574 2,4-diaminotoluene
575 1,2-dibromo-3-chloropropane
576 2,6-dichloro-4-nitroaniline
577 1,3-dichloro-2-propanol
578 2,3-dichloroaniline
579 2,3-dichloronitro-benzene
580 1,2:3,4-diepoxybutane
581 3,3'-dimethoxybenzidine
582 dimethyl sulfone
583 p-dimethylamino-azobenzene
584 7,12-dimethylbenz-(a)anthracene
585 N.N-dimethylformamide
586 3,6-dimethylphenanthrene
008 A d2 93951-72-5
001 A d^ 93951-73-6
002 A dj 93951-74-7
005 A cij 93951-77-0
004 A d2 93951-76-9
006 A d4 93951-75-1
007 A d, 93951-79-2
1-1 4
009 A °C, 85380-74-1
O
d2 93951-81-6
d2 93951-82-7
011 A dj 93951-80-5
BY REVERSE SEARCH AND QUANT I TAT ION
EPA- EGO
222 A
224 A
231 A
259 A
260 A
257 A
258 A
264 A
630 A
631 A
221 A
RESPONSE FACTORS, REFERENCE COMPOUND, AND MASS SPECTRA
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
99-30-9
96-23-1
608-27-5
3209-22-1
1464-53-5
119-90-4
67-71-0
60-11-7
57-97-6
68-12-2
1576-67-6
EGD
No.
587
588
589
590
591
592
593
594
595
596
597
598
599
900
901
902
903
904
905
906
907
908
909
910
911
912
913
914
915
916
917
918
Compound
1,4-di nitrobenzene
diphenyldisulfide
ethyl methanesulfonate
ethylenethiourea
ethynylestradiol3-methyl ether
hexach loropropene
2- isopropy (.naphthalene
isosafrole
longifolene
malachite green
methapyrilene
methyl methanesulfonate
2-methy I benzoth i oazo I e
3-methylcholanthrene
4,4'-methylene-bis(2-chloroaniline)
4,5-methylene-phenanthrene
1-methylf luorene
2-methylnaphthalene
1 -me thy I phenanth rene
2-(methylthio)-benzothiazole
1 ,5-naphthalenediamine
1 ,4-naphthoquinone
alpha-naphthylamine
5-nitro-o-toluidine
2-nitroani line
3-nitroaniline
4-nitroani line
4-nitrobiphenyl
N-nitrosodi-n-butylamine
N-nitrosodiethylamine
CAS
Registry
100-25-4
882-33-7
62-50-0
96-45-7
72-33-3
1888-71-7
2027-17-0
120-58-1
475-20-7
569-64-2
91-80-5
66-27-3
120-75-2
56-49-5
101-14-4
203-64-5
1730-37-6
91-57-6
832-69-9
615-22-5
2243-62-1
130-15-4
134-32-7
99-55-8
88-74-4
99-09-2
100-01-6
92-93-3
924-16-3
55-18-5
N-nitrosomethyl-ethylamine 10595-95-6
N-nitrosomethyl-phenylamine
614-00-6
-------
Table 3 (continued)
BASE/NEUTRAL EXTRACTABLE COMPOUNDS TO BE DETERMINED
BY REVERSE SEARCH AND QUANT I TAT IOM USING KNOWN
RETENTION TIMES, RESPONSE FACTORS, REFERENCE
COMPOUND, AND MASS SPECTRA
Table 4
ACID EXTRACTABLE COMPOUNDS TO BE DETERMINED BY
REVERSE SEARCH AND QUANT I TATIOM USING KNOWN RETENTION
TIMES, RESPONSE FACTORS, REFERENCE COMPOUND, AND MASS
SPECTRA
EGD
No.
919
920
921
922
923
924
925
926
927
928
929
930
931
932
933
934
935
936
937
938
939
940
941
942
Compound
N-nitrosomorpholine
N-nitrosopiperidine
pentach I orobenzene
pentach I oroethane
pentamethy I benzene
perylene
phenacetin
phenothiazine
1-phenylnaphthalene
2-phenylnaphthalene
pronamide
pyridine
safrole
squalene
1,2,4,5-tetra-chlorobenzene
thianaphthene(2,3-benzothiophene)
thioacetamide
thioxanthone
o-toluidine
1 ,2,3-trimethoxybenzene
2,4,5-trimethylaniline
triphenylene
t ripropy I eneglycol methyl ether
1,3,5-trithiane
CAS
Registry
59-89-2
100-75-4
608-93-5
76-01-7
700-12-9
198-55-0
62-44-2
92-84-2
605-02-7
612-94-2
23950-58-5
110-86-1
94-59-7
7683-64-9
95-94-3
95-15-8
62-55-5
492-22-8
95-53-4
634-36-6
137-17-7
217-59-4
20324-33-8
291-21-4
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
ultrasonically, 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. Each extract is
dried over sodium sulfate, concentrated to
a volume of five mL, cleaned up using gel
permeation chromatography (GPC), if
EGD
No.
943
944
945
946
947
948
Compound
benzoic acid
p-cresol
3,5-dibromo-
4-hydroxybenzonitri le
2,6-dichlorophenol
hexanoic acid
2,3,4,6-tetrachlorophenol
CAS
Registry
65-85-0
106-44-5
1689-84-5
87-65-0
142-62-1
58-90-2
necessary, and concentrated. Extracts are
concentrated to one ml if GPC is not
performed, and to 0.5 ml if GPC is
performed. 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 (see Section 13).
2.3 Quantitative analysis is performed in one
of four ways by GCMS using extracted ion
current profile (EICP) areas: (1) For
36
-------
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
Table 5
GAS CHROMATOGRAPHIC RETENTION TIMES AND DETECTION LIMITS FOR BASE/NEUTRAL EXTRACTABLE COMPOUNDS
EGD
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
Retention time
Coffloound
2,2'-difluorobiphenyl (int std)
pyridine
N-nitrosodimethylamine-d, (5)
N-nitrosodimethylamine (5)
N,N-dimethylformamide
1,2:3,4-diepoxybutane
alpha picoline-d-
alpha picoline
N-nitrosomethylethylamine
methyl methanesulfonate
styrene-dg
styrene
N-nitrosodiethylamine
1,3-dichloro-2-propanol
ethyl methanesulfonate
dimethyl sulfone
benzenethiol
pentach I oroethane
aniline
p-cymene-d..
p-cymene
phenol -ct
phenol
bis(2-chloroethyl) ether-dg
bis(2-chloroethyl) ether
n-CIO-d--
n-C10
,3-dichlorobenzene-d.
,3-dichlorobenzene
,4-dichlorobenzene-d.
,4-dichlorobenzene
,2-dichlorobenzene-d.
,2-dichlorobenzene
thioacetamide
benzyl alcohol
bis(2-chloroisopropyl) ether-d.-
bis(2-chloroisopropyl) ether
o-cresol
N-nitrosodi-n-propylamine-d., (5)
N-nitrosodi-n-propylamine (5)
acetophenone
hexach I oroethane- C
hexachloroethane
o-toluidine
N-nitrosomorpholine
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
EGD
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
Relative (2)
1.000 -
0.325
0.286 -
1.006 -
0.350
0.352
0.326 -
1.006 -
0.338
0.439
0.450 -
1.002 -
0.490
0.506
0.548
0.558
0.574
0.585
0.597
0.624 -
1.008 -
0.584 -
0.995 -
0.584 •
1.007 -
0.585 -
1.022 -
0.605 -
0.998 -
0.601 •
0.997 -
0.632 -
0.995 -
0.660
0.675
0.664 -
1.010 -
0.700
0.689 -
1.008 -
0.703
0.690 -
0.999 -
0.714
0.717
1.000
0.364
1.028
0.393
1.028
0.488
1.009
0.652
1.023
0.613
1.010
0.607
1.016
0.615
1.038
0.636
1.008
0.666
1.009
0.667
1.008
0.691
1.016
0.716
1.023
0.717
1.001
Mini-
mum
Method Detection
Limit (4)
Level low high
(3) solids solids
(uq/mL) (ug/ka) (ug/kg)
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
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
37
-------
Table 5 (continued)
GAS CHROMATOGRAPHIC RETENTION TIMES AND DETECTION LIMITS FOR BASE/NEUTRAL EXTRACTABLE COMPOUNDS
EGD
No.
<1i
575
256
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
Retention time
Compound
1 ,2-dibromo-3-chloropropane
nitrobenzene-dg
nitrobenzene
3- bromoch I orobenzene
2 - bromoch I orobenzene
tripropylene glycol methyl ether
isophorone-dg
isophorone
1,3,5-trithiane
N-nitrosopiperidine
2,4-dimethylphenol-oL
2,4-dimethylphenol
bis(2-chloroethoxy) methane-d, (5)
bis(2-chloroethoxy) methane (5)
1,2,4-trichlorobenzene-d_
1,2,4-trichlorobenzene
o-anisidine
naphthalene-dg
naphthalene
thianapthene
alpha-terpineol-dj
alpha-terpineol
n-C12-d
n-d2
1,2,3-trichlorobenzene-d, (5)
1,2,3-trichlorobenzene (5)
hexach lorobutadi ene- C,
hexachlorobutadiene
N-nitrosomethylphenylamine
hexach I oropropene
4-chloroani line
3-chloronitrobenzene
N-nitrosodi-n-butylamine
pentamethy I benzene
1,3-benzenediol
safrole
2,4,5-trimethylani I ine
2-methylnaphthalene
2-methylbenzothiazole
5-chloro-o-toluidine
1,2,3-trimethoxybenzene
1 . 2 , 4 . 5 - tet rach I orobenzene
13
hexach lorocyclopentadiene- C,
hexach lorocyc 1 open t ad i ene
isosafrole (cis or trans)
isosafrole (cis or trans)
2,3-dichloroaniline
2,4-diaminotoluene
2-chloronaphthalene-dy
2-chloronaphthalene
Mean
(sec)
839
845
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
1160
1187
1185
1200
EGD
Ref
164
164
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
Relative (2)
0.721
0.706 -
1.002 -
0.734
0.757
0.758
0.747 -
0.999 -
0.764
0.770
0.781 -
0.999 -
0.792 -
1.000 -
0.813 -
1.000 -
0.827
0.819 -
1.001 -
0.835
0.829 -
0.998 -
0.730 -
0.986 -
0.852 -
1.000 -
0.856 -
0.999 -
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 -
0.999 -
0.986
1.023
0.997
1.021
1.014 -
0.997 -
0.727
1.007
0.767
1.017
0.803
1.003
0.807
1.013
0.830
1.005
0.836
1.006
0.844
1.008
0.908
1.051
0.868
1.005
0.871
1.002
0.986
1.001
1.024
1.007
Mini-
mum
Level
(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
Method Detection
Limit (4)
lou
solids
(ug/kg)
39
8
26
26
49
62
nd
high
solids
(ug/ka)
28
5
13
23
24
42
nd
860* 3885*
260*
46
nd
80
164*
22
nd
59
38
-------
Table 5 (continued)
GAS CHROMATOGRAPHIC RETENTION TIMES AND DETECTION LIMITS FOR BASE/NEUTRAL EXTRACTABLE COMPOUNDS
EGD
No.
(1)
518
612
712
60S
708
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
Retention time
Compound
n-C14
biphenyl-d1Q
biphenyl
diphenyl ether-d^Q
diphenyl ether
2,3-dichloroni trobenzene
2-nitroaniline
1 , 4-naphthoquinone
longifolene
acenaphthylene-dg
acenaphthylene
2- isopropy (.naphthalene
1,4-di nitrobenzene
2,6-dichloro-4-nitroaniline
dimethyl phthalate-d.
dimethyl phthalate
2,6-di-t-butyl-p-benzoquinone
2,6-dinitrotoluene-dj
2,6-dinitrotoluene
3-nitroaniline
acenaphthene-d^
acenaphthene
dibenzofuran-dg
dibenzofuran
pentach I orobenzene
alpha-naphthylamine
2,4-dinitrotoluene-dj
2,4-dinitrotoluene
beta-naphthylamine-d-
beta-naphthylamine
ethylenethiourea
f luorene-d.g
f luorene
4-chlorophenyl phenyl ether-dg
4-chlorophenyl phenyl ether
diethyl phthalate-d.
diethyl phthalate
2-(methylthio)benzothiazole
4-chloro-2-nitroaniline
5-nitro-o-toluidine
4-nitroaniline
n-C16-d,4
n-C!6
1 ,2-diphenylhydrazine- .8
1,2-diphenylhydrazine (6)
di phenyl ami ne-d..
diphenylamine
N-nitrosodiphenylamine-d-
N-nitrosodiphenylamine (7)
4-bromophenyl phenyl ether-d- (5)
4-bromophenyl phenyl ether (5)
Mean
(sec)
1203
1195
1205
1211
1216
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
EGD
Ref
164
164
612
164
608
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
Relative
1.034
1.016 -
1.001 •
1.036 -
0.997 -
1.044
1.047
1.052
1.053
1.080 -
1.000 -
1.078
1.079
1.083
1.083 -
0.998 -
1.095
1.090 -
1.001 -
1.115
1.107 -
0.999 -
1.134 -
0.998 -
1.152
1.168
1.152 -
1.000 -
1.163 -
0.996 -
1.187
1.185 -
0.999 -
1.194 -
0.990 -
1.197 -
0;996 -
.217
.222
.223
.230
.010 -
.013 -
1.216 -
0.999 -
1.213 -
1.000 -
1.225 -
1.000 -
1.271 -
0.990 -
(2)
1.027
1.006
1.047
1.009
1.095
1.004
1.102
1.005
1.112
1.005
1.125
1.009
1.155
1.007
1.181
1.002
1.189
1.007
1.214
1.008
1.223
1.015
1.229
1.006
1.478
1.020
1.248
1.009
1.249
1.007
1.252
1.002
1.307
1.015
Mini-
mum
Level
(3)
(ug/mL)
10
10
10
10
10
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
Method Detection
Limit (4)
low
solids
(ug/kg)
256
67
44
57
62
55
64
77
65
49
69
73
52
116*
48
58
55
55
high
solids
(ug/kg)
3533
55
12
18
21
47
55
210*
209*
37
61
59
16
644*
27
54
36
17
39
-------
Table 5 (continued)
GAS CHROMATOGRAPHIC RETENTION TIMES AND DETECTION LIMITS FOR BASE/NEUTRAL EXTRACTA8LE COMPOUNDS
EGO
NO.
0)
925
903
209
309
556
929
281
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
Compound
phenacetin
1 -methyl fluorene
hexachlorobenzene- C6
hexach I orobenzene
4-aminobiphenyl
pronamide
phenanthrene-d1Q
n-C18
phenanthrene
anthracene-d10
anthracene
dibenzothiophene-dg
dibenzothiophene
diphenyldisulf ide
4-nitrobiphenyl
1-phenylnaphthalene
carbazole-d. (5)
carbazole (5)
n-C20-d,-
n-C20
1,5-naphthalenediamine
4,5-methylenephenanthrene
1 -methylphenanthrene
di-n-butyl phthalate-d.
di-n-butyl phthalate
2 -pheny I naphthalene
3, 6-dimethy I phenanthrene
methapyri lene
phenothiazine
f luoranthene-d^j.
f luoranthene
crotoxyphos
thioxanthone
pyrene-d1Q
pyrene
benzidine-dg
benzidine
n-C22
aramite
aramite
p-dimethylaminoazobenzene
2,3-benzof luorene
n-C24-d
n-C24
squalene
butylbenzyl phthalate-d, (5)
butylbenzyl phthalate (5)
chrysene-d^-
chrysene
4,4'methylenebis(2-chloroani line)
benzo(a)anthracene-d1-
Mean
(sec)
1512
1514
1521
1522
1551
1578
1578
1580
1583
1583
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
Retention
EGD
Ref
164
164
164
209
164
164
164
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
time
Relative
1.300
1.302
1.283 -
3.999 -
1.334
1.357
1.334 -
1.359
1.000 -
1.342 -
3.998 -
1.314 -
1.000 -
1.396
1.409
1.413
1.388 -
1.000 -
1.184 -
1.010 -
1.441
1.453
1.459
1.446 -
1.000 -
1.490
1.516
1.531
1.544
.522 -
1.000 -
.567
.579
.523 -
.001 -
.549 -
.000 -
.624
.635
.647
.653
.661
.671 -
.012 -
.753
.715 -
.000 -
.743 -
.000 -
.791
.735 -
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
(2)
.327
.001
.380
.005
.388
.006
.361
.006
.439
.006
.662
.021
.510
.003
.596
.004
.644
.003
.632
.002
.764
.015
.824
.002
.837
.004
.846
Mini-
mum
Level
(3)
(ug/mL)
10
10
10
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
Method Detection
Limit (4)
Ion high
sol ids sol ids
(ug/ka) (ug/kg)
51 48
134* 844*
42 22
52 21
72 71
47 24
83 229*
64 80
54 22
40 48
nd nd
432* 447*
..
60 65
51 48
40
-------
Table 5 (continued)
GAS CHROMATOGRAPHIC RETENTION TIMES AND DETECTION LIMITS FOR BASE/NEUTRAL EXTRACTABLE COMPOUNDS
EGD
No.
(1)
372
581
228
328
940
560
266
366
524
591
269
369
525
584
274
374
275
375
924
273
373
626
726
596
900
083
282
382
279
379
(1)
(2)
(3)
Retention time
Mean
Compound
benzo(a)anthracene
3,3'-dimethoxybenzidine
3,3'-dichlorobenzidine-d^
3,3'-dichlorobenzidine
triphenylene
benzan throne
bis(2-ethylhexyl) phthalate-d^
bis(2-ethylhexyl) phthalate
n-C26
ethynylestradiol 3-methyl ether
di-n-octyl phthalate-d.
di-n-octyl phthalate
n-C28
7, 12-dimethy lbenz(a)anthracene
benzo( b) f 1 uoranthene-d. -
benzo(b)f luoranthene
benzo(k)f 1 uoranthene-d. -
benzo( k ) f I uoranthene
perylene
benzo( a )pyrene-d. -
benzo(a)pyrene
n-C30-d62
n-C30
malachite green
3-methylcholanthrene
indeno(1,2,3-cd)pyrene
dibenzo(a,h)anthracene-d., (5)
d ibenzo( a, h) anthracene (5)
benzo(ghi )perylene-d..
benzo(ghi)perylene
Reference numbers beginning with 0,
method; reference numbers beginning
standard method; reference numbers
dilution.
1,
(sec)
2090
2090
2088
2086
2088
2106
2123
2124
2147
2209
2239
2240
2272
2284
2281
2293
2287
2293
2349
2351
2350
2384
2429
2382
2439
2650
2649
2660
2741
2750
5, or 9
EGD
Ref Relative (2)
272 0.999
164
164
228
164
164
164
266
164
164
164
269
164
164
164
274
164
275
164
164
273
164
626
.797
.744
.000
.795
.811
.771
.000
.846
.899
.867
.000
.954
.964
.902
.000
.906
.000
2.020
.954
.000
.972
.011
164 2.048
164 2.097
164 2.279
164 2.107
282 1.000
164 2.187
279 1.001
- 1.
- 1.
- 1.
- 1.
- 1.
- 1.
- 1.
- 2.
- 1.
- 2.
- 1.
- 2.
- 1.
- 2.
- 1.
- 2.
- 1.
- 2.
- 1.
007
848
001
880
002
982
002
025
005
033
005
088
004
127
028
445
007
524
006
indicate a pollutant
with 2 or 6 indicate a labeled
beginning uith 3 or 7 indicate
Single values in this column are based on single
This is a minimum level at which the analytical
corrected) and acceptable calibration
points.
Mini
mum
Method Detection
Limit (4)
Level low
(3)
high
solids solids
(ug/mL) (ug/kg) (ug/kg)
10
50
50
10
10
10
10
10
10
10
10
10
10
10
10
10
10
20
20
20
20
20
quantified
61
62
553*
609*
72
492*
54
95
52
252*
67
49
44
47
111
1310*
886*
62
1810*
30
20
15
658*
263*
125
nd
by the internal standard
compound quantified by the internal
a pollutant quantified
by isotope
laboratory data.
system shall
give recognizable
The concentration
mass spectra
(background
in the aqueous or solid phase
is
determined using the equations in section 14.
(4)
(5)
(6)
(7)
nd =
Method detection limits determined
solids).
in digested
sludge (low solids) and in filter cake or compost (high
Specification derived from related compound.
Detected as azobenzene
Detected as diphenylamine
not detected when spiked into the sludge
* Background levels of these compounds were
MDL's. The HDL for these compounds is
Column: 30 +/- 2 m x 0.25 +/- 0.02 mm i.d
Temperature program: 5 min at 30°C; 30
tested
present
in the sludge
tested
expected to be approximately 50
. 94% methyl,
- 280°C at 8
4% phenyl, 1%
, resulting
ug/kg with no
vinyl bonded phase
°C per min; isothermal at 280 °C
in higher than expected
interferences
fused silica
present .
capillary
until benzo(ghi)perylene
elutes
Gas
velocity: 30 +/- 5 cm/sec at 30°C
41
-------
Table 6
GAS CHROMATOGRAPHIC RETENTION TIMES AND DETECTION LIMITS FOR ACID EXTRACTABLE COMPOUNDS
EGD
No.
-------
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 The quality of the analysis is assured
through reproducible calibration and
testing of the extraction and GCHS
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 lot
(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 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 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, dibenzo(a,h)anthracene,
benzo(a)pyrene, N-nitrosodimethylamine,
and beta-naphthylamine. 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.
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.
5.1.1.4 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 Cap liners are washed with detergent and
water, rinsed with reagent water (see
Section 6.5.1) and then solvent, and then
baked for at least one hour at
approximately 200 °C.
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. Only
glass or Teflon tubing shall be used. If
the sampler uses a peristaltic pump, a
43
-------
minimum length of compressible silicone
rubber tubing may be used only in the
pump. 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 maintaining a temperature
of 110 ± 5 "C.
5.2.2 Oessicator
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--TefIon
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, borosilicate glass beakers
calibrated to one liter
5.4.2.2 400 - 500 ml borosilicate glass beakers
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 column equipped with
coarse glass frit or glass wool plug.
5.6 Concentration apparatus
5.6.1 Concentrator tube--Kuderna-Danish (K-D) 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.
5.6.2 Evaporation flask--Kuderna-Danish (K-D)
500 mL (Kontes K-570001-0500, or
equivalent), attached to concentrator tube
with springs (Kontes K-662750-0012).
5.6.3 Snyder column--Kuderna-Danish (K-D) three
ball macro (Kontes K-503000-0232, or
equivalent).
5.6.4 Snyder column--Kuderna-Danish (K-D) 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 hour minimum.
5.6.6 Nitrogen evaporation device—equipped with
a water bath that can be maintained at 35
- 40 "C. The N-Evap by Organomation
Associates, Inc., South Berlin, MA (or
equivalent) is suitable.
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
44
-------
5.11
5.11.1
5.12
5.13
Gas chromatograph—shall have splitless 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.
Co I urn--30 ±5 m x 0.25 i 0.02 mm i.d. 5%
phenyl, 94% methyl, 1% vinyl silicone
bonded phase fused silica capillary column
(J & W DB-5, or equivalent).
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
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 grade, 6N in
reagent water.
6.2 Sodium su I fate—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 from
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)
45
-------
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
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 redissolved 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 solution--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 DFTPP 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 5 aliquots
of 0.5 ml each 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).
46
-------
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 ± 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.
Examine the spectrum to determine that
only a single compound is present.
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 referen.ee 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 eonmon
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 in/?:
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, quantitat ion 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.
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 fc>e
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 diIution--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
-------
Table 8
CHARACTERISTIC M/Z'S AND RESPONSE FACTORS OF BASE/NEUTRAL EXTRACTABLE COMPOUNDS
Response
Labeled
Compound Analog
acenaphthene
acenaphthylene
acetophenone
4-aminobiphenyl
aniline
o-anisidine
anthracene
arami te
benzanthrone
1,3-benzenediol
benzene thiol
benzidine
benzo(a)anthracene
benzo(b)f luoranthene
benzo(k)f luoranthene
benzo(a)pyrene
benzoCghi )perylene
2,3-benzof luorene
benzole acid
benzyl alcohol
bi phenyl
bis(2-chloroethyl) ether
bis(2-chloroethoxy)methane
bis(2-chloroisopropyl) ether
bis(2-ethylhexyl) phthalate
2-bromoch lorobenzene
3-bromoch lorobenzene
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-chloroaniline
2-chloronaphthalene
3-chlorom'trobenzene
d10
d8
d10
d8
d12
d12
d12
d12
d12
d10
d8
d6
d12
d4
dg
d4
d22
"26
"34
d42
d50
d62
da
d7
Primary
m/z (1)
154/164
152/160
105
169
93
108
178/188
185
230
110
110
184/192
228/240
252/264
252/264
252/264
276/288
216
105
79
154/164
93/101
93/99
121/131
149/153
111
192
248/253
149/153
57/82
57/66
57
57/66
57
57/66
57
57/66
57
57
57/66
167/175
172
106
127
162/169
157
Factor
(2)
0.79
0.81
1.04
0.43
0.19
0.15
0.78
0.18
0.35
0.16
0.47
0.33
0.40
0.20
0.50
0.73
0.18
Response
Labeled
Compound Analog
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-diaminotoluene
dibenzo(a,h)anthracene
dibenzofuran
dibenzothiophene
1,2-dibromo-3-chloropropane
2,6-dichloro-4-ni troani I ine
1,3-dichloro-2-propanol
2,3-dichloroaniline
1,2-dichlorobenzene
1 ,3-dich lorobenzene
1,4-dichlorobenzene
3,3' -dichlorobenzidine
2,2'-difluorobiphenyl
(int std)
2,3-dichloronitrobenzene
1,2:3,4-diepoxybutane
diethyl phthalate
3,3'-dimethoxybenzidine
dimethyl phthalate
dimethyl sulfone
p-dimethylaminoazobenzene
7,12-dimethylbenz(a)
anthracene
N,N-dimethylformamide
3,6-dimethylphenanthrene
2,4-dimethylphenol
1 ,4-dini trobenzene
2,4-dinitrotoluene
2,6-dinitrotoluene
di-n-octyl phthalate
diphenylamine
di phenyl ether
diphenyldisulf ide
1,2-diphenylhydrazine (3)
ethyl methanesulfonate
ethylenethiourea
•%
d12
d14
d4
d14
d8
d8
d4
d4
d4
d6
d4
d4
dj
dj
dj
d4
d10
d10
d10
Primary
m/z (1)
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
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
Factor
(2)
0.42
0.59
0.017
0.078
0.059
0.22
0.019
0.68
0.47
0.11
0.27
0.19
0.40
0.23
0.58
0.51
0.72
0.24
0.25
0.28
0.22
48
-------
Compound
Table 8
CHARACTERISTIC M/Z'S AND RESPONSE FACTORS OF BASE/NEUTRAL EXTRACTABLE COMPOUNDS
Response
Labeled Primary Factor
Analog m/z (1) (2)
Response
ethynylestradiol 3-methyl
ether
fluoranthene
fluorene
hexachlorobenzene
hexachlorobutadiene
hexachloroethane
hexachIorocycIopentadi ene
hexachIoropropene
indeno(1,2,3-cd)pyrene
isophorone
2-isopropylnaphthalene
isosafrole
longifolene
malachite green
methapyrilene
methyl methanesulfonate
2-methylbenzothiazole
3-methylcholanthrene
4,4'-methylenebis
(2-chloroaniline)
4,5-methylenephenanthrene
1-methylfluorene
2-methylnaphthalene
1-methylphenanthrene
2-(methylthio)benzothiazole
naphthalene
1,5-naphthalenediamine
1,4-naphthoquinone
alpha-naphthylamine
beta-naphthylamine
5-nitro-o-toluidine
2-nitroaniline
3-nitroaniline
4-nitroaniline
ni trobenzene
4-nitrobiphenyl
N-nitrosodi-n-butylamine
N-nitrosodi-n-propylamine
N-nitrosodiethylamine
N-nitrosodimethylamine
N-nitrosodiphenylamine (4)
N-nitrosomethylethylamine
N-nitrosomethylphenylamine
N-nitrosomorpholine
10
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
0.28
0.23
0.32
0.33
0.14
0.43
0.20
0.59
0.59
d8
d7
d5
d14
d6
d6
88
106
56
231
190
180
142
192
181
128/136
158
158
143
143/150
152
138
138
138
123/128
199
84
70/78
102
74/80
169/175
0.33
0.024
0.49
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
0.45
Labeled
Compound Analog
N-nitrosopi peri dine
pentach I orobenzene
pentach I oroethane
pent amethy I benzene
perylene
phenacetin
phenanthrene d.g
phenol d.
phenothiazine
1-phenylnaphthalene
2-phenylnaphthalene
alpha-picoline d-.
pronamide
pyrene d1Q
pyridine
safrole
squalene
styrene d_
alpha-terpineol d.
1,2,4,5-tetrachlorobenzene
thianaphthene
thioacetamide
thioxpnthone
o-toluidine
1,2,3-trichlorobenzene d.
1,2,4-trichlorobenzene d.
1,2,3-trimethoxybenzene
2,4,5-trimethylaniline
triphenylene
tripropylene glycol methyl
ether
1,3,5-trithiane
Primary
m/z (1)
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
Factor
(2)
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) native/labeled
(2) referenced to 2,2'-difluorobiphenyl
(3) detected as azobenzene
(4) detected as diphenylamine
NOTE: Because the composition and purity of
commercially-supplied isotopically labeled standards
may vary, the primary m/z of the labeled analogs
given in this table should be used as guidance. The
appropriate m/z of the labeled analogs should be
determined prior to use for sample analysis. Devia-
tions from the m/z's listed here must be documented
by the laboratory and submitted with the data.
49
-------
Table 9 ..-,•..
CHARACTERISTIC M/Z'S AND RESPONSE FACTORS Of ACID
EXTRACTABLE COMPOUNDS
Labeled
Compound Analog
benzoic acid
4-chloro-3-methylphenol
2-ch lorophenol
p-cresol
3,5-dibromo-
4-hydroxybenzonitri le
2 ,4-dich lorophenol
2 ,6-dich lorophenol
2,4-dinitrophenol
hexanoic acid
2-methyl-4,6-dinitrophenol
2-nitrophenol
4-nitrophenol
pentach lorophenol
2,3,4,6-tetrachlorophenol
2, 3, 6- trich lorophenol
2, 4, 5- trich lorophenol
2, 4, 6- trich lorophenol
d2
d4
dj
"3
d2
4
d2
d2
d2
Primary
m/z (1)
105
107/109
128/132
108
277
162/167
162
184/187
60
198/200
65/109
65/109
266/272
232
196/200
196/200
196/200
Response
Factor
(2)
0.16
0.61
0.12
0.42
0.62
0.17
(1) native/labeled
(2) referenced to 2,2'-difluorobiphenyl
NOTE: Because the composition and purity of
commercially-supplied isotopically labeled standards
may vary, the primary m/z of the labeled analogs
given in this table should be used as guidance. The
appropriate m/z of the labeled analogs should be
determined prior to use for sample analysis. Devia-
tions from the m/z's listed here must be documented
by the laboratory and submitted with the data.
10-
ffl
IT
UJ
> 1.0-
.0.1-
10 20 50 100 200
CONCENTRATION (ug/mL)
FIGURE 1 Relative Response Calibration Curve
for Phenol. The Dotted Lines Enclose a ± 1O Per-
cent Error Window.
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 procedures
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
acquired data. Three isotope ratios are
used in this process:
RX = the isotope ratio measured for the
pure pollutant.
7.4.1
profile (EICP) area for the compound is in
the calibration range. Alternate labeled
compounds and quantitat ion 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.
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
7.4.3
R = the isotope ratio measured for the
labeled compound.
R = the isotope ratio of an analytical
mixture of pollutant and labeled
compounds.
The m/z's are selected such that R > R .
If R is not between 2R and 0.5R , 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,
50
-------
AREA AT
M2/Z
AREA AT
M,/Z
FIGURE 2 Extracted Ion Current Profiles for
Chromatographically Resolved Labeled (m2/z)
and Unlabeled (m,/z) Pairs.
(3A)
AREA = 46100
AREA = 4780
•<3B)
AREA = 2650
AREA = 43600
AREA =48300
R - Carea m./z (at RTQ]
1
[area m2/z (at RT.,)]
= [area n
(at R
TQ]
[area nu/z (at RT.)]
as measured in the mixture of the
pollutant and labeled compounds (Figure
2), and RR » Rm-
7.4.4 Special precautions are taken when the
pollutant- labeled pair is not separated,
or when another labeled compound uith
interfering spectral masses overlaps the
pollutant (a case which can occur uith
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:
(Rm-Rx>
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.
For the example,
7.4.5
7.4.6
46100
4780
9.644
FIGURE 3 Extracted Ion Current Profiles for (3A)
Unlabeled Compound, (3B) Labeled Com-
pound, and (3C) Equal Mixture of Unlabeled
and Labeled Compounds.
2650 = 0.06078
43600
49200 = 1.019
48300
RR = 1.115.
The data from these analyses are reported
to three significant figures (see Section
U.6). Therefore, in order to prevent
rounding errors from affecting the Values
to be reported, all calculations performed
prior to the final determination of
concentrations should be carried out using
at least four significant figures.
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.
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 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:
= (A
(A
x C. ). where
i x Cs)
AS is the area of the characteristic mass
for the compound in the daily standard
A-s
is the area of the characteristic
mass for the internal standard
C-s is the concentration of the internal
standard (ug/mL)
C is the concentration of the compound in
the daily standard (ug/mL)
7.5.1.1 The response factor is determined for at
least five concentrations appropriate to
the response of each compound (Section
6.13); nominally, 10, 20, 50, 100, and 200
ug/mL. The amount of internal standard
added to each extract is the same (100
ug/mL) so that C. remains constant. The
RF is plotted vs concentration for each
compound in the standard (Cs) to produce a
calibration curve.
7.5.1.2 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 using calibration
solutions (Section 6.13) containing the
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'brat ion is required only if
calibration verification (Section 12.5)
criteria cannot be met.
8 QUALITY ASSURANCE/QUALITY CONTROL
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.
8.1.3 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.
52
-------
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).
8.1.5 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.
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
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).
8.2 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.
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
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.
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.
P P
-------
Table 10
ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS
EGD
No.
(D
301
201
377
277
378
278
305
205
372
272
374
274
375
275
373
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
Labeled and
native compound
initi-al precision
and accuracy
(Sec 8.2.3) (ua/L)
Compound
aeenaphthene
aeenaphthene-d...
aeenaphthylene
aeenaphthylene-dg
anthracene
anthracene-d...
benzidine
benzidine-dg
benzo(a)antnracene
bemo(a)anthracene-d._
benzo(b)f luoranthene
benzo(b)f luoranthene-d...
b@nzo( k) f I uoranthene
benzo(k)f luoranthene-d.2
benzo(a)pyrene
benzo( a )pyrene-d. _
benzo(ghi )perylene
benzo(ghi )perylene-d._
biphenyl (Appendix C)
biphenyl-d1Q
bis(2-chloroethyl) ether
bis(2-ehloroethyl) ether-dg
bis(2-chloroethoxy)methane
bis(2-ehloroethoxy)roethane (3)
bis(2-chloroisopropyl) ether
bis(2-chloroisopropyl)ether-d1-
bis(2-ethylhexyl) phthalate
bis(2-ethylhexyl) phthalate-d.
4-bromophenyl phenyl ether
4-bromophenylphenyl ether-d-(3)
butyl benzyl phthalate
butyl beniyl phthalate-d, (3)
n-C10 (Appendix C)
n-ClO-d?.
n-C12 (Appendix C)
n-C12-d26
n-C14 (Appendix C) (3)
n-C16 (Appendix C)
n-d6-d,.
n-C18 (Appendix C) (3)
n-C20 (Appendix C)
n-C20-d,.
n-C22 (Appendix C) (3)
n-C24 (Appendix C)
n-C24-dgn
n-C26 (Appendix C) (3)
s
21
38
38
31
41
49
119
269
20
41
183
168
26
114
26
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
X
79 -
38 -
69 -
39 -
58 -
31 -
16 -
ns(2)
65 -
25 -
32 -
11 -
59 -
15 -
62 -
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 -
Labeled
compound
recovery
(Sec 8.3
and 14.2)
P (%)
134
147
186
146
174
194
518
ns
168
298
545
577
143
514
195
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
20 -
23 -
14 -
ns -
12 -
ns -
ns -
21 -
14 -
ns -
15 -
15 -
20 -
18 -
19 -
18 -
ns -
ns -
18 -
19 -
15 -
270
239
419
ns
605
ns
ns
290
529
ns
372
372
260
364
325
364
ns
ns
308
306
376
Calibration
verification
(Sec 12.5)
(ug/mL)
80 -
71 -
60 -
66 -
60 -
58 -
34 -
ns -
70 -
28 -
61 -
14 -
13 -
13 -
78 -
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 -
125
141
166
152
168
171
296
ns
142
357
164
ns
ns
ns
129
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
Labeled
and native
compound
on- going
accuracy
(Sec 12.7)
R (ug/L)
72
30
61
33
50
23
11
ns
62
22
20
ns
53
ns
59
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
.
-
-
-
-
-
-
.
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
™
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
144
180
207
168
199
242
672
ns
176
329
ns
ns
155
685
206
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
-------
Table 10 (continued)
ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS
EGD
No.
(1)
525
726
626
728
628
320
220
322
222
324
224
340
240
376
276
713
613
382
282
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
Labeled and
native compound
initial precision
and accuracy
(Sec 8.2.3) (ug/L)
Compound
n-C28 (Appendix C) (3)
n-C30 (Appendix C)
n-C30-d,2
carbazole (4c)
carbazole-dg (3)
2-chloronapnthalene
2-chloronaphthalene-d-
4-chloro-3-methylphenol
S-chloco-S-methylphenol-dj
2-chlorophenol
2-chlorophenol-d,
4-chlorophenyl phenyl ether
4-chlorophenyl phenyl ether-d.
chrysene
chrysene-d...
p-cymene (Appendix C)
p-cymene-d^
d ibenzo( a, h) anthracene
dibenzo(a,h)anthracene-d. , (3)
dibenzofuran (Appendix C)
dibenzofuran-dg
dibenzothiophene (Synfuel)
dibenzothiophene-d.
di-n-butyl phthalate
di-n-butyl phthalate-d.
1 ,2-dichlorobenzene
1 , 2 - d i ch I orobenzene- d.
1 ,3-dichlorobenzene
1 ,3-dichlorobenzene-d.
1 ,4-dichlorobenzene
1 , 4-dichlorobenzene-d,
3,3'-dichlorobenzidine
3,3'-dichlorobenzidine-d^
2,4-dichlocophenol
2,4-dichlocophenol-d-
diethyl phthalate
diethyl phthalate-d.
2 ,4-di methyl phenol
2,4-dimethylphenol-dj
dimethyl phthalate
dimethyl phthalate-d^
2,4-dinitrophenol
2,4-dinitrophenol-d_
2,4-dinitrotoluene
2,4-dinitrotoluene-d-,
2,6-dinitrotoluene
2,6-dinitrotoluene-d-
s
35
32
41
38
31
100
41
37
111
13
24
42
52
51
69
18
67
55
45
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
X
35 -
61 -
27 -
36 -
48 -
46 -
30 -
76 -
30 -
79 -
36 -
75 -
40 -
59 -
33 -
76 -
ns -
23 -
29 -
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 -
Labeled
compound
recovery
(Sec 8.3
and 14.2)
P (%)
193
200
242
165
130
357
168
131
174
135
162
166
161
186
219
140
359
299
268
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
13
29
15
ns
23
19
13
ns
14
28
29
13
ns
ns
ns
ns
24
ns
ns
ns
ns
10
17
- 479
- 215
- 324
- 613
- 255
• 325
- 512
- ns
- 529
- 220
- 215
- 346
- 494
- 550
- 474
- ns
- 260
- ns
- 449
- ns
- ns
- 514
- 442
Calibration
verification
(Sec 12.5)
(ug/mL)
26
66
24
44
69
58
72
85
68
78
55
71
57
70
24
79
66
13
13
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
- 392
- 152
- 423
- 227
- 145
- 171
- 139
- 115
- 147
- 129
- 180
- 142
• 175
- 142
- 411
- 127
- 152
- 761
- ns
- 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
Labeled
and native
compound
on- go ing
accuracy
(Sec 12.7)
R (ug/L)
31 -
56 -
23 -
31 -
40 -
35 -
24 -
62 -
14 -
76 -
33 -
63 -
29 •
48 -
23 -
72 -
ns -
19 -
25 -
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 -
212
215
274
188
156
442
204
159
314
138
176
194
212
221
290
147
468
340
303
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
-------
Table 10 (continued)
ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS
EGD
No.
(1)
369
269
707
607
708
608
337
237
339
239
380
280
309
209
552
252
312
212
353
253
083
354
254
360
260
355
255
702
602
356
256
357
257
358
258
361
261
363
263
362
262
364
264
381
281
365
265
Labeled and
native compound
initial precision
and accuracy
(Sec 8.2.3) (ug/L)
Compound
di-n-octyl phthalate
di-n-octyl phthalate-d.
diphenylamine (Appendix C)
diphenylamine-d^Q
diphenyl ether (Appendix C)
diphenyl ether-d..-
1,2-diphenylhydrazine
1 ,2-diphenylhydrazine-d.Q
f luoranthene
f luoranthene-d.g
fluorene
f luorene-d...
hexachlorobenzene
hexachlorobenzene- C,
Q
hexachlorobutadiene
hexachlorobutadiene- C,
hexachloroethane
hexachloroethane- C
hexach 1 orocyc I opent ad i ene
hexachlorocyclopentadiene- C,
ideno(1,2,3-cd)pyrene (3)
isophorone
isophorone-dg
2-methyl-4,6-dinitrophenol
2-methyl-4,6-dinitrophenol-d2
naphthalene
naphthalene-d-
beta-naphthylamine (Appendix C)
beta- naph thy lamine-d.
nitrobenzene
nitrobenzene-dg
2-nitrophenol
2-nitrophenol-d,
4-nitrophenol
4-nitrophenol-d,
N-nitrosodimethylamine
N-nitrosodimethylamine-d, (3)
N-nitrosodi-n-propylamine
N-nitrosodi-n-propylamine (3)
N-nitrosodiphenylamine
N-nitrosodiphenylamine-d^
pent ach I oropheno I
pentachlorophenol- C,
phenanthrene
phenanthrene-d...
phenol
phenol-dj
s
16
46
45
42
19
37
73
35
33
35
29
43
16
81
56
63
227
77
15
60
55
25
23
19
64
20
39
49
33
25
?8
15
23
42
188
49
33
45
37
45
37
21
49
13
40
36
161
X
77 •
12 -
58 -
27 -
82 -
36 -
49 -
31 -
71 -
36 -
81 -
51 -
90 -
36 -
51 -
ns -
21 -
ns -
69 -
ns -
23 -
76 -
49 -
77 -
36 -
80 -
28 -
10 -
ns -
69 -
18 -
78 -
41 -
62 -
14 -
10 -
ns -
65 -
54 -
65 -
54 -
76 -
37 -
93 -
45 -
77 -
21 -
Labeled
compound
recovery
(Sec 8.3
and 14.2)
P (%)
161
383
205
206
136
155
308
173
177
161
132
131
124
228
251
316
ns
400
144
ns
299
156
133
133
247
139
157
ns
ns
161
265
140
145
146
398
ns
ns
142
126
142
126
140
212
119
130
127
210
ns
11
19
17
20
27
13
ns
ns
ns
33
16
14
ns
ns
27
ns
ns
26
26
18
24
ns
- ns
- 488
- 281
- 316
- 278
- 238
- 595
- ns
- ns
- ns
- 193
- 527
- 305
- ns
- ns
- 217
- ns
- ns
- 256
- 256
- 412
- 241
- ns
Calibration
verification
(Sec 12.5)
(ua/mL)
71 -
21 -
57 -
59 -
83 -
77 -
75 -
58 -
67 -
47 -
74 -
61 -
78 -
38 -
74 -
68 -
71 -
47 -
77 -
47 -
13 -
70 -
52 -
69 -
56 -
73 -
71 -
39 -
44 -
85 -
46 -
77 -
61 -
55 -
35 -
39 -
44 -
68 -
59 -
68 -
59 -
77 -
42 -
75 -
67 -
65 -
48 -
140
467
176
169
120
129
134
174
149
215
135
164
128
265
135
148
141
212
129
211
761
142
194
145
177
137
141
256
230
115
219
129
163
183
287
256
230
148
170
148
170
130
237
133
149
155
208
Labeled
and native
compound
on- go ing
accuracy
(Sec 12.7)
R (ug/L)
74 -
10 -
51 -
21 -
77 -
29 -
40 -
26 -
64 -
30 -
70 -
38 -
85 -
23 -
43 -
ns -
13 -
ns -
67 •
ns -
19 -
70 -
44 -
72 -
28 -
75 -
22 -
ns -
ns -
65 -
15 -
75 -
37 -
51 -
ns -
ns -
ns -
53 -
40 -
53 -
40 -
71 -
29 -
87 -
34 -
62 -
ns -
166
433
231
249
144
186
360
200
194
187
151
172
132
321
287
413
ns
563
148
ns
340
168
147
142
307
149
192
ns
ns
169
314
145
158
175
ns
ns
ns
173
166
173
166
150
254
126
168
154
ns
56
-------
Table 10 (continued)
ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS
EGD
No.
(1)
703
603
334
234
710
610
709
609
729
629
308
208
530
731
631
321
221
(1)
Labeled and
native compound
initial precision
and accuracy
(Sec 8.2.3) (ug/L)
Compound
alpha-picoline (Synfuel)
alpha-picoline-d_
pyrene
pyrene-d.Q
styrene (Appendix C)
styrene-d-
alpha-terpineol (Appendix
alpha-terpineol-d.
C)
1,2,3-trichlorobenzene (4c)
1,2,3-trichlorobenzene-d,
1,2,4-trichlorobenzene
1 ,2,4-trichlorobenzene-o™
2,3,6-trichlorophenol (4c)
2,4,5-trichlorophenol (4c)
(3)
(3)
2,4,5-trichlorophenol-d2 (3)
2,4,6-trichlorophenol
2,4,6-trichlorophenol-d-
Reference numbers beginning
with 0,
method; reference numbers beginning
(2)
(3)
standard method; reference
dilution.
numbers
s
38
138
19
29
42
49
44
48
69
57
19
57
30
30
47
57
47
1 or 5
X
59 -
11 -
76 -
32 -
53 -
ns -
42 -
22 -
15 -
15 -
82 -
15 -
58 -
58 -
43 -
59 -
43 -
indicate
Labeled
compound
recovery Calibration
(Sec 8.3 verification
and 14.2) (Sec 12.5)
P (%) (us/ml)
149
380
152
176
221
281
234
292
229
212
136
212
137
137
183
205
183
ns -
18 -
ns -
ns -
ns -
ns -
; 21 -
21 -
ns
303
ns
672
592
592
363
363
60 -
31 -
76 -
48 -
65 -
44 -
54 -
20 -
60 -
61 -
78 -
61 -
56 -
56 -
69 -
81 -
69 -
a pollutant quantified by
with 2 or 6 indicate a
beginning
with 3
ns = no specification: limit is outside the range that
or 7
can
labeled
indicate
compound
165
324
132
210
153
228
186
502
167
163
128
163
180
180
144
123
144
Labeled
and native
compound
on- go ing
accuracy
(Sec 12.7)
R (ua/L)
50
ns
72
28
48
ns
38
18
11
10
77
10
51
51
34
48
34
the internal
quantified by
a pollutant
the
quantified by
- 174
- 608
- 159
- 196
- 244
- 348
- 258
- 339
- 297
- 282
- 144
- 282
- 153
- 153
- 226
- 244
- 226
standard
internal
isotope
be measured reliably.
This compound is to be determined by internal standard; specification
compound.
is
derived
from
related
For example, if P = 90% and s = 10% for 8.5.2
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
(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 8-6
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.
If any of the compounds of interest
(Tables 1 - 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 - 4), analysis of samples is
halted until the source of contamination
is eliminated and a blank shows no
evidence of contamination at this level.
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 12.5), and for
initial (Section 8.2) and on-going
(Section 12.7) precision and recovery
should be identical, so that the most
57
-------
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 semivolatiles 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
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).
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)
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
10.2.1
10.2.1.1
10.2.1.2
10.2.1.3
10.2.2
10.2.2.1
10.2.2.2
10.2.2.3
10.2.2.4
Preparation of samples for extraction
Samples containing one percent solids or
less—extract sample directly using
continuous liquid/liquid extraction
techniques.
Measure 1.00 ± 0.01 liter of sample into a
clean 1.5 - 2.0 liter beaker.
Dilute aliquot--for samples which are
expected to be difficult to extract,
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.
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).
Samples containing one to 30 percent
solids
Mix sample thoroughly.
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
% solids
grams
Discard all sticks, rocks, leaves and
other foreign material prior to weighing.
Place the weight determined in 10.2.2.2 in
a clean 1.5 - 2.0 liter beaker.
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
58
-------
[10.2.3.1]
[10.2.1.3, 10.2.3.2]
[10.2.3.3]
[10.2.4]
[10.3.2]
[10.3.4]
[10.5]
[10.6]
[11.3]
[11-4]
STANDARD
1 L REAGENT
WATER
SPIKE
1,0 mL
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
JL
INJECT
BLANK
1 LREAGENT
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
TO 1.0 mL
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 1625. Numbers in Brackets [ ] Refer to Section Numbers in the Method.
-------
1.5 - 2.0 liter beaker. When diluted to
1.0 liter, this dilute aliquot will
contain 0.1 percent solids.
10.2.2.5 Bring the sample aliquot(s) above to 100 -
200 mL volume with reagent water.
10.2.2.6 Spike 0.5 mL of the labeled compound
spiking solution (Section 6.8) 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 inch horn on the ultrasonic
probe approx 1/2 inch 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.
10.2.2.9 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 three 1.0 i 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 Discard all sticks, rocks, leaves and
other foreign material prior to weighing.
Weigh 30 ± 0.3 grams into a clean 400 -
500 ml beaker.
10.2.5.3 Dilute aliquot—for samples which are
expected to be difficult to extract,
concentrate, or clean-up, weigh 3 ± 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 i 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 and QC aliquots
per Sections 10.4 through 10.8.
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 QC 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.
60
-------
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 sulfate. 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
Haters 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.
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
semivolatile 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
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. 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 lower 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
10.6.1 Kuderna-Danish (K-D)--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
61
-------
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, to 1.0 ml if it does
not require clean-up, or to 0.5 ml if it
has been cleaned up.
10.6.2 Nitrogen blowdown--Place the concentrator
tube in a warm water bath (35 °C) and
evaporate the solvent volume using a
gentle stream of clean, dry nitrogen
(filtered through a column of activated
carbon). Caution: New plastic tubing
must not be used between the carbon trap
and the sample, since it may introduce
interferences. The internal wall of the
tube must be rinsed down several times
with methylene chloride during the
operation. During evaporation, the tube
solvent level must be kept below the water
level of the bath. The extract must never
be allowed to become dry. Adjust the
final volume to 5.0 ml if the extract is
to be cleaned up by GPC, to 1.0 ml if it
does not require clean-up, or to 0.5 ml if
it has been cleaned 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 fop analysis.
10.8 GPC setup and calibration
10.8.1 Column packing
10.8.1.1 Place 75 i 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 will be corn oil, bis(2-
ethylhexyl) phthalate, 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 85%. 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 to 5.0
mL 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
62
-------
blank shall be run through the system to
check for carry-over.
10.9.6 Concentrate the extract to 0.5 mL and
transfer to a screw-cap vial per Sections
10.6 and 10.7. Concentrating extracts
cleaned up by GPC to 0.5 ml Mill place the
analytes in the same part of the GCMS
calibration range as in samples not
subjected to GPC.
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
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
en-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 recall brat 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.
12.2 OFTPP spectrum validity-inject 1 uL of
the DFTPP solution (Section 6.11) either
separately or within a feu 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).
63
-------
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
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.
NQTE: 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
for each pollutant and labeled compound in
each matrix type by calculating the
average percent recovery (R) and the
standard deviation of percent recovery
(s ). Express the accuracy as a recovery
interval from R - 2sp to R + 2s . For
example, if R = 95% and sp = 5%, the
accuracy is 85 - 105%.
45,000
35,000
25,000
1 1 1 1 1 1 i 1 1
ANTHRACENE-D,,,
f - - '
, .
• • «
•
123456789 10
ANALYSIS NUMBER
"A*
go
8*5
mS
11
non -
i i t t t i t i i
ANTHRACENE
r « • * a t •
r 9 9
6/1 6/1 6/1 6/1 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 Anrhracene-d10 (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 1-4):
64
-------
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.
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 t 15 seconds (whichever is
greater) for compounds for which no window
is specified.
13.1.4 The system has not been calibrated for the
compounds listed in Tables 3 and 4,
however, the relative retention times and
mass spectra of these compounds are known.
Therefore, for a compound in Tables 3 or 4
to be identified, its retention time
relative to the internal standard 2,2'-
difluorobiphenyl must fall within a
retention time window of t 30 seconds, or
t 30 scans (whichever is greater) of the
nominal retention time of the compound
specified in Tables 5 or 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 i 1 scan.
13.3.2 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.
13.4 The 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.
14 QUANTITATIVE DETERMINATION
14.1 Isotope dilution--Because the pollutant
and its labeled analog exhibit the same
effects upon extraction, concentration,
and gas chromatography, correction for
recovery of the pollutant can be made by
adding a known amount of a labeled
compound to every sample prior to
extraction. Relative response (RR) values
for sample mixtures are used in
conjunction with the 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 ).
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:
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
in the extract and the weight of the
solids (Section 10), as follows:
-------
Concentration in solid (ug/kg)
where V is the extract volume in ml, and
Wg is the sample weight in kg.
14.4 Dilution of samples—if the EICP area at
the quantitation 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. For water samples, where the
base/neutral and acid extracts are not
combined, re-analysis is only required for
the extract (8/M or A) in which the
compound exceeds the calibration range.
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 ± 0.003 g
of sample per Section 10.2.5.
14.5 Dilution of samples containing high
concentrations of compounds to be
identified per Section 13.3 -- When the
EICP area of the quant i tat ion m/z of a
compound to be identified per Section 13.3
exceeds the linear range of the GCMS
system, or when any peak is saturated,
dilute the sample per Section 14.4.1-
14.4.3.
14.6 Results are reported to three significant
figures for all pollutants, labeled
compounds, and tentatively identified
compounds found in all standards, blanks,
and samples. For aqueous samples, the
units are ug/L, and for samples containing
one percent solids or greater (soils,
sediments, filter cake, compost), the
units are ug/kg, based on the dry weight
of the solids.
14.6.1 Results for samples which have been
diluted are reported at the least dilute
level at which the area at the
quantitat 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
quant itat ion 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
than one-half of its respective area, then
loss of the internal standard in the
66
-------
extract has occurred. In this case, 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 14.4. 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 apply to 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.
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).
3 "Working with Carcinogens," DHEU, PHS,
CDC, NIOSH, Publication 77-206, (Aug
1977).
4 "OSHA Safety and Health Standards, General
Industry" OSHA 2206, 29 CFR 1910 (Jan
1976).
5 "Safety in Academic Chemistry
Laboratories," ACS Committee on Chemical
Safety (1979).
6 "Interlaboratory Validation of U. S.
Environmental Protection Agency Method
1625A, Addendum Report", SRI
International, Prepared for Analysis and
Evaluation Division (UH-557), USEPA, 401 M
St SU, Washington DC 20460 (January
1985).
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, ASTH,
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
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 (UH-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-CUBED Division
of Maxwell Laboratories, Inc., Prepared
for W. A. Telliard, Industrial Technology
Division (UH-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
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).
67
-------
RIC DATft: ABNID1166 #1
83/13/84 5:24:00 CftLl: ABH1D1166 #1
SAMPLE: AB,G,UER,80100.00,C,NA:HA..HAS
CONDS.: 1625A, 30(1,0.25(111,5630,30-28068,158280,30CIVSJ
RANGE: G 1,3208 LABEL: N 2, 3.0 QUAN: A 2, 2.0 J
SCANS 1 TO 3200
3 8ASE: U 20, 3
100.0-1
RIC
I
715776.
580
7:55
1000
15:50
1500
23:45
2000
31:40
2500
39:35
3006
47:30
SCAN
TIME
FIGURE 6 Chromatogram of Combined Acid/Base/Neutral Standard.
68
-------
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
SO
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
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
MJ.
524
13
12
m/z
52
74
91
int.
75
64
22
4-aminobiphenyl
int.
55
65
aniline
int.
65
47
59
10
o-anisidine
int.
22
286
142
915
1000
arami te
M..
606
155
143
270
benz ant h rone
int.
69
278
762
m/z
63
141
m/z
41
52
64
92
m/z
41
54
66
81
109
m/z
57
91
175
334
m/z
75
150
203
int.
65
132
int.
66
54
33
136
int.
43
39
20
41
55
jnL,
758
339
182
137
int..
71
58
126
m/z
72
167
m/z
42
53
65
93
m/z
42
61
76
92
122
m/z
59
105
185
m/z
87
174
230
int.
82
163
int.
16
12
226
1000
int.
10
12
13
47
123
int.
328
153
1000
int.
97
67
1000
m/z
83
168
m/z
46
54
66
94
m/z
50
62
77
93
844
m/z
63
107
187
m/z
88
199
231
int.
73
280
IT&L
11
40
461
73
int.
60
25
36
14
124
int.
782
239
328
int.
160
63
177
m/z
85
169
m/z
47
61
74
m/z
51
63
68
94
56
m/z
65
121
191
m/z
99
200
int.
163
1000
int..
75
17
11
int.
106
43
32
18
int.
285
107
346
int.
69
350
m/z
115
170
m/z
50
62
78
m/z
52
64
79
105
m/z
74
123
197
m/z
100
201
int.
142
216
IHts.
40
28
14
int.
202
24
25
18
int.
113
120
191
int.
215
236
1,3-benzenediol
int.
64
54
27
16
51
benzenethiol
int.
128
161
m/z
41
52
63
81
m/z
50
84
int.
19
29
74
201
int.
149
259
m/z
52
53
64
82
m/z
51
109
int.
42
184
61
251
int.
205
316
m/z
43
54
65
95
m/z
65
110
int.
36
89
13
13
int.
175
1000
m/z
49
55
68
109
m/z
66
111
jnt^
11
97
56
11
jnt^
505
102
m/z
50
61
69
110
fflZz
69
int.
43
15
119
1000
int.
114
69
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
563 2,3-benzof luorene
m/z int. m/z
74 52 81
108 491 187
216 1000 217
943 benzoic acid
m/z int. m/z
45 29 50
75 25 76
564 benzyl alcohol
m/z int. m/z
40 17 59
61 11 62
75 13 76
89 65 90
108 737 109
565 2-bromochlorobenzene
m/z int. m/z
49 237 50
76 202 111
566 3-bromochlorobenzene
m/z int. m/z
49 201 50
76 197 111
567 4-chloro-2-nitroaniline
m/z int. m/z
49 119 50
63 1000 64
76 127 78
126 766 128
568 5-chloro-o-toluidine
m/z int. m/z
50 115 51
79 140 89
143 313
569 4-chloroani line
m/z int. m/z
41 60 62
91 63 92
129 292
570 3-chloronitrobenzene
m/z int. m/z
50 619 51
85 101 99
int.
69
75
166
int.
221
81
int.
16
31
18
64
43
int.
890
961
Ink
834
1000
int.
174
315
152
234
ink
261
152
int.
55
186
Int..
189
258
m/z
94
189
m/z
51
77
m/z
50
63
77
91
m/z
51
113
m/z
51
113
m/z
51
65
90
142
m/z
52
106
m/z
63
99
m/z
73
111
ink
143
90
ink
413
778
int.
155
70
565
125
int.
183
287
int.
174
301
int.
260
192
724
211
int.
257
1000
int.
~147
67
int.
144
851
m/z
95
213
m/z
52
78
m/z
51
64
78
105
m/z
73
190
m/z
73
190
m/z
52
73
91
172
m/z
53
140
m/z
64
100
m/z
74
113
Ink
253
233
ink
45
76
int.
319
12
116
38
int.
158
638
int.
169
625
int.
531
290
253
915
int.
137
599
int.
135
115
int.
330
266
m/z
106
214
m/z
66
105
m/z
52
65
79
106
m/z
74
192
m/z
74
192
m/z
61
74
101
174
m/z
77
141
m/z
65
127
m/z
75
157
int.
60
60
int.
11
1000
int.
78
75
1000
18
ink
506
809
int.
509
802
int.
205
105
232
289
int.
420
964
int.
329
1000
int.
1000
424
m/z
107
215
m/z
74
122
m/z
53
74
80
107
m/z
75
194
m/z
75
194
m/z
62
75
114
m/z
78
142
m/z
73
128
m/z
76
159
int.
205
987
int.
53
868
int.
84
35
73
523
int.
1000
193
int.
914
191
int.
394
156
312
int.
134
265
int.
51
81
ink
169
137
70
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
571
m/z
50
89
944
ITl/Z
50
80
572
m/z
40
105
573
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
o-cresol
int.
102
114
p-cresol
int.
136
145
crotoxyphos
int.
633
484
m/z int.
51 181
90 231
m/z int.
51 224
90 122
m/z int.
44 448
109 21
m/z
53
107
m/z
52
107
m/z
67
127
int.
144
783
int.
106
822
int.
42
1000
m/z
77
108
m/z
53
108
m/z
77
166
int.
358
1000
int.
196
1000
int.
70
180
m/z
79
m/z
77
m/z
79
193
int..
380
int.
420
JDS*
41
401
m/z
80
m/z
79
m/z
104
194
int.
159
int.
308
int..
100
20
2,6-di-t-butyl-p-benzoquinone
int.
392
376
538
410
m/z i nt .
53 586
79 308
136 240
m/z
55
91
149
int.
325
456
429
m/z
57
95
163
int.
668
322
292
m/z
65
107
177
int..
416
248
1000
m/z
67
121
205
int.
927
255
203
2,4-diaminotoluene
int.
70
50
134
m/z int.
42 55
77 147
106 67
m/z
51
78
121
int.
76
69
958
m/z
52
93
122
int.
70
63
1000
m/z
53
94
123
int.
51
224
79
m/z
61
104
int.
91
128
1,2-dibromo-3-chloropropane
int.
38
331
17
204
m/z int.
59 341
81 43
119 74
187 10
m/z
51
93
121
int.
104
117
66
m/z
61
95
155
int.
38
106
635
m/z
75
97
157
int.
1000
12
784
m/z
76
105
158
int.
75
67
20
3, 5-dibromo-4-hydroxybenzonitri le
int.
148
141
m/z int.
61 193
275 489
m/z
62
277
int.
222
1000
m/z
88
279
int..
632
451
m/z
117
int.
137
m/z
168
int.
152
2,6-dichloro-4-m'troaniline
int.
206
137
218
m/z i nt .
52 1000
89 218
160 401
m/z
61
90
176
int.
523
443
431
m/z
62
97
178
int.
828
458
134
m/z
63
124
206
int.
588
954
378
m/z
73
126
int.
470
401
1,3-dichloro-2-propanol
int.
14
113
11
m/z int.
42 55
50 15
79 1000
m/z
43
51
80
ink
503
37
25
m/z
44
57
81
jnt^
22
10
310
m/z
47
61
int.
12
12
m/z
58
75
Int.
15
14
71
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
578
m/z
52
73
163
579
m/z
49
74
110
161
946
m/z
49
126
580
m/z
40
57
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
2,3-dichloroaniline
int. m/z
138 61
130 90
626 165
int.
151
460
101
m/z
62
99
int.
265
202
m/z
63
125
int.
455
108
m/z
64
126
int.
142
149
m/z
65
161
int.
105
1000
2,3-dichloronitrobenzene
int. m/z
220 50
976 75
204 111
190 163
2,6-dichlorophenol
i nt . m/z
111 62
260 162
1,2:3,4-diepoxy butane
i nt . m/z
37 41
155 58
3,3'-dimethoxybenzidine
int. m/z
44 79
115 158
152
dimethyl sulfone
int. m/z
10 45
69 64
23
int.
257
743
303
121
Mi
160
1000
int.
29
16
int.
222
154
int..
94
22
m/z
61
84
133
191
m/z
63
164
m/z
42
85
m/z
85
186
m/z
46
65
int.
150
351
701
411
int.
714
613
int.
83
13
Mi
69
144
int.
29
19
m/z
62
85
135
193
m/z
73
166
m/z
43
m/z
93
201
m/z
47
79
int.
120
166
435
263
Mi
132
101
int.
60
int.
84
552
int.
18
1000
WLL
63
86
145
m/z
98
m/z
55
m/z
107
229
m/z
48
81
int.
173
125
580
int.
293
M^
1000
Mi
46
162
Mi
69
36
m/z
73
109
147
m/z
99
m/z
56
m/z
115
244
m/z
62
94
int.
336
1000
368
int.
117
Mi
67
int.
110
1000
int.
14
528
p-dimethylaminoazobenzene
int. m/z
483 51
142 105
Mi
181
190
m/z
77
120
int.
447
1000
m/z
78
148
int.
120
160
m/z
79
225
int.
147
676
m/z
91
int.
109
7,12-dimethylbenzo(a)anthracene
int. m/z
24 112
46 126
23 239
68 253
N,N-dimethylformamide
int. m/z
58 41
17 58
int.
34
81
313
33
Mi
79
83
m/z
113
127
240
255
m/z
42
72
Mi
112
60
230
84
int.
497
89
m/z
114
128
241
256
m/z
43
73
int.
38
76
433
1000
int.
115
994
m/z
119
215
242
257
m/z
44
74
Mi
212
24
61
180
int.
1000
35
m/z
120
226
250
m/z
45
Mi
296
47
32
int.
19
72
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
586
m/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
947
m/z
41
56
73
593
m/z
51
128
170
594
m/z
50
104
3,6-dimethylphenanthrene
int. m/z. int.
113 89
193 191
1,4-dinitrobenzene
int. m/z
1000 51
664 92
diphenyldisulf ide
int. m/z
153 51
132 154
ethyl methanesulfonate
int. m/z
16 43
22 65
206 109
ethylenethiourea
int. m/z
46 42
151 102
ethynylestradiol 3-meth
int. m/z
155 53
115 173
516
hexachloropropene
int. m/z
131 71
206 143
hexanoic acid
int. m/z
627 42
90 57
412 74
2- isopropylnaphthalene
int. m/z
100 63
216 152
368
isosafrole
int. m/z
110 51
441 131
129
430
int.
131
240
Inti
293
191
int.
72
93
579
int.
126
1000
lyl ether
int.
101
199
inii
333
196
int.
535
102
56
int.
111
133
int.
222
371
m/z
94
205
m/z
63
122
m/z
65
185
m/z
45
79
111
m/z
45
m/z
91
174
m/z
106
211
m/z
43
60
87
m/z
76
153
m/z
63
132
int.
179
246
int.
228
166
int.
671
117
int.
208
1000
18
int.
97
int.
157
313
int.
334
631
int.
214
1000
98
int.
157
184
int.
127
107
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
m/z
45
61
m/z
77
154
m/z
77
135
int.
142
1000
int.
218
399
int.
282
418
int.
40
127
15
int.
42
int.
143
1000
int.
200
1000
int.
186
66
int.
129
114
int.
277
129
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
m/z
46
69
m/z
115
155
m/z
78
161
int.
151
159
int.
311
int.
141
int.
19
42
33
int.
14
int.
226
149
int.
329
623
Iflti.
19
21
iDU
147
1000
int.
208
250
m/z
189
m/z
75
m/z
109
m/z
63
96
m/z
72
m/z
159
242
m/z
119
217
m/z
55
70
m/z
127
156
m/z
103
162
int.
388
Mi
623
int.
1000
int.
23
16
int.
89
int.
132
153
int.
320
186
int.
128
20
int.
131
139
int.
355
1000
73
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
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
903
m/z
50
76
139
166
181
longifolene
Mi
438
1000
394
m/z
55
93
133
int.
719
611
338
m/z
65
94
161
Mi
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
methapyri line
int.
72
54
m/z
126
253
m/z
45
79
Mi
313
1000
int.
47
48
m/z
165
254
m/z
53
97
int.
369
160
int.
40
516
m/z
208
329
m/z
58
190
MA
135
189
int.
1000
40
m/z
209
330
m/z
71
191
int.
233
775
Mi
188
67
m/z
210
331
m/z
72
int.
181
170
.MA
225
methyl methanesulfonate
int.
178
285
137
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
Mi
133
392
m/z
58
109
int.
153
102
m/z
62
148
int.
106
279
m/z
63
149
Mi
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
MA
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
Mi
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
Mi
145
213
m/z
86
188
int.
53
137
1-methylf luorene
int.
66
196
54
136
99
m/z
51
83
151
176
MA
87
135
73
96
ffl/Z
62
87
152
177
int.
57
53
124
52
m/z
63
88
163
178
Mi
137
78
57
202
m/z
74
89
164
179
int.
64
203
58
182
m/z
75
90
165
180
int.
85
58
1000
686
74
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
904 2-methylnaphthalene
m/z int. m/z
50 29 51
65 19 69
76 14 77
114 13 115
141 748 142
905 1-methylphenanthrene
m/z int. m/z
51 54 63
96 132 163
193 152
int.
39
56
15
303
1000
int.
86
55
m/z
57
70
86
116
143
m/z
70
165
int.
28
25
13
25
105
int..
62
217
m/z
58
71
87
126
m/z
74
189
int.
47
126
18
13
int.
51
165
m/z
62
74
89
139
m/z
81
191
int.
26
25
42
98
int.
52
532
m/z
63
75
113
140
m/z
83
192
int.
65
23
19
24
Ms.
164
1000
906 2-(methylthio)benzothiazole
m/z int. m/z
45 790 50
136 239 148
907 1,5-naphthalenediamine
m/z i nt . m/z
51 48 65
130 262 131
908 1,4-naphthoquinone
m/z i nt . m/z
50 445 51
76 590 101
158 1000 159
909 alpha-naphthylamine
m/z int. m/z
50 25 51
65 27 71
115 401 116
910 5-nitro-o-toluidine
m/z int. m/z
51 194 52
94 168 104
911 2-nitroaniline
m/z i nt . m/z
41 64 50
63 181 64
92 566 108
912 3-nitroaniline
m/z i nt . m/z
41 101 52
65 1000 66
108 87 138
int.
212
938
int.
83
40
int.
62
51
100
int.
31
58
212
int.
159
120
int.
51
155
170
int.
120
114
717
m/z
63
180
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.
383
250
int.
75
43
int.
52
613
int.
36
104
53
int.
121
691
int.
89
960
1000
int.
59
169
51
m/z
69
181
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.
578
1000
int.
111
89
int.
69
52
int.
46
62
1000
int.
766
1000
int.
207
96
63
int.
58
62
m/z
82
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.
233
int.
86
1000
int.
189
550
int.
28
22
101
int.
176
int.
74
212
int.
143
764
m/z
108
m/z
118
159
m/z
75
130
m/z
63
114
m/z
79
m/z
62
91
m/z
64
93
int.
627
int.
52
117
int.
205
433
mti
59
34
Mi
619
int.
58
86
int.
121
62
75
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
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
4-nitroaniline
int.
228
124
m/z
53
80
int.
160
266
m/z
62
92
Mi
110
300
m/z
63
108
int.
216
636
m/z
64
138
int.
164
520
m/z
65
int.
1000
4-nitrobiphenyl
int..
131
902
m/z
63
153
int.
104
284
m/z
76
169
int.
179
374
m/z
115
199
int.
134
1000
m/z
141
200
int.
277
125
m/z
151
Mi
259
N-nitroso-di-n-butylamine
int.
1000
994
161
m/z
42
84
int.
536
985
m/z
43
86
int.
570
103
m/z
44
99
int..
313
197
m/z
55
115
jnt^.
129
158
m/z
56
116
int.
167
237
N-nitrosodiethylamine
int.
170
525
807
m/z
42
57
103
int.
079
492
35
m/z
43
70
int.
69
24
m/z
44
71
int.
1000
28
m/z
45
85
int.
20
25
m/z
54
87
int.
18
31
N-nitrosomethylethylamine
int.
117
99
m/z
42
59
int.
1000
13
m/z
43
71
Mi
667
60
m/z
44
73
int.
26
57
m/z
54
88
int.
17
772
m/z
56
89
int.
189
20
N-nitrosomethytphenylamine
int.
181
331
m/z
51
104
int.
434
147
m/z
52
106
int.
104
673
m/z
63
107
int.
110
220
m/z
77
212
int.
1000
137
m/z
78
int.
194
N-nitrosomorpholine
int.
181
1000
m/z
42
57
int.
192
49
m/z
43
85
int.
52
13
m/z
44
86
int.
17
333
m/z
54
87
int.
85
14
m/z
55
116
int.
95
337
N-nitrosopiperidine
int.
320
58
28
m/z
42
55
84
int.
1000
444
47
m/z
43
56
114
int.
43
224
491
m/z
51
57
115
int.
14
17
26
m/z
52
67
int.
12
21
m/z
53
82
int.
32
26
pentach I orobenzene
int.
160
106
m/z
108
248
int.
239
648
m/z
125
250
int.
102
1000
m/z
178
252
Mi
102
642
m/z
213
254
int.
179
199
m/z
215
int.
218
pentach I oroethane
int.
203
165
716
m/z
60
117
167
int.
398
1000
901
m/z
62
119
169
int.
119
979
422
m/z
83
121
Mi
378
306
m/z
85
130
ML
218
293
m/z
94
132
int.
114
272
76
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
923
m/z
51
91
147
924
m/z
74
126
252
925
m/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
pentamethylbenzene
int. m/z
126 53
218
60
perylene
int.
33
243
1000
phenacetin
int.
443
47
50
phenothiazine
int.
145
240
1-phenylnaphth
int.
132
101
144
105
148
m/z
111
224
253
m/z
51
79
137
m/z
51
167
lalene
m/z
51
88
201
2-phenylnaphthalene
int. m/z
108 63
188
pronamide
int.
270
334
102
pyridine
int.
45
112
151
safrole
int.
132
477
109
202
m/z
66
147
257
m/z
48
54
79
m/z
51
105
int.
84
128
420
Mi
43
49
219
int.
33
31
461
int.
120
607
int.
156
183
136
int.
101
398
int.
109
198
122
int.
11
12
1000
int.
369
130
m/z
63
115
m/z
112
248
m/z
52
80
138
m/z
63
198
m/z
63
89
202
m/z
76
203
m/z
74
173
m/z
49
55
80
m/z
63
131
int.
61
120
int.
70
75
int.
112
179
40
int.
134
186
int.
148
162
643
int.
136
270
int.
112
1000
int.
62
16
101
int.
108
437
m/z
65
117
m/z
113
249
m/z
53
31
179
m/z
69
199
m/z
74
100
203
m/z
88
204
m/z
75
175
m/z
50
75
81
m/z
77
132
int.
99
91
int.
111
52
int.
164
154
672
int.
190
1000
int.
124
155
1000
int.
133
1000
int.
137
615
int.
324
21
58
int.
391
166
m/z
77
133
m/z
124
250
m/z
63
108
180
m/z
100
200
m/z
75
101
204
m/z
89
205
m/z
84
254
m/z
51
76
m/z
78
161
Inti
145
1000
int.
132
284
int.
39
1000
64
int.
128
143
int.
142
527
999
int.
158
157
int.
194
133
int.
414
19
int.
228
298
m/z
79
134
m/z
125
251
m/z
64
109
m/z
154
m/z
76
102
205
m/z
101
m/z
109
255
m/z
52
77
m/z
103
162
inr,
64
105
int.
251
86
int.
30
196
int.
149
int.
136
111
159
int.
333
int.
186
211
int.
879
22
int.
348
1000
77
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
932 squalene
m/z int. m/z int.
53 62 55 94
79 43 81 465
109 47 121 46
933 1,2,4, 5-tetrachlorobenzene
m/z int. m/z int.
47 125 49 176
84 197 108 284
181 224 214 791
948 2,3,4,6-tetrachlorophenol
m/z int. m/z int.
61 234 65 167
97 107 131 463
196 164 230 793
934 thianaphthene
m/z int. m/z int.
45 80 50 91
69 139 74 55
135 104 136 52
935 thioacet amide
m/z int. m/z int.
40 225 42 485
59 165 60 437
936 thioxanthone
m/z int. m/z int.
50 262 63 180
92 188 108 129
185 137 212 1000
937 o-toluidine
m/z int. m/z int.
40 51 41 38
52 164 53 192
65 59 66 24
78 113 79 243
104 45 106 1000
938 1,2,3-trimethoxybenzene
m/z int. m/z int.
50 257 51 459
67 114 77 246
107 190 108 144
939 2,4,5-trimethylaniline
m/z int. m/z int.
41 80 52 58
79 62 91 167
120 1000 121 87
m/z
67
82
137
m/z
61
109
216
m/z
66
133
232
m/z
51
89
m/z
43
75
m/z
69
139
213
m/z
42
53
74
80
107
m/z
52
79
110
m/z
51
93
134
Int..
105
52
41
int.
127
231
1000
inti
105
270
1000
int.
65
191
int.
44
1000
int.
320
385
145
int.
35
86
19
80
90
int.
139
132
898
Int..
63
51
670
m/z
68
93
m/z
72
143
218
m/z
83
166
234
m/z
62
90
m/z
46
76
m/z
74
152
m/z
49
62
65
89
m/z
53
82
125
m/z
53
117
135
int.
119
70
int.
183
194
482
int.
134
298
471
int.
82
136
int.
18
25
int.
116
227
int.
10
26
14
107
int.
276
117
578
int.
66
54
978
m/z
69
95
m/z
73
145
220
m/z
84
168
m/z
63
108
m/z
57
77
m/z
69
183
m/z
50
63
76
90
m/z
63
93
153
m/z
65
118
136
int.
1000
104
int.
332
117
101
int.
178
273
int.
162
82
int.
36
43
int.
176
112
int.
88
68
21
76
int.
112
483
759
int.
150
65
99
m/z
70
107
m/z
74
179
m/z
96
194
m/z
67
134
m/z
58
m/z
82
184
m/z
51
64
77
91
m/z
65
95
168
m/z
67
119
int.
57
43
int.
448
237
int.
202
168
int.
78
1000
int.
93
int.
121
951
int.
169
30
313
52
int.
341
801
1000
int.
74
93
78
-------
Appendix A (continued)
Mass Spectra in the Form of Mass/Intensity Lists
940 triphenylene
m/z int.
74 52
114 181
227 132
m/z
87
200
228
int.
55
67
1000
m/z
100
202
229
int.
107
56
184
m/z
101
224
int.
108
84
m/z
112
225
int.
131
56
m/z
113
226
int.
244
313
941 tripropylene glycol methyl ether
m/z int. m/z int. m/z int. m/z int. m/z int. m/z int.
45 492 46 15 47 19 55 17 57 68 58 43
59 1000 60 34 71 16 72 44 73 363 74 232
103 57 117 92 161 21
942 1,3,5-trithiane
m/z int. m/z int. m/z int. m/z int. m/z int. m/z jrrt..
46 1000 47 150 48 98 59 93 60 76 64 136
73 102 91 92 92 111 110 58 138 Z59
79
-------
80
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EPA METHOD 1618
ORGANO-HALIDE PESTICIDES, ORGANO-PHOSPHORUS
PESTICIDES, AND PHENOXY-ACID HERBICIDES BY WIDE
BORE CAPILLARY COLUMN GAS CHROMATOGRAPHY
WITH SELECTIVE DETECTORS
81
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82
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Introduction
Method 1618 Mas developed by the Industrial Technology
Division (ITD) within the United States Environmental
Protection Agency's (USEPA) Office of Water Regulations and
Standards (OWRS) 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.
Method 1618 is a wide bore capillary column gas chromatography
method for analysis of organo-halide and organo-phosphorus
pesticides, 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 method or its 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: July 1989
83
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84
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Method 1618 Revision A July 1989
Organo-haiide Pesticides, Organo-phosphorus Pesticides, and
Phenoxy-acid Herbicides by Wide Bore Capillary Column
Gas Chromatography with Selective Detectors
1 SCOPE AND APPLICATION
1.1 This method Is designed to meet the survey
requirements of the USEPA ITD. The method
is used to determine the organo-halide
pesticides and polychlorinated biphenyls
(PCB's), the organo-phosphorus pesticides,
and the phenoxy-acid herbicides and
herbicide esters, associated with the
Clean Water Act (as amended 1987); the
Resource Conservation and Recovery Act (as
amended 1986); the Comprehensive
Environmental Response, Compensation and
Liability Act (as amended 1986); and other
compounds amenable to extraction and
analysis by automated, wide bore capillary
column gas chromatography (GC) with
halogen specific and organo-phosphorus
detectors.
1.2 The chemical compounds listed in Tables 1
- 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 require-
ments in Section 8.6 of this method give
the steps necessary to determine its
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 (GCHS)
can be used to confirm compounds in
extracts produced by this method when
analyte levels are sufficient.
1.4 The detection limits of this method are
usually dependent on the level of
interferences rather than instrumental
limitations. The limits in Tables 4 - 6
typify the minimum quantities 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 chromatographic
data. Each laboratory that uses this
method must demonstrate the ability to
generate acceptable results using the
procedure in Section 8.2.
2 SUMMARY OF METHOD
2.1 Extraction
2.1.1 The percent solids content of a sample is
determined.
2.1.2 Aqueous samples containing 1 - 30 percent
solids -- The sample is diluted to one
percent solids, if necessary. The
pesticides and PCB's are extracted from a
one liter sample with methylene chloride
using continuous extraction techniques.
For the herbicides, the pH of the sample
is raised to 12 - 13 to hydrolyze esters,
the sample is back-extracted to remove
basic and neutral species, the pH is then
reduced to less than 2, and the sample is
extracted with diethyl ether using
separatory funnel techniques.
2.1.3 Samples containing greater than 30.percent
solids -- The sample is extracted with
acetonitrile and then methylene chloride
using ultrasonic techniques. The extract
is back extracted with two percent (w/v)
sodium sulfate in reagent water to remove
water soluble interferences and residual
acetonitrile. 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-Danish
evaporator, cleaned up (if necessary)
using gel permeation chromatography (GPC),
85
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and/or adsorption chromatography, and/or
solid phase extraction, and then 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 dried over
acidified sodium sulfate and the acids are
derivatized to form the methyl esters.
The solution containing the methyl esters
is cleaned up (if necessary) using
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
compounds, 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.
2.4 Identification of a pollutant (qualitative
analysis) is performed by (1) comparing
the GC retention times of the compound on
two dissimilar columns with the respective
retention times of an authentic standard,
and (2) comparing the concentrations of
the compound determined on the primary and
confirmatory GC systems. Compound
identity is confirmed when the retention
times and amounts agree within their
respective windows.
2.5 Quantitative analysis is performed by
using an authentic 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 The quality of the analysis 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 chroma-
tog rams. 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.
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 ph thai ate 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 4-6.
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SAFETY
5 APPARATUS AND MATERIALS
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'-ODD, 4,4'-DDT, 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
stimers; 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.
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 wi11 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.
5.1 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, 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.
5.1.1.4 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 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 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
5.2.3 Crucibles, porcelain
87
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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-Wolf Extractor,
Cal-Glass, Costa Mesa, 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).
5.3.3 Separatory funnels -- 250, 500, 1000, and
2000 ml, with Teflon stopcocks.
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
5.4.1.1
5.4.1.2
5.4.1.3
5.4.1.4
5.4.1.5
5.4.1,5.1
5.4.1.5.2
5.4.2
5.4.3
5.4.4
5.5
5.5.1
5.5.2
5.6
5.6.1
5.6.1.1
Kuderna-Danish (K-D) apparatus
Evaporation flask -- 500 ml (Kontes K-
570001-0500, or equivalent), attached to
concentrator tube with springs (Kontes K-
662750-0012).
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.
Snyder column -- Three ball macro (Kontes
K-503000-0232, or equivalent).
Snyder column -- Two ball micro (Kontes K-
469002-0219, or equivalent).
Boiling chips
Glass or silicon carbide -- Approx 10/40
mesh, extracted with methylene chloride
and baked at 450 "C for one hr minimum.
Teflon (optional)
methylene chloride.
Extracted with
Water bath -- Heated, with concentric ring
cover, capable of temperature control (±2
°C), installed in a fume hood.
Nitrogen evaporation device -- Equipped
with heated bath that can be maintained at
35 - 40 °C (N-Evap, Organomation
Associates, Inc., or equivalent).
Sample vials -- Amber glass, 1 - 5 mL with
Teflon-lined screw or crimp cap, to fit GC
autosampler.
Balances
Analytical -- Capable of weighing 0.1 mg.
Top loading -- Capable of weighing 10 mg.
Apparatus for sample cleanup.
Automated gel permeation chromatograph
(Analytical Biochemical Labs, Inc,
Columbia, MO, Model GPC Autoprep 1002, or
equivalent).
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
Acrodisc-CR, 1 - 5 micron, 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 mm
path; Pharmacia UV-1, 3 mm flow cell; LDC
Milton-Roy UV-3, monitor #1203; or
equivalent).
5.6.2 Vacuum system and cartridges for solid
phase extraction (SPE)
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 (Analytichem
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 50 ml volumetric flasks
in the manifold.
5.6.2.5 Column -- Mega Bond Elut, Non-polar, C18
Octadecyl, 10 g/60 ml (Analytichem
International Cat. No. 607H060, or
equivalent).
5.6.3 Chromatographic column -- 400 mm x 22 mm
i.d., with Teflon stopcock 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
5.7.4 Funnel, Buchner, 15 cm.
5.7.4.1 Flask, filter, for use with Buchner funnel
5.7.4.2 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 Scintillation vials, glass, 20 - 50 ml,
with Teflon-lined screw caps.
5.10 Gas chromatographs -- Two GC's shall be
employed. Both shall have split I ess 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 14.
5.10.1 GC columns -- Bonded phase fused silica
capillary
5.10.1.1 Primary for organo-halide compounds -- 30
±3 m x 0.5 ±0.05 mm i.d. DB-608, or
equivalent).
5.10.1.2 Primary for organo-phosphate compounds --
DB-1 (or equivalent) with same dimensions
as column for organo-halide compounds.
5.10.1.3 Confirmatory .-- DB-1701, or equivalent,
with same dimensions-^as primary column.
5.10.2 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
89
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(Section 7). Computations of relative
standard deviation (coefficient of
variation) are used for testing
calibration linearity. Statistics on
initial (Section 8.2) and ongoing (Section
14.6) performance shall be computed and
maintained.
5.10.2.3 Data processing -- 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, O.I., or equivalent), capable of
detecting 8 pg of aldrin under the
analysis conditions given in Table 4.
5.10.3.2 Flame photometric -- Capable of detecting
11 pg of ma lath ion under the analysis
conditions given in Table 5.
5.10.4 Chromatographs may be configured in one of
two ways: (1) Two halide specific
detectors (HSD's) in one GC; two flame
photometric detectors (FPD's) 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 (0.1M) -- 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 H.SO^
(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 and back extraction
6.3.1 Sodium sulfate, reagent grade, granular
anhydrous (Baker 3375, or equivalent),
rinsed with methylene chloride (20 ml/3),
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
H2SO^ 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 ±5 °C.
6.3.3 Sodium sulfate solution -- Two percent
(w/v) in reagent water, pH adjusted to 8.5
- 9.0 with KOH or H-SO,.
6.3.4 Sodium sulfate, reagent grade, powdered
anhydrous (Baker 73898, 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.
NOTE: The powdered sodium sulfate is used
only in ultrasonic extraction of samples
containing 30 percent solids or greater,
and not for drying of sample extracts.
Use of granular sodium sulfate during
ultrasonic extraction may lead to poor
recovery of analytes.
6.4 Solvents -- Methylene chloride, hexane,
ethyl ether, acetone, acetonitrile,
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 EM 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.
6.5 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
90
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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 °C for 16
hours minimum immediately prior to use.
Alternatively, 500 mg cartridges (J.T.
Baker, or equivalent) may be used.
6.6.2 Solid phase extraction
6.6.2.1 SPE cartridge calibration solution --
2,4,6-trichlorophenol, 0.1 ug/mL in
acetone.
6.6.2.2 SPE elution solvent -- Methylene
chloride:acetonitrile:hexane (50:3:47).
6.6.3 Alumna, neutral, Brockman Activity I, SO
- 200 mesh (Fisher Scientific Certified,
or equivalent). Heat for 16 hours at 400
- 450 °C. Seal and cool to room
temperature. Add 7 percent w/w reagent
water and mix for 10 - 12 hours. Keep
bottle tightly sealed.
6.6.4 Silicic acid, 100 mesh
6.6.5 Sulfur removal -- Mercury (triple
distilled), copper powder (bright, non-
oxidized), or TBA sodium sulfite. If
mercury is used, observe the handling
precautions in Section 4.
6.7 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.
May 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
6.10
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
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.
6.10.1 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.
6.10.2 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 USEPA, Environmental
Monitoring and Support Laboratory,
Cincinnati, Ohio 45268.
6.10.3 Stock standard solutions shall be replaced
after six months, or sooner if comparison
with quality control check standards
indicates a change in concentration.
6.11 Secondary mixtures -- Using stock
solutions (Section 6.10), prepare mixtures
at the levels required for calibration and
calibration verification (Sections 7.3 and
14.5), for initial and ongoing precision
and recovery (Sections 8.2 and 14.6), and
for spiking into the sample matrix
(Section 8.4).
6.12 Surrogate spiking solutions
6.12.1 Chlorinated pesticides -- Prepare dibutyl
chlorendate at a concentration of 2 ug/mL
in acetone.
6.12.2 Phosphorus containing pesticides
Prepare tributyl phosphate and triphenyl
phosphate each at a concentration of 2
ug/mL in acetone.
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6.12.3 Phenoxyacid herbicides -- Prepare 2,4-
dichlorophenylacetic acid at a
concentration of 2 ug/mL in acetone.
6.13 DOT and endrin decomposition solution --
Prepare a solution containing endrin at a
concentration of 1 ug/mL and DDT at a
concentration of 2 ug/mL.
6.14 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 ±15 percent of the area obtained in
the initial analysis of the standard.
7 SETUP AND CALIBRATION
7.1 Configure the GC systems in one of the two
ways given in Section 5.10.4 and establish
the operating conditions in Tables 4-5.
7.2 Attainment of Method Detection Limit (HDL)
and DDT/Endrin decomposition requirements
-- Determine that each column/detector
system meets the HDL's (Tables 4 - 6) and
that the organohalide systems meet the DOT
and Endrin decomposition test (Section
14.4).
7.3 Calibration
7.3.1 Calibration solutions -- Prepare
calibration standards at a minimum of
three concentration levels for each
compound of interest by adding volumes of
one or more stock standards to a
volumetric flask and diluting to volume
with hexane or isooctane. The lowest
concentration solution should be at a
concentration near, but above, the HDL's
(Tables 4-6). The highest concentration
solution should be near, but below, the
maximum linear range of the analytical
system. The other concentration(s) should
be ideally equally spaced on a logarithmic
scale between the lowest and highest
concentration solutions. The ratio
between the highest and lowest
concentration should be 100 or greater.
Note: the GC retention time overlap
between analytes requires that the
compounds separated and analyzed as
groups. Divide the single component
analytes into three or four calibration
groups each for the organo-halide and
organo-phosphorus compounds with an
approximately equal number of analytes per
group. The compound pairs specified for
GC resolution (Section 14.3) shall be in
the same group. PCS 1254 or 1260 and
Toxaphene are calibrated separately.
7.3.2 Inject the calibration solutions 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 each column/detector system (primary
and confirmatory). For the multicomponent
analytes (PCB's, toxaphene), store the
retention time and peak area for the five
largest peaks.
7.3.3 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.
7.3.3.1 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.
7.3.3.2 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.
7.3.4 Calibration factor (ratio of area to
amount injected)
7.3.4.1 Compute the coefficient of variation
(relative standard deviation) of the
calibration factor over the calibration
range for each compound on each
column/detector system.
7.3.4.2 Linearity -- If the calibration factor for
any compound is constant (Cv < 20 percent)
over the 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.
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7.4 Combined QC standards -- To preclude
periodic analysis of all of the individual
calibration groups of compounds (Section
7.3.1), the GC systems are calibrated with
combined solutions 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 factors are verified for the
compounds that are resolved, and
calibration factors are obtained for the
unresolved peaks. These combined QC
standards are prepared at the level of the
mid-range calibration standard (7.3.1).
7.4.1 Analyze the combined QC standards on their
respective column/detector pairs.
7.4.1.1 For those compounds that exhibit a single,
resolved GC peak, the retention time shall
be within ±5 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 (Section 7.3.4).
7.4.1.2 For the peaks containing two or more
compounds, compute and store the retention
times at the peak maxima on both columns
(primary and confirmatory), and also
compute and store the calibration factors
on both columns. These results will be
used for calibration verification (Section
14.2 and 14.5) and for precision and
recovery studies (Section 14.6).
7.5 Florisil calibration -- The cleanup
procedure in Section 11 utilizes florisil
column chromatography. Florisil from
different batches or sources may vary in
adsorptive capacity. To standardize the
amount of florisil that is used, the use
of the lauric acid value (Reference 4) is
suggested. The referenced procedure
determines 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
matrix spike, and matrix spike duplicate
(MS/MSD) samples to assess accuracy and
precision. 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 separations 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 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).
8.1.4 The laboratory shall, on an ongoing basis,
demonstrate through calibration
verification and the analysis of the
combined QC standard (Section 7.4) that
the analysis system is in control. These
procedures are described in Sections 14.1,
14.5, and 14.6.
8.1.5 The laboratory shall maintain records to
define the quality of data that is
generated. Development of accuracy
statements is described in Section 8.4.
8.1.6 Analyses of blanks are required to
demonstrate freedom from contamination.
93
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The procedures and criteria for analysis
of a blank are described in Section 8.5.
8.1.7 Other analytes may be determined by this
method. The procedure for establishing a
preliminary quality control limit for a
neu 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 analysis of samples containing low
solids (aqueous samples), extract,
concentrate, and analyze one set of four
one-liter aliquots of reagent water spiked
with the combined QC standard (Section
7.4) according to the procedure in Section
10. Alternatively, sets of four
replicates of the individual calibration
groups (Section 7.3) may be used. For
samples containing high solids, sets 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 percent recovery (X)
and the coefficient of variation (Cv) of
percent recovery (s) for each compound.
8.2.3 For each compound, compare s and X with
the corresponding limits for initial
precision and accuracy in Tables 7-9.
For coeluting compounds, use the coeluted
compound with the least restrictive
specification (largest Cv and widest
range). 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. In this case, correct the
problem and repeat the test.
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).
8.3.3 The recovery of the surrogate compound
shall be within the limits of 40 to 120
percent. If the recovery of any surrogate
falls outside of these limits, method
performance is unacceptable for 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 and precision -- The
laboratory shall spike (matrix spike) at
least ten percent of the samples from a.
given site type (e.g., influent to
treatment, treated effluent, produced
water, river sediment) in duplicate
(MS/HSD). If only one sample from a given
site type is analyzed, two aliquots of
that sample shall be spiked.
8.4.1 The concentration of the analytes spiked
into the HS/HSD 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
spiking level 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 matrix
spike shall be at the concentration of the
combined QC standard (Section 7.4) or at
one to five times higher than the
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 matrix spike concentration
shall be 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 7.4).
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 two
additional sample aliquots with the
94
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standard solution and analyze them to
determine the concentrations after spiking
(A) of each analyte. Calculate the
percent recovery (P) of each analyte in
each aliquot:
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 7-9. 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 Determine the precision of the MS/MSD
analyses by comparing the recoveries
calculated in 8.4.2 of each spiked analyte
in both aliquots. Calculate the relative
percent difference (RPD) of the recoveries
(not the concentrations) of each analyte
with MS/MSD aliquots as:
RPD
P - P
MS HSD
/2
x 100
8.4.5 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 = 90%
and sp = 10% for five analyses of compost,
the accuracy interval is expressed as 70 -
110%. 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 anal/zed 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-hour
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 - 3) or any potentially inter-
fering compound is found in an aqueous
blank at greater than 0.05 ug/L, or in a
high solids reference matrix blank at
greater than 1 ug/kg (assuming the same
calibration factor as aldrin and diazinon
for compounds not listed in Tables 1 - 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. If the analyte coelutes with
an analyte in the QC standard, prepare a
new QC standard without the coeluting
component(s). 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; otherwise, the
analytical problem is corrected and the
test is repeated. Establish a preliminary
quality control limit of A ±2sn for the
new analyte and add the limit to Table 7,
8, or 9.
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 ongoing (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 require-
ments, field replicates and field spikes
95
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of the analytes of interest into samples
may be required to assess the precision
and accuracy of the sampling and sample
transporting techniques.
9 SAHPLE 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
Figure 1 outlines the extraction and
concentration steps. Samples containing
one percent solids or less are extracted
directly using continuous liquid/liquid
extraction techniques (Section 10.2.1).
Samples containing one through 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 may be combined
with the organo-chlorine extract for gas
chromatography.
10.1 Determination of percent solids
10.1.1 Weigh 5 - 10 g of sample into a tared
beaker. Record the weight to three
significant 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:
% solids = "ei3ht of dry sample x 10Q
weight of wet sample
10.2 Preparation of samples for extraction
10.2.1 Samples containing one percent solids or
less -- Pesticides and PCB samples are
extracted directly using continuous
liquid/liquid extraction techniques;
herbicides are extracted using separatory
funnel techniques.
10.2.1.1 Shake the samples to ensure thorough
mixing and measure 1.00 ±0.01 liter of
each sample into a separate clean 1.5 -
2.0 liter beaker. Measure a separate one
liter aliquot for each sample to be tested
for the phenoxy-acid herbicides.
10.2.1.2 Spike 0.5 mL of the surrogate spiking
solution (Section 6.12) 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 -- Samples are diluted to one
percent solids and then extracted.
10.2.2.1 Mix sample thoroughly.
10.2.2.2
10.2.2.3
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 grams
% solids
Place the weight of sample as 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.
96
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10.2.2.4 Bring the sample aliquot(s) to 100 - 200
ml volume with reagent water.
10.2.2.5 Spike 0.5 ml of the appropriate surrogate
spiking solution (Section 6.12) into each
sample aliquot.
10.2.2.6 Using a clean metal spatula, break any
solid portions of the sample into small
pieces.
10.2.2.7 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.
10.2.2.8 Bring the sample volume to 1.0 ±0.1 liter
with reagent water.
10.2.3 Preparation of QC aliquots for samples
containing <30 percent solids.
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
7.4) into a reagent water aliquot. For
the herbicides, spike the herbicide
standard into the remaining reagent water
aliquot.
10.2.3.4 If a matrix spike is required, prepare an
aliquot at the concentrations specified in
Section 8.4.
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 ml 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
amount of acid required to acidify the
sample to pH <2. Discard this aliquot.
The volume of H,SO, will be used during
the extraction of the samples in Section
10.4.6.
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 0.5 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 the combined QC standard (Section
7.4) 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 Extraction of low solids (aqueous) samples
10.3.1 Continuous extraction of pesticides/PCB's
-- Place 100 - 150 mL methylene chloride
in each continuous extractor and 200 - 300
ml in each distilling flask.
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10.3.1.1
10.3.1.2
10.3.1.3
10.3.1.4
10.3.2
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.
Extraction -- Adjust the pH of the waters
in the extractors to 5 - 9 with NaOH or
H.SO, 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 Mill 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.
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 (acidified sodium sulfate for the
herbicides). 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, and
concentrate per Sections 10.5 through
10.7.
Hydrolysis
herbicides
and
back-extraction
of
10.3.2.1 Pour the sample and QC aliquot s into
separate 1.5 - 2 L separatory funnels.
Add 250 g NaCl and shake to dissolve.
10.3.2.2 Add 17 mL of 6 N NaOH to each separatory
funnel and shake to mix thoroughly. Check
the pH of the sample and QC aliquots and
adjust to >12 if required. Periodically
shake the aliquots during a 1 - 2 hour
hydrolysis period.
10.3.2.3 Rinse each beaker used for measurement of
the sample and QC aliquots with 60 ml of
methylene chloride, add to its respective
separatory funnel, and extract the sample
by shaking the funnel for two minutes with
periodic venting to release excess
pressure. Allow the organic layer to
separate from the water phase for a
minimum of 10 minutes. If the emulsion
interface between layers is more than one
third the volume of the solvent layer, the
analyst must employ mechanical techniques
to complete the phase separation. The
optimum technique depends upon the sample,
but may include stirring, filtration of
the emulsion through glass wool,
centrifugation, or other physical methods.
Discard the methylene chloride phase. If
the emulsion cannot be broken, continuous
liquid/liquid extraction techniques may be
used. Check and adjust the pH of the
sample to >12 with NaOH if required.
10.3.2.4 Add a second 60 ml volume of methylene
chloride to the sample bottle and repeat
the extraction procedure a second time,
combining the extracts in the Erlenmeyer
flask. Perform a third extraction in the
same manner.
10.3.3 Extraction of the herbicides
10.3.3.1 Add 17 mL of 12 N H-S04 to the sample and
QC aliquots. Seaf and shake to mix.
Caution: some samples require
acidification in a hood because of the
potential for generating hydrogen sulfide.
Check and adjust the pH of the sample to
<2 if required.
10.3.3.2 Add 120 ml ethyl ether to the sample and
QC aliquots. Seal and extract per Section
10.3.2. Drain the aqueous phase
completely into the respective beaker used
for measurement of aliquot volume. Drain
the ether phase into 500 ml round-bottom
flask containing approx 10 g of acidified
sodium sulfate making certain that the
amount of water drained into the flask is
minimized. Periodically, shake the round-
bottom flask to mix the ether solution and
the drying agent.
10.3.3.3 Return the aqueous phase to the separatory
funnel, add a 60 ml volume of ether, and
repeat the extraction a second time.
Drain the aqueous phase completely into
the beaker used for measurement of aliquot
volume and the ether phase into the round-
bottom flask.
10.3.3.4 Repeat the extraction a third time,
combining the ether with the other
extracts in the round-bottom flask. Allow
the sodium sulfate to remain in contact
with the ether solution for a minimum of
two hours, periodically shaking the round-
bottom flask to mix the ether and the
98
-------
drying agent. Concentrate the extract to
5 mL per Sections 10.5 through 10.7.
10.4 Ultrasonic extraction of high solids
aliquots
10.4.1 Add 60 g powdered (not granular) anhydrous
sodium sulfate to the sample and the QC
aliquots. Add 100 ±10 mL acetonitrile to
each of the aliquots (Section 10.2.5) and
mix thoroughly, to produce a free-flowing
mixture.
10.4.2 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.
10.4.3 Decant the pesticide and herbicide
extracts through filter paper into 1000 -
2000 ml separatory funnels.
10.4.4 Repeat the extraction and filtration steps
(Sections 10.4.2 - 10.4.3) using a second
100 ±10 ml of acetonitrile.
10.4.5 Repeat the extraction step (Section
10.4.2) using 100 ±10 mL of methylene
chloride. On this final extraction, swirl
the sample or QC aliquot, pour into its
respective filter paper, and rinse with
methylene chloride. Record the total
extract volume.
10.4.6 For each extract, prepare 1.5 - 2 liters
of reagent water containing two percent
sodium sulfate. For the pesticide
extracts, adjust the pH of the water to
6.0 - 9.0 with NaOH or H2S04- For the
herbicide extracts, adjust the pH of the
water to <2.
10.4.7 Back extract each extract three times
sequentially with 500 ml of the aqueous
sodium sulfate solution, returning the
bottom (organic) layer to the separatory
funnel the first two times while
discarding the top (aqueous) layer. On
the final back extraction, filter each
pesticide extract through a prerinsed
drying column containing 7 to 10 cm
anhydrous sodium sulfate into a 500 - 1000
ml graduated cylinder. Filter the
herbicide extracts similarly using
acidified sodium sulfate. Record the
final extract volume.
10.4.8 Filter the extracts through Whatman #41
paper into 500 mL K-D evaporator flasks
equipped with 10 ml concentrator tubes.
Rinse the graduated cylinder or centrifuge
tube with 30 - 50 ml of methylene chloride
and pour through the filter to complete
the transfer. Concentrate the extracts
per Sections 10.5 through 10.7.
10.5 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 Florisil or silica gel
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.
99
-------
10.6.2 Remove the Snyder column 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 Florisil,
silica gel, and/or sulfur removal
procedures in Section 11.
10.7 Herbicide extracts -- These extracts are
concentrated to 5 - 10 ml and the
herbicides are derivatized per Section 12.
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 shall first repeat
the tests in Section 8.2 to demonstrate
that the requirements of Section 8.2 can
be met using the cleanup procedure(s) as
an integral part of the method. Figure 1
outlines the cleanup steps.
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 solid phase extraction cartridge
(Section 11.3) removes polar organic
compounds such as phenols. It is used for
cleanup of organo-chlorine and organo-
phosphate extracts.
11.1.3 The Florisil column (Section 11.4) allows
for selected fractionation of the organo-
chlorine compounds and will also eliminate
polar interferences.
11.1.4 Alumina column cleanup (Section 11.5) may
also be used for cleanup of the organo-
chlorine compounds.
11.1.5 Elemental sulfur, which interferes with
the electron capture gas chromatography of
some of the pesticides and herbicides, is
removed using GPC, mercury, or activated
copper. Sulfur removal (Section 11.6)
from extracts containing organo-chlorine
is required when sulfur is known or
suspected to be present. Mercury and
copper should not be used for sulfur
removal from extracts expected to contain
the organo-phosphorus pesticides because
some of these analytes are also removed
(Reference 8).
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
recovery of the pentachlorophenol is
greater than 85 percent. If calibration
is not verified, the system shall be
recalibrated using the calibration
100
-------
< 30% SOLIDS
Percent Solids
> 30% SOLIDS
Dilute to 1% Solids
ACN and CH2CL2 Sonication
CH2CL2 Liquid/Liquid
Extraction
Back Extraction
Concentrate
Concentrate
To Cleanup To Cleanup
Method 1618 - Extraction and Concentration Steps
Gel Permeation Cleanup
ORGANaPHOSPHORUS
Solid Phase Extraction
ORGANOCHLORINE
GCFPD
Horisil
Remove Sulfur
GCHSD
Method 1618 - Cleanup and Analysis Steps
FIGURE 1 Method 1618 - Extraction, Cleanup, and Analysis
101
-------
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
gravimetricly by evaporating the solvent
from a 50 uL aliquot.
11.2.3.1 Filter the extract or load through the
filter holder to remove participates.
Load the 5.0 mL extract onto the column.
11.2.3.2 Elute the extract using the calibration
data determined in Section 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 extracts per Sections 10.5
- 10.7.
11.3 Solid phase extraction (SPE)
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 SPE 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 sequentially with
10 ml portions each of hexane, methanol,
and water using vacuum for 30 seconds
after each eluant. Follow this pre-
elution with 1 ml methylene chloride and
three 10 ml portions of the elution
solvent (6.6.2.2) using vacuum for five
minutes after each eluant. Tap the
cartridge lightly while under vacuum to
dry between eluants. The three portions
of elution solvent may be collected and
used as a blank if desired. Finally,
elute the cartridge with 10 ml each of
methanol and water, using the vacuum for
30 seconds after each eluant.
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 test mixture, add the
trichlorophenol solution (Section 6.6.2.1)
to the combined calibration standard
(Section 7.4). Elute the mixture using
the procedure in 11.3.4.
11.3.3.2 Concentrate the eluant to 1.0 ml and
inject 1.0 uL of the concentrated eluant
into the GC using the procedure in Section
13. The recovery of all organo-chlorine
or organo-phosphorus analytes (including
the unresolved GC peaks) shall be within
the ranges for recovery specified in
Tables 7-8, and the peak for
trichlorophenol shall not be detectable;
otherwise the SPE cartridge is not
performing properly and the cartridge lot
shall be 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 50 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 the SPE
cartridge. Apply vacuum for five minutes
to dry the cartridge. Tap gently to aid
in drying.
11.3.4.3 Elute each cartridge into its volumetric
flask sequentially with three 10 mL
portions of the elutions solvent
(6.6.2.2), using vacuum for five minutes
after each portion. Collect the eluants
in the 50 mL volumetric flasks.
11.3.4.4 Release the vacuum and remove the 50 mL
volumetric flasks.
11.3.4.5 Concentrate the eluted extracts to 1.0 mL
using the nitrogen blow-down apparatus.
Adjust the final volume to 5 or 10 mL (per
Section 10.6), depending on whether or not
102
-------
the extract was subjected to GPC cleanup,
and proceed to Section 13 for GC analysis.
11.4 Florisil column
11.4.1 Place a weight of Florisil (nominally 20
g) predetermined by calibration (Section
7.5) 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 percent
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).
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 alumina.
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 extrapt 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 i sopropanoI.
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 sodium 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.
11.6.3 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.
11.6.4 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 ESTERIFICATION OF PHENOXY-ACID HERBICIDES
12.1 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.
103
-------
12.2 Esterification -- Observe the safety
precautions regarding diazomethane in
Section 4.
12.2.1 Set up the diazomethane generation
apparatus as given in the instructions in
the Diazald kit.
12.2.2 Transfer one mL of the isooctane solution
(Section 12.1} 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 to account for the loss.
12.2.3 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.
12.2.4 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 11.3 for
SPE 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 through 6 summarize the
recommended operating conditions for the
gas chromatographs. 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 confirmatory columns are shown in
Figures 2 through 6.
13.1 Calibrate the system as described in
Section 7.
13.2 Combining pesticide and herbicide extracts
13.2.1 Pesticide extracts cleaned up by solid
phase extraction -- Combine the 1.0 mL
final organo-chlorine pesticide extract
(Section 11.3.4.5 or 11.5.4) with the 1.0
mL final herbicide extract (Section
11.3.4.5 or 11.5.4 if the herbicide
extract required cleanup; Section 12.2.4
if it did not).
13.2.2 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 11.5.4 i,f the
herbicide extract required cleanup;
Section 12.2.4 if it did not).
13.3 Set the injection volume on the
autosampler to inject 1.0 uL of all
standards and extracts of blanks and
samples.
13.4 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 after the
last analyte is expected to elute 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
verified for all pollutants and surrogates
on all column/detector systems. For these
tests, analysis of the combined QC
standard (Section 7.4) shall be used to
verify all performance criteria.
Adjustment and/or recall brat 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 -- The absolute retention
times of the peak maxima shall be within
±10 seconds of the retention times in the
initial calibration (Section 7.4.1).
14.3 GC resolution -- Resolution is acceptable
if the valley height between two peaks (as
measured from the baseline) is less than
10 percent of the taller of the two peaks.
104
-------
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FIGURE 5 Organophosphate Mix B [(A) DB-1 and (B) DB-1701].
108
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-------
14.3.1 Organo-halide compounds
14.3.1.1 Primary column (DB-608) -- DDT and endrin
aldehyde.
14.3.1.2 Confirmatory column (DB-1701) -- Alpha and
gamma chlordane.
14.3.2 Organo-phosphorus compounds
14.3.2.1 Primary column (DB-1) -- Malathion and
ethyl parathion.
14.3.2.2 Confirmatory column (DB-1701) -- Terbufos
and diazinon.
14.4 Decomposition of DDT and endrin
14.4.1 Analyze a total of 2 ng DDT and 1 ng
endrin on each organo-chlorine column
using the analytical conditions specified
in Table 4.
14.4.2 Measure the total area of all peaks in the
chromatogram.
14.4.3 The area of peaks other than the sun of
the areas of the DDT and endrin peaks
shall be less than 20 percent the sum of
the areas of these two peaks. If the area
is greater than this sum, the system is
not performing acceptably for DDT and
endrin. In this case, the GC system that
failed shall be repaired and the
performance tests (Sections 14.1 - K.4)
shall be repeated until the specification
is met. Note: DDT and endrin
decomposition are usually caused by
accumulations of particulates in the
injector and in the front end of the
column. Cleaning and silanizing the
injection port liner, and breaking off a
short Section of the front end of the
column will usually eliminate the
decomposition problem.
14.5 Calibration verification -- Calibration is
verified for the combined QC standard
only.
14.5.1 Inject the combined QC standard (Section
7.4)
14.5.2 Compute the percent recovery of each
compound or coeluting compounds, based on
the calibration data (Section 7.4).
14.5.3 For each compound or coeluted compounds,
compare this calibration verification
recovery with the corresponding limits for
ongoing accuracy in Tables 7 - 9. For
coeLuting compounds, use the coeluted
compound with the least restrictive
specification (the widest range). If the
recoveries for all compounds meet the
acceptance criteria, system performance is
acceptable and analysis of blanks and
samples may begin. If, however, any
recovery falls outside the calibration
verification range, system performance is
unacceptable for that compound. In this
case, correct the problem and repeat the
test, or recalibrate (Section 7). If
verification requirements are met, the
calibration is assumed to be valid for the
mult{component analytes (PCB's and
toxaphene).
14.6 Ongoing precision and recovery
14.6.1 Analyze the extract of the precision and
recovery standard extracted with each
sample lot (Sections 10.2.3.3 and
10.2.5.7).
14.6.2 Compute the percent recovery of each
analyte and coeluting compounds.
14.6.3 For each compound or coeluted compounds,
compare the percent recovery with the
limits for ongoing recovery in Tables 7 -
9. For coeluted compounds, use the
coeluted compound with the least
restrictive specification (widest range).
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 ongoing precision and recovery
test.
14.6.4 Add results which pass the specifications
in Section 14.6.3 to initial and previous
ongoing data. Update QC charts to form a
graphic representation of continued
laboratory performance. Develop a
statement of 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 = 95%
and sr = 5%, the accuracy is 85 - 105%.
110
-------
15 QUALITATIVE DETERMINATION
16 QUANTITATIVE 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 14.2), and
with data stored in the retention time and
calibration libraries (Section 7.3.3 and
7.3.4.1). Identification is confirmed
when retention time and amounts agree per
the criteria below.
15.2 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 7.3.3). For compounds that have
a retention time curve (Section 7.3.3.2),
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 7.3.3).
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 7.3.3.2), and
(2) the computed amounts (Section 16) on
each system (primary and confirmatory)
agree within a factor of three.
16.1
16.2
16.3
16.A
16.5
16.6
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).
Liquid samples
concentration in the
following equation:
Compute
sample using
the
the
Cs =
where,
Cs =
10 =
Cex =
Vs =
10 (Cex)
(Vs)
the concentration in the sample
in ug/L.
extract total volume in mL.
concentration in the extract in
ug/mL.
volume of sample extracted in
liters.
Solid samples -- Compute the concentration
in the solid phase of the sample using the
following equation:
Cs =
where,
Cs
10
Cex
1000
Us
% solids
10 (Cex)
1000 (Us) (% solids)
concentration in the sample
in ug/kg.
extract total volume in mL.
concentration in the extract
in ug/mL.
used to convert grams to
kilograms.
sample weight in grams.
percent solids as determined
in Section 10.1.3.
If the concentration of any analyte
exceeds the calibration range of the
system, the extract is diluted by a factor
of 10, and a one uL aliquot of the diluted
extract is analyzed.
Two or more PCB's in a given sample are
quantitated and reported as total PCB.
Report results for all pollutants found in
all standards, blanks, and samples to
three significant figures. Results for
samples that have been di luted are
reported at the least dilute level at
which the concentration is in the
calibration range.
Ill
-------
17 ANALYSIS OF COMPLEX SAMPLES
17.1 Some samples may contain high levels
(>1000 ng/L) of the compounds 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 11.2), and the
SPE cartridge (Section 11.3), and samples
for the organo-halide compounds by
florisil (Section 11.4) or alumina (11.5),
and sulfur removal (Section 11.6). If
these techniques do not remove the
interfering compounds, the extract is
diluted by a factor of 10 and reanalyzed
(Section 16.4).
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 specified in
Section 8.3, the sample shall be
reextracted and reanalyzed. If the
surrogate recovery is still outside this
range, the sample is diluted by a factor
of 10 and reanalyzed (Section 16.4).
17.4 Recovery of matrix spikes -- In most
samples, matrix spike recoveries will be
similar to those from reagent water or
from the high solids reference matrix. If
the matrix spike recovery is outside the
range specified in Tables 7-9, the
sample shall be diluted by a factor of 10,
respiked, and reanalyzed. If the matrix
spike recovery is still outside the range,
the method does not apply to the sample
being analyzed and the result may not be
reported for regulatory compliance
purposes.
18 METHOD PERFORMANCE
18.1
REFERENCES
1
Development of this method is detailed in
Reference 10.
"Working with Carcinogens," DHEW, PHS,
CDC, NIOSH, Publication 77-206, (August
1977).
"OSHA Safety and Health Standards, General
Industry" OSHA 2206, 29 CFR 1910 (January
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 Wastewater Laboratories," USEPA,
EMSL, Cincinnati, OH 45268, EPA-600/4-79-
019 (March 1979).
6 "Standard Practice for Sampling Water,"
ASTH Annual Book of Standards, ASTH,
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 "Method Development and Validation, EPA
Method 1618, Cleanup Procedures", Colorado
State University, Department of
Environmental Health, Colorado Pesticide
Center, November 1988 and January 1989.
9 Goerlitz, D.F., and Law, L.M. "Bulletin
for Environmental Contamination and
Toxicology,1' 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 Jolla, CA, 92038-1620
(September 1986).
112
-------
Table 1
ORGANO-HALIDE PESTICIDES DETERMINED BY WIDE BORE,
FUSED SILICA CAPILLARY COLUMN GAS CHROMATOGRAPHY
WITH HALIDE SPECIFIC DETECTOR
Table 2
ORGANO-PHOSPHORUS PESTICIDES DETERMINED BY WIDE BORE,
FUSED SILICA CAPILLARY COLUMN GAS CHROMATOGRAPHY
WITH FLAME PHOTOMETRIC DETECTOR
EGD
No.
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
Chlorobenzf late
4,4' -ODD
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 (TDK)
PCB-1016
PCB-1221
PCS- 1232
PCS- 1242
PCS- 1248
PCS- 1254
PCB-1260
PCNB (pentachloronitrobenzene)
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
EGD
No.
468
453
461
469
443
479
471
460
450
455
449
452
458
467
463
446
454
447
464
474
475
456
444
470
459
448
457
465
473
477
476
472
466
445
451
462
Compound
Azinphos ethyl
Azinphos methyl
Chlorfevinphos
Chlorpyrifos
Coumaphos
Crotoxyphos
Demeton
Diazinon
Dichlorvos
Dicrotophos
Dimethoate
Dioxathion
Disulfoton
EPN
Ethion
Famphur
Fensulfothion
Fenthion
Hexamethylphosphoramide
Leptophos
Ma lath ion
Methyl parathion
Mevinphos
Monocrotophos
Naled
Parathion
Phorate
Phosmet
Phosphamidon
Sulfotepp
TEPP
Terbufos
Tetrachlorvinphos
Trichlorofon
Tricresylphosphate
Trimethylphosphate
NON-ITD ORGANO-PHOSPHATE COMPOUNDS
CAS Registry
2642-71-9
86-50-0
470-90-6
2921-88-2
56-72-4
7700-17-6
8065-48-3
333-41-5
62-73-7
141-66-2
60-51-5
78-34-2
298-04-4
2104-64-5
563-12-2
52-85-7
115-90-2
55-38-9
680-31-9
21609-90-5
121-75-5
298-00-0
7786-34-7
6923-22-4
300-76-5
56-38-2
298-02-2
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
THAT CAN BE
ANALYZED BY THIS METHOD
NON-ITD ORGANO-HALIDE COMPOUNDS THAT CAN BE
ANALYZED BY THIS METHOD
Compound
Chloroneb
Chloropropylate
DBCP
Dicofol
Etridiazole
Perthane (Ethylan)
Propachlor
Strobane
CAS Registry
2675-77-6
5836-10-2
96-12-8
115-32-2
2593-15-9
72-56-0
1918-16-7
8001-50-1
Compound
Bolstar
D i chIorofenth i on
Ethoprop
Merphos
Methyl chlorpyrifos
Methyl trithion
Ronnel
Sulprofos
Tokuthion
Trichloronate
CAS Registry
35400-43-2
97-17-6
13194-48-4
150-50-5
5598-13-0
953-17-3
299-84-3
35400-43-2
34643-46-4
327-98-0
113
-------
Table 3
PHENOXYACID HERBICIDES DETERMINED BY WIDE BORE,
FUSED SILICA CAPILLARY COLUMN GAS CHROMATOGRAPHY
WITH HALIDE SPECIFIC DETECTOR
EGD
No.
481
480
482
483
Compound
2,4-D
Dinoseb
2,4,5-T
2,4,5-TP
CAS Registry
94-75-7
88-85-7
93-76-5
93-72-1
NON-ITD PHENOXYACIO HERBICIDES THAT CAN BE
ANALYZED BY THIS METHOD
Compound CAS Registry
Dalapon 75-99-0
2,4-DB (Butoxon) 94-82-6
Oicamba 1918-00-9
Dichlorprop 120-36-5
MCPA 94-74-6
MCPP 93-65-2
114
-------
Table 4
GAS CHROMATOGRAPHY OF ORGANO-HALIDE PESTICIDES
EGO
No.
442
432
102
440
104
103
100
478
105
089
437
101
091
095
093
090
433
431
098
436
439
094
096
092
441
099
097
434
438
Compound
Trif luralin
Diallate-A
Diallate-B
alpha-BHC
PCNB
gamma-BHC (Lindane)
beta-BHC
Heptachlor
Dichlone
delta-BHC
Aldrin
Isodrin
Keptachlor epoxide
gamma-Chlordane
alpha-Chlordane
Endosulfan I
4, 4' -DDE
Dieldrin
Captan
Chlorobenzilate
Endrin
Nitrofen (TOK)
Kepone
4,4' -ODD
Endosulfan II
4,4'-DDT
Carbophenothion
Endrin aldehyde
Endosulfan sulfate
Captafol
Mi rex
Retention
DB-608
5.16
7.15
7.42
8.14
9.03
9.52
9.86
10.66
10.80
11.20
11.84
13.47
13.97
14.63
15.24
15.25
16.34
16.41
16.83
17.58
17.80
17.86
17.92
18.43
18.45
19.48
19.65
19.72
20.21
22.51
22.75
Time (1)
DB-1701
8.58
8.05
8.58
9.45
9.91
10.84
13.58
11.56
(3)
14.39
12.50
13.93
15.03
16.20
16.48
15.96
16.76
17.32
17.32
18.97
18.06
19.14
25.03
19.56
19.72
20.10
20.21
21.18
22.36
23.11
21.82
HDL (2)
(ng/L)
50 est
45
32
6
6
11
7
5
(4)
5
8
13
12
9
8
11
10
6
(4)
25
4
13
(4)
5
8
12
50
11
7
(4)
4
EGD
No.
430
435
106
109
112
108
110
107
111
113
(1)
Compound
Methoxychlor
Endrin ketone
PCB-1242
PCB-1232
PCB-1016
PCB-1221
PCB-1248
PCB-1254
PCB-1260
Toxaphene
Columns: 30 m x
micron; DB-1701: 1.
Conditions: 150 °
Retention Time (1)
DB-608 DB-1701
22.80 22.34
23.00 23.71
15.44 14.64
15.73 15.36
16.94 16.53
17.28 18.70
19.17 19.92
16.60 16.60
17.37 17.52
18.11 17.92
19.46 18.73
19.69 19.00
HDL (2)
(ng/L)
30
8
140
910
0.53 ma i.d.; DB-608: 0.83
0 micron.
C for 0.5 min, 150 -
°C per minute, 270 °C until endrin
elutes.
270 3 5
ketone
Carrier gas flow rate: approximately 7 mL/min.
(2)
(3)
(4)
40 CFR Part 136,
Detection limits
estimated to be 30
Appendix B (49 FR 43234).
for soils (in ng/kg) are
- 100 times this level.
Does not elute from DB-1701 column at level
tested.
Not recovered from
water at levels tested
•
115
-------
Table 5
GAS CHROMATOGRAPHY OF ORGANO-PHOSPHORUS PESTICIDES
EGO
NO.
450
444
445
471
459
455
470
477
457
449
452
472
473
458
460
456
475
447
448
469
Retention
Compound
Dichlorvos
Mevinphos
Trichlorofon
Demeton-A
Ethoprop
Naled
Oicrotophos
Monocrotophos
Sulfotepp
Phorate
Dimethoate
Demeton-B
Dioxathion
Terbufos
Phosphamidon-E
Disulfoton
Diazinon
Tributyl phosphate
(surr)
Phosphamidon-Z
Methyl parathion
D i ch lorof enth i on
Methyl chlorpyrifos
Ronnel
Ma lath ion
Fenthion
Parathion (ethyl)
Chlorpyrifos
Trichloronate
DB-608
6.
11.
12.
56
85
69
17.70
18.
18.
19.
19.
20.
20.
20.
21.
22.
22.
23.
23.
24.
24.
25.
25.
26.
26.
27.
28.
29.
29.
29.
30.
49
92
33
62
04
12
59
40
24
97
70
89
03
50
88
98
11
29
33
87
14
29
48
44
Time
(1)
DB-1701
9.
16.
18.
20.
21.
23.
26.
29.
23.
23.
29.
25.
26.
24.
29.
27.
26.
17.
32.
32.
28.
29.
30.
33.
32.
34.
32.
32.
22
20
85
57
43
00
30
24
68
08
29
52
70
55
89
01
10
20
62
12
66
53
09
49
16
61
15
12
MDL (2)
(ng/L)
4
74
150 (3)
19
7
18
81
85
6
10
27
21
121
26
28
32
38
-
116
18
6
13
11
11
22
10
4
14
EGO
No.
461
479
466
454
463
446
465
467
453
474
468
443
(1)
(2)
(3)
Compound
Chlorfevinphos
Crotoxyphos
Tokuthion
Tetrach lorvi nphos
Merphos-B
Fensulfothion
Methyl trithion
Ethion
Sulprofos (Bolstar)
Famphur
Phosmet
EPN
Azi nphos methyl
Leptophos
Azi nphos ethyl
Triphenyl phosphate
(surr)
Coumaphos
Retention
DB-608
32.05
32.65
33.30
33.40
35.16
36.58
36.62
37.61
38.10
38.24
41.24
41.94
43.33
44.32
45.55
47.68
48.02
Columns: 30 m x 0.53 mm i.d.;
DB-1701: 1.0 micron.
Conditions: 110 °C
°C per minute, 250 °C
Carrier gas flow rate
Time (1) MDL
(2)
DB-1701 (ng/L)
36.08
37.58
37.17
37.85
37.37
43.86
40.52
41.67
41.74
46.37
48.22
47.52
50.26
47.36
51.88
40.43
56.44
2
81
2
12
18
104
10
13
6
27
14
9
9
14
22
-
24
DB-1: 1.5 micron;
for 0.5 min, 110 - 250
a 3
until coumaphos elutes.
: approximately 7 mL/min.
40 CFR Part 136, Appendix B (49
FR 43234).
Estimated: Detection limits for soils
ng/kg) are estimated
level.
to be 30
- 100 times
(in
this
116
-------
Table 6
GAS CHROMATOGRAPHY OF PHENOXY-ACID
HERBICIDES
EGD
No.
481
480
482
483
Compound
2,4-D
Dinoseb
2,4,5-T
2,4,5-TP (Silvex)
Dalapon
2,4-DB (Butoxon)
Dicamba
Dichlorprop
MCPA
MCPP
Retention
DB-608
5.85
7.92
6.97
8.74
4.39
5.15
4.74
4.24
Time (1)
DB-1701
6.05
8.20
7.37
9.02
4.39
5.46
4.94
4.55
MDL (2)
(ng/L)
100
100 est
50
40
1000 est
50
110
40
90
56
(1) Columns: Same as for the organo-chlorine
pesticides. See Table 4.
Conditions: 175 "C for 0.5 min. 175 - 270 a 5
"C per minute.
Carrier gas flow rate: approximately 7 mL/min.
(2) 40 CFR Part 136, Appendix B (49 FR 43234).
Detection limits for soils (in ng/kg) are
estimated to be 30 - 100 times this level.
117
-------
Table 7
ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS FOR ORGANO-HALIDE COMPOUNDS
Acceptance Criteria
EGD
No.(D Compound
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
(1)
(2)
Aldrin
alpha-BHC
beta-BHC
delta-BHC
gamma-BHC (Lindane)
Captafol (2)
Captan (2)
Carbophenoth i on
Chlordane-alpha
Chlordane- gamma
Chlorobenzilate
4,4'-DDD
4,4'-DDE
4,4'-DDT
Diallate
Dichlone (2)
Dieldrin
Endosulfan I
Endosulfan II
Endosulfan sulfate
Endrin
Endrin aldehyde
Endrin ketone
Heptachlor
Heptachlor epoxide
Isodrin
Kepone (2)
Methoxychlor
Mi rex
Nitrofen (TDK)
PCB-1016
PCB-1221
PCB-1232
PCB-1242
PCB-1248
PCB-1254
PCB-1260
PCNB
Toxaphene
Trif luralin
Reference numbers beginning
Not recovered.
Spike
level
(ng/L)
100
100
100
100
100
1000
100
100
500
100
200
200
250
100
200
200
100
100
200
100
100
100
100
200
100
200
1000
100
5000
200
with 0 or 1 indicate a
Initial
precision
and accuracy
Sec 8.2 (%)
s
12
10
10
24
10
10
10
13
19
12
13
19
16
11
14
19
17
13
13
25
12
13
15
19
23
22
20
11
20
12
pollutant
X
82 -
57 -
66 -
60 -
66 -
63 -
79 -
32 -
58 -
69 -
66 -
86 -
44 -
79 -
66 -
41 -
78 -
50 -
17 -
0 -
36 -
78 -
63 -
69 -
50 -
25 -
15 -
82 -
49 -
82 -
32 -
Recovery
Sec 8.4
Calibration Ongoing
verification accuracy
Sec 14.5 Sec 14.6
(ug/mL)
108
135
130
122
112
141
122
140
118
117
114
112
120
110
140
133
142
130
149
149
126
104
117
113
136
155
139
112
129
112
148
quantified by
79
69
85
79
75
70
49
79
73
79
54
77
81
77
70
48
78
76
70
5
86
68
80
79
71
47
47
78
59
79
78
68
47
the
- 113
- 108
- 102
- 103
- 119
- 107
- 114
- 102
- 102
- 113
- 129
- 109
- 121
- 118
- 124
- 115
- 119
- 119
- 109
- 117
- 117
- 135
- 114
- 117
- 126
- 134
- 128
- 114
- 142
- 126
- 101
- 134
- 134
internal
R <%)
76 -
38 -
50 -
45 -
55 -
43 -
69 -
4 -
43 -
57 -
54 -
79 -
24 -
48 -
18 -
62 -
31 -
0 -
0 -
14 -
71 -
49 -
45 -
28 -
0 -
0 -
75 -
29 -
76 -
3 -
114
154
146
136
123
161
133
169
133
129
126
119
139
158
156
158
149
182
190
148
111
131
127
158
188
170
119
149
122
177
standard method.
118
-------
Table 8
ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS FOR ORGANO-PHOSPHORUS COMPOUNDS
Acceptance Criteria
EGD
No.
468
453
461
469
443
479
471
460
450
455
449
452
458
467
463
446
454
447
464
474
475
456
444
470
459
448
457
465
473
477
476
472
466
445
451
462
Compound
Azinphos ethyl
Azinphos methyl
Chlorfevinphos
Chlorpyrifos
Counaphos
Crotoxyphos
Demston-S
Diazinon
Dichlorvos
Dicrotophos (1)
Dimethoate
Dioxathion
Disulfoton
EPN
Ethion
Famphur
Fensulfothion
Fenthion
Hexamethylphosphoramide (1)
Leptophos
Ma lath ion
Methyl para th ion
Mevinphos
Monocrotophos (1)
Naled
Parathion
Phorate
Phosmet
Phosphamidon-Z
Sulfotepp
TEPP (1)
Terbufos
Tet rach lorvi nphos
Trichlorofon (1)
Tricresylphosphate
Trimethylphosphate (1)
D i ch I orof enth i on
Ethoprop
Merphos-B
Methyl chlorpyrifos
Methyl trithion
Ronnel
Sulprofos (Bolstar)
Tokuthion
Trichloronate
Spike
level
(ng/L)
100
100
50
50
50
200
200
100
50
100
600
100
100
100
200
200
100
100
100
100
100
100
100
100
200
330
50
100
100
300
100
100
200
100
100
100
50
100
100
Initial
precision
and accuracy
Sec 8.2 (%)
s
10
10
11
10
10
46
23
10
18
89
22
30
13
11
12
65
13
10
10
15
23
10
10
19
39
45
10
23
11
10
10
14
10
10
20
10
10
17
10
X
71 -
52 -
56 -
61 -
78 -
28 -
33 -
70 -
52 -
27 -
59 -
46 -
74 -
72 -
81 -
13 -
69 -
85 -
75 -
72 -
24 -
0 -
71 -
54 -
44 -
0 -
70 -
60 -
48 -
81 -
75 -
79 -
68 -
88 -
21 -
79 -
75 -
73 -
82 -
117
112
132
112
104
116
101
110
106
100
101
98
124
134
101
115
101
105
109
112
100
148
111
100
119
100
120
110
110
101
115
103
102
108
137
111
100
105
102
Calibration
verification
Sec 14.5
(ug/mL)
77 -
83 -
83 -
80 -
82 -
68 -
64 -
86 -
77 -
73 -
79 -
70 -
81 -
70 -
81 -
42 -
73 -
85 -
82 -
89 -
73 -
77 -
79 -
70 -
61 -
81 -
75 -
82 -
73 -
70 -
80 -
84 -
72 -
81 -
78 -
78 -
81 -
70 -
80 -
127
119
114
119
120
136
123
114
103
127
121
118
108
118
113
139
137
112
108
114
135
114
110
118
159
102
115
111
119
130
110
108
118
114
122
113
118
130
113
Recovery
Sec 8.4
Ongoing
accuracy
Sec 14.6
R (%)
59 -
37 -
37 -
48 -
72 -
6 -
16 -
60 -
39 -
78 -
22 -
49 -
33 -
62 -
47 -
76 -
0 -
61 -
70 -
80 -
66 -
61 -
7 -
19 -
0 -
61 -
43 -
25 -
0 -
58 -
70 -
47 -
32 -
70 -
74 -
70 -
65 -
73 -
59 -
83 -
0 -
71 -
70 -
65 -
77 -
129
127
151
125
110
138
118
120
119
122
100
111
111
136
149
106
141
109
130
110
118
123
107
206
176
121
109
138
100
132
130
123
126
130
114
130
125
109
111
113
166
119
100
113
107
(1) Not recovered.
119
-------
Table 9
ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS FOR PHENOXY-ACID COMPOUNDS
Acceptance Criteria
EGO
No. Compound
Spi ke
level
(ng/L)
Initial
precision
and accuracy
Sec 8.2 (%)
s X
Calibration
verification
Sec 14.5
(ug/mL)
Recovery
Sec 8.4
Ongoing
accuracy
Sec 14.6
R (%)
481 2,4-D
480 Dinoseb
482 2,4,5-T
483 2,4,5-TP (Si Ivex)
Dalapon
2,4-DB (Butoxon)
Dicamba
Dichlorprop
HCPA
HCPP
200
100
100
100
200
100
200
400
16
41 - 107
17 30 - 132
14 36 - 120
16
18
14
14
14
22-118
37 - 145
49 - 133
46 - 130
65 - 149
70 - 130
70 - 130
70 - 130
70 - 130
70 - 130
70 - 130
70 - 130
70 - 130
23 - 131
5 - 158
15 - 141
0 - 142
10 - 172
28 - 154
25 - 151
42 - 170
120
-------
EPA METHOD 1613
TETRA- THROUGH OCTA- CHLORINATED DIOXINS
AND FURANS BY ISOTOPE DILUTION HRGC/HRMS
121
-------
122
-------
Introduction
Method 1613 was developed by the Industrial Technology
Division (ITD) within the United States Environmental
Protection Agency's (USEPA) Office of Water Regulations and
Standards (OWRS) 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
effluent limitation guidelines for pollutant levels in
industrial and municipal discharges.
Method 1613 is a high resolution capillary column gas
chromatography (HRGO/high resolution mass spectrometry (HRMS)
method for analysis of tetra- through octa- chlorinated
dibenzo-p-dioxins and dibenzofurans using isotope dilution.
Specificity is provided for determination of the seventeen
2,3,7,8-substituted polychorinated dibenzo-p-dioxins (PCDD)
and polychlorinated dibenzofurans (PCDF).
Questions concerning the method or its application should be
addressed to:
W. A. Telliard, Chief
Analytical Methods Staff (WH-552)
USEPA Office of Water Regulations and Standards
401 M Street, S.W.
Washington, DC 20460
202/382-7120
OR
USEPA OWRS
Sample Control Center
P.O. Box 1407
Alexandria, Virginia 22313
703/557-5040
Revision A Publication Date - April 1990; Updated - June 1990
123
-------
124
-------
Method 1613 Revision A April 1990
Tetra- through Oeta™ Chlorinated Dioxins and Furans
by Isotope Dilution HRGC/HRMS
1 SCOPE AND APPLICATION
1.1 This method is designed to meet the survey
requirements of the USEPA ITD. The method
is used to determine the tetra- through
octa- chlorinated dibenzo-p-dioxins and
dibenzofurans associated with the Clean
Water Act (as amended 1987); the Resource
Conservation and Recovery Act (as amended
1986); and the Comprehensive Environmental
Response, Compensation and Liability Act
(as amended 1986); and other dioxin and
furan compounds amenable to high resolu-
tion capillary column gas chromatography
(HRGC)/high resolution mass spectrometry
(HRMS). Specificity is provided for de-
termination of the 17 2,3,7,8-substituted
polychlorinated dibenzo-p-dioxins (PCDD)
and polychlorinated dibenzofurans (PCDF).
1.2 The method is based on EPA, industry, com-
mercial laboratory, and academic methods
(References 1-6).
1.3 The compounds listed in Table 1 may be
determined in waters, soils, sludges, and
other matrices by this method.
1.4 The detection limits of the method are
usually dependent on the level of inter-
ferences rather than instrumental limita-
tions. The levels in Table 2 typify the
minimum quantities that can be determined
in environmental samples using the method.
1.5 The GCMS portions of the method are for
use only by analysts experienced with
HRGC/HRMS or under the close supervision
of such qualified persons. Each labora-
tory that uses this method must demon-
strate the ability to generate acceptable
results using the procedure in Section
8.2.
2 SUMMARY OF METHOD
2.1 Stable isotopically labeled analogs of 15
of the PCDDs and PCDFs are added to each
sample prior to extraction. Samples con-
taining coarse solids are prepared for
extraction by grinding or homogenization.
Water samples are filtered and then
extracted with methylene chloride using
separatory funnel procedures; the particu-
lates from the water samples, soils, and
other finely divided solids are extracted
using a combined Soxhlet extraction/Dean-
Stark azeotropic distillation (Reference
7). Prior to cleanup and analysis, the
extracts of the filtered water and the
particulates are combined.
2.2 After extraction, 37Cl4-labeled 2,3,7,8-
TCDD is added to each extract to measure
the efficiency of the cleanup process.
Samples cleanup may include back extrac-
tion with acid and/or base, and gel perme-
ation, alumina, silica gel, and activated
carbon chromatography. High performance
liquid chromatography (HPLC) can be used
for further isolation of the 2,3,7,8-
isomers or other specific isomers or
congeners.
2.3 After cleanup, the extract is concentrated
to near dryness. Immediately prior to
injection, two internal standards are
added to each extract, and a 1 uL aliquot
of the extract is injected into the gas
chromatograph. The analytes are separated
by the GC and detected by a high resolu-
tion (>10,000) mass spectrometer. Two
exact masses (m/z's) are monitored for
each analyte. The isotopically labeled
compounds serve to correct for the
variability of the analytical technique.
2.4 Dioxins and furans are identified by
comparing GC retention times and the ion
abundance ratios of the m/z's with the
corresponding retention time ranges of
authentic standards and the theoretical
ion abundance ratios of the exact m/z's.
Isomers and congeners are identified when
the retention times and m/z abundance
ratios agree within pre-defined limits.
By using a GC column or columns capable of
resolving the 2,3,7,8-substituted isomers
from all other tetra- isomers, the
2,3,7,8-substituted isomers are identified
when the retention time and m/z abundance
ratios agree within pre-defined limits of
the retention times and exact m/z ratios
of authentic standards.
2.5 Quantitative analysis is performed by GCMS
using selected ion current profile (SICP)
areas, in one of two ways.
2.5.1 For the 15 2,3,7,8-substituted isomers for
which labeled analogs are available (see
125
-------
Table 1), the GCMS system is calibrated
and the compound concentration is deter-
mined using an isotope dilution technique.
Although a labeled analog of the octa-
chlorinated dibenzofuran (OCDF) is avail-
able, using high resolution mass spectrom-
etry it produces an m/z that may interfere
with the identification and quantitat ion
of the unlabeled octachlorinated dibenzo-
p-dioxin (OCDD). Therefore, this labeled
analog has not been included in the cali-
bration standards, and the unlabeled OCDF
is quantitated against the labeled OCDD.
Because the labeled analog of 1,2,3,7,8,9-
HxCDD is used as an internal standard
(i.e., not added before extraction of the
sample), it cannot be used to quantitate
the unlabeled compound by strict isotope
dilution procedures. Therefore, the unla-
beled 1,2,3,7,8,9-HxCDD is quantitated
using the average of the responses of the
labeled analogs of the other two 2,3,7,8-
substituted HxCDD's, 1,2,3,4,7,8-HxCDD and
1,2,3,6,7,8-HxCDD. As a result, the con-
centration of the unlabeled 1,2,3,7,8,9-
HxCDD is corrected for the average
recovery of the other two HxCDD's.
2.5.2 For non-2,3,7,8-substituted isomers and
the total concentrations of all isomers
within a level of chlorination (i.e.,
total TCDD), concentrations are determined
using response factors from the calibra-
tion of labeled analogs at the same level
of chlorination.
2.6 The quality of the analysis is assured
through reproducible calibration and test-
ing of the extraction, cleanup, and GCMS
systems.
3 CONTAMINATION AND INTERFERENCES
3.1 Solvents, reagents, glassware, and other
sample processing hardware may yield arti-
facts and/or elevated baselines causing
misinterpretation of chromatograms (Ref-
erences 8-9). Specific selection of
reagents and purification of solvents by
distillation in all-glass systems may be
required. Where possible, reagents are
cleaned by extraction or solvent rinse.
3.2 Proper cleaning of glassware is extremely
important because glassware may not only
contaminate the samples, but may also
remove the analytes of interest by
adsorption on the glass surface.
3.2.1 Glassware should be rinsed with solvent
and washed with a detergent solution as
soon after use as is practical. Sonica-
tion of glassware containing a detergent
solution for approximately 30 seconds may
aid in cleaning. Glassware with removable
parts, particularly separately funnels
with teflon stopcocks, must be
disassembled prior to detergent washing.
3.2.2 After detergent washing, glassware should
be immediately rinsed first with methanol,
then with hot tap water. The tap water
rinse is followed by another methanol
rinse, then acetone, and then methylene
chloride.
3.2.3 Do not bake reusable glassware in an oven
as a routine part of cleaning. Baking may
be warranted after particularly dirty
samples are encountered, but should be
minimized, as repeated baking of glassware
may cause active sites on the glass
surface that will irreversibly adsorb
PCDDs/PCDFs.
3.2.4 Immediately prior to use, Soxhlet extrac-
tion glassware should be pre-extracted
with toluene for approximately 3 hours.
See Section 11.1.2.3. Separatory funnels
should be shaken with methylene
chloride/toluene (80/20 mixture) for 2
minutes, drained, and then shaken with
pure methylene chloride for 2 minutes.
3.3 All materials used in the analysis shall
be demonstrated to be free from interfer-
ences by running reference matrix blanks
initially and with each ^sample set
(samples started through the extraction
process on a given 12-hour shift, to a
maximum of 20 samples). The reference
matrix blank must simulate, as closely as
possible, the sample matrix under test.
Reagent water (Section 6.6.1) is used, to
simulate water samples; playground sand
(Section 6.6.2) or white quartz sand
(Section 6.3.2) can be used to simulate
soils; filter paper (Section 6.6.3) is
used to simulate papers and similar
materials; other materials (Section 6.6.4)
can be used to simulate other matrices.
3.4 Interferences coextracted from samples
will vary considerably from source to
source, depending on the diversity of the
site being sampled. Interfering compounds
may be present at concentrations several
orders of magnitude higher than the PCDDs
and PCDFs. The most frequently encoun-
tered interferences are chlorinated-
biphenyls, methoxy biphenyls, hydroxy-
126
-------
diphenyl ethers, benzylphenyl ethers,
polynuclear aromatics, and pesticides.
Because very low levels of PCDDs and PCDFs
are measured by this method, the elimina-
tion of interferences is essential. The
cleanup steps given in Section 12 can be
used to reduce or eliminate these inter-
ferences and thereby permit reliable
determination of the PCDDs and PCDFs at
the levels shown in Table 2.
3.5 Each piece of reusable glassware should be
numbered in such a fashion that the
laboratory can associate all reusable
glassware with the processing of a parti-
cular sample. This will assist the
laboratory in: 1) tracking down possible
sources of contamination for individual
samples, 2) identifying glassware assoc-
iated with highly contaminated samples
that may require extra cleaning, and 3)
determining when glassware should be
discarded.
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.
4.1.1 The 2,3,7,8-TCDD isomer has been found to
be acnegenic, carcinogenic, and terato-
genic in laboratory animal studies. It is
soluble in water to approximately 200 ppt
and in organic solvents to 0.14 percent.
On the basis of the available
toxicological and physical properties of
2,3,7,8-TCDD, all of the PCDDs and PCDFs
should be handled only by highly trained
personnel thoroughly familiar with
handling and cautionary procedures, and
who understand the associated risks.
4.1.2 It is recommended that the laboratory pur-
chase dilute standard solutions of the
analytes in this method. However, if
primary solutions are prepared, they shall
be prepared in a hood, and a NIOSH/MESA
approved toxic gas respirator shall be
worn when high concentrations are handled.
4.2 The laboratory is responsible for main-
taining 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 10-13.
The references and bibliography at the end
of Reference 13 are particularly compre-
hensive in dealing with the general
subject of laboratory safety.
4.3 The PCDDs and PCDFs and samples suspected
to contain these compounds are handled
using essentially the same techniques
employed in handling radioactive or
infectious materials. Uell-ventilated,
controlled access laboratories are
required. Assistance in evaluating the
health hazards of particular laboratory
conditions may be obtained from certain
consulting laboratories and from State
Departments of Health or Labor, many of
which have an industrial health service.
The PCDDs and PCDFs are extremely toxic to
laboratory animals. Each laboratory must
develop a strict safety program for
handling the PCDDs and PCDFs. The follow-
ing practices are recommended (References
2 and 14).
4.3.1 Facility -- When finely divided samples
(dusts, soils, dry chemicals) are handled,
all operations (including removal of
samples from sample containers, weighing,
transferring, and mixing), should be
performed in a glove box demonstrated to
be leak tight or in a fume hood demon-
strated to have adequate air flow. Gross
losses to the laboratory ventilation
system must not be allowed. Handling of
the dilute solutions normally used in
analytical and animal work presents no
inhalation hazards except in the case of
an accident.
4.3.2 Protective equipment -- Throwawa'y plastic
gloves, apron or lab coat, safety glasses
or mask, and a glove box or fume hood
adequate for radioactive work should be
utilized. During analytical operations
which may give rise to aerosols or dusts,
personnel should wear respirators equipped
with activated carbon filters. Eye
protection equipment (preferably full face
shields) must be worn while working with
exposed samples or pure analytical
standards. Latex gloves are commonly used
to reduce exposure of the hands. When
handling samples suspected or known to
contain high concentrations of the PCDDs
or PCDFs, an additional set of gloves can
also be worn beneath the latex gloves.
127
-------
4.3.3 Training -- Workers must be trained in the
proper method of removing contaminated
gloves and clothing without contacting the
exterior surfaces.
4.3.4 Personal hygiene -- Thorough washing of
hands and forearms after each manipulation
and before breaks (coffee, lunch, and
shift).
4.3.5 Confinement -- Isolated work area, posted
with signs, segregated glassware and
tools, plastic absorbent paper on bench
tops.
4.3.6 Effluent vapors -- The effluents of sample
splitters for the gas chromatograph and
roughing pumps on the GC/HS should pass
through either a column of activated char-
coal or be bubbled through a trap contain-
ing oil or high-boiling alcohols.
4.3.7 Waste Handling and Disposal
4.3.7.1 Handling -- Good technique includes
minimizing contaminated waste. Plastic
bag liners should be used in waste cans.
Janitors and other personnel must be
trained in the safe handling of waste.
4.3.7.2 Disposal
4.3.7.2.1 The PCDDs and PCDFs decompose above 800
°C. Low-level waste such as absorbent
paper, tissues, animal remains, and
plastic gloves may be burned in an appro-
priate incinerator. Gross quantities
(milligrams) should be packaged securely
and disposed through commercial or govern-
mental channels., which are capable of
handling extremely toxic wastes.
4.3.7.2.2 Liquid or soluble waste should be dis-
solved in methanol or ethanol and
irradiated with ultraviolet light with a
wavelength greater than 290 nm for several
days. (Use F 40 BL lamps or equivalent.)
Analyze liquid wastes and dispose of the
solutions when the PCDDs and PCDFs can no
longer be detected.
4.3.8 Decontamination
4.3.8.1 Personal decontamination -- Use any mild
soap with plenty of scrubbing action.
4.3.8.2 Glassware, tools, and surfaces
Chlorothene NU Solvent (Trademark of the
Dow Chemical Company) is the least toxic
solvent shown to be effective. Satis-
factory cleaning may be accomplished by
rinsing with Chlorothene, then washing
with any detergent and water. If glass-
ware is first rinsed with solvent, then
the dish water may be disposed of in the
sewer. Given the cost of disposal, it is
prudent to minimize solvent wastes.
4.3.9 Laundry -- Clothing known to be contami-
nated 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 of
the potential problem. The washer should
be run through a cycle before being used
again for other clothing.
4.3.10 Wipe tests -- A useful method of determin-
ing cleanliness of work surfaces and tools
is to wipe the surface with a piece of
filter paper. Extraction and analysis by
GC can achieve a limit of detection of 0.1
ug per wipe. Less than 0.1 ug per wipe
indicates acceptable cleanliness; anything
higher warrants further cleaning. More
than 10 ug on a wipe constitutes an acute
hazard and requires prompt cleaning before
further use of the equipment or work
space, and indicates that unacceptable
work practices have been employed.
4.3.11 Accidents -- Remove contaminated clothing
immediately, taking precautions not to
contaminate skin or other articles. Wash
exposed skin vigorously and repeatedly
until medical attention is obtained.
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 simi-
lar materials containing five percent
solids or less) -- Sample bottle, amber
glass, 1.1 L minimum, with screw cap.
5.1.1.2 Solid samples (soils, sediments, sludges,
paper pulps, 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.
5.1.1.4 Bottle caps -- Threaded to fit sample
bottles. Caps shall be lined with Teflon.
128
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5.1.1.5 Cleaning
5.1.1.5.1 Bottles are detergent water washed, then
solvent rinsed before use.
5.1.1.5.2 Liners are detergent water washed, then
rinsed with reagent Mater (Section 6.6.1)
and then solvent, and baked at
approximately 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. 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 to minimize sample
contamination. An integrating flow meter
is used to collect proportional composite
samples.
5.2 Equipment for glassware cleaning
5.2.1 Laboratory sink with overhead fume hood
5.3 Equipment for sample preparation
5.3.1 Laboratory fume hood of sufficient size to
contain the sample preparation equipment
listed below
5.3.2 Glove box (optional)
5.3.3 Tissue homogenizer -- VirTis Model 45
Macro homogenizer (American Scientific
Products H-3515, or equivalent) with
stainless steel Macro-shaft and Turbo-
shear blade.
5.3.4 Meat grinder -- Hobart, or equivalent,
with 3-5 mm holes in inner plate.
5.3.5 Equipment for determining percent moisture
5.3.5.1 Oven, capable of maintaining a temperature
of 110 ±5 "C.
5.3.5.2 Dessicator
5.3.6 Balances
5.3.6.1 Analytical -- Capable of weighing 0.1 mg.
5.3.6.2 Top loading -- Capable of weighing 10 mg.
5.4 Extraction apparatus
5.4.1 Water samples
5.4.1.1 pH meter, with combination glass
electrode.
5.4.1.2 pH paper, wide range (Hydrion Papers, or
equivalent).
5.4.1.3 Graduated cylinder, 1 L capacity
5.4.1.4 1 L filtration flasks with side arm, for
use in vacuum filtration of water samples.
5.4.1.5 Separatory funnels -- 250, 500, and 2000
mL, with Teflon stop cocks.
5.4.2 Soxhlet/Dean-Stark
(Figure 1)
(SDS)
extractor
FIGURE 1 Soxhlet/Dean-Stark Extractor
129
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5.4.2.1 Soxhlet -- 50 mm i.d., 200 ml capacity
with 500 mL flask (Cal-Glass LG-6900, or
equivalent, except substitute 500 ml round
bottom flask for 300 mL flat bottom
flask).
5.4.2.2 Thimble -- 43 x 123 to fit Soxhlet (Cal-
Glass LG-6901-122, or equivalent).
5.4.2.3 Moisture trap -- Dean Stark or Barret with
Teflon stopcock, to fit Soxhlet.
5.4.2.4 Heating mantle -- Hemispherical, to fit
500 ml round bottom flask (Cal-Glass LG-
8801-112, or equivalent).
5.4.2.5 Variable transformer -- Powerstat (or
equivalent), 110 volt, 10 amp.
5.4.3 Beakers, 400-500 ml
5.4.4 Spatulas -- Stainless steel
5.5 Fi Itration apparatus
5.5.1 Pyrex glass wool -- Solvent extracted by
SDS for three hours minimum. (NOTE: Bak-
ing glass wool may cause active sites that
will irreversibly adsorb PCDDs/PCDFs.)
5.5.2 Glass funnel -- 125-250 mL
5.5.3 Glass fiber filter paper (Whatman GF/D, or
equivalent)
5.5.4 Drying column -- 15 to 20 mm i.d. Pyrex
chromatographic column equipped with
coarse glass frit or glass wool plug.
5.5.5 Buchner funnel, 15 cm.
5.5.6 Glass fiber filter paper for above.
5.5.7 Pressure filtration apparatus -- Hillipore
YT30 142 HW, or equivalent.
5.6 Centrifuge apparatus
5.6.1 Centrifuge -- Capable of rotating 500 ml
centrifuge bottles or 15 ml centrifuge
tubes at 5,000 rpm minimum
5.6.2 Centrifuge bottles -- 500 ml, with screw
caps, to fit centrifuge
5.6.3 Centrifuge tubes -- 12-15 ml, with screw
caps, to fit centrifuge
5.7 Cleanup apparatus
5.7.1 Automated gel permeation chromatograph
(Analytical Biochemical Labs, Inc,
Columbia, MO, Model GPC Autoprep 1002, or
equivalent).
5.7.1.1 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).
5.7.1.2 Syringe, 10 mL, with Luer fitting.
5.7.1.3 Syringe filter holder, stainless steel,
and glass fiber or Teflon filters (Gelman
4310, or equivalent).
5.7.1.4 UV detectors -- 254-nm, 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 mm
path; Pharmacia UV-1, 3 mm flow cell; LDC
Milton-Roy UV-3, monitor #1203; or
equivalent).
5.7.2 Reverse phase high performance liquid
chromatograph
5.7.2.1 Column oven and detector -- Perkin-Elmer
Model LC-65T (or equivalent) operated at
0.02 AUFS at 235 nm.
5.7.2.2 Injector -- Rheodyne 7120 (or equivalent)
with 50 uL sample loop.
5.7.2.3 Column -- Two 6.2 x 250 mm Zorbax-ODS
columns in series (DuPont Instruments
Division, Wilmington, DE, or equivalent),
operated at 50 °C with 2.0 mL/min methanol
isocratic effluent.
5.7.2.4 Pump -- Altex 110A (or equivalent).
5.7.3 Pipets
5.7.3.1 Disposable, Pasteur, 150 mm x 5 mm i.d.
(Fisher Scientific 13-678-6A, or
equivalent).
5.7.3.2 Disposable, serological, 10 mL (6 mm
i.d.).
5.7.4 Chromatographic columns
5.7.4.1 150 mm x 8 mm i.d., (Kontes K-420155, or
equivalent) with coarse glass frit or
glass wool plug and 250 mL reservoir.
130
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5.7.4.2
5.7.5
5.8
5.8,1
5.8.1.1
5.8.1.2
200 mm x 15 mm
frit or glass
reservoir.
i.d., with
wool plug
coarse glass
and 250 ml
5.8.1.3
Oven -- For storage of adsorbents, capable
of maintaining a temperature of 130 ±5 °C.
Concentration apparatus
Rotary evaporator •• Buchi/Brinkman-
American Scientific No, E5045-10 or
equivalent, equipped with a variable
temperature water bath.
A vacuum source is required for use of the
rotary evaporator. It must be equipped
with a shutoff valve at the. evaporator,
and preferably, have a vacuum gauge.
A recirculating water pump and chiller are
recommended, as use of tap water for cool-
ing the evaporator wastes large volumes of
water and can lead to inconsistent
performance as water temperatures and
pressures vary.
Round bottom flasks -- 100 ml and 500 ml
or larger, with ground glass fitting
compatible with the rotary evaporator.
5.8.2 Kuderna-Danish (K-D)
5.8.2.1 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.
5.8.2.2 Evaporation flask--500 ml (Kontes K-
570001-0500, or equivalent), attached to
concentrator tube with springs (Kontes K-
662750-0012).
5.8.2.3 Snyder column—three ball macro (Kontes K-
503000-0232, or equivalent).
5.8.2.4 Boiling chips
5.8.2.4.1 Glass or silicon carbide--approx 10/40
mesh, extracted with methylene chloride
and baked at 450 °C for one h minimum.
5.8.2.4.2 Teflon (optional)--extracted with
methylene chloride.
5.8.2.5 Water bath--heated, with concentric ring
cover, capable of maintaining a tempera-
ture within +/- 2 °C, installed in a fume
hood.
5.8.3 Nitrogen blowdown apparatus -- Equipped
with water bath control ted at 35-40 °C (N-
Evap, Organomation Associates, Inc., South
Berlin, MA, or equivalent), installed in a
fume hood.
5.8.4 Sample vials -- Amber glass, 2-5 ml with
Teflon-lined screw cap.
5.9 Gas chromatograph -- Shall have splitless
or on-column injection port for capillary
column, temperature program with
isothermal hold, and shall meet all of the
performance specifications in Section 7.
5.9.1 GC Column for PCDDs and PCDFs and for
isomer specificity for 2,3,7,8-TCDD -- 60
±5 m x 0.32 ±0.02 mm i.d.; 0.25 urn 5%
phenyl, 94% methyl, 1% vinyl silicone
bonded phase fused silica capillary column
(J & W DB-5, or equivalent).
5.9.2 GC Column for isomer specificity for
2,3,7,8-TCDF -- 30 ±5 m x 0.32 ±0.02 mm
i.d,; 0.25 urn bonded phase fused silica
capillary column (J & U DB-225, or
equivalent).
5.10 Mass spectrometer -- 28-40 eV electron
impact ionization, shall be capable of
repetitively selectively monitoring 12
exact m/z's minimum at high resolution
(>10,000) during a period of approximately
1 second, and shall meet all of the
performance specifications in Section 7.
5.11 GCMS interface -- The mass spectrometer
(MS) shall be interfaced to the GC such
that the end of the capillary column
terminates within 1 cm of the ion source
but does not intercept the electron or ion
beams.
5.12 Data system -- Capable of collecting,
recording and storing MS data.
6 REAGENTS AND STANDARDS
6.1 pH adjustment and back extraction
6.1.1 Potassium hydroxide -- Dissolve 20 g
reagent grade KOH in 100 ml reagent water.
6.1.2 Sulfuric acid -- Reagent grade (specific
gravity 1.84).
6.1.3 Sodium chloride -- Reagent grade, prepare
a five percent (w/v) solution in reagent
water.
131
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6.2 Solution drying and evaporation
6.2.1 Solution drying -- Sodium sulfate, reagent
grade, granular anhydrous (Baker 3375, or
equivalent), rinsed with methylene
chloride (20 mL/g), baked at 400 °C for
one hour minimum, cooled in a dessicator,
and stored in a pre-cleaned glass bottle
with screw cap that prevents moisture from
entering. If after heating the sodium
sulfate develops a noticeable grayish cast
(due to the presence of carbon in the
crystal matrix), that batch of reagent is
not suitable for use and should be
discarded. Extraction with methylene
chloride (as opposed to simple rinsing)
and baking at a lower temperature may
produce sodium sulfate that is suitable
for use.
6.2.2 Prepurified nitrogen
6.3 Extraction
6.3.1 Solvents -- Acetone, toluene, cyclohexane,
hexane, nonane, methanol, methylene
chloride, and nonane: distiIled-in-glass,
pesticide quality, lot certified to be
free of interferences.
6.3.2 White quartz sand, 60/70 mesh -- For
Soxhlet/Dean-Stark extraction, (Aldrich
Chemical Co, Milwaukee WI Cat No.
27,437-9, or equivalent). Bake at 450 °C
for four hours minimum.
6.4 GPC calibration solution -- Solution con-
taining 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 Adsorbents for sample cleanup
6.5.1 Silica gel
6.5.1.1 Activated silica gel -- Bio-Sil A, 100-200
mesh (Bio-Rad 131-1340, or equivalent),
rinsed with methylene chloride, baked at
180 °C for one hour minimum, cooled in a
dessicator, and stored in a pre-cleaned
glass bottle with screw cap that prevents
moisture from entering.
6.5.1.2 Acid silica gel (30 percent w/w) --
Thoroughly mix 44.0 g of concentrated
sulfuric acid with 100.0 g of activated
silica gel in a clean container. Break up
aggregates with a stirring rod until a
uniform mixture is obtained. Store in a
screw-capped bottle with Teflon-lined cap.
6.5.1.3 Basic silica gel -- Thoroughly mix 30 g of
1N sodium hydroxide with 100 g of
activated silica gel in a clean container.
Break up aggregates with a stirring rod
until a uniform mixture is obtained.
Store in a screw-capped bottle with
Teflon-Iined cap.
6.5.2 Alumina -- Either one of two types of
alumina, acid or basic, may be used in the
cleanup of sample extracts, provided that
the laboratory can meet the performance
specifications for the recovery of labeled
compounds described in Section 8.3. The
same type of alumina must be used for all
samples, including those used to demon-
strate initial precision and accuracy
(Section 8.2) and ongoing precision and
accuracy (Section 14.5).
6.5.2.1 Acid alumina -- Bio-Rad Laboratories 132-
1340 Acid Alumina AG 4 {or equivalent).
Activate by heating to 130 °C for 12 hours
minimum.
6.5.2.2 Basic alumina -- Bio-Rad Laboratories 132-
1240 Basic Alumina AG 10 (or equivalent).
Activate by heating to 600 °C for 24 hours
minimum. Alternatively, activate by heat-
ing alumina in a tube furnace at 650-700
°C under an air flow of approximately 400
cc/min. Do not heat over 700 °C, as this
can lead to reduced capacity for retaining
the analytes. . Store at 130 °C in a
covered flask. Use within five days of
baking.
6.5.3 AX-21/Celite
6.5.3.1 Activated carbon --' AX-21 (Anderson
Development Company, Adrian, HI, or equiv-
alent). Prewash with methanol and dry in
vacuo at 110 °C.
6.5.3.2 Celite 545 -- (Supelco 2-0199, or equiva-
lent).
6.5'.3.3 Thoroughly mix 5.35 g AX-21 and 62.0 g
Celite 545 to produce a 7.9% w/w mixture.
Activate the mixture at 130 °C for six
hours minimum. Store in a dessicator.
6.6 Reference matrices
6.6.1 Reagent water -- Water in which the PCDDs
and PCDFs and interfering compounds are
not detected by this method.
132
-------
6.6.2 High solids reference matrix -- Playground
sand or similar material in which the
PCDDs and PCDFs and interfering compounds
are not detected by this method. May be
prepared by extraction with methylene
chloride and/or baking at 450 °C for four
hours minimum.
6.6.3 Filter paper '-- Gelman type A (or equiva-
lent) glass fiber filter paper in which
the PCDDs and PCDFs and interfering com-
pounds are not detected by this method.
Cut the paper to simulate the surface area
of the paper sample being tested.
6.6,4 Other matrices -- This method may be
verified on any matrix by performing the
tests given in Section 8.2. Ideally, the
matrix should be free of the PCDDs and
PCDFs, but in no case shall the background
level of the PCDDs and PCDFs in the
reference matrix exceed three times the
minimum levels given in Table 2. If low
background levels of the PCDDs and PCDFs
are present in the reference matrix, the
spike level of the analytes used in
Section 8.2 should be increased to provide
a spike-to-background ratio in the range
of 1/1 to 5/1 (Reference 15).
6.7 Standard solutions -- Purchased as solu-
tions or mixtures with certification to
their purity, concentration, and authen-
ticity, or prepared from materials of
known purity and composition. If compound
purity is 98 percent or greater, the
weight may be used without correction to
compute the concentration of the standard.
When not being used, standards are stored
in the dark at room temperature in screw-
capped vials with Teflon-lined caps. A
mark is placed on the vial at the level of
the solution so that solvent evaporation
loss can be detected. If solvent loss has
occurred, the solution should be replaced.
6.8 Stock solutions
6.8.1 Preparation -- Prepare in nonane per the
steps below or purchase as dilute solu-
tions (Cambridge Isotope Laboratories,
Cambridge, HA, or equivalent). Observe
the safety precautions in Section 4, and
the recommendation in Section 4.1.2.
6.8.2 Dissolve an appropriate amount of assayed
reference material in solvent. For
example, weigh 1-2 mg of 2,3,7,8-TCDD to
three significant figures in a 10 mL
ground glass stoppered volumetric flask
and fill to the mark with nonane. After
the TCDD is completely dissolved, transfer
the solution to a clean 15 ml vial with
Teflon-lined cap.
6.8.3 Stock standard solutions should be checked
for signs of degradation prior to the
preparation of calibration or performance
test standards. Reference standards that
can be used to determine the accuracy of
calibration standards are available from
Cambridge Isotope Laboratories.
6.9 Secondary standard -- Using stock solu-
tions (Section 6.8), prepare secondary
standard solutions containing the com-
pounds and concentrations shown in Table 4
in nonane.
6.10 Labeled compound stock standard -- From
stock standard solutions prepared as
above, or from purchased mixtures, prepare
this standard to contain the labeled com-
pounds at the concentrations shown in
Table 4 in nonane. This solution is
diluted with acetone prior to use (Section
10.3.2).
6.11 Cleanup standard - Prepare Cl,-2,3,7,8-
TCDD at the concentration shown in Table 4
in nonane.
6.12 Internal standard -- Prepare at the con-
centration shown in Table 4 in nonane.
6.13 Calibration standards (CS1 through CSS) --
Combine the solutions in Sections 6.9,
6.10, 6.11, and 6.12 to produce the five
calibration solutions shown in Table 4 in
nonane. These solutions permit the rela-
tive response (labeled to unlabeled) and
response factor to be measured as a func-
tion of concentration. The CS3 standard
is used for calibration verification
(VER).
6.14 Precision and recovery standard (PAR) --
Used for determination of initial (Section
8.2) and ongoing (Section 14.5) precision
and accuracy. This solution contains the
analytes and labeled compounds at the con-
centrations listed in Table 4 in nonane.
This solution is diluted with acetone
prior to use (Section 10.3.4).
6.15 GC retention time window defining solu-
tions --Used to define the beginning and
ending retention times for the dioxin and
furan isomers.
133
-------
6.15.1 DB-5 column window defining standards -•
Cambridge Isotope Laboratories ED-1732-A
(dioxins) and ED-1731-A (furans), or
equivalent, containing the compounds
listed in Table 5.
6.16 Isomer specificity test standards -- Used
to demonstrate isomer specificity for the
2,3,7,8-tetra- isomers of dioxin and
furan.
6.16.1 Standards for the DB-5 column -- Cambridge
Isotope Laboratories ED-908, ED-908-C, or
ED-935, or equivalent, containing the com-
pounds listed in Table 5.
6.16.2 Standards for the DB-225 column
Cambridge Isotope Laboratories EF-937 or
EF-938, or equivalent, containing the
compounds listed in Table 5.
6.17 Stability of solutions -- Standard solu-
tions used for quantitative purposes
(Sections 6.9-6.14) 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 quanti-
tation m/z remains within ±15 percent of
the area obtained in the initial analysis
of the standard. Any standards failing to
meet this criterion should be assayed
against reference standards, as in Section
6.8.3., before further use.
7 CALIBRATION
7.1 Assemble the GCMS and establish the
operating conditions necessary to meet the
relative retention time specifications in
Table 2.
7.1.1 The following GC operating conditions may
be used for guidance and adjusted as
needed to meet the relative retention time
specifications in Table 2:
Injector temp: 270 °C
Interface temp: 290 °C
Initial temp and time: 200 °C, 2 min
Temp Program: 200-220 °C at 5 °C/min
220 °C for 16 min
220-235 °C at 5 °C/min
235 °C for 7 min
235-330 °C at 5 °C/min
NOTE: All portions of the column which
connect the GC to the ion source shall
remain at the interface temperature
specified above during analysis, to pre-
clude condensation of less volatile
compounds.
7.1.2 Mass spectrometer (MS) resolution
Obtain a selected ion current profile
(SICP) of each analyte in Table 4 at the
two exact masses specified in Table 3 and
at >10,000 resolving power by injecting an
authentic standard of the PCDDs and PCDFs
either singly or as part of a mixture in
which there is no interference between
closely eluted components, using the pro-
cedure in Section 13.
7.1.2.1 The analysis time for PCDDs and PCDFs may
exceed the long-term mass stability of the
mass spectrometer. Because the instrument
is operated in the high-resolution mode,
mass drifts of a few ppm (e.g., 5 ppm in
mass) can have serious adverse effects on
instrument performance. Therefore, a
mass-drift correction is mandatory. A
lock-mass ion from the reference compound
(PFK) is used for tuning the mass
spectrometer. The lock-mass ion is depen-
dent on the masses of the ions monitored
within each descriptor, as shown in Table
3. 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 time
lost in cleaning the source.
7.1.2.2 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
-------
calibration solution (Table 4) per the
procedure in Section 13 and the conditions
in Table 2.
7.2.1 Measure the SICP areas for each analyte
and compute the ion abundance ratios
specified in Table 3A. Compare the
computed ratio to the theoretical ratio
given in Table 3A.
7.2.1.1 The groups of m/z's to be monitored are
shown in Table 3. Each group or
descriptor shall be monitored in succes-
sion as a function of GC retention time to
ensure that all PCDDs and PCDFs are
detected. The theoretical abundance
ratios for the m/z's are given in Table
3A, along with the control limits of each
ratio.
7.2.1.2 The mass spectrometer shall be operated in
a mass drift correction mode, using per-
fluorokerosene (PFK) to provide lock
masses. The lock mass for each group of
m/z's is shown in Table 3. Each lock mass
shall be monitored and shall not vary by
more than ±10 percent throughout its
respective retention time window. Varia-
tions of the lock mass by more than 10
percent indicate the presence of coeluting
interferences that may significantly
reduce the sensitivity of the mass
spectrometer. Re-injection of another
aliquot of the sample extract will not
resolve the problem. Add-itional cleanup
of the extract may be required to remove
the interferences.
7.2.2 All PCDDs and PGDFs shall be within their
respective ratios; otherwise, the mass
spectrometer shall be adjusted and this
test repeated until the m/z ratios fall
within the limits specified. If the
adjustment alters the resolution of the
mass spectrometer, resolution shall be
verified (Section 7.1) prior to repeat of
the test.
7.2.3 Verify that the HRGC/HRHS instrument meets
the minimum levels in Table 2. The peaks
representing both unlabeled and labeled
analytes in the calibration standards must
have a signal-to-noise ratio (S/N) greater
than or equal to 10; otherwise, the mass
spectrometer shall be adjusted and this
test repeated until the minimum levels in
Table 2 are met.
7.2.4 The absolute retention time of
12.
1,2,3,4-TCDD (Section 6-.12) shall exceed
25.0 minutes on the DB-5 column, and the
retention time of C.2-1,2,3,4-TCDD shall
exceed 15.0 minutes on the DB-225 column;
otherwise, the GC temperature program
shall be adjusted and this test repeated
until the above-stated minimum retention
time criteria are met.
7.3 Retention time windows — Analyze the
window defining mixtures (Section 6.15)
using the procedure in Section 13 (Figures
2A-2D). Table 5 gives the elution order
(first/last) of the compound pairs.
7.4 Isomer specificity
7.4.1 Analyze the isomer specificity test
standards (Section 6.16) using the
procedure in Section 13.
7.4.2 Compute the percent valley between the GC
peaks that elute most closely to the
2,3,7,8- TCDD and TCDF isomers, on their
respective columns, per Figure 3.
7.4.3 Verify that the height of the valley
between the most closely eluted isomers
and the 2,3,7,8- isomers is less than 25
percent (computed as 100 x/y in Figure 3).
If the valley exceeds 25 percent, adjust
the analytical conditions and repeat the
test or replace the GC column and recali-
brate (Section 7.2 through 7.4).
7.5 Calibration with isotope dilution
Isotope dilution is used for the 15
2,3,7,8-substituted PCDDs and PCDFs with
labeled compounds added to the samples
prior to extraction, and for 1,2,3,7,8,9-
HxCDD and OCDF (see Section 16.1). The
reference compound for each unlabeled
compound is shown in Table 6.
7.5.1 A calibration curve encompassing the
concentration range is prepared for each
compound to be determined. The relative
response (RR) (unlabeled to labeled) vs.
concentration in standard solutions is
plotted or computed using a linear regres-
sion. Relative response is determined
according to the procedures described
below. A minimum of five data points are
employed for calibration.
7.5.2 The relative response of each unlabeled
PCDD/PCDF and its labeled analog is
determined using the area responses of
135
-------
6-MAY-88 Sir: Voltage 705 Sys: DB5US
Sample 1 Injection 1 Group 2 Mass 303.9016
100
80
60
40-
20-
1,3,6,8-TCDF
Norm: 3044
1,2,8,9-TCDF
25:20 26:40 28:00 29:20 30:40 32:00 33:20 34:40 36:00 37:20 38:40
6-MAY-88 Sir: Voltage 705 Sys: DB5US
Sample 1 Injection 1 Group 2 Mass 319.8965
100i
80-
60
40
20
1,3,6,8-TCDD
Norm: 481
1,2,8,9-TCDD
2,3,7,8-TCDD
1,2,3,7/1,2,3,8-TCDD -
25:20 26:40 28:00 29:20 30:40- 32:00^x33:20 34:40 36:00 37:20 38:40
FIGURE 2A First and Last Eluted Tetra- Dioxin and Furan Isomers
136
-------
6-MAY-88 Sir: Voltage 705 Sys: DB5US
Sample 1 Injection 1 Group 2 Mass 339.8597
100
80
60
40
20-
1,3,4,6,8-PeCDF
Norm: 652
1,2,3,8,9-PeCDF
29:20 30:40 32:00 33:20 34:40 36:00 37:20 38:40
6-MAY-88 Sir: Voltage 705 Sys: DB5US
Sample 1 Injection 1 Group 2 Mass 355.8546
80-
60-
40-
20-
OJ-,
1,2,4,7,9-PeCDD
Norm: 503
1,2,3,8,9-PeCDD
\
"T———T
29:20 30:40 32:00 33:20 34:40 36:00 37:20 38:40
FIGURE 28 First and Last Eluted Penta- Dioxin and Furan Isomers
137
-------
6-MAY-88 Sir: Voltage 705 Sys: DB5US
Sample 1 Injection 1 Group 3 Mass 373.8208
100
80
60
40-
20-
Norm: 560
1,2,3,4,6,8-HxCDF
1,2,3,4,8,9-HxCDF
39:30 40:00 40:30 41:00 41:30 42:00 42:30 43:00 43:30 44:00 44:30
6-MAY-88 Sir: Voltage 705 Sys: DB5US
Sample 1 Injection 1 Group 3 Mass 389.8156
100
00'
60'
40-
20-
1,2,4,6,7,9/1,2,4,6,8,9-HxCDD .
Norm: 384
1,2,3,4,6,7-HxCDD
39:30 40:00 40:30 41:00 41:30 42:00 42:30 43:00 43:30 44:00 44:30
FIGURE 2C First and Last Eluted Hexa- Dioxin and Furan Isomers
138
-------
6-MAY-88 Sir: Voltage 705 Sys: DB5US
Sample 1 Injection 1 Group 4 Mass 407.7818
100 1.2.3.4.6.7.8-HPCDF
60
40
20
1,2,3.4,7,8,9-HpCDF
45:20 46:40 48:00 49:20 50:40 52:00 53:20 54:40 56:00 57:20
6-MAY-88 Sir: Voltage 705 Sys: DB5US
Sample 1 Injection 1 Group 4 Mass 423.7766
1, 2,3,4,6, 7,9-HpCDD
100-, .* - - - Norm: 282
80-
60-
40
20
1.2.3.4.6.7,8-HpCDD
45:20 46:40 48:00 49:20 50:40 52:00 53:20 54:40 56:00 57:20
6-MAY-88 Sir: Voltage 705 Sys: DB5US
Sample 1 Injection 1 Group 4 Mass 441.7428
100 Norm: 13
OCDF
80-1
60.
40
20-I
45:20 46:40 48:00 49:20 50:40 52:00 53:20 54:40 56:00 57:20
6-MAY-88 Sir: Voltage 705 Sys: DB5US
Sample 1 Injection 1 Group 4 Mass 457.7377
1001 I OCDD Norm:
80-1
60
40
20^
45:20 46:40 48:00 49:20 50:40 52:00 53:20 54:40 56:00 57:20
FIGURE 20 First and Last Eluted Hepta- Dioxin and Furan Isomers
139
-------
3A DB225 Column
21-APR-88 Sir: Voltage 705 Sys: DB225
Sample 1 Injection 1 Group 1 Mass 305.8987
Text: COLUMN PERFORMANCE
20-
2,3,7,8-TCDF Norm: 3466
1,2,3,9-TCDF
16:10 16:20 16:30 16:40 16:50 17:00 17:10 17:20 17:30 17:40 17:50 18:00
o
o
3B DBS Column
100
22:30 24:00 25:30
Time
27:00
FIGURE 3 Valley between 2,3,7,8- Tetra Dioxin and Furan Isomers and Other Closely Eluted Isomers
140
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both the primary and secondary m/z's
specified in Table 3, for each calibration
standard, as follows:
RR =
l
where,
1 2
A and A are the areas of the primary
and secondary m/z's for the unlabeled
compound.
1 2
A^ and A^ are the areas of the primary
and secondary m/z's for the labeled
compound.
C. is the concentration of the labeled
compound in the calibration standard.
C is the concentration of the unlabeled
compound in the calibration standard.
7.5.3 To calibrate the analytical system by
isotope dilution, inject a 1.0 uL aliquot
of calibration standards CS1 through CS5
(Section 6.13 and Table 4) using the
procedure in Section 13 and the conditions
in Table 2. Compute the relative response
(RR) at each concentration.
7.5.4 Linearity -- If the relative response for
any compound is constant (less than 20
percent coefficient of variation) over the
5-point calibration range, an averaged
relative response may be used for that
compound; otherwise, the complete calibra-
tion curve for that compound shall be used
over the 5-point calibration range.
7.6 Calibration by internal standard -- The
internal standard method is applied to
determination of non-2,3,7,8-substituted
compounds having no labeled analog in this
method, and to measurement of labeled
compounds for intralaboratory statistics
(Sections 8.4 and 14.5.4).
7.6.1 Response factors -- Calibration requires
the determination of response factors (RF)
defined by the following equation:
Cis
RF = V"S
where,
1 2
A and A are the areas of the primary
and secondary m/z's for the compound
7.6.2
7.6.3
7.7
7.8
7.8.1
7.8.2
to be calibrated. (NOTE: There is
only one m/z for 7Cl,-2,3,7,8-TCDD.
See Table 3.)
A.g and A.g are the areas of the primary
and secondary m/z's for the GCMS
internal standard.
C.g is the concentration of the GCMS
internal standard (Section 6.12 and
Table 4).
C is the concentration of the compound in
the calibration standard.
To calibrate the analytical system by
internal standard, inject a 1.0 uL aliquot
of calibration standards CS1 through CSS
(Section 6.13 and Table 4) using the
procedure in Section 13 and the conditions
in Table 2. Compute the response factor
(RF) at each concentration.
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 calibra-
tion curve for that compound shall be used
over the 5-point range.
Combined calibration -- By using calibra-
tion solutions (Section 6.13 and Table 4)
containing the unlabeled and labeled
compounds, and the internal standards, 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 14.3) by analyzing the calibra-
tion verification standard (VER, Table 4).
Recall'brat ion is required if calibration
verification criteria (Section. 14.3.4)
cannot be met.
Data storage -- MS data shall
collected, recorded, and stored.
be
Data acquisition -- The signal at each
exact m/z shall be collected repetitively
throughout the monitoring period and
stored on a mass storage device.
Response factors and multipoint
calibrations -- The data system shall be
used to record and maintain lists of
response factors (response ratios for
141
-------
isotope dilution) and multipoint
calibration curves. Computations of
relative standard deviation (coefficient
of variation) shall be used to test
calibration linearity. Statistics on
initial performance (Section 8.2) and
ongoing performance (Section 14.5) shall
be computed and maintained.
8 QUALITY ASSURANCE/QUALITY CONTROL
8.1 Each laboratory that uses this method is
required to operate a formal quality
assurance program (Reference 16). 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 characteris-
tics 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) or to
an alternate matrix, the high solids
reference matrix (Section 6.6.2) or the
alternate matrix (Section 6.6.4) is sub-
stituted for the reagent water matrix
(Section 6.6.1) in all performance tests.
8.1.1 The analyst shall make an initial demon-
stration 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 that all
performance specifications are met. Each
time a modification is made to the method,
the analyst is required to repeat the pro-
cedures in Sections 7.2 through 7.4 and
Section 8.2 to demonstrate method
performance.
8.1.3 Analyses of blanks are required to demon-
strate freedom from contamination (Section
3.2). 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. Procedures for dilutions are
given in Section 16.4.
8.1.5 The laboratory shall, on an ongoing basis,
demonstrate through calibration verifica-
tion and the analysis of the precision and
recovery standard that the analytical sys-
tem is in control. These procedures are
described in Sections 14.1 through 14.5.
8.1.6 The laboratory shall maintain records to
define the quality of data that is gener-
ated. Development of accuracy statements
is described in Section 8.4.
8.2 Initial precision and accuracy -- To
establish the ability to generate accept-
able precision and accuracy, the analyst
shall perform the following operations.
8.2.1 For low solids (aqueous samples), extract,
concentrate, and analyze four 1-liter
aliquots of reagent water spiked with the
diluted precision and recovery standard
(PAR) (Sections 6.14 and 10.3.4) according
to the procedures in Sections 10 through
13. For an alternate sample matrix, four
aliquots of the alternate matrix are used.
All sample processing steps, including
preparation (Section 10), extraction
(Section 11), and cleanup (Section 12)
that are to be used for processing samples
shall be included in this test.
8.2.2 Using results of the set of four analyses,
compute the average concentration (X) of
the extracts in ng/mL and the standard
deviation of the concentration (s) in
ng/mL for each compound, by isotope
dilution for PCDDs and PCDFs with a
labeled analog, and by internal standard
for labeled compounds. Calculate the
recovery of the labeled compounds.
8.2.3 For each unlabeled and labeled compound,
compare s and X with the corresponding
limits for initial precision and accuracy
in Table 7. 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
142
-------
performance is unacceptable for that
compound. Correct the problem and repeat
the test (Section 8.2). The concentration
limits in Table 7 for labeled compounds
are based on the requirement that the
recovery of each labeled compound be in
the range of 25-150%.
8.3 The laboratory shall spike all samples and
QC aliquots with the diluted labeled
compound spiking solution (Sections 6.10
and 10.3.2) to assess method performance
on the sample matrix.
8.3.1 Analyze each sample according to the
procedures in Sections 10 through 13.
8.3.2 Compute the percent recovery (R) of the
labeled compounds in the labeled compound
spiking standard and the cleanup standard
using the internal standard method
(Section 7.6).
8.3.3 The recovery of each labeled compound must
be within 25-150%. If the recovery of any
compound falls outside of these limits,
method performance is unacceptable for
that compound in that sample. To overcome
such difficulties, water samples are
diluted and smaller amounts of soils,
sludges, sediments and other matrices are
reanalyzed per Section 17.
8.4 Method accuracy for samples shall be
assessed and records shall be maintained.
8.4.1 After the analysis of five samples of a
given matrix type (water, soil, sludge,
pulp, etc) for which the labeled compound
spiking standards pass the tests in
Section 8.3, compute the average percent
recovery (R) and the standard deviation of
the percent recovery (S_) for the labeled
compounds only. Express the accuracy
assessment as a percent recovery interval
from R - 2SR to R + 2SR for each matrix.
For example, if R = 90% and SR = 10% for
five analyses of pulp, the accuracy
interval is expressed as 70-110%.
8.4.2 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 --' Reference matrix blanks are
analyzed to demonstrate freedom from
contamination (Section 3.2).
8.5.1 Extract and concentrate a 1-liter reagent
water blank (Section 6.6.1), high solids
reference matrix blank (Section 6.6.2),
paper matrix blank (Section 6.6.3) or
alternate reference matrix blank (Section
6.6.4) with each sample set (samples
started through the extraction process on
the same 12-hour shift, to a maximum of 20
samples). Analyze the blank immediately
after analysis of the precision and
recovery standard (Section 14.5) to
demonstrate freedom from contamination.
8.5.2 If any of the PCDDs or PCDFs (Table 1) or
any potentially interfering compound is
found in blank at greater than the minimum
level (Table 2), assumina a response
factor of 1 relative to the C,--!,2,3,4-
TCDD internal standard for compounds not
listed in Table 1, analysis of samples is
halted until the source of contamination
is eliminated and a blank shows no
evidence of contamination at this level.
NOTE: All samples associated with a
contaminated method blank must be re-
extracted and reanalyzed before the
results may be reported for regulatory
compliance purposes.
8.6 The specifications contained in this
method can be met if the apparatus used is
calibrated properly and then maintained in
a calibrated state. The standards used
for calibration (Section 7), calibration
verification (Section 14.3), and for
initial (Section 8.2) and ongoing (Section
14.5) precision and recovery should be
identical, so that the most precise
results will be obtained. A GCMS instru-
ment will provide the most reproducible
results if dedicated to the settings and
conditions required for the analyses of
PCDDs and PCDFs 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,
HANDLING
PRESERVATION, AND
9.1 Collect samples in amber glass containers
following conventional sampling practices
(Reference 17). Aqueous samples which
flow freely are collected in refrigerated
143
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bottles using automatic sampling equip-
ment. Solid samples are collected as grab
samples using wide mouth jars.
9.2 Maintain samples at 0-4 °C in the dark
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 18).
9.3 Perform sample analysis within 40 days of
extraction.
10 SAMPLE PREPARATION
The sample preparation process involves
modifying the physical form of the sample
so that the PCDDs and PCDFs can be
extracted efficiently. In general, the
samples must be in a liquid form or in the
form of finely divided solids in order for
efficient extraction to take place. Table
8 lists the phase(s) and quantity
extracted for various sample matrices.
Samples containing a solid phase and
samples containing particle sizes larger
than 1 mm require preparation prior to
extraction. Because PCDDs/PCDFs are
strongly associated with particulates, the
preparation of aqueous samples is depen-
dent on the solids content of the sample.
Aqueous samples containing one percent
solids or less are extracted in a
separatory funnel. A smaller sample
aliquot is used for aqueous samples
containing more than one percent solids.
For samples expected or known to contain
high levels of the PCDDs and/or PCDFs, the
smallest sample size representative of the
entire sample should be used, and the
sample extract should be diluted, if
necessary, per Section 16.4.
10.1 Determine percent solids
10.1.1 Weigh 5-10 g of sample (to three signifi-
cant figures) into a tared beaker. NOTE:
This aliquot is used only for determining
the solids content of the sample, not for
analysis of PCDDs/PCDFs.
10.1.2 Dry overnight (12 hours minimum) at 110 ±5
°C, and cool in a dessicator.
10.1.3 Calculate percent solids as follows:
x 100
% solids =
weight of sample after drying
weight of sample-before drying
10.2 Determine particle size
10.2.1 Spread the dried sample from Section
10.1.2 on a piece of filter paper or
aluminum foil in a fume hood or glove box.
10.2.2 Estimate the size of the particles in the
sample. If the size of the largest
particles is greater than 1 mm, the
particle size must be reduced to 1 mm or
less prior to extraction.
10.3 Preparation of aqueous samples containing
one percent solids or less -- The
extraction procedure for aqueous samples
containing less than or equal to one
percent solids involves filtering the
sample, extracting the particulate phase
and the filtrate separately, and combining
the extracts for analysis. The aqueous
portion is extracted by shaking with
methylene chloride in a separatory funnel.
The particulate material is extracted
using the SDS procedure.
10.3.1 Mark the original level of the sample on
the sample bottle for reference. Weigh
the sample in the bottle on a top loading
balance to ±1 g.
10.3.2 Dilute a sufficient volume of the labeled
compound stock solution by a factor of 50
with acetone to prepare the labeled
compound spiking solution. 1.0 rnL of the
diluted solution is required for each
sample, but no more solution should be
prepared than can be used in one day.
Spike 1.0 mL of the diluted solution into
the sample bottle. Cap the bottle and mix
the sample by careful shaking. Allow the
sample to equilibrate for 1-2 hours, with
occasional shaking.
10.3.3 For each sample or sample set (to a
• maximum of 20 samples) to be extracted
during the same 12-hour shift, place two
1.0 liter aliquots of reagent water in
clean 2 liter separatory flasks.
10.3.4 Spike 1.0 mL of the diluted labeled
compound spiking standard (Section 6.10)
into one reagent water aliquot. This
aliquot will serve as the blank. Dilute
10 uL of the precision and recovery
144
-------
standard (Section 6.14) to 2.0 mL with
acetone. Spike 1.0 mL of the diluted
precision and recovery standard into the
remaining reagent water aliquot. This
aliquot will serve as the PAR (Section
14.5).
10.3.5 Assemble a Buchner funnel on top of a
clean 1 L filtration flask. Apply a
vacuum to the flask, and pour the entire
contents of the sample bottle through a
glass fiber filter (Section 5.5.4) in the
Buchner funnel, swirling the sample
remaining in the bottle to suspend any
particulates.
10.3.6 Rinse the sample bottle twice with 5 mL of
reagent water to transfer any remaining
particulates onto the filter.
10.3.7 Rinse the any particulates off the sides
of the Buchner funnel with small quanti-
ties of reagent water.
10.3.8 Weigh the empty sample bottle on a top-
loading balance to ±1 g. Determine the
weight of the sample by difference. Do
not discard the bottle at this point.
10.3.9 Extract the filtrates using the procedures
in Section 11.1.1.
10.3.10 Extract the particulates using the proce-
dures in Section 11.1.2.
10.4 Preparation of samples containing greater
than one percent solids
10.4.1 Weigh a well-mixed aliquot of each sample
(of the same matrix type) sufficient to
provide 10 g of dry solids (based on the
solids determination in 10.1.3) into a
clean beaker or glass jar.
10.4.2 Spike 1.0 mL of the diluted labeled
compound spiking solution (Section 10.3.2)
into the sample aliquot(s).
10.4.3 For each sample or sample set (to a
maximum of 20 samples) to be extracted
during the same 12-hour shift, weigh two
10 g aliquots of the appropriate reference
matrix (Section 6.6) into clean beakers or
glass jars.
10.4.4 Spike 1.0 mL of the diluted labeled
compound spiking Solution into one
reference matrix aliquot. This aliquot
will serve as the blank. Spike 1.0 mL of
the diluted precision and recovery
standard (Section 10.3.4) into the
remaining reference matrix aliquot. This
aliquot will serve as the PAR (Section
14.5).
10.4.5 Stir or tumble and equilibrate the
aliquots for 1-2 hours.
10.4.6 Extract the aliquots using the procedures
in Section 11.
10.5 Multiphase samples
10.5.1 Pressure filter the sample, blank, and PAR
aliquots through Whatman GF/D glass fiber
filter paper. If necessary, centrifuge
these aliquots for 30 minutes at greater
than 5000 rpm prior to filtration.
10.5.2 Discard any aqueous phase (if present).
Remove any non-aqueous liquid (if present)
and reserve for recombination with the
extract of the solid phase (Section
11.1.2.5). Prepare the filter papers of
the sample and QC aliquots for particle
size reduction and blending (Section
10.6).
10.6 Sample grinding, homogenization, or blend-
ing -- Samples with particle sizes greater
than 1 mm (as determined by Section
10.2.2) are subjected to grinding, homo-
genization, or blending. The method of
reducing particle size to less than 1 mm
is matrix dependent. In general, hard
particles can be reduced by grinding with
a mortar and pestle. Softer particles can
be reduced by grinding in a Wiley mill or
meat grinder, by homogenization, or by
blending.
10.6.1 Each size reducing preparation procedure
on each matrix shall be verified by run-
ning the tests in Section 8.2 before the
procedure is employed routinely.
10.6.2 The grinding, homogenization, or blending
procedures shall be carried out in a glove
box or fume hood to prevent particles from
contaminating the work environment.
10.6.3 Grinding -- Tissue samples, certain papers
and pulps, slurries, and amorphous solids
can be ground in a Wiley mill or heavy
duty meat grinder. In some cases, reduc-
ing the temperature of the sample to
freezing or to dry ice or liquid nitrogen
temperatures can aid in the grinding
process. Grind the sample aliquots from
Section 10.4.5 or 10.5.2 in a clean
145
-------
grinder. Do not allow the sample tempera-
ture to exceed 50 °C. Grind the blank and
reference matrix aliquots using a clean
grinder.
10.6.4 Homogenization or blending -- Particles
that are not ground effectively, or
particles greater than 1 mm in size after
grinding, can often be reduced in size by
high speed homogenization or blending.
Homogenize and/or blend the sample, blank,
and PAR aliquots from Section 10.4.5,
10.5.2, or 10.6.3.
10.6.5 Extract the aliquots using the procedures
in Section 11.
11 EXTRACTION AND CONCENTRATION
11.1 Extraction of filtrates -- extract the
aqueous samples, blanks, and PAR aliquots
according to the following procedures.
11.1.1 Pour the filtered aqueous sample from the
filtration flask into a 2-L separatory
funnel. Rinse the flask twice with 5 ml
of reagent water and add these rinses to
the separatory funnel. Add 60 ml methy-
lene chloride to the sample bottle
(Section 10.3.8), seal,and shake 60
seconds to rinse the inner surface.
11.1.2 Transfer the solvent to the separatory
funnel and extract the sample by shaking
the funnel for 2 minutes with periodic
venting. Allow the organic layer to
separate from the water phase for a
minimum of 10 minutes. If the emulsion
interface between layers is more than one-
third the volume of the solvent layer,
employ mechanical techniques to complete
the phase separation (e.g., a glass stir-
ring rod). Drain the methylene chloride
extract into a solvent-rinsed glass funnel
approximately one-half full of clean
sodium sulfate. Set up the glass funnel
so that it will drain directly into a
solvent-rinsed 500-mL K-D concentrator
fitted with a 10 ml concentrator tube.
NOTE: Experience with aqueous samples
high in dissolved organic materials (e.g.,
paper mill effluents) has shown that acid-
ification of the sample prior to
extraction may reduce the formation of
emulsions. Paper industry methods suggest
that the addition of up to 400 mL of
ethanol to a 1 L effluent sample may also
reduce emulsion formation. However,
studies by the Agency to date suggest that
the effect may be a result of the dilution
of the sample, and that the addition of
reagent water may serve the same function.
Mechanical techniques may still be neces-
sary to complete the phase separation. If
either of these techniques is utilized,
the laboratory must perform the startup
tests described in Section 8.2 using the
same techniques.
11.1.3 Extract the water sample two more times
using 60 ml of fresh methylene chloride
each time. Drain each extract through the
funnel containing the sodium sulfate into
the K-D concentrator. After the third
extraction, rinse the separatory funnel
with at least 20 ml of fresh methylene
chloride, and drain this rinse through the
sodium sulfate into the concentrator.
Repeat this rinse at least twice.
11.1.4 The extract of the filtrate must be
concentrated before it is combined with
the extract of the particulates for
further cleanup. Add one or two clean
boiling chips to the receiver and attach a
three-ball macro Snyder column. Pre-wet
the column by adding approximately 1 ml of
hexane 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.
11.1.5 Adjust the vertical position of the
apparatus and the water temperature as
required to complete the concentration in
15-20 minutes. At the proper rate of
distillation, the balls of the column will
actively chatter but the chambers will not
flood.
11.1.6 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 lower joint into the concen-
trator tube with 1-2 mL of hexane. A 5 mL
syringe is recommended for this operation.
11.1.7 The concentrated extracts of the filtrate
and the particulates are combined using
the procedures in Section 11.2.13.
11.2 Soxhlet/Dean-Stark extraction of solids --
Extract the solid samples, particulates,
blanks, and PAR aliquots using the follow-
ing procedure.
11.2.1 Charge a clean extraction thimble with 5.0
g of 100/200 mesh silica (Section 6.5.1.1)
146
-------
and 100 g of quartz sand (Section 6.3.2).
NOTE: Do not disturb the silica layer
throughout the extraction process.
11.2.2 Place the thimble in a clean extractor.
Place 30-40 ml of toluene in the receiver
and 200-250 ml of toluene in the flask.
11.2,3 Pre-extract the glassware by heating the
flask until the toluene is boiling. When
properly adjusted, 1-2 drops of toluene
per second will fall from the condenser
tip into the receiver. Extract the
apparatus for three hours minimum.
11.2.4 After pre-extraction, cool and disassemble
the apparatus. Rinse the thimble with
toluene and allow to air dry.
11.2.5 Load the wet sample from Sections 10.4.6,
10.5.2, 10.6.3, or 10.6.4, and any non-
aqueous liquid from Section 10.5.2 into
the thimble and manually mix into the sand
layer with a clean metal spatula carefully
breaking up any large lumps of sample. If
the material to be extracted is the
particulate matter from the filtration of
an aqueous sample, add the filter paper to
the thimble also.
11.2.6 Reassemble the pre-extracted SDS apparatus
and add a fresh charge of toluene to the
receiver and reflux flask.
11.2.7 Apply power to the heating mantle to begin
refluxing. Adjust the reflux rate to
match the rate of percolation through the
sand and silica beds until water removal
lessens the restriction to toluene flow.
Check the apparatus for foaming frequently
during the first 2 hours of extraction.
If foaming occurs, reduce the reflux rate
until foaming subsides.
11.2.8 Drain the water from the receiver at 1-2
hours and 8-9 hours, or sooner if the
receiver fills with water. Reflux the
sample for a total of 16-24 hours. Cool
and disassemble the apparatus. Record the
total volume of water collected.
11.2.9 Remove the distilling flask. Drain the
water from the Dean Stark receiver and add
any toluene in the receiver to the extract
in the flask.
11.2.10 For solid samples, the extract must be
concentrated to approximately 10 ml prior
to back extraction. For the participates
filtered from an aqueous sample, the
extract must be concentrated prior to
combining with the extract of the
filtrate. Therefore, add one or two clean
boiling chips to the round bottom flask
and attach a three-ball macro Snyder
column. Pre-wet the column by adding
approximately 1 ml of toluene through the
top. Place the round bottom flask in a
•i heating mantle and apply heat as required
to complete the concentration in 15-20
minutes. At the proper rate of distilla-
tion, the balls of the column will
actively chatter but the chambers will not
flood.
11.2.11 When the liquid has reached an apparent
volume of 10 mL, remove the round bottom
flask from the heating mantle and allow
the solvent to drain and cool for at least
10 minutes. Remove the Snyder column.
11.2,12 If the extract is from a solid sample, not
the particulates from an aqueous sample,
transfer the concentrated extract to a 250
ml separator/ funnel. Rinse the flask
with toluene and add the rinse to the
separator/ funnel. Proceed with back
extraction per Section 11.3.
11.2.13 If the extract is from the particulates
from an aqueous sample, it must be com-
bined with the concentrated extract of the
filtrate (Section 11.1.7) prior to back
extraction. Assemble the glass funnel
filled approximately one-half full with
sodium sulfate from Section 11.1.2 such
that the funnel will drain into the K-D
concentrator from Section 11.1.7 contain-
ing the concentrated methylene chloride
extract of the filtrate. Pour the concen-
trated toluene extract of the particulates
through the sodium sulfate into the K-D
concentrator. Rinse the round-bottom
flask with three 15-20 ml volumes of
hexane, and pour each rinse through the
sodium sulfate into the K-D concentrator.
Add one or two fresh boiling chips to the
receiver and attach the three-ball macro
Snyder column to the K-D concentrator.
Pre-wet the column by adding approximately
1 mL of hexane to the top of the column.
Concentrate the combined extract to
approximately 10 ml (the volume of the
toluene). 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. Transfer the contents
of the K-D concentrator to a pre-rinsed
250 ml separatory funnel. Rinse the flask
147
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and lower joint with three 5 ml volumes of
hexane, and add each rinse* to the separa-
tory funnel. Proceed with back extraction
per Section 11.3.
11.3 Back extraction with base and acid
11.3.1 Spike 1.0 mL of the cleanup standard
(Section 6.11) into the separatory funnels
containing the sample and QC extracts
(Section 11.2.12 or 11.2.13).
11.3.2 Partition the extract against 50 ml of
potassium hydroxide solution (Section
6.1.1). Shake for 2 minutes with periodic
venting into a hood. Remove and discard
the aqueous layer. Repeat the base wash-
ing until no color is visible in the
aqueous layer, to a maximum of four wash-
ings. Minimize contact time between the
extract and the base to prevent degrada-
tion of the PCDDs and PCDFs. Stronger
potassium hydroxide solutions may be
employed for back extraction, provided
that the laboratory meets the specifica-
tions for labeled compound recovery and
demonstrates acceptable performance using
the procedures in Section 8.2.
11.3.3 Partition the extract against 50 ml of
sodium chloride solution (Section 6.1.3)
in the same way as with base. Discard the
aqueous layer.
11.3.4 Partition the extract against 50 ml of
sulfuric acid (Section 6.1.2) in the same
way as with base. Repeat the acid washing
until no color is visible in the aqueous
layer, to a maximum of four washings.
11.3.5 Repeat the partitioning against sodium
chloride solution and discard the aqueous
layer.
11.3.6 Pour each extract through a drying column
containing 7 to 10 cm of anhydrous sodium
sulfate. Rinse the separatory funnel with
30-50 ml of toluene and pour through the
drying column. Collect each extract in a
500 ml round bottom flask. Concentrate
and clean up the samples and QC aliquots
per Sections 11.4 and 12.
11.4 Macro-concentration -- Concentrate the
extracts in separate 100-mL round bottom
flasks on a rotary evaporator.
11.4.1 Assemble the rotary evaporator according
to manufacturer's instructions, and warm
the water bath to 45 °C. On a daily
basis, preclean the rotary evaporator by
concentrating 100 ml of clean extraction
solvent through the system. Archive both
the concentrated solvent and the solvent
in the catch flask for contamination check
if necessary. Between samples, three 2-3
ml aliquots of toluene should be rinsed
down the feed tube into a waste beaker.
11.4.2 Attach the round bottom flask containing
the sample extract to the rotary evapora-
tor. Slowly apply vacuum to the system,
and begin rotating the sample flask.
11.4.3 Lower the flask into the water bath and
adjust the speed of rotation and the
temperature as required to complete the
concentration in 15-20 minutes. At the
proper rate of concentration, the flow of
solvent into the receiving flask will be
steady, but no bumping or visible boiling
of the extract will occur. NOTE: If the
rate of concentration is too fast, analyte
loss may occur.
11.4.4 When the liquid in the concentration flask
has reached an apparent volume of 2 ml,
remove the flask from the water bath and
stop the rotation. Slowly and carefully,
admit air into the system. Be sure not to
open the valve so quickly that the sample
is blown out of the flask. Rinse the feed
tube with approximately 2 ml of hexane.
11.4.5 Transfer the extract to a vial using three
2-3 ml rinses of hexane. Proceed with
micro-concentration and solvent exchange.
11.5 Micro-concentration and solvent exchange
11.5.1 Toluene extracts to be subjected to GPC or
HPLC cleanup are exchanged into methylene
chloride. Extracts that are to be cleaned
up, using silica gel, alumina, and/or AX-
21/Celite are exchanged into hexane.
11.5.2 Transfer the vial containing the sample
extract to a nitrogen evaporation device.
Adjust the flow of nitrogen so that the
surface of the solvent is just visibly
disturbed. NOTE: A large vortex in the
solvent may cause analyte loss.
11.5.3 Lower the vial into a 45 °C water bath and
continue concentrating.
11.5.4 When the volume of the liquid is approxi-
mately 100 uL, add 2-3 mL of the desired
solvent (methylene chloride or hexane) and
148
-------
continue concentration to approximately
100 uL. Repeat the addition of solvent
and concentrate once more.
11.5.5 If the extract is to be cleaned up by GPC
or HPLC, adjust the volume of the extract
to 5.0 ml with methylene chloride.
Proceed with GPC cleanup (Section 12.2).
11.5.6 If the extract is to be cleaned up by
column chromatography (alumina, silica
gel, AX-21/Celite), bring the final volume
to 1.0 ml with hexane. Proceed with
column cleanups (Sections 12.3-12.5).
11.5.7 For extracts to be concentrated for
injection into the GCMS -- add 10 ul_ of
nonane to the vial. Evaporate the solvent
to the level of the nonane. Evaporate the
hexane in the vial to the level of the
nonane.
11.5.8 Seal the vial and label with the sample
number. Store in the dark at room temper-
ature until ready for GCMS analysis.
12 EXTRACT CLEANUP
12.1 Cleanup may not be necessary for
relatively clean samples (e.g., 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.
Before using a cleanup procedure, the
analyst must demonstrate that the require-
ments of Section 8.2 can be met using the
cleanup procedure.
12.1.1 Gel permeation chromatography (Section
12.2) removes many high molecular weight
interferences that cause GC column
performance to degrade. It may be 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).
12.1.2 Acid, neutral, and basic silica gel, and
alumina (Sections 12.3 and 12.4) are used
to remove nonpolar and polar
interferences.
12.1.3 AX-21/Celite (Section 12.5) is
remove nonpolar interferences.
used to
12.1.4 HPLC (Section 12.6) is used to provide
specificity for the 2,3,7,8-substituted
and other PCDD and PCDF isomers.
12.2 Gel permeation chromatography (GPC)
12.2.1 Column packing
12.2.1.1 Place 70-75 g of SX-3 Bio-beads in a 400-
500 mL beaker.
12.2.1.2 Cover the beads with methylene chloride
and allow to swell overnight (12 hours
minimum).
12.2.1.3 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.
12.2.1.4 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.
12.2.2 Column calibration
12.2.2.1 Load 5 mL of the calibration solution
(Section 6.4) into the sample loop.
12.2.2.2 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.
12.2.2.3 Set the "dump time" to allow >85 percent
removal of the corn oil and >85 percent
collection of the phthalate.
12.2.2.4 Set the "collect time" to the peak minimum
between perylene and sulfur.
12.2.2.5 Verify the calibration with the calibra-
tion solution after every 20 extracts.
Calibration is verified if the 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.
12.2.3 Extract cleanup -- GPC requires that the
column not be overloaded. The column
specified in this method is designed to
handle a maximum of 0.5 g of high
molecular weight material in a 5 mL
149
-------
12.2.3.1
extract. If the extract is known or
expected to contain more than 0.5 g, the
extract is split into aliquots for GPC and
the aliquots are combined after elution
from the column. The residue content of
the extract may be obtained gravimetri-
cally by evaporating the solvent from a 50
uL aliquot.
Filter the extract or load through the
filter holder to remove particulates.
Load the 5.0 ml extract onto the column.
12.2.3.2 Elute the extract using the calibration
data determined in Section 12.2.2.
Collect the eluate in a clean 400-500 ml
beaker.
12.2.3.3 Rinse the sample loading tube thoroughly
with methylene chloride between extracts
to prepare for the next sample.
12.2.3.4 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.
12.2.3.5 Concentrate the eluate per Section 11.2.1,
11.2.2, and 11.3.1 or 11.3.2 for further
cleanup or for injection into the GCHS.
12.3 Silica gel cleanup
12.3.1 Place a glass wool plug in a 15 mm i.d.
chromatography column. Pack the column in
the following order (bottom to top): 1 g
silica gel (Section 6.5.1.1), four g basic
silica gel (Section 6.5.1.3), 1 g silica
gel, 8 g acid silica gel (Section
6.5.1.2), 2 g silica gel. Tap the column
to settle the adsorbents.
12.3.2 Pre-rinse the column with 50-100 ml of
hexane. Close the stopcock when the
hexane is within 1 mm of the sodium
sulfate. Discard the eluate. Check the
column for channeling. If channeling is
present, discard the column and prepare
another.
12.3.3 Apply the concentrated extract to the
column. Open the stopcock until the
extract is within 1 mm of the sodium
sulfate.
12.3.4 Rinse the receiver twice with 1 ml
portions of hexane and apply separately to
the column. Elute the PCDDs/PCDFs with
100 ml hexane and collect the eluate.
12.3.5 Concentrate the eluate per Section 11.4 or
11.5 for further cleanup or for injection
into the HPLC or GCMS.
12.3.6 For extracts of samples known to contain
large quantities of other organic
compounds (such as paper mill effluents)
it may be advisable to increase the
capacity of the silica gel column. This
may be accomplished by increasing the
strengths of the acid and basic silica
gels. The acid silica gel (Section
6.5.1.2) may be increased in strength to
as much as 44% w/w (7.9 g sulfuric acid
added to 10 g silica gel). The basic
silica gel (Section 6.5.1.3) may be
increased in strength to as much as 33%
w/w (50 ml 1N NaOH added to 100 g silica
gel). NOTE: The use of stronger acid
silica gel (44% w/w) may lead to charring
of organic compounds in some extracts.
The charred material may retain some of
the analytes and lead to lower recoveries
of PCDDs/PCDFs. Increasing the strengths
of the acid and basic silica gel may also
require different volumes of hexane than
those specified above, to elute the
analytes off the column. Therefore, the
performance of the method after such
modifications must be verified by the
procedures in Section 8.2.
12.4 Alumina cleanup
12.4.1 Place a glass wool plug in a 15 mm i.d.
chromatography column.
12.4.2 If using acid alumina, pack the column by
adding 6 g acid alumina (Section 6.5.2.1).
If using basic alumina, substitute 6 g
basic alumina (Section 6.5.2.2). Tap the
column to settle the adsorbents.
12,4.3 Pre-rinse the column with 50-100 ml of
hexane. Close the stopcock when the
hexane is.within 1 mm of the alumina.
12.4.4 Discard the eluate. Check the column for
channeling. If channeling is present,
discard the column and prepare another.
12.4.5 Apply the concentrated extract to the
column. Open the stopcock until the
extract is within 1 mm of the alumina.
12.4.6 Rinse the receiver twice with 1 ml
portions of hexane and apply separately to
the column. Elute the interfering
compounds with 100 ml hexane and discard
the eluate.
150
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12.4.7 The choice of eluting solvents will depend
on the choice of alumina (acid or basic)
made in Section 12.4.2.
12.4.7.1 If using acid alumina, elute the PCDDs and
PCDFs from the column with 20 ml methylene
chloride:hexane (20:80 v/v). Collect the
eluate.
12.4.7.2 If using basic alumina, elute the PCDDs
and PCDFs from the column with 20 ml
methylene chloride:hexane (50:50 v/v).
Collect the eluate.
12.4.8 Concentrate the eluate per Section 11.4 or
11.5 for further cleanup or for injection
into the HPLC or GCHS.
12.5 AX-21/Celite
12.5.1 Cut both ends from a 10 mL disposable
serological pi pet to produce a 10 cm
column. Fire polish both ends and flare
both ends if desired. Insert a glass wool
plug at one end, then pack the column with
1 g of the activated AX-21/Celite to form
a 2 cm long adsorbent bed. Insert a glass
wool plug on top of the bed to hold the
adsorbent in place.
12.5.2 Pre-rinse the column with five ml of
toluene followed by 2 mL methylene
chloride.-methanol:toluene (15:4:1 v/v), 1
mL methylene chloride:cyclohexane (1:1
v/v), and five mL hexane. If the flow
rate of eluate exceeds 0.5 mL per min,
discard the column.
12.5.3 When the solvent is within 1 mm of the
column packing, apply the sample extract
to the column. Rinse the sample container
twice with 1 mL portions of hexane and
apply separately to the column. Apply 2
mL of hexane to complete the transfer.
12.5.4 Elute the interfering compounds with 2 mL
of hexane, 2 mL of methylene
chloride:cyclohexane (1:1 v/v), and 2 mL
of methylene chloride:methanol:toluene
(15:4:1 v/v). Discard the eluate.
12.5.5 Invert the column and elute the PCDDs and
PCDFs with 20 mL of toluene. If carbon
particles are present in the eluate,
filter through glass fiber filter paper.
12.5.6 Concentrate the eluate per Section 11.4 or
11.5 for further cleanup or for injection
into the HPLC or GCHS.
12.6 HPLC (Reference 6)
12.6.1 Column calibration
12.6.1.1 Prepare a calibration standard containing
the 2,3,7,8- isomers and/or other isomers
of interest at a concentration of approxi-
mately 500 pg/uL in methylene chloride.
12.6.1.2 Inject 30 uL of the calibration solution
into the HPLC and record the signal from
the detector. Collect the eluant for re-
use. The elution order will be the tetra-
through octa-isomers.
12.6.1.3 Establish the collect time for the tetra-
isomers and for the other isomers of
interest. Following calibration, flush
the injection system with copious
quantities of methylene chloride, includ-
ing a minimum of five 50-uL injections
while the detector is monitored, to ensure
that residual PCDDs and PCDFs are removed
from the system.
12.6.1.4 Verify the calibration with the calibra-
tion solution after every 20 extracts.
Calibration is verified if the recovery of
the PCDDs and PCDFs from the calibration
standard (Section 12.6.1.1) is 75-125
percent compared to the calibration
(Section 12.6.1.2). 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
system.
12.6.2 Extract cleanup -- HPLC requires that the
column not be overloaded. The column
specified in this method is designed to
handle a maximum of 30 uL of extract. If
the extract cannot be concentrated to less
than 30 uL, it is split into fractions and
the fractions are combined after elution
from the column.
12.6.2.1 Rinse the sides of the vial twice with 30
uL of methylene chloride and reduce to 30
uL with the blowdown apparatus.
12.6.2.2 Inject the 30 uL extract into the HPLC.
12.6.2.3 Elute the extract using the calibration
data .determined in 12.6.1. Collect the
fraction(s) in a clean 20 mL concentrator
tube containing 5 mL of hexane:acetone
(1:1 v/v).
151
-------
12.6.2.4 If an extract containing greater than 100
ng/mL of total PCDD or PCDF is encoun-
tered, a 30 uL methylene chloride blank
shall be run through the system to check
for carry-over.
12.6.2.5 Concentrate the eluate per Section 11.5
for injection into the GCMS.
13 HRGC/HRMS ANALYSIS
13.1 Establish the operating conditions given
in Section 7.1.
13.2 Add 10 uL of the internal standard solu-
tion (Section 6.12) to the sample extract
immediately prior to injection to minimize
the possibility of loss by evaporation,
adsorption, or reaction. If an extract is
to be reanalyzed and evaporation has
occurred, do not add more instrument
internal standard solution. Rather, bring
the extract back to its previous volume
(e.g., 19 uL) with pure nonane only.
13.3 Inject 1.0 uL of the concentrated extract
containing the internal standard solution,
using on-column or splitless injection.
Start the GC column initial isothermal
hold upon injection. Start MS data
collection after the solvent peak elutes.
Stop data collection after the octachloro-
dioxin and furan have eluted. Return the
column to the initial temperature for
analysis of the next extract or standard.
14 SYSTEM AND LABORATORY PERFORMANCE
14.1 At the beginning of each 12-hour shift
during which analyses are performed, GCMS
system performance and calibration are
verified for all unlabeled and labeled
compounds. For these tests, analysis of
the CS3 calibration verification (VER)
standard (Section 6.13 and Table 4) and
the isomer specificity test standards
(Sections 6.16 and Table 5) 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.
14.2 MS resolution -- A static resolving power
of at least 10,000 (10 percent valley
definition) must be demonstrated at appro^
priate masses before any analysis is
performed. Static resolving power checks
14.3.1
14.3.2
14.3.3
14.3.4
14.3.5
must be performed at the beginning and at
the end of each 12-hour shift according to
procedures in Section 7.1.2. Corrective
actions must be implemented whenever the
resolving power does not meet the
requirement.
14.3 Calibration verification
Inject the VER standard
procedure in Section 13.
using the
The m/z abundance ratios for all PCDDs and
PCDFs shall be within the limits in Table
3A; otherwise, the mass spectrometer shall
be adjusted until the m/z abundance ratios
fall within the limits specified, and the
verification test (Section 14.3.1)
repeated. If the adjustment alters the
resolution of the mass spectrometer, reso-
lution shall be verified (Section 7.1.2)
prior to repeat of the verification test.
The peaks representing each unlabeled and
labeled compound in the VER standard must
be present with a S/N of at least 10;
otherwise, the mass spectrometer shall be
adjusted and the verification test
(Section 14.3.1) repeated.
Compute the concentration of each
unlabeled compound by isotope dilution
(Section 7.5) for those compounds that
have labeled analogs (Table 1). Compute
the concentration of the labeled compounds
by the internal standard method. These
concentrations are computed based on the
averaged relative response and averaged
response factor from the calibration data
in Section 7.
For each compound, compare the concentra-
tion with the calibration verification
limit in Table 7. If all compounds meet
the acceptance criteria, calibration has
been verified. 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 resolution
(Section 14.2) and verification (Section
14.3.1) tests, or recalibrate (Section 7).
14.4 Retention times and GC resolution
14.4.1 Retention times
152
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14.4.1.1 Absolute -- The absolute retention times
of the 13C12-1,2,3,4-TCDD and C^-
1,2,3,7,8,9-HxCDD GCMS internal standards
shall be within +15 seconds of the
retention times obtained during calibra-
tion (Section 7.2.4).
14.4.1.2 Relative -- The relative retention times
of unlabeled and labeled PCDDs and PCDFs
shall be within the limits given in Table
2.
14.4.2 GC resolution
14.4.2.1 Inject the isomer specificity standards
(Section 6.16) on their respective
columns.
14.4.2.2 The valley height between 2,3,7,8-TCDD and
the other tetra- dioxin isomers at m/z
319.8965, and between 2,3,7,8-TCDF and the
other tetra- furan isomers at m/z 303.9016
shall not exceed 25 percent on their
respective columns (Figure 3).
14.4.3 If the absolute retention time of any
compound is not within the limits
specified or the 2,3,7,8- isomers are not
resolved, the GC is not performing
properly. In this event, adjust the GC
and repeat the verification test (Section
14.3.1) or recalibrate (Section 7).
14.5 Ongoing precision and accuracy
14.5.1 Analyze the extract of the diluted
precision and recovery standard (PAR)
(Section 10.3.4 or 10.4.4) prior to
analysis of samples from the same set.
14.5.2 Compute the concentration of each PCDD and
PCDF by isotope dilution for those
compounds that have labeled analogs
(Section 7.5). Compute the concentration
of each labeled compound by the internal
standard method.
14.5.3 For each unlabeled and labeled compound,
compare the concentration with the limits
for ongoing accuracy in Table 7. 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, the extraction/concentration
processes are not being performed properly
for that compound. In this event, correct
the problem, re-extract the sample set
(Section 10) and repeat the ongoing
precision and recovery test (Section
14.5). The concentration limits in Table
7 for labeled compounds are based on the
requirement that the recovery of each
labeled compound be in the range of 25-
150%.
14.5.4 Add results which pass the specifications
in Section 14.5.3 to initial and previous
ongoing data for each compound in each
matrix. Update QC charts to form a
graphic representation of continued
laboratory performance. Develop a state-
ment of laboratory accuracy for each PCDD
and PCDF in each matrix type by calculat-
ing 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 = 95% and SR = 5%, the
accuracy is 85-105%.
15 QUALITATIVE DETERMINATION
For a gas chromatographic peak to be
identified as a PCDD or PCDF (either a
unlabeled or a labeled compound), it must
meet all of the criteria in Sections 15.1-
15.4.
15.1 The signals for the two exact m/z's being
monitored (Table 3) must be present, and
must maximize within + 2 seconds of one
another.
15.2 The signal-to-noise ratio (S/N) of each of
the two exact m/z's must be greater than
or equal to 2.5 for a sample extract, and
greater than or equal to 10 for a calibra-
tion standard (see Sections 7.2.3 and
14.3.3).
15.3 The ratio of the integrated ion currents
of both the exact m/z's monitored must be
within the limits in Table 3A.
15.4 The relative retention time of the peaks
representing a unlabeled 2,3,7,8-
substituted PCDD or PCDF must be within
the limits given in Table 2. The
retention time of peaks representing non-
2,3, 7,8-substituted PCDDs or PCDFs must be
within the retention time windows
established in Section 7.3.
15.5 Confirmatory analysis -- Isomer
specificity for all of the 2,3,7,8-substi-
tuted analytes cannot be attained by
analysis on the DB-5 (or equivalent) GC
column alone. The lack of specificity is
153
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of greatest concern for the unlabeled
2,3,7,8-TCDF. Therefore, any sample in
which 2,3,7,8-TCDF is identified by
analysis on a DB-5 (or equivalent) GC
column must have a confirmatory analysis
performed on a DB-225, SP-2330, or equiva-
lent GC column. The operating conditions
in Section 7.1.1 may be adjusted for
analyses on the second GC column, but the
GCMS must meet the mass resolution and
calibration specifications in Section 7.
15.6 If any gas chromatographic peak meets the
identification criteria in Sections 15.1,
15.2, and 15.4, but does not meet the ion
abundance ratio criterion (Section 15.3),
and is not a labeled analog, that sample
must be analyzed on a second GC column, as
in Section 15.5 above. Interferences co-
eluting in either of the two m/z's may
cause the ion abundance ratio to fall out-
side of the limits in Table 3A. If the
ion abundance ratio of the peak fails to
meet the criteria on the second GC column,
then the peak does not represent a PCDD or
PCDF. If the peak does meet all of the
criteria in Sections 15.1-15.4 on the
second GC column, then calculate the
concentration of that peak from the
analysis on the second GC column, accord-
ing to the procedures in Section 16.
15.7 If any gas chromatographic peak that
represents a labeled analog does not meet
all of the identification criteria in
Sections 15.1-15.4 on the second GC
column, then the results may not be
reported for regulatory compliance
purposes and a new aliquot of the sample
must be extracted and analyzed.
16 QUANTITATIVE DETERMINATION
16.1 Isotope dilution -- By adding a known
amount of a labeled compound to every
sample prior to extraction, correction for
recovery of the unlabeled compound can be
made because the unlabeled compound and
its labeled analog exhibit similar effects
upon extraction, concentration, and gas
chromatography. Relative response (RR)
values are used in conjunction with cali-
bration data described in Section 7.5 to
determine concentrations directly, so long
as labeled compound spiking levels are
constant, using the following equation:
where, C is the concentration at the
unlabeled compound in the extract and the
other terms are as defined in Section
7.5.2.
16.1.1 Because of a potential interference, the
labeled analog of OCDF is not added to the
sample. Therefore, this unlabeled analyte
is quantitated against the labeled OCDD.
As a result, the concentration of
unlabeled OCDF is corrected for the
recovery of the labeled OCDD. In
instances where OCDD and OCDF behave
differently during sample extraction,
concentration, and cleanup procedures,
this may decrease the accuracy of the OCDF
results. However, given the low toxicity
of this compound relative to the other
dioxins and furans, the potential decrease
in accuracy is not considered significant.
16.1.2 Because the labeled analog of 1,2,3,7,8,9-
HxCDD is used as an internal standard
(i.e., not added before extraction of the
sample), it cannot be used to quantitate
the unlabeled compound by strict isotope
dilution procedures. Therefore, the
unlabeled 1,2,3,7,8,9-HxCDD is quantitated
using the average of the responses of the
labeled analogs of the other two 2,3,7,8-
substituted HxCDD's, 1,2,3,4,7,8-HxCDD and
1,2,3,6,7,8-HxCDD. As a result, the
concentration of the unlabeled
1,2,3,7,8,9-HxCDD is corrected for the
average recovery of the other two HxCDD's.
16.1.3 Any peaks representing non-2,3,7,8-substi-
tuted dioxins or furans are quantitated
using an average of the response factors
from all of the labeled 2,3,7,8- isomers
in the same level of chtori nation.
16.2 Internal standard -- Compute the concen-
trations of the C-labeled analogs and
the C-labeled cleanup standard in the
extract using the response factors deter-
mined from calibration data (Section 7.6)
and the following equation:
Cex (ng/mL) =
Cis
A.s) RF
Cex (ng/mL) =
Gis
where, C is the concentration of the
compound in the extract and the other
terms are as defined in Section 7.6.1.
(NOTE: There is only one m/z for the
37Cl-labeled standard.)
RR
154
-------
16.3
16.4
16.5
16.5.1
16.5.2
16.5.3
16.6
The concentration of the unlabeled
compound in the solid phase of the sample
is computed using the concentration of the
compound in the extract and the weight of
the solids (Section 10), as follows:
/r
ex
Concentration
in solid (ng/Kg) u
s
where,
V is the extract volume in ml.
6X
W is the sample weight in Kg.
The concentration of the unlabeled
compound in the aqueous phase of the
sample is computed using the concentration
of the compound in the extract and the
volume of water extracted (Section 10.3),
as follows:
(Cex *
Vs
Concentration
in aqueous phase
(P9/L)
where,
V is the extract volume in ml.
ex
V is the sample volume in liters.
S
If the SICP areas at the quantisation
m/z's for any compound exceed the calibra-
tion range of the system, a smaller sample
aliquot is extracted.
For aqueous samples containing one percent
solids or less, dilute 100 ml, 10 ml,
etc., of sample to 1 liter with reagent
water and extract per Section 11.
For samples containing greater than one
percent solids, extract an amount of
sample equal to 1/10, 1/100, etc., of the
amount determined in Section 10.1.3.
Extract per Section 10.4.
If a smaller sample size will not be
representative of the entire sample,
dilute the sample extract by a factor of
10, adjust the concentration of the
instrument internal standard to 100 pg/uL
in the extract, and analyze an aliquot of
this diluted extract by the internal
standard method.
Results are reported to three significant
figures for the unlabeled and labeled
isomers found in all standards, blanks,
and samples. For aqueous samples, the
units are pg/L; for samples containing
greater than one percent solids (soils,,
sediments, filter cake, compost), the
units are ng/Kg based on. the dry weight of
the sample.
16.6.1 Results for samples which have been
diluted are reported at the least dilute
level at which the areas at the quantita-
tion m/z's are within the calibration
range (Section 16.5).
16.6.2 Fpr unlabeled compounds having a labeled
analog, results are reported at the least
dilute level at which the area at the
quantisation m/z is within the calibration
range (Section 16.5) and the labeled
compound recovery is within the normal
range for the method (Section 17.4).
16.6.3 Additionally, the total concentrations of
all isomers in an individual level of
chlorination (i.e., total TCDD, total
PeCDD, etc.) are reported to three signi-
ficant figures in units of pg/L, for both
dioxins and furans. The total or ng/Kg
concentration in each level of chlorina-
tion is the sum of the concentrations of
all isomers identified in that level,
including any non-2,3,7,8-substituted
isomers.
17 ANALYSIS OF COMPLEX SAMPLES
17.1 Some samples may contain high levels (>10
ng/L; >1000 ng/Kg) of the compounds of
interest, interfering compounds, and/or
polymeric materials. Some extracts will
not concentrate to 10 uL (Section 11);
others may overload the GC column and/or
mass spectrometer.
17.2 Analyze a smaller aliquot of the sample
(Section 16.4) when the extract will not
concentrate to 20 uL after all cleanup
procedures have been exhausted.
17.3 Recovery of labeled compound spiking
standards -- In most samples, recoveries
of the labeled compound spiking standards
will be similar to those from reagent
water or from the alternate matrix
(Section 6.6). If recovery is outside of
the 25-150% range, a diluted sample
(Section 16.4) shall be analyzed. If the
recoveries of the labeled compound spiking
standards in the diluted sample are
outside of the limits (per the criteria
above), then the verification standard
(Section 14.3) shall be analyzed and
calibration verified (Section 14.3.4). If
the calibration cannot be verified, a new
155
-------
calibration must be performed and the
original sample extract reanalyzed. If
the calibration is verified and the
diluted sample does not meet the limits
for labeled compound recovery, then the
method does not apply to the sample being
analyzed and the result may not be
reported for regulatory compliance
purposes.
18 METHOD PERFORMANCE
The performance specifications in this
method are based on the analyses of more
than 400 samples, representing matrices
from at least five industrial categories.
These specifications will be updated
periodically as more data are received,
and each time the procedures in the method
are revised.
REFERENCES
1 Tondeur, Yves, "Method 8290: Analytical
Procedures and Quality Assurance for
Multimedia Analysis of Polychlorinated
Dibenzo-p-dioxins and Dibenzofurans by
High-Resolution Gas Chromatography/High-
Resolution Mass Spectrometry", USEPA EMSL,
Las Vegas, Nevada, June 1987.
2 "Measurement of 2,3,7,8-Tetrachlorinated
Dibenzo-p-dioxin (TCDD) and 2,3,7,8-Tetra-
chlorinated Dibenzofuran (TCDF) in Pulp,
Sludges, Process Samples and Waste-waters
from Pulp and Paper Hills", Wright State
University, Dayton, OH 45435, June 1988.
3 "NCASI Procedures for the Preparation and
Isomer Specific Analysis of Pulp and Paper
Industry Samples for 2,3,7,8-TCDD and
2,3,7,8- TCDF", National Council of the
Paper Industry for Air and Stream Improve-
ment, 260 Madison Avenue, New York, NY
10016, Technical Bulletin No. 551, Pre-
release Copy, July 1988.
4 "Analytical Procedures and Quality
Assurance Plan for the Determination of
PCDD/PCDF in Fish", USEPA, Environmental
Research Laboratory, 6201 Congdon
Boulevard, Duluth, MN 55804, April 1988.
5 Tondeur, Yves, "Proposed GC/HS Methodology
for the Analysis of PCDDs and PCDFs in
Special Analytical Services Samples",
Triangle Laboratories, Inc., 801-10
Capitola Dr, Research Triangle Park, NC
27713, January 1988; updated by personal
communication September 1988.
6 Lamparski, L.L., and Nestrick, T.J.,
"Determination of Tetra-, Hexa-, Hepta-,
and Octachlorodibenzo-p-dioxin Isomers in
Particulate Samples at Parts per Trillion
Levels", Analytical Chemistry. 52: 2045-
2054, 1980.
7 Lamparski, L.L., and Nestrick, T.J.,
"Novel Extraction Device for the
Determination of Chlorinated Dibenzo-p-
dioxins (PCDDs) and Dibenzofurans (PCDFs)
in Matrices Containing Water",
Chemosphere. 19:27-31, 1989.
8 Patterson, D.G., et. al. "Control of
Interferences in the Analysis of Human
Adipose Tissue for 2,3,7,8-Tetra-
chlorodibenzo-p-dioxin", Environmental
Toxicological Chemistry. 5: 355-360, 1986.
9 Stanley, John S., and Sack, Thomas M.,
"Protocol for the Analysis of 2,3,7,8-
Tetrachlorodibenzo-p-dioxin by High-
Resolution Gas Chromatography/High-
Resolution Mass Spectrometry", USEPA EMSL,
Las Vegas, Nevada 89114, EPA 600/4-86-004,
January 1986.
10 "Working with Carcinogens", DHEW, PHS,
CDC, NIOSH, Publication 77-206, August
1977.
11 "OSHA Safety and Health Standards, General
Industry" OSHA 2206, 29 CFR 1910, January
1976.
12 "Safety in Academic Chemistry
Laboratories", ACS Committee on Chemical
Safety, 1979.
13 "Standard Methods for the Examination of
Water and Wastewater", 16th edition and
later revisions, American Public Health
Association, 1015 15th St, N.W.,
Washington, DC 20005, 46: Section 108
(Safety), 1985.
14 "Method 613 -- 2,3,7,8-Tetrachlorodibenzo-
p-dioxin", 40 CFR 136 (49 FR 43234),
October 26, 1984, Section 4.1.
15 Provost, L.P., and Elder, R.S.,
"Interpretation of Percent Recovery Data",
American Laboratory. 15: 56-83, 1983.
16 "Handbook of Analytical Quality Control in
Water and Wastewater Laboratories", USEPA
EMSL, Cincinnati, OH 45268, EPA-600/4-79-
019, March 1979.
156
-------
17 "Standard Practice for Sampling Water",
ASTH Annual Book of Standards, ASTM, 1916
Race Street, Philadelphia, PA 19103-1187,
1980.
18 "Methods 330.4 and 330.5 for Total
Residual Chlorine", USEPA, EHSL, Cincin-
nati, OH 45268, EPA 600/4-70-020, March
1979.
19 Barnes, Donald G., Kutz, Frederick W., and
Bottimore, David P., "Update of Toxicity
Equivalency Factors (TEFs) for Estimating
Risks Associated with Exposures to
Mixtures of Chlorinated Dibenzo-p-Dioxins
and Dibenzofurans (CDDs/CDFs)", Risk
Assessment Forum, USEPA, Washington, DC
20460, February 1989.
157
-------
Table 1
POLYCHLORINATED DIBENZODIOXINS AND FURANS DETERMINED BY ISOTOPE DILUTION AND INTERNAL STANDARD
HIGH RESOLUTION GAS CHROHATOGRAPHY (HRGO/HIGH RESOLUTION MASS SPECTROMETRY (HRMS)
PCDDs/PCDFs (1)
Isomer/Congener CAS Registry
2,3,7,8-TCDD
Total-TCDD
2,3,7,8-TCDF
Total-TCDF
1,2,3,7,8-PeCDD
Total-PeCDD
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
Total-PeCDF
1,2,3,4,7,8-HxCDD
1',2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
Total-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
Total-HxCDF
1,2,3,4,6,7,8-HpCDD
Total-HpCDD
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
Total-HpCDF
OCDD
OCDF
(1) Polychlorinated dioxins and
1746-01-6
41903-57-5
51207-31-9
55722-27-5
40321-76-4
36088-22-9
57117-41-6
57117-31-4
30402-15-4
39227-28-6
57653-85-7
19408-74-3
34465-4608
70648-26-9
57117-44-9
72918-21-9
60851-34-5
55684-94-1
35822-46-9
37871-00-4
67562-39-4
55673-89-7
38998-75-3
3268-87-9
39001-02-0
furans
TCDD = Tetrachlorodibenzo-p-dioxin
PeCDD = Pentachlorodibenzo-p-dioxin
HxCDD = Hexachlorodibenzo-p-dioxin
HpCDD = Heptachlorodibenzo-p-dioxin
OCDD = Octachlorodibenzo-p-dioxin
Labeled
13C -2 3 7
12
37Cl4-2,3,7
13C12-2,3,7
13C12-1,2,3
!IC12-1'2'3
C12-2,3,4
13C -1 ? 3
13 12
C -123
13 12 ' '
C -123
13C -1 ? 1
13 12
C -123
•IT 12 ' '
r -1 ? 7
13C12 1'2'3
C,,-2 3 4
13C12-1,2,3
!3C12-1'2'3
c -1 ? 7
13C12-OCDD
none
TCDF
PeCDF =
HxCDF =
HpCDF =
OCDF
Analog
,8-TCDD
,8-TCDD
,8-TCDF
,7,8-PeCDD
,7,8-PeCDF
,7,8-PeCDF
4 7 8-HxCDD
,6,7,8-HxCDD
7 8 9-HxCDD(2)
4 7 8-HxCDF
,6,7,8-HxCDF
,7,8,9-HxCDF
6 7 8-HxCDF
,4,6,7,8-HpCDD
,4,6,7,8-HpCDF
,4,7,8,9-HpCDF
Tetrachlorodibenzofuran
Pentachlorodibenzofuran
Hexachlorodibenzofuran
Heptachlorodibenzofuran
Octachlorodibenzofuran
CAS Registry
76523-40-5
85508-50-5
89059-46-1
109719-79-1
109719-77-9
116843-02-8
109719-80-4
109719-81-5
109719-82-6
114423-98-2
116843-03-9
116843-04-0
116843-05-1
109719-83-7
109719-84-8
109719-94-0
114423-97-1
(2) Labeled analog is used as an internal standard and therefore is not used for quantitation of the native
compound.
158
-------
Table 2
RETENTION TIMES AND MINIMUM LEVELS FOR PCDDs AND PCDFs
Minimum Level (1)
Compound
Compounds using C.,-1,2,
Native Compounds
2,3,7,8-TCDF
2,3,7,8-TCDD
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,7,8-PeCDD
Labeled Compounds
13Cl2-2,3,7,8-TCDF
13C12-1, 2, 3,4-TCDD
13Cl2-2,3,7,8-TCDD
37Cl4-2,3,7,8-TCDD
13Cl2-1,2,3,7,8-PeCDF
13Cl2-2,3,4,7,8-PeCDF
13Cl2-1,2,3,7,8-PeCDD
Compounds using C-2-1,2,
Native Compounds
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
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
1,2,3,4,6,7,8-HpCDF
1,2,3,4,6,7,8-HpCDD
1,2,3,4,7,8,9-HpCDF
OCDD
OCDF
Labeled Compounds
13C._-1,2,3,4,7,8-HxCDF
l:>Cl2-1,2,3,6,7,8-HxCDF
13Cl2-1,2,3,7,8,9-HxCDF
13Cl2-2,3,4,6,7,8-HxCDF
13Cl2-1,2,3,4,7,8-HxCDD
13Cl2-1,2,3,6,7,8-HxCDD
13C12-1,2,3,7,8,9-HxCDD
13C..,-1,2,3,4,6,7,8-HpCDF
' C.0-1,2,3,4,6,7,8-HpCDD
'^2-1,2,3,4,7 8,9-HpCDF
C12-OCDD
Retention
Time
Reference
3,4-TCDD as internal standard
|3C12-2,3,7,8-TCDF
3C12-2,3,7,8-TCDD
13Cl2-1,2,3,7,8-PeCDF
]3C12-2,3,4,7,8-PeCDF
13Cl2-1,2,3,7,8-PeCDD
13C12-1, 2, 3,4-TCDD
]3C12-1,2,3,4-TCDD
13C12-1,2,3,4-TCDD
3C12-1,2,3,4-TCDD
JC12-1,2,3,4-TCDD
3C12-1,2,3,4-TCDD
13C12-1,2,3,4-TCDD
3,7,8,9-HxCDD as internal standard
13C12-1,2,3,4,7,8-HxCDF
13Cl2-1,2,3,6,7,8-HxCDF
13Cl2-1,2,3,7,8,9-HxCDF
13Cl2-2,3,4,6,7,8-HxCDF
13Cl2-1,2,3,4,7,8-HxCDD
13Cl2-1,2,3,6,7,8-HxCDD
13Cl2-1,2,3,6,7,8-HxCDD
13C12-1,2,3,4,6,7,8-HPCDF
13C,,-1,2,3,4,6,7,8-HpCDD
C.,-1,2,3,4,7,8,9-HpCDF
C..-OCDD
13 "
C12-OCDD
13C12-1, 2, 3,7,8,9-HXCDD
13Cl2-1,2,3,7,8,9-HxCDD
13Cl2-1,2,3,7,8,9-HxCDD
13C.,-1,2,3,7,8,9-HxCDD
1-7 12 ' ' ' ' '
C12-1,2,3,7,8,9-HxCDD
13C12-1,2,3,7,8,9-HxCDD
13C12-1,2,3,7,8,9-HxCDD
13C12-1,2,3,7,8,9-HxCDD
13C..>-1,2,3,7,8,9-HxCDD
C12-1,2,3,7,8,9-HxCDD
13C.,-1,2,3,7,8,9-HxCDD
Relative Water Solid
Retention pg/L ng/kg
Time ppq ppt
0.993 -
0.993 -
0.918 -
0.999 -
0.987 -
0.931 -
1.000 -
0.993 -
1.002 -
1.091 -
1.123 -
1.134 -
0.986 -
0.973 -
0.937 -
0.999 -
0.999 -
0.992 -
0.986 -
0.930 -
0.986 -
0.896 -
0.996 -
0.995 -
0.947 -
0.940 -
0.993 -
0.971 -
0.974 -
0.975 -
1.000 -
0.953 -
1.023 -
1.024 -
1.050 -
1.009 10 1
1.009 10 1
1.076 50 5
1.001 50 5
1.016 50 5
0.994
1.000
1.036
1.013
1.371
1.408
1.428
1.015 50 5
1.025 50 5
1.068 50 5
1.001 50 5
.001 50 5
.009 50 5
.016 50 5
.022 50 5
.016 50 5
.079 50 5
1.005 100 10
1.013 100 10
0.992
1.006
1.017
1.000
1.002
1.006
1,000
1.172
1.125
1.148
1.275
Extract
pg/uL
ppb
0.5
0.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
5.0
5.0
(1) Level at which the analytical system will give acceptable SICP and calibration.
159
-------
DESCRIPTORS,
Table 3
MASSES, M/Z TYPES, AND ELEMENTAL COMPOSITIONS OF THE CDDs AND CDFs (1)
Descriptor Accurate
Number m/z (2)
1 292.9825
303.9016
305.8987
315.9419
317.9389
319.8965
321 .8936
327.8847
330.9792
331.9368
333.9339
375.8364
_
^ 339.8597
341.8567
351.9000
353.8970
354.9792
355.8546
357.8516
367.8949
369.8919
409.7974
3 373.8208
375.8178
383.8639
385.8610
389.8157
391.8127
392.9760
401.8559
403.8529
430.9729
445.7555
m/z
Type
Lock
M
M+2
M
M+2
M
M+2
M
QC
M
M+2
M+2
M+2
M+4
M+2
M+4
Lock
M+2
M+4
M+2
M+4
M+2
M+2
M+4
M
M+2
M+2
M+4
Lock
M+2
M+4
QC
M+4
Elemental Composition
C7F11
C._ H, 35Cl. 0
124 4
35 37
C._ H, "clJ^Cl 0
124 3
13C15 H, 35Cl, 0
\i 4 4
13C12 H4 35C13 37Cl 0
C12H435CI402
35 37
CM J-'r 1 ' r 1 n
*— n , "^7 "^ *^T
C12 H4 37cl4 °2
C7F13
13C12 H4 35C14 02
13 35 37
C12 H4 C13 Cl °2
C H 35Cl 37Cl 0
C12 H4 C15 Cl 0
35 37
Cu p i r 1 n
._n7 LI . LIU
123 4
35 37
C12 H, *5C13 ^C12 0
13C12 H, 35C14 37Cl 0
13 35 37
C12 H3 C13 C12 °
C9F13
35 37
C12 H, "Cl, ^Cl O,
C.- H3 35Cl 37Cl 0?
13 35 . 37 .
C12 H3 C13 C12 02
35 37
C12 H3 C16 Cl °
C._ H, 35Cl,. 37Cl 0
122 5
C12 H, 35C14 37C12 0
13C. H 35CU 0
\i i o
13C12 H, 35C15 37Cl 0
35 37
C., H- CU Cl 0_
12 2 5 2
35 37
C12 H2 C14 C12 02
C9F15
13C12 H2 35C15 37Cl 02
13C H 35cl 37cl Q
C9F13
C12 H2 35cl6 37cl2 °
Compound
(3)
PFK
TCDF
TCDF
TCDF(4)
TCDF(4)
TCDD
TCDD
TCDD(5)
PFK
TCDD(4)
TCDD(4)
HxCDPE
PeCDF
PeCDF
PeCDF(4)
PeCDF(4)
PFK
PeCDD
PeCDD
PeCDD(4)
PeCDD(4)
HpCDPE
HxCDF
HxCDF
HxCDF(4)
HxCDF(4)
HxCDD
HxCDD
PFK
HxCDD(4)
HxCDD(4)
PFK
OCDPE
Primary
m/z?
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
160
-------
Table 3 (continued)
DESCRIPTORS, MASSES, M/Z TYPES, AND ELEMENTAL COMPOSITIONS OF THE CDDs AND CDFs (1)
Descriptor
Number
(1)
(2)
4
5
Accurate
m/z (2)
407.
409.
417.
419.
423.
425.
430.
435.
437.
479.
441.
442.
443.
457.
459.
469.
471.
513.
From Reference
Nuclidic
7818
7789
8253
8220
7766
7737
9729
8169
8140
7165
7428
9728
7399
7377
7348
7779
7750
6775
5
m/z
Type
M+2
M+4
M
M+2
M+2
M+4
Lock
M+2
M+4
M+4
M+2
Lock
M+4
M+2
M+4
M+2
M+4
M+4
Elemental
C12
C12
13C
13c
C12
C12
C9
13C
13C
C12
C12
C10
C12
C12
C12
13C
13C
C12
H
H
12
12
H
H
35
35
H
H
35
35
Cl
Cl
35
35
Cl
Cl
Composition
6 37cl °
ci7o
F17
12
12
H
35
H
H
35
Cl
35
35
Cl
7
37CIO
F17
35
35
35
12
12
35
Cl
Cl
Cl
35
35
Cl
6
7
6
Cl
Cl
8
37ci2 o
37cl o2
6 37C12 °2
37ci2 o
Compound
(3)
HpCDF
HpCDF
HpCDF(4)
HpCDF(4)
HpCDD
HpCDD
PFK
HpCDD(4)
HpCDD(4)
NCDPE
OCDF
PFK
OCDF
OCDD
OCDD
OCDD (4)
OCDD(4)
DCDPE
Primary
m/z?
Yes
Yes
Yes
Yes
Yes
Yes
Yes
masses used:
H = 1.007825
0 = 15.994915
(3)
Compound
C = 12.00000
35Cl = 34.968853
13C = 13.003355
F = 18.9984
37Cl = 36.965903
abbreviations:
Chlorinated dibenzo-p-dioxins
TCDD
PeCDD
HxCDD
HpCDD
OCDD
Chlorinated diphenyl ethers
= Tetrachlorodibenzo-p-dioxin
= Pentachlorodibenzo-p-dioxin
= Hexachlorodibenzo-p-dioxin
= Heptachlorodibenzo-p-dioxin
= Oetachlorodibenzo-p-dioxin
HXCDPE =
HpCDPE =
OCDPE
NCDPE
DCDPE
Hexachlorodiphenyl ether
Heptachlorodiphenyl ether
Octachlorodiphenyl ether
Nonachlorodiphenyl ether
Decachlorodiphenyl ether
Chlorinated dibenzofurans
TCDF = Tetrachlorodibenzofuran
PeCDF = Pentachlorodibenzofuran
HxCDF = Hexachlorodibenzofuran
HpCDF = Heptachlorodibenzofuran
Lock mass and QC compound
PFK = Perfluorokerosene
(4) Labeled.compound
37,
(5) There is only one m/z for Cl^-2,3,7,8-TCDD (cleanup standard).
161
-------
Table 3A
THEORETICAL ION ABUNDANCE RATIOS AND CONTROL LIMITS
No. of
Chlorine
Atoms
4 (2)
5
6
6 (3)
7
7 (4)
8
m/z's
Forming
Ratio
H/H+2
M+2/H+4
M+2/M+4
M/M+2
H+2/M+4
M/M+2
M+2/M+4
Theoretical
Ratio
0.77
1.55
1.24
0.51
1.05
0.44
0.89
Control
Lower
0.65
1.32
1.05
0.43
0.88
0.37
0.76
Limitsd)
Upper
0.89
1.78
1.43
0.59
1.20
0.51
1.02
(1) Represent + 15% windows around the theoretical ion
abundance ratios.
(2) Does not apply to 37Cl,-2,3,7,8-TCDD (cleanup
standard).
(3) Used for C-HxCDF only.
(4) Used for 13C-HpCDF only.
162
-------
Table 4
CONCENTRATIONS OF SOLUTIONS CONTAINING LABELED AND UNLABELED PCDDS AND PCDFS -.-
STOCK AND SPIKING SOLUTIONS
Labeled
Compound
Stock
Labeled
Compound
Spiking
Solution (1) Solution (2)
Compound
Native CDDs and CDFs
2,3,7,8-TCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDD
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
OCDD
OCDF
Labeled CDDs and CDFs
13Cl2-2,3,7,8-TCDD
13Cl2-2,3,7,8-TCDF
13C12-1,2,3,7,8-PeCDD
13C._-1,2,3,7,8-PeCDF
"IT 1 C.
l:>C12-2,3,4,7,8-PeCDF
13C10-1,2,3,4,7,8-HxCDD
1-7 12
C12-1,2,3,6,7,8-HxCDD
13C.2-1,2,3,4,7,8-HxCDF
13C12-1,2,3,6,7,8-HxCDF
13C12-1,2,3,7,8,9-HxCDF
13C12-2,3,4,6,7,8-HxCDF
13C -1,2,3,4,6,7,8-HpCDD
1-7 \e.
'^2-1,2,3,4,6,7,8-HpCDF
13C12-1,2,3,4,7,8,9-HPCDF
13C12-OCDD
Cleanup Standard
37Cl4-2,3,7,8-TCDD
Internal Standards
13C12-1,2,3,4-TCDD
Cl2-1,2,3,7,8,9-HxCDD
(1) Section 6.10 - prepared in
(2) Section 10.3.2 - prepared
(ng/mL)
-
-
-
-
-
-
-
.
-
-
-
-
-
-
-
-
-
100
100
100
100
100
100
100
100
100
100
100
100
100
100
200
-
-
-
nonane and diluted
from stock solution
(ng/mL)
-
-
-
-
-
.
-
.
-
-
-
-
-
-
.
-
-
2
2
2
2
2
2
2
2
2
2
2
2
2
2
4
-
-
-
to prepare
daily.
PAR
Stock
Solution (3)
(ng/mL)
40
40
200
200
200
200
200
200
200
200
200
200
200
200
200
400
400
-
-
-
-
-
-
-
.
.
_
-
-
-
-
-
-
-
-
spiking solution.
(3) Precision and Recovery (PAR) standard. Section 6.14 - prepared in nonane and
solution in Section 10.3.4
(4) Section 6.11 - prepared in
(5) Section 6.12 - prepared in
.
nonane.
nonane.
Cleanup Internal
Standard Standard
Spiking Spiking
Solution (4) Solution (5)
(ng/mL) (ng/mL)
-
-
-
-
.
.
.
.
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
.
.
.
-
-
-
-
-
0.8
200
200
diluted to prepare spiking
163
-------
Table 4 (continued)
CONCENTRATIONS OF SOLUTIONS CONTAINING LABELED AND UNLABELED PCDDS AND PCDFS
CALIBRATION AND VERIFICATION SOLUTIONS
Compound
Native CDDs and CDFs
2,3,7,8-TCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDD
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
OCDD
OCDF
Labeled CDDs and CDFs
13Cl2-2,3,7,8-TCDD
13Cl2-2,3,7,8-TCDF
13C12-1,2,3,7,8-PeCDD
3C12-1,2,3,7,8-PeCDF
l3C12-2,3,4,7,8-PeCDF
13C12-1,2,3,4,7,8-HxCDD
13Cl2-1,2,3,6,7,8-HxCDD
13C12-1,2,3,4,7,8-HxCDF
13Cl2-1,2,3,6,7,8-HxCDF
13C12-1,2,3,7,8,9-HxCDF
13Cl2-2,3,4,6,7,8-HxCDF
13C12-1,2,3,4,6,7,8-HPCDD
1 C.,-1,2,3,4,6,7,8-HpCDF
' C12-1, 2,3,4, 7^8,9-HpCDF
C12-OCDD
Cleanup Standard
37Cl4-2,3,7,8-TCDD
Internal Standards
13C12-1,2,3,4-TCDD
Cl2-1,2,3,7,8,9-HxCDD
CS1
(ng/mL)
0.5
0.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
2.5
5.0
5.0
100
100
100
100
100
100
100
100
100
100
100
100
100
100
200
0.5
100
100
CS2
(ng/mL)
2
2
10
10
10
10
10
10
10
10
10
10
10
10
10
20
20
100
100
100
100
100
100
100
100
100
100
100
100
100
100
200
2
100
100
VER(6)
CS3
(ng/mL)
10
10
50
50
50
50
50
50
50
50
50
50
50
50
50
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
200
10
100
100
CS4
(ng/mL)
40
40
200
200
200
200
200
200
200
200
200
200
200
200
200
400
400
100
100
100
100
100
100
100
100
100
100
100
100
100
100
200
40
100
100
CS5
(ng/mL)
200
200
1000
1000
1000
1000
1000
1000
1000
1000
1000
1000
1000
1000
1000
2000
2000
100
100
100
100
100
100
100
100
100
100
100
100 .
100
100
200
200
100
100
(6) Section 14.3 - calibration verification (VER) solution.
164
-------
Table 5
GC RETENTION TIME. WINDOW DEFINING STANDARD MIXTURES AND
ISOMER SPECIFICITY TEST STANDARD MIXTURES
DB-5 Column GC Retention Time Window Defining Standard
(Section 6.15)
Congener First Eluted Last Eluted
TCDF
TCDD
PeCDF
PeCDD
UyrnP
n AUU r
HxCDD
HpCDF
HpCDD
1,3,6,8-
1,3,6,8-
1,3,4,6,8-
1,2,4,7,9-
1 2,3,4,6,8-
1,2,4,6,7,9-
1,2,3,4,6,7,8-
1,2,3,4,6,7,9-
1,2,8,9-
1,2,8,9-
1,2,3,8,9-
1,2,3,8,9-
1,2,3,4,8,9-
1,2,3,4,6,7-
1,2,3,4,7,8,9-
1,2,3,4,6,7,8-
DB-5 TCDD Isomer Specificity Test Standard
(Section 6.16.1)
1,2,3,4-TCDD 1,2,3,7-TCDD
1,2,7,8-TCDD 1,2,3,8-TCDD
1,4,7,8-TCDD 2,3,7,8-TCDD
DB-225 Column TCDF Isomer Specificity Test Standard
(Section 6.16.2)
2,3,4,7-TCDF
2,3,7,8-TCDF
1,2,3,9-TCDF
164a
-------
Table 6
REFERENCE COMPOUNDS FOR QUANTITATION OF NATIVE AND LABELED PCDDS AND PCDFS
Native PCDDs and PCDFs
2,3,7,8-TCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDD
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
OCDD
OCDF
(1) 1,2,3,7,8,9-HxCDD is
1,2,3,6,7,8-HxCDD.
Reference Compound
13C12-2,3,7,8-TCDD
13C12-2,3,7,8-TCDF
13C12-1,2,3,7,8-PeCDD
13C -1,2,3, 7,8-PeCDF
13
C -2 3
L1P <-,->,
13
^C.,-1,2,3,
13C1?-1,2,3,
13C 1 2 3
., 12 ' ' '
13C,,- 1,2,3,
17 12 ' '
C12-1,2,3,
C -234
C -1234
,, 12 ' ' ' '
I3C,,-1,2,3,<4,
l:>C12-1,2,3,4,
quantified using
4,7,8-PeCDF
4,7,8-HxCDD
6,7,8-HxCDD
(1)
6,7,8-HxCDF
7,8,9-HxCDF
6 7 8-HxCDF
6 7 8-HpCDD
6,7,8-HpCDF
7,8,9-HpCDF
13
1 C,, -OCDD
13C -OCDD
the average
Labeled PCDDs and PCDFs
13
13
'cl2-2,3,7,8-TCDD
13
13
13
L12"
13
13
'cl2-1,2,3,7,8-PeCDD
'c12-1,2,3,7,8-PeCDF
'c12-2,3,4,7,8-PeCDF
'C12-1,2,3,4,7,8-HXCDD
'C12-1,2,3,6,7,8-HxCDD
13 I*
l:>C12-1,2,3,7,8,9-HxCDD
'C12-1,2,3,4,7,8-HXCDF
13
13
13
13
13
13
'C12-1,2,3,6,7,8-HXCDF
'C12-1,2,3,7,8,9-HXCDF
'C12~2,3,4,6,7,8-HxCDF
13,
Reference Compound
13C12-1,2,3,4-TCDD
13C12-1,2,3,4-TCDD
13C -1,2,3,4-TCDD
17C12-1,2,3,4-TCDD
C12-1,2,3,4-TCDD
'Cl2-1,2,3,7,8,9-HxCDD
C12-1,2,3,7,8,9-HxCDD
.--1,2,3,7,8,9-HxCDD
'C12-1,2,3,7,8,9-HxCDD
C12-1,2,3,7,8,9-HxCDD
C12-1,2,3,7,8,9-HXCDD
Cl2-1,2,3,7,8,9-HxCDD
13
'c12-1,2,3,4,6,7,8-HpCDD C12'1,2,3,7,8,9-HxCDD
'C12-1,2,3,4,6,7,8-HPCDF
'c,--1,2,3,4,7,8,9-HpCDF
13
C12-OCDD
37Cl4-2,3,7,8-TCDD
13
C12-1,2,3,7,8,9-HxCDD
C12-1,2,3,7,8,9-HxCDD
C12-1,2,3,7,8,9-HxCDD
13C12-1,2,3,4-TCDD
responses for the C 2-1,2,3,4,7,8-HxCDD and C
12
164b
-------
Table 7
ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS
Compound
2,3,7,8-TCDD
2,3,7,8-TCDF
1,2,3,7,8-PeCDD
1,2,3,7,8-PeCDF
2,3,4,7,8-PeCDF
1,2,3,4,7,8-HxCDD
1,2,3,6,7,8-HxCDD
1,2,3,7,8,9-HxCDD
1,2,3,4,7,8-HxCDF
1,2,3,6,7,8-HxCDF
1,2,3,7,8,9-HxCDF
2,3,4,6,7,8-HxCDF
1,2,3,4,6,7,8-HpCDD
1,2,3,4,6,7,8-HpCDF
1,2,3,4,7,8,9-HpCDF
OCDD
OCDF
13C12-2,3,7,8-TCDD
13Cl2-2,3,7,8-TCDF
13C12-1,2,3,7,8-PeCDD
13Cl2-1,2,3,7,8-PeCDF
13C12-2,3,4,7,8-PeCDF
13C,.,-1,2,3,4,7,8-HxCDD
,12 ' '
'C12-1, 2,3,6, 7,8-HxCDD
13C., -1,2,3,4,7,8-HxCDF
17
IJC -1,2, 3,6,7, 8-HxCDF
13
13C.--2,3,4,6,7,8-HxCDF
13 12
'•*(:,., -1,2, 3, 4,6, 7,8-HpCDD
13C._-1,2,3,4,6,7,8-HpCDF
l:>Cl2-1,2,3,4,7,8,9-HpCDF
13C.--OCDD
37 12
Cl4-2,3,7,8-TCDD
Test
Cone. (1) s
(ng/mL) (ng/mL)
• 10
10
50
50
50
50
50
50
50
50
50
50
50
50
50
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
100
200
10
1.5
2.0
4.2
4.6
4.2
5.5
5.5
9.5
6.3
4.0
4.0
5.0
6.4
3.6
4.2
13.0
45.0
.
-
-
-
-
_
-
-
-
„
_
_
-
-
_
-
IPR (2)
X
(ng/mL)
3.9 -
3.2 -
47.5 -
44.2 -
45.3 -
30.9 -
33.2 -
22.7 -
25.2 -
39.1 -
37.9 -
27.4 -
27.4 -
39.5 -
36.6 -
69.4 -
46.1 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
50.0 -
2.5 -
20.6
26.8
50.5
54.0
50.3
70.2
65.9
90.9
92.0
54.4
62.9
85.5
76.5
62.1
64.9
154.6
139.8
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
300.0
15.0
OPR(2)
(ng/mL)
5.9 -
6.6 -
35.6 -
36.7 -
37.8 -
35.1 -
33.3 -
31.8 -
36.9 -
34.8 -
37.1 -
35.7 -
37.5 -
37.4 -
36.9 -
75.6 -
69.5 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
25.0 -
50.0 -
2.5 -
14.2
12.7
58.1
57.3
56.9
60.4
64.4
61.2
58.8
58.8
55.7
60.0
56.8
60.5
60.6
118.7
127.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
150.0
300.0
15.0
VER
(ng/mL)
8.6 -
8.8 -
44.2 -
46.7 -
47.2 -
37.6 -
39.7 -
42.6 -
41.5 -
40.5 -
45.7 -
44.1 -
41.6 -
43.1 -
43.6 -
87.5 -
83.9 -
90.0 -
87.7 -
80.6 -
81.8 -
83.0 -
76.1 -
84.0 -
85.2 -
85.0 -
89.5 -
85.7 -
82.2 -
88.5 -
89.0 -
164.2 -
6.1 -
11.6
11.3
56.6
53.5
53.0
66.5
63.0
58.7
60.2
61.7
54.5
56.7
60.2
58.0
57.3
114.4
119.2
111.2
114.0
124.0
122.3
120.5
131.3
119.1
117.4
117.7
111.7
116.7
121.6
113.1
112.4
243.6
11.6
(1) All specifications are given as concentrations in the final extract or standard solution.
(2) s = standard deviation of the concentration; X = average concentration. Concentration limits for labeled
compounds in IPR and OPR aliquots are based on requirements for labeled compound recovery of 25-150% (Sections
8.2.3 and 14.5.3).
164c
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Table 8
SAMPLE PHASE .AND QUANTITY EXTRACTED FOR VARIOUS MATRICES
Sample Matrix (1)
SINGLE PHASE
Aqueous
Solid
Organic
Example
Drinking water
Groundwater
Treated wastewater
Dry soi I
Compost
Ash
Waste solvent
Waste oil
Organic polymer
Percent
Solids Phase
<1 (2)
>20 Solid
<1 Organic
Quantity
Extracted
1000 mL
10 g
10 g
MULTIPHASE
Liquid/Solid
Aqueous/solid
Organic/solid
Liquid/Liquid
Aqueous/organic
Aqueous/organic/
solid
Wet soil 1-30
Untreated effluent
Digested municipal sludge
Filter cake
Paper pulp
Tissue
Industrial sludge 1-100
Oily waste
In-process effluent <1
Untreated effluent
Drum waste
Untreated effluent >1
Drum waste
Solid
Both
Organic
Organic
& solid
10 g
10 g
10 g
10 g
(1) The exact matrix may be vague for some samples. In general, when the CDDs and CDFs are in contact with a
multiphase system in which one of the phases is water, they will be preferentially dispersed in or adsorbed
on the alternate phase, because of their low solubility in water.
(2) Aqueous samples are filtered after spiking with labeled analogs. The filtrate and the material trapped on
the filter are extracted separately, and then the extracts are combined for cleanup and analysis.
164d
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EPA METHOD 1620
METALS BY INDUCTIVELY COUPLED
PLASMA ATOMIC EMISSION SPECTROSCOPY
AND ATOMIC ABSORPTION SPECTROSCOPY
165
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166
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Introduction
Method 1620 was developed by the Industrial Technology
Division (ITD) within the United States Environmental
Protection Agency's (USEPA) Office of Hater Regulations and
Standards (OURS) 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.
Method 1620 includes inductively coupled plasma atomic
emission (ICP) spectroscopy, graphite furnace atomic
absorption (GFAA) spectroscopy, and cold vapor atomic
absorption (CVAA) spectroscopy techniques for analysis of 30
specified metals. The method also includes an ICP technique
for use as a semiquantitative screen for 42 specified
elements.
Questions concerning the method or its application should be
addressed to:
W. A. Telliard
USEPA
Office of Water Regulations and Standards
401 M Street SU
Washington, DC 20460
202/382-7131
OR
USEPA OURS
Sample Control Center
P.O. Box 1407
Alexandria, Virginia 22313
703/557-5040
Publication date: July 1989 DRAFT
167
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168
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Method 1620 DRAFT July 1989
Metals by inductively Coupled Plasma Atomic Emission
Spectroscopy and Atomic Absorption Spectroscopy
1 SCOPE AND APPLICATION
1.1 This method is designed to meet the survey
requirements of the USEPA I TO. It is used
to determine specified elements associated
with the Clean Water Act (as amended
1987); the Resource Conservation and
Recovery Act (as amended 1986); and the
Comprehensive Environmental Response,
Compensation and Liability Act (as amended
1986); and other elements amenable to
analysis by inductively coupled plasma
(ICP) atomic emission Spectroscopy,
graphite furnace atomic absorption (GFAA)
Spectroscopy, and cold vapor atomic
absorption (CVAA) Spectroscopy.
1.2 The method is a consolidation of USEPA
Methods 200.7 (ICP for trace elements),
204.2 (Sb), 206.2 (As), 210.2 (Be), 213.2
(Cd), 218.2 (Cr), 239.2 (Pb), 270.2 (Se),
279.2 (Tl), 245.5 (Hg). 245.1 (Hg), and
245.2 (Hg). The method is used for
analysis of trace elements by ICP atomic
emission Spectroscopy and GFAA
Spectroscopy, for analysis of mercury by
CVAA Spectroscopy, and as a semi-
quantitative ICP screen for specified
elements.
1.3 The elements listed in Tables 1-4 may be
determined in Maters, soils, sediments,
and sludges by this method.
1.4 The recommended wavelengths and instrument
detection limits of this method are shown
in Tables 1-3. Actual sample detection
limits are dependent on the sample matrix
rather than instrumental limitations. The
levels given typify the minimum quantities
that can be detected with no interferences
present. Table 3 also lists the optimum
concentration range.
1.5 Table 4 lists the wavelengths and loner
threshold limits (LTD for the 42 elements
for semiquantitative ICP screen.
1.6 The ICP and AA portions of this method are
for use only by analysts experienced with
the instrumentation or under the close
supervision of such qualified persons.
Each laboratory that uses this method must
demonstrate the ability to generate
acceptable results using the procedure in
Section 8.2.
2 SUMMARY OF METHOD
2.1 ICP-Atomic Emission Spectrometric Method
for Analysis of Water and Soil/Sediment
Samples
2.1.1 The method describes a technique for the
simultaneous or sequential inuiti-element
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.
2.1.2 A background correction technique is
required to compensate for variable
background contribution to the
determination of trace elements.
Background must be measured adjacent to
analyte lines during sample 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 Section 3.1.1 (and tests for
their presence as described in Section
3.1.2) should also be recognized and
appropriate corrections made.
2.1.3 Dissolved elements (those which Mill pass
through a 0.45 urn membrane filter) are
determined in samples that have been
filtered and acidified. Appropriate steps
169
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must be taken in all analyses to ensure
that potential interferences are taken
into account. This is especially true
when dissolved solids exceed 1500 mg/L.
(See Section 3.1.)
2.1.4 Total elements (total concentration in an
unfiltered sample) 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 the effects of
potential interferences. (See Section
3.1.)
2.1.5 Table 1 lists elements that may be
analyzed by this method 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.
2.1.6 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.
2.1.7 The semiquantitative screening procedure
requires a sequential ICP instrument (2
channel minimum) interfaced with a
computerized data system capable of the
short sampling times and the narrow survey
windows necessary to perform a
semiquantitative ICP screen.
2.1.7.1 Table 4 lists the wavelengths to be used
in the semiquantitative ICP screen for
each analyte, and the lower threshold
limits (LTD. The LTL for each analyte is
highly dependent upon sample matrix and
subject to change on a sample-by-sample
basis.
2.1.8 Sludge samples having less than 1% solids
are treated as water samples. Those
having between 1% to 30% solids should be
diluted to less than 1% solids, and then
treated as water samples. Sludge samples
having greater than 30% solids should be
treated as soil samples.
2.2 GFAA Spectroscopy for Analysis of Water
and Soil/Sediment Samples
2.2.1 This method describes a technique for
multi-element determination of trace
elements in solution. A few microliters
of the sample are first evaporated at a
low temperature (sufficient heat to remove
the solvent from the sample) and then
ashed at a higher temperature on an
electrically heated surface of carbon,
tantalum, or other conducting material.
The conductor can be formed as a hollow
tube, a strip, a rod, a boat, or a trough.
After ashing, the current is rapidly
increased to several hundred amperes,
which causes the temperature to rise to
2000-3000 "C; atomization of the sample
occurs in a period of a few milliseconds
to seconds. The absorption or
fluorescence of the atomized particles can
then be measured in the region above the
heated conductor. At the wavelength at
which absorbance (or fluorescence) occurs,
the detector output rises to a maximum
after a few seconds of ignition, followed
by a rapid decay back to zero as the
atomization products escape into the
surroundings. The change is rapid enough
to require a high speed recorder.
2.2.2 The matrix interference problem is one of
the major causes of poor accuracy
encountered with this method. It has been
found empirically that some of the sample
matrix effects and poor reproducibiIity
associated with graphite furnace
atomization can be alleviated by reducing
the natural porosity of the graphite tube.
A background correction technique is
required to compensate for variable
background contribution to the
determination of trace elements.
2.2.3
2.2.4
2.3
2.3.1
2.3.1.1
Table 3 lists the recommended instrumental
operating parameters.
For treatment of sludge samples, see
Section 2.1.8.
Cold Vapor AA (CVAA)
Analysis of Mercury
Techniques for
Manual CVAA Technique for Analysis of
Mercury in Water Samples
The flameI ess AA procedure is a method
based on the absorption of radiation at
253.7 nm by mercury vapor. Mercury
compounds are oxidized and the mercury is
reduced to the elemental state and aerated
from solution in a closed system. The
mercury vapor passes through a cell
170
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positioned in the light path of an atomic
absorption spectrophotometer. Absorbance
(peak height) is measured as a function of
mercury concentration.
2.3.1.2 In addition to inorganic forms of mercury,
organic mercurials may also be present.
These organo-mercury compounds will not
respond to the cold vapor atomic
absorption technique unless they are first
broken down and converted to mercuric
ions. Potassium permanganate oxidizes
many of these compounds, but recent
studies have shown that , a number of
organic mercurials, including phenyl
mercuric acetate and methyl mercuric
chloride, are only partially oxidized by
this reagent. Potassium persulfate has
been found to give approximately 100%
recovery when used as the oxidant with
these compounds. Therefore, a persulfate
oxidation step following the addition of
the permanganate has been included to
ensure that organo-mercury compounds, if
present, will be oxidized to the mercuric
ion before measurement. A heating step is
required for methyl mercuric chloride when
present in or spiked into a natural
system. The heating step is not necessary
for distilled water.
2.3.1.3 The working range of the method may be
varied through instrument and/or recorder
expansion. Using a 100 ml sample, a
detection limit of 0.2 ug Hg/L can be
achieved (see Section 7.2.3).
2.3.1.4 For treatment of sludge samples, see
Section 2.1.8.
2.3.2 Automated CVAA Technique for Analysis of
Mercury in Water Samples
2.3.2.1 See Section 2.3.1.1.
2.3.2.2 See Section 2.3.1.2.
2.3.2.3 The working range of the method is 0.2 to
20.0 ug Hg/L.
2.3.2.4 For treatment of sludge samples, see
Section 2.1.8.
2.3.3 Manual CVAA Technique for Analysis of
Mercury in Soil/Sediment Samples
2.3.3.1 A weighed portion of the sample is
digested in acid for 2 minutes at 95 °C,
followed by oxidation with potassium
permanganate and potassium persulfate.
2.3.3.2
2.3.3.3
3.1
3.1.1
Mercury in the digested sample is then
measured by the conventional cold vapor
technique. An alternate digestion
involving the use of an autoclave is
described in Section 10.5.2.
The working 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/or recorder expansion.
For treatment of sludge samples, see
Section 2.1.8.
INTERFERENCES
Interferences Observed with
Emission Spectrometric Method
ICP-Atomic
Three types of interference effects may
contribute to inaccuracies in the
determination of trace elements:
spectral, physical, and chemical. These
are summarized as follows.
3.1.1.1 Spectral interferences
3.1.1.1.1 Spectral interferences can be categorized
as: 1) overlap of a spectral line from
another element, 2) 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 for 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 for by a
background correction adjacent to the
analyte line. In addition, users of
simultaneous multi-element 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.
3.1.1.1.2 Listed in Table 5 are some interference
effects for the recoirmended wavelengths
given in Table 1. The data in Table 5 are
intended for use only as a rudimentary
guide for the indication of potential
spectral interferences. For this purpose,
linear relations between concentration and
171
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intensity for the analytes and the
interferents can be assumed. The
interference information, which was
collected at the Ames Laboratory (USOOE,
Iowa State University, Ames, Iowa 50011)
is expressed as analyte concentration
equivalents (i.e., false analyte
concentrations) arising from 100 mg/L of
the interferent element.
3.1.1.1.3 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 5, 100 mg/L
of aluminum Mould 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 0.13 mg/L.
The reader is cautioned that other
analytical systems may exhibit somewhat
different levels of interference than
those shown in Table 5, and that the
interference effects must be evaluated for
each individual system. Only those
interferents listed were investigated, and
the blank spaces in Table 5 indicate that
measurable interferences were not observed
from the interferent concentrations listed
in Table 6. Generally, interferences were
discernible if they produced peaks or
background shifts corresponding to 2-5% of
the peak heights generated by the analyte
concentrations also listed in Table 6.
3.1.1.1.4 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 766.491 nm.
3.1.1.2 Physical interferences
3.1.1.2.1 Physical interferences are generally
considered to be effects associated with
the sample nebulization and transport
processes. Changes in properties such as
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 aerosol flow rate and causes
instrumental drift. Internal standards
may also be used to compensate for
physical interferences.
3.1.1.2.2 Wetting the argon prior to nebulization,
the use of a tip washer, or sample
dilution techniques 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. Nebulizers specifically
designed for use with solutions containing
high concentration of dissolved solids may
be used.
3.1.1.3 Chemical interferences -- These interfer-
ences are characterized by molecular
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.
3.1.2 The ICP Serial Dilution Analysis must be
performed on 10% of the samples, or at
least once for each set or Episode of
samples. Samples identified as field
blanks cannot be used for serial dilution
analysis. If the analyte concentration is
sufficiently high (minimally a factor of
50 above the instrumental detection limit
in the original sample), the serial
dilution (a five-fold dilution) must then
agree within 10% of the original
determination after correction for
dilution. If the dilution analysis for
one or more analytes is not within 10%, a
chemical or physical interference effect
must be suspected, and the data for all
affected analytes in the samples
associated with that serial dilution must
be flagged.
3.2 Interferences Observed
Spectroscopic Method
with
GFAA
3.2.1 Interferences of three types are
encountered in atomic absorption methods
using electrothermal atomization:-
172
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spectral, chemical, and physical. These
interferences are summarized as follows.
3.2.1.1 Spectral interferences
3.2.1.1.1 Spectral interferences arise when the
absorption of an interfering species
either overlaps or lies close to the
analyte absorption. Then resolution by
the monochromator becomes impossible.
This effect can be compensated for by
monitoring the presence of the interfering
element.
3.2.1.1.2 Spectral interferences could also arise
because of the presence of combustion
products that exhibit broad band
absorption or participate products that
scatter radiation. This problem can also
originate in the sample matrix itself. If
the source of interference is known, an
excess of the interfering substance can be
added to both the sample and standards.
Provided that the excess is large with
respect to the concentration from the
sample matrix, the contribution from the
sample matrix will become insignificant.
3.2.1.1.3 The matrix interference problem is greatly
exacerbated with electrothermal
atomization; this is one of the major
causes for poor accuracy. Scattering by
incompletely decomposed organic particles
also occurs commonly. As a consequence,
the need for background correction
techniques is encountered with
electrothermal atomization.
3.2.1.2 Chemical interferences are more common
than spectral ones. Their effects can be
minimized by a suitable choice of
operating conditions. These interferences
can be categorized as: 1) formation of
compounds of low volatility which reduces
the rate at which the sample is atomized,
2) lonization of atoms and molecules, and
3) solute vaporization effects. These
interferences can be minimized by varying
the temperature and addition of ionization
suppressor or by standard addition
technique. These interferences can be
highly dependent on the matrix type and
the specific analyte element.
3.2.1.3 Physical interferences are pronounced with
samples containing high dissolved solids
and/or acid concentration resulting in
-change in viscosity and surface tension.
If these types of interferences are
operative, they can be reduced by dilution
of the sample.
3.2.2 Possible interferences observed during
analysis of trace elements by GFAA
spectroscopic methods and certain
recommended instrumental parameters -- All
furnace elements must be analyzed by
method of standard addition (Section
8.15). The use of background correction
is also required for all of these
elements.
3.2.2.1 Antimony
3.2.2.1.1 Nitrogen may also be used as the purge
gas.
3.2.2.1.2 If chloride concentration presents a
matrix problem or causes a loss previous
to atomization, add an excess 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.
3.2.2.2 Arsenic
3.2.2.2.1 The use of background correction is
required. Background correction made by
the deuterium arc method does not
adequately compensate for high levels of
certain interferents (ie., Al, Fe). If
conditions occur where significant
interference is suspected, the laboratory
must switch to an alternate wavelength or
take other appropriate action to
compensate for the interference effects.
3.2.2.2.2 The use of an edlectrodeless discharge
lamp (EDL) for the light source is
recommended.
3.2.2.3 Beryllium
3.2.2.3.1 Because of possible chemical interaction,
nitrogen should not be used as a purge
gas.
3.2.2.4 Cadmium
3.2.2.4.1 Contamination from the work area is
critical in cadmium analysis. Use pipette
tips which are free of cadmium.
3.2.2.5 Chromium
3.2.2.5.1 Hydrogen peroxide is added to the
acidified solution to convert all chromium
to the trivalent state. Calcium is added
173
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to the solution at a level of at least 200
mg/L where its suppress]'ve effect becomes
constant up to 1000 mg/L.
3.2.2.5.2 Nitrogen should not be used as a purge gas
because of possible CN band interference.
3.2.2.5.3 Pipette tips have been reported to be a
possible source of contamination.
3.2.2.6 Lead
3.2.2.6.1 Greater sensitivity can be achieved using
the 217.0 rtm line, but the optimum
concentration range is reduced. The use
of a lead Electrodeless Discharge Lamp at
this lower wavelength has been found to be
advantageous. Also, a lower atomization
temperature (2400 °C) may be preferred.
3.2.2.6.2 To suppress sulfate interference (up to
1500 ppm) lanthanum nitrate is added to
both samples and calibration standards.
(Atomic Absorption Newsletter Vol. 15, No.
3, p. 71, Hay-June 1976).
3.2.2.6.3 Since glassware contamination is a severe
problem in lead analysis, all glassware
should be cleaned immediately prior to
use, and once cleaned, should not be open
to the atmosphere except when necessary.
3.2.2.7 Selenium
3.2.2.7.1 The use of background correction is
required. Background correction made by
the deuterium arc method does not
adequately compensate for high levels of
certain interferents (i.e., Al, Fe). If
conditions occur where significant
interference is suspected, the laboratory
must switch to an alternate wavelength or
take other appropriate actions to
compensate for the interference effects.
3.2.2.7.2 Selenium analysis suffers interference
from chlorides (>800 mg/L) and sulfate
(>200 mg/L). For the analysis of
industrial effluents and samples with
concentrations of sulfate from 200 to 2000
mg/L, both samples and standards should be
prepared to contain 1% nickel.
3.2.2.7.3 The use of an electrodeless discharge lamp
(EDL) for the light source is recommended.
3.2.2.8 Thallium
3.2.2.8.1 Nitrogen may also be used as the purge
gas.
3.3
3.3.1
Interferences Observed with Cold Vapor AA
(CVAA) Techniques for Analysis of Mercury
Manual CVAA technique
mercury in water
for analysis of
3.3.1.1 Possible interference from sulfide is
eliminated by the addition of potassium
permanganate. Concentrations as high as
20 mg/l of sulfide as sodium sulfide do
not interfere with the recovery of added
inorganic mercury from distilled water.
3.3.1.2 Copper may interfere in the analysis of
mercury; however, copper concentrations as
high as 10 rog/L had no effect on recovery
of mercury from spiked samples.
3.3.1.3 Seawaters, brines and industrial effluents
high in chlorides require additional
permanganate (as much as 25 mL). During
the oxidation step, chlorides are
converted to free chlorine which will also
absorb radiation of 253 nm. Care must be
taken to assure that free chlorine is
absent before the mercury is reduced and
swept into the cell. This may be
accomplished by using an excess of
hydroxylamine sulfate reagent (25 ml).
Both inorganic and organic mercury spikes
have been quantitatively recovered from
the seawater using this technique.
3.3.1.4 While the possibility of absorption from
certain organic substances actually being
present in the sample does exist, EPA
laboratories have not encountered such
samples to date. This is mentioned only
to caution the analyst of the possibility.
3.3.2 Automated CVAA technique for analysis of
mercury in water
3.3.2.1 Some seawaters and wastewaters high in
chlorides have shown a positive
interference, probably due to the
formation of free chlorine.
3.3.2.2 Formation of a heavy precipitate, in some
wastewaters and effluents, has been
reported upon addition of concentrated
sulfuric acid. If this is encountered,
the problem sample cannot be analyzed by
this method.
3.3.2.3 If total mercury values are to be
reported, samples containing solids must
be blended and then mixed while being
sampled.
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3.3.3 Manual CVAA technique for analysis of
mercury in soiI
3.3.3.1 The same types of interferences that may
occur in water samples are also possible
with soils/sediments, i.e., sulfides, high
copper, high chlorides, etc.
3.3.3.2 Samples containing high concentrations of
oxidizable organic materials, as evidenced
by high chemical oxygen demand values, may
not be completely oxidized by this
procedure. When this occurs, the recovery
of organic mercury will be low. The
problem can be eliminated by reducing the
weight of the original sample or by
increasing the amount of potassium
persulfate (and consequently stannous
chloride) used in the digestion.
3.3.3.3 Volatile materials which absorb at 253.7
ran will cause a positive interference. In
order to remove any interfering volatile
material, purge the dead air space in the
BOO bottle before the addition of stannous
sulfate.
4 SAFETY
4.1 The toxicity or carcinogenicity of each
reagent used in these methods has not been
precisely defined; however, each chemical
compound should be treated as a potential
health hazard. 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 data
sheets should be made available to all
personnel involved in the chemical
analysis.
5 APPARATUS AND EQUIPMENT
5.1 ICP-Atomic Emission Spectrometer
5.1.1 Sequential ICP instruments (2 channel
minimum) interfaced with a computerized
data system capable of short sampling
times and narrow survey windows necessary
for the semiquantitative ICP screening
procedure and facility for background
correction.
5.1.2 Radio frequency generator.
5.1.3 Argon gas supply, welding grade or better.
5.2 GFAA Spectrometer.
5.2.1 Computer-controlled atomic absorption
spectrometer with background correction.
5.2.2 Argon gas supply, welding grade or better.
5.3 For ICP-Atomic Emission and GFAA, the
following is also required.
5.3.1 250 ml beaker or other appropriate vessel.
5.3.2 Hatch glasses.
5.3.3 Thermometer that covers range of 0 - 200
"C.
5.3.4 Whatman No. 42 filter paper or equivalent.
5.4 Apparatus for manual CVAA mercury analysis
in water
5.4.1 Atomic absorption spectrophotometer: 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: 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.
5.4.2 Mercury hollow cathode lamp: Westinghouse
UL-22847, argon-filled, or equivalent.
5.4.3 Recorder: Any multirange variable speed
recorder that is compatible with the UV
detection system is suitable.
5.4.4 Absorption cell: Standard spectrophoto-
meter 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. The
cell is strapped to a burner for support
and aligned in the light beam by use of
two 2" x 2" cards. One-inch diameter
holes are cut in the middle of each card;
the cards are then placed over each end of
the cell. The cell is then positioned and
adjusted vertically and horizontally to
find the maximum transmittance.
5.4.5 Air pump: Any peristaltic pump capable of
delivering 1 liter of air per minute may
be used. A Masterflex pump with
175
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electronic speed control has been found to
be satisfactory.
5.4.6 Floumter: Capable of measuring an air
flow of 1 liter per minute.
5.4.7 Aeration tubing: A straight glass fit
having a coarse porosity. Tygon tubing is
used for passage of the mercury vapor from
the sample bottle to the absorption cell
and return.
5.4.8 Drying tube: 6" X 3/4" diameter tube
containing 20 g of magnesium perch Iorate.
The apparatus is assembled as shown in
Figure 1. NOTE: 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.
5.5.2 Vapor-liquid separator (Figure 2).
AIR
OUT
7/25 T
MR AND
sm i irinu 9*
IN
1
I
0.4 cm ID
0.7 cm ID
f
1
• SOLUTION
OUT
FIGURE 2 Vapor Liquid Separator
DESICCANT
ABSORPTION
CELL
SAMPLE SOLUTION
IN BOD BOTTLE
.SCRUBBER
CONTAINING
MERCURY
ABSORBING
MEDIA
FIGURE 1 Apparatus for Flameless Mercury
Determination
5.5 Apparatus for automated CVAA mercury
analysis in water
5.5.1 Technicon auto analyzer or equivalent
instrumentation consisting of:
5.5.1.1 Sampler II with provision for sample
mixing.
5.5.1.2 Manifold.
5.5.1.3 Proportioning pump II or III.
5.5.1.4 High temperature heating bath with two
distillation coils (Technicon Part #116-
0163) in series.
5.5.3 Absorption cell, 100 mm long, 10 mm
diameter with quartz windows.
5.5.4 Atomic absorption spectre-photometer (see
Section 5.4.1).
5.5.5 Mercury hollow cathode lamp (see Section
5.4.2).
5.5.6 Recorder (see Section 5.4.3).
5.6 Apparatus for manual CVAA mercury analysis
in soiI/sediment
5.6.1 Atomic Absorption Spectrophotometer (see
Section 5.4.1).
5.6.2 Mercury Hollow Cathode Lamp (see Section
5.4.2).
5.6.3 Recorder (see Section 5.4.3).
5.6.4 Absorption Cell (see Section 5.4.4).
5.6.5 Air Pump (see Section 5.4.5).
5.6.6 Flowmeter (See Section 5.4.6.).
5.6.7 Aeration tubing (see Section 5.4.7).
5.6.8 Drying tube: 6" X 3/4" diameter tube
containing 20 g of magnesium perch I orate
(see NOTE in Section 5.4.8).
176
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6 REAGENTS AND STANDARDS
6.1 ICP-Atomic Absorption Spectrometry
Quantitative screening of 21 elements
6.1.1 Acids used in the preparation of standards
and for sample processing must be ultra-
high purity grade or equivalent.
Redistilled acids are acceptable.
6.1.1.1 Acetic acid, cone, (sp gr 1.06).
6.1.1,2 Hydrochloric acid, cone, (sp gr 1.19).
6.1.1.3 Hydrochloric acid, (1+1): Add 500 mL
cone. HCl (sp gr 1.19) to 400 ml
deionized distilled water and dilute to 1
liter.
6.1.1.4 Nitric acid, cone, (sp gr 1.41).
6.1.1.5 Nitric acid, (1+1): Add 500 mL cone.
HNO- (sp gr 1.41) to 400 mL de ionized
distilled water and dilute to 1 liter.
6.1.2 Deionized distilled water: Prepare by
passing distilled water through a mixed
bed of cation and anion exchange resins.
Use deionized distilled water for the
preparation of all reagents, calibration
standards and as dilution water. The
purity of this water must be equivalent to
ASTH Type II reagent water of
Specification D 1193.
6.1.3 Standard stock solutions may be purchased
or prepared from ultra high purity grade
chemicals or metals. All salts must be
dried for one hour at 105 °C unless
otherwise specified. (CAUTION: Many
metal salts are extremely toxic and may be
fatal if swallowed. Wash hands thoroughly
after handling.) Typical stock solution
preparation procedures follow.
6.1.3.1 Aluminum solution, stock, 1 ml = 100 ug
Al: Dissolve 0.100 g aluminum metal in an
acid mixture of 4 mL of (1*1) HCl and 1 mL
of cone. HNO, in a beaker. Warm gently to
effect solution. When solution is
complete, transfer quantitatively to a
one-liter flask, add an additional 10 mL
(1+1) HCl, and dilute to 1000 mL with
deionized distilled water.
6.1.3.2 Antimony solution stock, 1 mL = 100 ug Sb:
Dissolve 0.2669 g K(SbO)C,H406 in
deionized distilled water, add 10 mL (1+1)
HCl and dilute to 1000 mL with deionized
distilled water.
6.1.3.3 Arsenic solution, stock, 1 mL ~ 100 ug As:
Dissolve 0.1320 g As^O, in 100 mL
deionized distilled water containing 0.4 g
NaOH. Acidify the solution with 2 mL
cone. HNOj and dilute to 1000 mL with
deionized distilled water.
6.1.3.4 Barium solution, stock, 1 mL - 100 ug Ba:
Dissolve 0.1516 g BaCl2 (dried at 250 °C
for 2 hours) in 10 mL deionized distilled
water with 1 mL (1 + 1) HCl. Add 10.0 mL
(1+1) HCl and dilute to 1000 mL with
deionized distilled water.
6.1.3.5 Beryllium solution, stock, 1 mL = 100 ug
Be: Do not dry. Dissolve 1.966 g
BeSO,"4H-0, in deionized distilled water,
add lO.O mL cone. HNOj and dilute to 1000
mL with deionized distilled water.
6.1.3.6 Boron solution, stock, 1 mL - 100 ug B:
Do not dry. Dissolve 0.5716 g anhydrous
HjBOj in deionized distilled water and
dilute to 1000 mL. Use a reagent meeting
ACS specifications, keep the bottle
tightly stoppered, and store in a
desiccator to prevent the entrance of
atmospheric moisture.
6.1.3.7 Cadmium solution, stock, 1 mL = 100 ug Cd:
Dissolve 0.1142 g CdO in a minimum amount
of (1+1) HNOj. Heat to increase rate of
dissolution. Add 10.0 mL cone. HNO, and
dilute to 1000 mL with deionized distilled
water.
6.1.3.S Calcium solution, stock, 1 mL = 100 ug Ca:
Suspend 0.2498 g CaCO, (dried at 180 °C
for one hour before weighing) in deionized
distilled water, and dissolve cautiously
with a minimum amount of (1+1) HNO,. Add
10.0 mL cone. HNO, and dilute to 1000 mL
with deionized distilled water.
6.1.3.9 Chromium solution, stock, 1 mL = 100 ug
Cr: Dissolve 0.1923 g CrO, in deionized
distilled water. When solution is
complete, acidify with 10 mL cone. HNO,
and dilute to 1000 mL with deionized
distilled water.
6.1.3.10 Cobalt solution stock, 1 mL = 100 ug Co:
Dissolve 0.1000 g of cobalt metal in a
minimum amount of (1+1) HNOj. Add 10.0 mL
(1+1) HCl and dilute to 1000 mL with
deionized distilled water.
6.1.3.11 Copper solution, stock, 1 mL = 100 ug Cu:
Dissolve 0.1252 g CuO in a minimum amount
177
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6.1.3.12
6.1.3.13
6.1.3.14
6.1.3.15
6.1.3.16
6.1.3.17
6.1.3.18
6.1.3.19
6.1.3.20
6.1.3.21
of (1+1) HNO,. Add 10.0 mL cone. HNO, and
dilute to 1000 mL with deionized distilled
water.
Iron solution, stock, 1 ml = 100 ug Fe:
Dissolve 0.1430 g Fe.O, in a warm mixture
of 20 mL (1+1) HCl and 2 ml cone.
Cool, add an additional 5 mL cone.
and dilute to 1000 ml with deionized
distilled water.
Lead solution, stock, 1 mL = 100 ug Pb:
Dissolve 0.1599 g Pb(NO,)_ in a minimum
amount of (1+1) HNOj. Add 10.0 ml of
cone. HNOj and dilute to 1000 ml with
deionized distilled water.
Magnesium solution, stock, 1 ml = 100 ug
Mg: Dissolve 0.1658 g MgO in a minimum
amount of (1+1) HNO,. Add 10.0 ml cone.
HN03 and dilute to 1000 mL with deionized
distilled water.
Manganese solution, stock, 1 mL = 100 ug
Mn: Dissolve 0.1000 g manganese metal in
10 ml cone. HCl and 1 ml cone. HNO., and
dilute to 1000 ml with deionized distilled
water.
Molybdenum solution, stock, 1 ml = 100 ug
Mo: Dissolve 0.2043 g (NH4)2Mo04 in
deionized distilled water and dilute to
1000 ml.
Mickel solution, stock, 1 mL = 100 ug Ni:
Dissolve 0.1000 g of nickel metal in 10 mL
hot cone. HNO,, cool and dilute to 1000 mL
with deionized distilled water.
Potassiun solution, stock, 1 mL = 100 ug
<: Dissolve 0.1907 g KCl, dried at 110
°C, in deionized distilled water. Dilute
to 1000 mL.
Selenium solution, stock, 1 mL = 100 ug
Se: Do not dry. Dissolve 0.1727 g H-SeO,
(actual assay 94.6%) in deionized
distilled water and dilute to 1000 mL.
Silica solution, stock, 1 mL = 100 ug
SiO.: Do not dry. Dissolve 0.4730 g
Na-jSiOj^HjO in deionized distilled water.
Add 10.0 mL cone. HN03 and dilute to 1000
mL with deionized distilled water.
Silver solution, stock, 1 mL = 100 ug Ag:
Dissolve 0.1575 g AgNO, in 100 mL
deionized distilled water and 10 mL cone.
HNO,. Dilute to 1000 mL with deionized
distilled water.
6.1.3.22 Sodium solution, stock, 1 ml = 100 ug Na:
Dissolve 0.2542 g NaCl in deionized
distilled water. Add 10.0 ml cone. HNO,
and dilute to 1000 ml with deionized
distilled water.
6.1.3.23
Thalliun solution, stock, 1 ml = 100 ug
Tl: Dissolve 0.1303 g TlNO, in deionized
"' Add 10.0 ml cone. HNOj
distilled water.
and dilute to
distilled water.
1000 ml with deionized
6.1.3.24 Vanadium solution, stock, 1 ml = 100 ug V:
Dissolve 0.2297 NH.VOj in a minimum amount
of cone. HNO,. Heat to increase rate of
dissolution. Add 10.0 mL cone. HNO, and
dilute to 1000 mL with deionized distilled
water.
6.1.3.25 Zinc solution, stock, 1 ml = 100 ug Zn:
Dissolve 0.1245 g ZnO in a minimum amount
of dilute HNOj. Add 10.0 mL cone. HNO,
and dilute to 1000 ml with deionized
distilled water.
6.1.4 Mixed calibration standard solutions
6.1.4.1 Prepare mixed calibration standard
solutions by combining appropriate volumes
of the stock solutions in volumetric
flasks. (Recommended solutions are given
in Sections 6.1.4.4.1-6.1.4.4.5.). Add 2
ml (1+1) HN03 and 10 ml (1+1) HCl, and
dilute to 100 ml with deionized distilled
water. (See NOTE in Section 6.1.4.4.5.)
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 mixed standards that the
elements are compatible and stable.
Transfer the mixed standard solutions to a
FEP fluorocarbon or unused polyethylene
bottle for storage.
6.1.4.2 The calibration standards must contain the
same acid concentration as the prepared
sample. Fresh mixed standards should be
prepared as needed, recognizing that
concentration can change over time.
6.1.4.3 Calibration standards must be initially
verified using an ICV standard and
monitored weekly for stability (see
Section 8.4.1.1).
6.1.4.4 Typical calibration standard combinations
are given in Sections 6.1.4.4.1 through
6.1.4.4.5. Although not specifically
178
-------
required, these combinations are
appropriate when using the specific
wavelengths listed in Table 1.
6.1.4.4.1 Mixed standard solution I - Manganese,
beryllium, cadmium, lead, and zinc.
6.1.4.4.2 Mixed standard solution II -- Barium,
copper, iron, vanadium, and cobalt.
6.1.4.4.3 Mixed standard solution III -- Molybdenum,
silica, arsenic, and selenium.
6.1.4.4.4 Nixed standard solution IV -- Calcium,
sodium, potassium, aluminum, chromium and
nickel.
6.1.4.4.5 Nixed standard solution V -- Antimony,
boron, magnesium, silver, and thallium.
NOTE: 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 concentration 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 HCl.
6.1.5 Initial calibration verification (ICV)
standard solutions -- Prepared in the same
acid matrix as the calibration standards
(see Section 6.1.4) and in accordance with
the instructions provided by the supplier.
Certified ICV standard solutions should be
obtained from an outside source. If the
certified solution of the ICV standard is
not available from any source, analyses
shall be conducted on an independent
standard (defined as a standard composed
of the analytes from a different source
than those used in the standards for the
instrument calibration) at a concentration
other than that used for instrument
calibration but within the calibration
range. NOTE: ICV standards for
semiquantitative ICP screen elements are
not available commercially at this time
and should be prepared by the laboratory.
The standards used must be traceable to
EPA or NIST materials.
6.1.6 Continuing calibration verification (CCV)
standard solutions -- Prepared by
combining compatible elements at a
concentration equivalent to the midpoints
of their respective calibration curves.
The aggregated CCV standard solutions must
contain all analytes. The CCV standard
may be an outside standard of NIST or EPA
materials, NIST SRM 1643a, or laboratory-
prepared standards traceable to EPA or
NIST.
6.1.7 ICP interference check sample (ICS) -- The
ICP ICS consists of two solutions:
Solution A (interferents) and Solution AB
(analytes mixed with the interferents).
The materials used in the ICS must be
traceable to NIST or EPA material.
6.1.7.1 If the ICP ICS is not available from any
source, the laboratory must prepare
independent ICP check samples with
interferent and analyte concentrations at
the levels specified in Table 11.
6.1.7.2 The mean value and standard deviation of
independent ICP check samples must be
established by initially analyzing the
check samples at least five times
repetitively for each parameter in Table
11. Results must fall within the control
limit of +20% of the established mean
value.
6.1.8 Blanks -- Two types of blanks are
required. Initial and continuing
calibration blanks are used in
establishing the analytical curve; the
preparation (reagent) blank is used to
correct for possible contamination
resulting from varying amounts of the
acids used in the sample processing.
6.1.8.1 Initial and continuing calibration blanks
-- Prepared by diluting 2 mL of (1+1) UNO,
and 10 ml of (1+1) HCl to 100 mL with
deionized distilled water. Prepare a
sufficient quantity to be used to flush
the system between standards and samples.
The calibration blank must contain the
same acid concentration as the prepared
sample solution.
6.1.8.2 Preparation (reagent) blank -- Must
contain all the reagents and in the same
volumes as used in the processing of the
samples. The preparation 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.
6.1.9 Laboratory control sample -- Should be
obtained from an outside source. If
unavailable, the ICV standard solutions
179
-------
may be used. The laboratory control
sample must contain all analytes of
interest. Standards used must be
traceable to NIST or EPA material.
6.2 ICP-Atomic Absorption Spectrometry
Semiquantitative screening of 42 elements
6.2.1 Individual stock solution (1000 mg/L) for
the elements listed in Table 4 may be
prepared by the laboratory or purchased
from a comnercial source. These solutions
are available from J.T. Baker Alfa
Products and other suppliers.
6.2.1.1 Osmium stock solution: Osmium stock
solution can be prepared from osmium
chloride (available from Alfa Products or
other suppliers). Dissolve 1.559 g OsCl,
in 6 ml, cone. HCl + 2 mL cone. HNO^, and
dilute to 1 liter to yield 1000 mg/L stock
solution.
6.2.1.2 Sulfur stock solution: Can be prepared
from ammonium sulfate (available from J.
T. Baker or other suppliers). Dissolve
4.122 g of anhydrous ammonium sulfate in
deionized water and dilute to 1 liter to
yield 1000 mg/L stock solution.
6.2.1.3 Uranium stock solution: Hade from uranyl
nitrate (available from Alfa Products or
other suppliers). Dissolve 2.110 g uranyl
nitrate hexahydrate in 6 mL cone. HCl + 2
mL cone. HNO, and dilute to 1 liter to
give 1000 mg/L.
6.2.2 Mixed calibration solution -- Prepare a
mixed working (calibration) standard
directly from the individual stock
solutions to give final concentrations for
each analyte as listed in Table 7. It is
recommended that a micro-pipette with
disposable plastic tips be used to
transfer each stock solution to the
volumetric flask. The stability of this
solution is limited, but can be extended
by storing it in a dark brown plastic
bottle away from light. Care should be
taken to include analyte contribution from
other stock standards. For example: a
number of the stock standards are prepared
from potassium salts. If alternative
solutions are not available, the final
solution (Section 6.2.2) must be analyzed
quantitatively by ICP to derive its true
concentration. The resulting calibration
standard must contain the same acid
concentration as the prepared sample
solution.
6.2.3
6.2.4
6.3
6.3.1
6.3.1.1
6.3.1.2
6.3.1.3
6.3.2
6.3.2.1
6.3.2.2
6.3.2.3
6.3.2.4
ICV standard solutions (see Section
6.1.5), CCV standard solutions (see
Section 6.1.6), and interference check
samples (see Section 6.1.7) are also
required.
Two types of blanks are required --
Initial and continuing calibration blanks
and the preparation blank (see Section
6.1.8).
GFAA Spectrophotometric Method
Antimony
Stock solution: Carefully weigh 2.669 g
of antimony potassium tartrate (analytical
reagent grade) and dissolve in deionized
distilled water. Dilute to 1 liter with
deionized water. 1 mL = 1 mg Sb (1000
mg/L).
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."
The calibration standards must be prepared
using the same type of acid and at the
same concentration as will result in the
sample to be analyzed after sample
preparation.
Arsenic
Stock solution: Dissolve 1.320 g arsenic
trioxide, As,0, (analytical reagent grade)
in 100 mL deionized distilled water
containing 4 g NaOH. Acidify the solution
with 20 mL cone. HNO, and dilute to 1
liter. 1 mL = 1 mg As (1000 mg/L).
Nickel nitrate solution, 5%: Dissolve
24.770 g ACS reagent grade Ni(N03)2-6H20
in deionized distilled water and make up
to 100 mL.
up
Nickel nitrate solution, 1%: Dilute 20 mL
of the 5% nickel nitrate to 100 mL with
deionized distilled water.
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 cone.
HNO,, 2 mL 30% H-Oj, and 2 mL of the 5%
nickel nitrate solution. Dilute to 100 mL
with deionized distilied water.
180
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6.3.3
6.3.3.1
6.3.3.2
6.3.3.3
6.3.4
6.3.4.1
6.3.4.2
6.3.4.3
6.3.5
6.3.5.1
6.3.5.2
Beryllium
Stock solution: Dissolve 11.6586 g
beryllium sulfate, BeSO,, in deionized
distilled water containing 2 ml cone.
nitric acid and dilute to 1 liter. 1 ml =
1 mg Be (1000 mg/L).
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."
The calibration standards must be prepared
using the same type of acid and at the
same concentration as will result in the
sample to be analyzed after sample
preparation.
Cadmium
Stock solution: Carefully weigh 2.282 g
cadmium sulfate, 3CdS04'8H20 (analytical
reagent grade) and dissolve in deionized
distilled water. Hake up to 1 liter with
deionized distilled water. 1 ml = 1 mg Cd
(1000 mg/L).
Ammonium phosphate solution (40%):
Dissolve 40 g ammonium phosphate,
(NH4)2HP04 (analytical reagent grade) in
deionized distilled water, and dilute to
100 ml.
Prepare dilutions of stock cadmium
solution to be used as calibration
standards at the time of analysis. To
each 100 ml of standard and sample alike
add 2.0 ml of the ammonium phosphate
solution. The calibration standards must
be prepared using the same type of acid
and at the same concentration as will
result in the sample to be analyzed after
sample preparation.
Chromium
Stock solution: Dissolve 1.923 g chromium
trioxide (CrO_ reagent grade) in deionized
distilled water.
When solution is
complete, acidify with redistilled HNO,
and dilute to 1 liter with deionized
distilled water 1
mg/L).
1 mg Cr (1000
Calcium nitrate solution: Dissolve 11.784
g calcium nitrate, Ca(N03)2'4H20
(analytical reagent grade) in deionized
distilled water and dilute to 100 mL. 1
mL = 20 mg Ca.
6.3.5.3 Prepare dilutions of the stock chromium
solution to be used as calibration
standards at the time of analysis. The
calibration standards must be prepared
using the same type of acid and at the
same concentration as will result in the
sample to be analyzed after sample
preparation. To each 100 mL of standard
and sample alike, add 1 mL 30% H202 and 1
mL of the calcium nitrate solution.
6.3.6 Lead
6.3.6.1 Stock solution: Carefully weigh 1.599 g
lead nitrate, PbOJO,),, (analytical reagent
grade), and dissolve in deionized
distilled water. When solution is
complete, acidify with 10 mL redistilled
HN03 and dilute to 1 liter with deionized
distilled water. 1 mL = 1 mg Pb (1000
mg/L).
6.3.6.2 Lanthanum nitrate solution: Dissolve
58.639 g of ACS reagent grade La^O, in 100
mL cone. UNO, and dilute to 1000 mL with
deionized distilled water. 1 mL = 50 mg
La.
6.3.6.3 Working lead solution: Prepare dilutions
of stock lead solution to be used as
calibration standards at the time of
analysis. The calibration standards must
be prepared using the same type of acid
and at the same concentration as will
result in the sample to be analyzed after
sample preparation. To each 100 mL of
diluted standard, add 10 mL of the
lanthanum nitrate solution.
6.3.7 Selenium
6.3.7.1 Stock selenium solution: Dissolve 0.3453
g selenous acid (actual assay 94.6%
HjSeOj) in deionized distilled water and
make up to 200 mL. 1 mL = 1 mg Se (1000
mg/L).
6.3.7.2 Nickel nitrate solution, 5%: Dissolve
24.770 g ACS reagent grade Ni(NO,) '6H.O
in deionized distilled water and make up
to 100 mL.
6.3.7.3 Nickel nitrate solution, 1%: Dilute 20 mL
of the 5% nickel nitrate to 100 mL with
deionized distilled water.
6.3.7.4 Working selenium solution: Prepare
dilutions of the stock solution to be used
as calibration standards at the time of
analysis. The calibration standards must
181
-------
be prepared using the same type of acid
and at the same concentration as will
result in the sample to be analyzed after
sample preparation. Withdraw appropriate
aliquots of the stock solution, add 1 ml
cone. HHOj, 2 ml 30% H,0,, and 2 ml of the
5% nickel nitrate solution. Dilute to 100
ml with deionized distilled water.
6.3.8 Silver
6.3.8.1 Stock solution: Dissolve 1.575 g AgNO,
(analytical reagent grade) in deionized
distilled water. Add 10 ml cone. HNOj and
make up to 1 liter. 1 mL = 1 mg Ag (1000
mg/L).
6.3.8.2 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."
6.3.8.3 The calibration standards must be prepared
using the same type of acid and at the
same concentration as will result in the
sample to be analyzed after sample
preparation.
6.3.9 Thallium
6.3.9.1 Stock solution: Dissolve 1.303 g thallium
nitrate, TlNO, (analytical reagent grade)
in deionized distilled water. Add 10 ml
cone, nitric acid and dilute to 1 liter
with deionized distilled water. 1 ml = 1
mg Tl (1000 mg/L).
6.3.9.2 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."
6.3.9.3 The calibration standards must be prepared
using the same type of acid and at the
same concentration as will result in the
sample to be analyzed after sample
preparation.
6.4 Mercury Analysis in Water by Manual Cold
Vapor Technique
6.4.1 Sulfuric acid, cone: Reagent grade.
6.4.1.1 Sulfuric acid, 0.5 N: Dilute 14.0 ml
cone, sulfuric acid to 1.0 liter.
6.4.2 Nitric acid, cone: Reagent grade of low
mercury content. NOTE: If a high reagent
blank is obtained, it may be necessary to
distill the nitric acid.
6.4.3 Stannous sulfate: Add 25 g stannous
sulfate to 250 ml 0.5 N sulfuric acid.
This mixture is a suspension and should be
stirred continuously during use. NOTE:
Stannous chloride may be used in place of
stannous sulfate.
6.4.4 Sodium chloride-hyroxylamine sulfate
solution: Dissolve 12 g sodium chloride
and 12 g hydroxylamine sulfate in
deionized distilled water, and dilute to
100 ml. NOTE: Hydroxylamine
hydrochloride may be used in place of
hydroxylamine sulfate.)
6.4.5 Potassium permanganate: 5% solution, w/v.
Dissolve 5 g potassium permanganate in 100
ml distilled water.
6.4.6 Potassium persulfate: 5% solution, w/v.
Dissolve 5 g potassium persulfate in 100
ml distilled water.
6.4.7 Stock mercury solution: Dissolve 0.1354 g
mercuric chloride in 75 ml deionized
distilled water. Add 10 ml cone, nitric
acid and adjust the volume to 100.0 ml. 1
ml - 1 mg Hg.
6.4.8 Working mercury solution: Hake successive
dilutions of the stock mercury solution to
obtain a working standard containing 0.1
ug per ml. This working standard and the
dilutions of the stock mercury solution
should be prepared fresh daily. Acidity
of the 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.
6.5 Mercury Analysis in Water by Automated
Cold Vapor Technique
6.5.1 Sulfuric acid, cone: Reagent grade.
6.5.1.1 Sulfuric acid, 2 N: Dilute 56 ml cone.
sulfuric acid to 1 liter with deionized
distilled water.
6.5.1.2 Sulfuric acid, 10%: Dilute 100 ml cone.
sulfuric acid to 1 liter with deionized
distilled water.
6.5.2 Nitric acid, cone:
mercury content.
Reagent grade of low
6.5.2.1 Nitric acid, 0.5% wash solution: Dilutes
mL cone, nitric acid to 1 liter with
deionized distilled water.
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6.5.3 Stannous sulfate: Add 50 g stannous
sulfate to 500 ml 2N sulfuric acid
(Section 6.5.1.1). This mixture is a
suspension and should be stirred
continuously during use. NOTE: Stannous
chloride may be used in place of stannous
sulfate.
6.5.4 Sodium chloride-hydroxylamine sulfate
solution: Dissolve 30 g sodium chloride
and 30 g hydroxylamine sulfate in
deionized distilled water and dilute to 1
liter. NOTE: Hydroxylamine hydrochloride
may be used in place of hydroxylamine
sulfate.
6.5.5 Potassium permanganate: See Section
6.4.5.
6.5.6 Potassium permanganate, 0.1N: Dissolve
3.16 g potassium permanganate in deionized
distilled water and dilute to 1 liter.
6.5.7 Potassium persulfate: See Section 6.4.6.
6.5.S Stock mercury solution: See Section
6.4.7.
6.5.9 Working mercury solution: See Section
6.4.S. From this solution, prepare
standards containing 0.2, 0.5, 1.0, 2.0,
5.0, 10.0, 15.0, and 20.0 ug Hg/L.
6.5.10 Air scrubber solution: Mix equal volumes
of 0.1 N potassium permanganate (Section
6.5.6) and 10% sulfuric acid (Section
6.5.1.2).
6.6 Mercury Analysis in Soil/Sediments by
Manual Cold Vapor Technique
6.6.1 Sulfuric acid, cone: Reagent grade of low
mercury content.
6.6.2 Nitric acid, cone: See Section 6.4.2.
6.6.3 Stannous sulfate: See Section 6.4.3.
6.6.4 Sodium chloride-hydroxylmine sulfate: See
Section 6.4.4.
6.6.5 Potassium permanganate: See Section
6.4.5.
6.6.6 Potassium persulfate: See Section 6.4.6.
6.6.7 Stock mercury solution: See Section
6.4.7.
6.6.8 Working mercury solution: See Section
6.4.S.
7 CALIBRATION
7,1 ICP and GFAA Spectroscopic Methods
7.1.1 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 dynamic range, and
interference effects must be investigated
and established for each individual
analyte line on that particular
instrument. All measurements must be
within the instrument linear range where
correction factors are valid.
7.1.2 It is the 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.
7.2 Analysis of Mercury in Water by Cold Vapor
Technique
7.2.1 Transfer 0, 0.5, 1.0, 5.0 and 10.0 ml
aliquots of the working mercury solution
containing 0 to 1.0 ug mercury to a series
of 300 ml BOD bottles. Add enough
distilled water to each bottle to make a
total volume of 100 ml. Mix thoroughly
and add 5 ml cone, sulfuric acid (Section
6.4.1) and 2.5 mL cone, nitric acid
(Section 6.4.2) to each bottle. Add 15 ml
KMn04 (Section 6.4.5) solution to each
bottle and allow to stand at least 15
minutes. Add 8 ml potassium persulfate
(Section 6.4.6) to each bottle, and heat
for 2 hours in a water bath maintained at
95 °C. Alternatively, cover the BOD
bottles with foil and heat in an autoclave
for 15 minutes at 120 °C and 15 psi. Cool
and add 6 ml of sodium chloride-
hydroxylamine sulfate solution (Section
6.4.4) to reduce the excess permanganate.
When the solution has been decolorized,
wait 30 seconds, add 5 ml of the stannous
sulfate solution (Section 6.4.3), and
immediately attach the bottle to the
aeration apparatus forming a closed
system. At this point, the sample is
allowed to stand quietly without manual
agitation.
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7.2.2 The circulating pump, which has previously
been adjusted to a rate of 1 liter per
minute, is allowed to run continuously
(see NOTE 1). The absorbance trill
increase and reach maximum within 30
seconds. As soon as the recorder pen
levels off, approximately 1 minute, open
the bypass valve and continue the aeration
until the absorbance returns to its
minimum value (see NOTE 2). Close the
bypass valve, remove the stopper and frit
from the BOO bottle and continue the
aeration. Proceed with the standards and
construct a standard curve by plotting
peak height versus micrograms of mercury.
MOTE 1: An open system (where the mercury
vapor is passed through the absorption
cell only once) may be used instead of the
closed system.
NOTE 2: . Because of the toxic nature of
mercury vapor, precautions 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 M KMnO, and 10% H^SO^, or
b) 0.25% iodine in a 3% KI solution. A
specially treated charcoal that will
adsorb mercury vapor is available.
7.2.3 If additional sensitivity is required, a
200 ml sample with recorder expansion may
be used provided the instrument does not
produce undue noise.
7.3 Analysis of Mercury in Soil/Sediments by
Cold Vapor Technique
7.3.1 Transfer 0, 0.5, 1.0, 5.0, and 10 mL
aliquots of the working mercury solutions
(Section 6.6.8) containing 0-1.0 ug
mercury to a series of 300 mL BOO bottles.
Add enough deionized distilled water to
each bottle to make a total volume of 10
mL. Add 5 mL cone. H2S04 (Section 6.6.1)
and 2.5 mL cone. HNO, (Section 6.6.2), and
heat for 2 minutes in a water bath at 95
°C. Allow the sample to cool. Add 50 mL
deionized distilled water, 15 mL KMnO^
solution (Section 6.6.5), and 8 mL
potassium persulfate solution (Section
6.6.6) to each bottle and return bottles
to the water bath for 30 minutes. Cool
and add 6 mL sodium chloride-hydroxylamine
sulfate solution (Section 6.6.4) to reduce
the excess permanganate. Add 50 mL
deionized distilled water. Treating each
bottle individually, add 5 mL stannous
sulfate solution (Section 6.6.3) and
immediately attach the bottle to the
aeration apparatus. At this point, the
sample is allowed to stand quietly without
manual agitation.
7.3.2 The circulating pump, which has previously
been adjusted to a rate of 1 liter per
minute, is allowed to run continuously
(see NOTE 1 in Section 7.2.2). 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 valve and continue the aeration
until the absorbance returns to its
minimum value (see NOTE 2 in Section
7.2.2). Close the bypass valve, remove
the fritted tubing from the BOO bottle and
continue the aeration- Proceed with the
standards and construct a standard curve
by plotting peak height versus micrograms
of mercury.
8 QUALITY ASSURANCE/QUALITY CONTROL
8.1 Each laboratory that uses this method is
required to operate a formal quality
assurance program. The minimum require-
ments of this program consist of: 1) an
initial demonstration of laboratory
capability, 2) analysis of samples spiked
with the analytes of interest to evaluate
and document data quality, and 3) 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 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.
8.2 Initial Precision and Accuracy -- To
establish the ability to generate
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acceptable precision and accuracy, the
analyst shall perform the following
operations.
8.2.1 For analysis of samples containing low
solids (aqueous samples), prepare four 500
mL aliquots of reagent water spiked with
the 27 elements listed in Tables 1-3 at
concentrations at or near the aqueous
detection limits given in the tables.
Digest these samples according to the
procedures in Section 10.1.1 and analyze
the samples according to the ICP, GFAA and
Hg procedures in Sections 10.1.3, 10.3,
and 10.4, respectively.
8.2.2 For analysis of samples containing high
solids, prepare four aliquots of reagent
water containing the 27 elements at
concentrations at or near the detection
limits given in Tables 1-3 for aqueous
samples. Digest these samples according
to the procedures for water samples in
Section 10.1.1, but analyze them as if
they were soil samples according to
Sections 10.1.3, 10.3, and 10.4, and
calculate the concentrations of the
analytes as if the original sample weight
was 1 g of soil.
8.2.3 Using the results of the set of four
analyses (from Section 8.2.1 or 8.2.2),
compute the average percent recovery (x)
and the coefficient of variation (Cv) of
the percent recovery(ies) for each
element.
8.2.4 For each element, compare s and x with the
corresponding limits in Table 8. If s and
x for all elements 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 the element. In this
case, correct the problem and repeat the
test.
8.3 Instrument Calibration
8.3.1 Guidelines for instrumental calibration
are given in EPA 600/4-79-020 and/or
Section 7. Instruments must be calibrated
daily or once every 24 hours and each time
the instrument is set up (Section 10).
8.3.2 For atomic absorption systems, calibration
standards are prepared by diluting the
stock metal solutions at the time of
analysis.
8.3.3 Calibration standards
8.3.3.1 For ICP systems, calibrate the instrument
according to instrument manufacturer's
recommended procedures. At least two
standards must be used for ICP
calibration. One of the standards must be
a blank.
8.3.3.2 AA Systems
8.3.3.2.1 Calibration standards for AA procedures
must be prepared by dilution of the stock
solutions (Section 6.3).
8.3.3.2.2 Calibration standards must be prepared
fresh each time an analysis is to be made
and discarded after use. Prepare a blank
and at least three calibration standards
in graduated amounts in the appropriate
range. One atomic absorption calibration
standard must be at the minimum level (see
Table 9), except for mercury. The
calibration standards must be prepared
using the same type of acid or combination
of acids and at the same concentration as
will result in the samples following
sample preparation.
8.3.3.2.3 Beginning with the blank, aspirate or
inject the standards and record the
readings. If the AA instrument
configuration prevents the required four-
point calibration, calibrate according to
instrument manufacturer's recommendations,
and analyze the remaining required
standards immediately after calibration.
Results for these standards must be within
± 5% of the true value. Each standards
concentration and the calculations to show
that the ± 5% criterion has been met, must
be given in the raw data. If the values
do not fall within this range,
recalibration is necessary. NOTE: The ±
5% criteria does not apply to the atomic
absorption calibration standard at the
minimum level.
8.3.3.2.4 Baseline correction is acceptable as long
as it is performed after every sample or
after the continuing calibration
verification and blank check; resIoping is
acceptable as long as it is immediately
preceded and immediately followed by
continuing calibration verification and
continuing calibration blank analyses.
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8.3.4 Mercury analysis techniques -- Follow the
calibration procedures outlined in Section
7.
8.4 Initial Calibration Verification (ICV) and
Continuing Calibration Verification (CCV)
8.4.1 Initial Calibration Verification (ICV)
8.4.1.1 The accuracy of the initial calibration
shall be verified and documented for every
analyte by the analysis of an ICV standard
(Sections 6.1.5 and 6.2.3) at each
wavelength used for analysis. If the
results are not within ±10% of the true
value, the analysis must be terminated,
the problem corrected, the instrument
recalibrated, and the calibration
reverified. NOTE: For semiquantitative
ICP analysis, 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.
8.4.1.2 ICV standard solutions must be run
immediately after each of the ICP and AA
systems have been calibrated and each time
the system is set up. The ICV standard
solution(s) must be run for each analyte
at each wavelength used for analysis.
8.4.2 Continuing Calibration Verification (CCV)
8.4.2.1 To ensure calibration accuracy during each
analysis run, a CCV standard (Sections
6.1.6 and 6.2.3) is to be used for
continuing calibration verification and
must be analyzed and reported for every
wavelength used for the analysis of each
analyte, at a frequency of 10% or every 2
hours during an analysis run, whichever is
more frequent. The CCV standard must also
be analyzed and reported for every
wavelength used for analysis of each
analyte at the beginning of the run and
after the last analytical sample.
8.4.2.2 The same continuing calibration standard
must be used throughout the analysis run
for each set or Episode of samples
received.
8.4.2.3 Each CCV standard analysis must reflect
the conditions of analysis of all
associated analytical samples (all
preceding analytical samples up to the
previous CCV standard analysis). The
duration of analysis, rinses and other
related operations that may affect the CCV
measured result may not be applied to the
CCV standard to a greater extent than the
extent applied to the associated
analytical samples. For instance, the
difference in time between a CCV standard
analysis and the blank immediately
following it, as well as the difference in
time between the CCV standard analysis and
the analytical sample immediately
preceding it, may not exceed the lowest
difference in time between analysis of any
two consecutive analytical samples
associated with the CCV.
8.4.2.4 If the deviation of the continuing
calibration verification is greater than
the control limits specified in Table 10,
the analysis must be stopped, the problem
corrected, the instrument recalibrated,
the calibration verified, and the
preceding samples analyzed since the last
good calibration verification reanalyzed
for the analytes affected.
8.5 Minimum Level (ML) Standards for ICP (CRI)
and AA (CRA)
8.5.1 To verify linearity near the ML for ICP
analysis, analyze an ICP standard (CRI) at
2x ML (Table 9) or 2x IDL, whichever is
greater, at the beginning and end of each
sample analysis run, or a minimum of twice
per 8-hour working shift, whichever is
more frequent, but not before initial
calibration verification. This standard
must be run by ICP for every wavelength
used for analysis, except those for Al,
Ba, Ca, Fe, Mg, Na and K.
8.5.2 To verify linearity near the ML for AA
analysis, analyze an AA standard (CRA) at
the ML or the IDL, whichever is greater,
at the beginning of each sample analysis
run, but not before the initial
calibration verification.
8.5.3 The laboratory shall determine the ML by
performing 3 replicate analyses and
determining a 99% confidence level (3x
standard deviation). If any GFAA element
exceeds the ICP ML by 2x, it can be
analyzed by ICP rather than GFAA.
8.5.4 Specific acceptance criteria for CRI and
CRA standards will be set by EPA in the
future.
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8.6 Initial Calibration Blank (ICB),
Continuing Calibration Blank (CCB), and
Preparation Blank (PB) Analyses
8.6.1 Initial and continuing calibration blank
analyses -- A calibration blank (Section
6.1.8.1 and 6.2.4) must be analyzed at
each wavelength used for analysis,
immediately after every initial and
continuing calibration verification, at a
frequency of 10% or each time the
instrument is calibrated, whichever is
more frequent. The blank must be analyzed
at the beginning of the run and after the
last analytical sample. NOTE: A CCB
must be run after the last CCV that was
run after the last analytical sample of
the run.
8.6.1.1 For quantitative ICP analysis, if the
absolute value blank result exceeds the ML
(Table 9), terminate analysis, correct the
problem, recalibrate, verify the
calibration, and reanalyze the preceding
10 analytical samples or all analytical
samples analyzed since the last acceptable
calibration blank analysis.
8.6.1.2 For semiquantitative ICP analysis, the
result should be within ± values of the
lower threshold limits (Table 4). If the
result is not within the control limits,
terminate the analysis, correct the
problem, and recalibrate the instrument.
8.6.2 Preparation blank analysis -- At least one
preparation (reagent) blank (Sections
6.1.8.2 and 6.2.4) must be prepared and
analyzed with each batch of samples (group
of samples prepared at the same time)
digested. This blank is to be reported
for each batch of samples and used in all
analyses to ascertain whether sample
concentrations reflect contamination.
8.6.2.1 If the absolute value of the concentration
of the blank is less than or equal to the
ML (Table 9), no correction of sample
results is performed.
8.6.2.2 If any analyte concentration in the blank
is above the ML (Table 9), the lowest
concentration of that analyte in the
associated samples must be 10x the blank
concentration. Otherwise, all samples
associated with the blank with the
analyte's concentration less than 10x the
blank concentration and above the ML, must
be redigested and reanalyzed for that
analyte. The sample concentration is not
to be corrected for the blank value.
8.6.2.3 If the concentration of the blank is below
the negative ML, then all samples reported
below 10x ML associated with the blank
must be redigested and reanalyzed.
8.7 ICP Interference Check Sample (ICS)
Analysis
8.7.1 To verify inter-element and background
correction factors, analyze and report the
results for the ICP ICS (Sections 6.1.7
and 6.2.3) at the beginning and end of
each analysis run or a minimum of twice
per 8-hour working shift, whichever is
more frequent, but not before initial
calibration verification.
8.7.2 The ICP ICS consists of two solutions:
Solution A (interferents) and Solution AB
(analytes mixed with the interferents).
An ICS analysis consists of analyzing both
solutions consecutively (starting with
Solution A) for all wavelengths used for
each analyte reported by ICP.
8.7.3 Results for the ICP analyses of Solution
AB during the analytical runs must fall
within the control limit of +20% of the
true value for the analytes included in
the ICS. If not, terminate the analysis,
correct the problem, recalibrate the
instrument, and reanalyze the analytical
samples analyzed since the last acceptable
ICS. If true values for analytes
contained in the ICS and analyzed by ICP
are not supplied with the ICS, the mean
must be determined by initially analyzing
the ICS at least five times repetitively
for the particular analyte(s). This mean
determination must be made during an
analytical run where the results for the
previously-analyzed ICS met all method
specifications. Additionally, the result
of this initial mean determination is to
be used as the true value for the lifetime
of that solution (i.e., until the solution
is exhausted).
8.8 Spike Sample Analysis (Matrix Spike)
8.8.1 The spike sample analysis is designed to
provide information about the effect of
the sample matrix on the digestion and
measurement methodology. The spike is
added before the digestion (i.e., prior to
the addition of other reagents) and prior
to any distillation steps. Spike sample
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S.S.2
8.8.3
8.8.4
8.8.5
8.8.6
8.8.7
analyses shall be performed on 10% of the
samples analyzed, or at least one spike
sample analysis (matrix spike) shall be
performed for each set or Episode of
samples, whichever is more frequent.
If the spike analysis is performed on the
same sample that is chosen for the
duplicate sample analysis, spike
calculations must be performed using the
results of the sample designated as the
"original sample" (see Section 8.9). The
average of the duplicate results cannot be
used for the purpose of determining
percent recovery. NOTE: Samples
identified as field blanks cannot be used
for the spike sample analysis. EPA may
require that a specific sample be used for
the spike sample analysis.
Analyze an aliquot of the sample by the
ICP parameters for all elements listed in
Table 1 to determine the background
concentration of each element.
Using these concentrations, prepare a QC
spike standard containing the analytes.
The standard shall produce a concentration
in the sample of 1x - 5x the background
level determined above. For not-detected
analytes, the spike Shall be in the range
of 5x - 50x the detection limit.
Spike a second sample aliquot with the QC
spike concentrate and analyze it to
determine the concentration in the sample
after spiking of each analyte.
Calculate the percent recovery of each
analyte as follows:
analyte that falls within the
percent recovery limits.
75-125
Where,
A =
T =
100
Concentration of element in the
sample after spiking. NOTE: When
A is less than the instrument
detection limit, use A=0 only for
the purpose of calculation.
Background concentration of each
element in the sample.
Known true value of the spike.
The acceptable range for recovery of the
predigested spike is 75-125 percent for
all analytes. EPA will develop recovery
limits based on single or inter laboratory
data when sufficient data have been
accumulated. Report the result for each
8.8.8 If the recovery limit is not met for any
analyte, proceed as follows.
8.8.8.1 For ICP elements, repeat the test. If the
recovery is still outside the range, the
instrument conditions should be verified
by running the CCV. If the calibration
criteria are not met, the instrument
should be recalibrated and the spike
recovery test repeated. If after
recall bration, the spike recovery remains
outside of 75-125% limits, the sample
should be diluted by a factor of 10 and
the test repeated. Report and qualify the
results.
8.8.8.2 For AA elements, analyze the sample by the
method of standard addition (MSA) (Section
8.15). If the correlation coefficient
meets method requirements (Section
8.15.7), report and qualify the results.
If these specifications are not met,
dilute an aliquot of the original sample
by a factor of 10 and repeat the analysis
by MSA.
8.8.8.3 If correlation coefficient of the diluted
analysis meets specifications, report and
qualify the results. If these
specifications are not met, recalibrate
the instrument and repeat the analysis by
MSA on the diluted sample. If the
correlation coefficient specifications are
not met, report and qualify the results.
8.9 Duplicate Spike Sample Analysis (Matrix
Spike Duplicate)
8.9.1 Duplicate spike analyses (matrix spike
duplicate) shall be performed on 10% of
the samples analyzed, or at least one
duplicate analyses shall be performed for
each set or Episode of samples, whichever
is more frequent.
8.9.2 Repeat the spiking and analysis of a third
aliquot of the same sample as used for
determination of spike recovery (Section
8.8.5), using the same analysis scheme as
used for analysis of the sample. For
example, if an analyte determined by AA
required dilute MSA analysis in order to
meet the spike recovery limits, determine
that metal in the duplicate spike analysis
by dilute MSA analysis.
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8,9.3
8.9.4
8.9.5
Samples identified as field blanks cannot
be used for duplicate spike sample
analysis. EPA may require that a specific
sample be used for duplicate spike sample
analysis.
The acceptable range for precision of the
spike recovery is less than twenty percent
relative percent difference (<20% RPD) for
all analytes. EPA will develop precision
limits based on a single or inter-
laboratory data when sufficient data have
been accumulated. Report and qualify the
result for each analyte that fails the
RPD.
The relative percent differences (RPD) for
each component are calculated as follows:
RPD
Where,
RPD
S
IS " P|
(S + D)/2
100
Relative percent difference
First spike sample value (matrix
spike)
Second spike sample value
(matrix spike duplicate)
8.10 Laboratory Control Sample (LCS) Analysis
8.10.1 Laboratory control samples (Section 6.1.9)
must be analyzed for each analyte using
the same sample preparation technique,
analytical methods, and QA/OC procedures
as employed in sample analysis. An LCS
must be analyzed for each set or Episode
of samples or for each standard stock
batch.
8.10.2 If the percent recovery for the LCS falls
outside the control limits of 80-120%
(with the exception of Ag and Sb), the
analyses must be terminated, the problem
corrected, and the samples associated with
that LCS redigested and reanalyzed.
8.11 ICP Serial Dilution Analysis
8.11.1 For quantitative ICP analysis, prior to
reporting concentration data for the
analyte elements, analyze and report the
results of the ICP serial dilution
analysis. The ICP serial dilution
analysis must be performed on 10% of the
samples analyzed, or at least one serial
dilution analysis shall be performed for
each set or Episode of samples, whichever
is more frequent. NOTE: Samples
8.11.2
8.11.3
8.11.4
identified as field blanks cannot be used
for serial dilution analysis.
If the analyte concentration is
sufficiently high (minimally a factor of
50 above the instrumental detection limit
in the original sample), the serial
dilution (a five-fold dilution) must then
agree within 10% of the original
determination after correction for
dilution. If the dilution analysis for
one or more analytes is not at or within
10%, a chemical or physical interference
effect must be suspected and the data for
all sample analyses associated with that
serial dilution must be flagged.
The percent differences for each component
are calculated as follows:
% Difference =
100
I
Where,
I = Initial Sample Result
S = Serial Dilution Result (Instrument
Reading x 5)
In the instance where there is more than
one serial dilution per sample set or
Episode, if one serial dilution result is
not within method specifications (see
Section 8.11.2), flag all samples in the
set or Episode that are associated with
that serial dilution.
8.12 Instrument Detection
Determination
Limit
(IDL)
8.12.1 Before any field samples are analyzed
under this method, the instrument
detection limits (in ug/L) must be
determined for each instrument used,
within 30 days of the start of analyses
under this method and at least quarterly
(every three calendar months), and must
meet the levels specified in the method.
8.12.2 The instrument detection limits (in ug/L)
shall be determined by multiplying by
three, the average of the standard
deviations obtained on three
nonconsecutive days from the analysis of a
standard solution (each analyte in reagent
water) at a concentration 3-5x the
instrument manufacturer's suggested IDL,
with seven consecutive measurements per
day. Each measurement must be performed
as though it were a separate analytical
sample (i.e., each measurement must be
189
-------
followed by a rinse and/or any other
procedure normally performed between the
analysis of separate samples). IDL's must
be determined and reported for each
wavelength used in the analysis of the
samples.
8.12.3 The quarterly determined IOL for an
instrument must always be used as the IOL
for that instrument during that' quarter.
If the instrument is adjusted in any way
that may affect the IOL, the IOL for that
instrument must be redetermined and the
results submitted for use as the
established IOL for that instrument for
the remainder of the quarter.
8.12.4 IDLs must be reported for each instrument
used. If multiple AA instruments are used
for the analysis of an element within a
sample set or Episode, the highest IOL for
the AAs must be used for reporting
concentration values for that sample set.
The same reporting procedure must be used
for multiple ICPs.
8.13 Inter-element Corrections for ICP
8.13.1 Prior to the start of analysis under this
method and at least annually thereafter,
the ICP inter-element correction factors
must be determined. Correction factors
for spectral interference due to Al, Ca,
Fe, and Hg must be determined for all ICP
instruments at all wavelengths used for
each analyte reported by ICP. Correction
factors for spectral interference due to
analytes other than Al, Ca, Fe, and Mg
must be reported if they were applied.
8.13.2 If the instrument Has adjusted in any way
that may affect the ICP interelament
correction factors, the factors must be
redetermined and the results submitted for
use.
8.14 Linear Range Analysis (LRA) -- For all
quantitative ICP analyses, a linear range
verification check standard must be
analyzed and reported quarterly (every
three calendar months) for each element.
The standard must be analyzed during a
routine analytical run performed under
this method. The analytically determined
concentration of this standard must be
within ±5% of the true value. This
concentration is the upper limit of the
ICP linear range beyond which results
should not be used without dilution of the
analytical sample.
8.15 Method of standard addition (MSA) -- All
GFAA elements must be analyzed by method
of standard addition in all samples.
8.15.1 The standard addition technique 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
compensates 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 interferences which
cause a baseline shift. The simplest
version of this technique is the single-
addition method. The procedure is as
follows.
8.15.1.1 Two identical aliquots of the sample
solution, each of volume V , are taken.
To the first (labeled A) is added a small
volume V of a standard analyte solution
of concentration Cg. To the second
(labeled B) is added the same volume Vg of
the solvent. The analytical signals of A
and B are measured and corrected for non-
ana I yte signals. The unknown sample
concentration C is calculated:
c » Ws
-------
the calibration curve generated at
beginning of the analytical run.
the
8.15.3 The sample and three spikes must be
analyzed consecutively for MSA
quantitat ion (the "initial" spike run data
is specifically excluded from use in the
MSA quantisation). Only single injections
are required for MSA quantitation.
8.15.4 Each full MSA counts as two analytical
samples towards determining 10% QC
frequency (i.e., five full MSAs can be
performed between calibration
verifications).
8.15.5 For analytical runs containing only MSAs,
single injections can be used for QC
samples during that run. For instruments
that operate in an MSA mode only, MSA can
be used to determine QC samples during
that run.
8.15.6 Spikes must be prepared such that:
8.15.6.1 Spike 1 is approximately 50% of the sample
absorbance.
8.15.6.2 Spike 2 is approximately 100X of the
sample absorbance.
8.15.6.3 Spike 3 is approximately 150X of the
sample absorbance.
8.15.7 The data for each MSA analysis should be
clearly identified in the raw data using
added concentration as the x-variable and
absorbance as the y-variable, along with
the slope, x-intercept, y-intercept, and
correlation coefficient (r) for the least
squares fit of the data. If the
correlation coefficient (r) for a
particular analysis is less than 0.995,
the MSA analysis must be repeated once.
If the correlation coefficient is still
less than 0.995, flag the result.
8.16 Quality control requirements for ICP
semiquantitative screen of 42 elements --
Instrument calibration (Section 8.3) and
performance of ICV (Section 8.4.1), CCV
(Section 8.4.2), ICB and CCB (Section
8.6.1), PB (Section 8.6.2), and ICS
(Section 8.7) analyses are required.
9 SAMPLE COLLECTION, PRESERVATION, AND
STORAGE
9.1 ICP and GFAA Spectroscopic Methods
9.1.1 Water sample preservation
9.1.1.1 Samples should be stored in polyethylene
or glass containers.
9.1.1.2 Samples are filtered immediately on site
by the sampler before adding preservative
for dissolved'metals.
9.1.1.3 Sample preservation is performed by the
sampler immediately following sample
collection. The sample should be
preserved with nitric acid to pH of less
than 2.
9.1.1.4 Samples should be maintained at 4 °C (±2
°C) until analysis.
9.1.1.5 Sample analysis should be completed within
six months of sample collection.
9.1.2 Soil/sediment sample preservation
9.1.2.1 The preservation required for soil samples
is maintenance at 4 °C (±2 °C) until
analysis.
9.1.2.2 Sample analysis should be completed within
six months of sample collection.
9.2 Mercury Analysis by CVAA
9.2.1 Analysis of Mercury in Water by Manual or
Automated CVAA
9.2.1.1 Until more conclusive data are obtained,
samples are preserved at the time of
collection by acidification with nitric
acid to a pH of 2 or lower.
9.2.1.2 Analysis for mercury should be completed
within 28 days after collection of the
sample.
9.2.2 Analysis of Mercury in Soil/Sediment by
Manual CVAA
9.2.2.1 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.
9.2.2.2 Refrigerate soil samples at 4 °C (+2 °C)
upon collection until analysis.
9.2.2.3 The sample should be analyzed without
drying. A separate percent solids
191
-------
determination is required (Section
11.1.1).
9.2.2.4 Analysis should be completed within 28
days after sample collection.
10 PROCEDURES FOR SAMPLE PREPARATION AND
ANALYSIS
10.1 ICP and GFAA Spectroscopi'c Techniques
10.1.1 Water Sample Preparation
10.1.1.1 Acid digestion procedure for GFAA -- Shake
sample and transfer 100 ml of well-mixed
sample to a 250-mL beaker, add 1 ml (1+1)
HNOj and 2 ml 30% HjO. to the sample.
Cover with watch glass or similar cover
and heat on a steam bath or hot plate for
2 hours at 95 °C or until sample volume is
reduced to between 25 and 50 ml, making
certain sample does not boil. Cool sample
and filter to remove insoluble material.
(NOTE: In place of filtering the sample,
after dilution and mixing the sample may
be centrifuged or allowed to settle by
gravity overnight to remove insoluble
material.) Adjust sample volume to 100 ml
with deionized distilled water. The
sample is now ready for analysis. NOTE:
If Sb is to be determined by furnace AA,
use the digestate prepared for ICP
analysis.
10.1.1.2 Acid digestion procedure for ICP analysis
-- Shake sample and transfer 100 ml of
well-mixed sample to a 250-mL beaker, add
2 mL (1+1) HN03 and 10 mL (1+1) HCl to the
sample. Cover with watch glass or similar
cover and heat on a steam bath or hot
plate for 2 hours at 95 °C or until sample
volume is reduced to between 25 and 50 mL,
making certain sample does not boil. Cool
sample and filter to remove insoluble
material. (NOTE: In place of filtering
the sample, after dilution and mixing the
sample may be centrifuged or allowed to
settle by gravity overnight to remove
insoluble material.) Adjust sample volume
to 100 mL with deionized distilled water.
The sample is now ready for analysis.
10.1.1.3 Sludge samples having less than 1% solids
should be treated by the above method.
Sludge samples having between 1 to 30%
solids should be diluted to less than 1%
solids and then treated by the above
method.
10.1.2 Soil Sample Preparation—This method is an
acid digestion procedure used to prepare
soils, sediments, and sludge samples
containing more than 30% solids, for
analysis by 6FAA or by ICP. A
representative 1 g (wet weight) sample is
digested in nitric, acid and hydrogen
peroxide. The digestate is then refluxed
with either nitric acid or hydrochloric
acid. Hydrochloric acid is used as the
final reflux acid for the furnace AA
analysis of Sb, the ICP analysis of Al,
Sb, Ba, Be, Ca, Cd, Cr, Co, Cu, Fe, Pb,
Hg, Mn, Ni, K, Ag, Na, Tl, V and Zn.
Nitric acid is employed as the final
reflux acid for the furnace AA analysis of
As, Be, Cd, Cr, Co, Cu, Fe, Pb, Mn, Mi,
Se, Ag, Tl, V, and Zn. A separate sample
shall be dried for a percent solids
determination (Section 11.1.1).
10.1.2.1 Nix the sample thoroughly to achieve homo-
geneity. For each digestion procedure,
weigh (to the nearest 0.01 g) a 1.0 - 1.5
g portion of sample and transfer it to a
beaker.
10.1.2.2 Add 10 mL of 1:1 nitric acid (HN03), mix
the slurry, and cover with a watch glass.
Heat the sample to 95 °C and reflux for 10
minutes without boiling. Allow the sample
to cool, add 5 mL of cone. HNO,, replace
the watch glass, and reflux for 30
minutes. Do not allow the volume to be
reduced to less than 5 mL, while
maintaining a covering of solution over
the bottom of the beaker.
10.1.2.3 After the second reflux step has been
completed and the sample has cooled, add 2
mL of deionized distilled water and 3 mL
of 30% H-0-. Return the beaker to the hot
plate for warming 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, then cool
the beaker.
10.1.2.4 Continue to add 30% H20. in 1 mL aliquots
with warming until the effervescence is
minimal or until the general sample
appearance is unchanged. NOTE: Do not
add more than a total of 10 mL 30% H.O..
10.1.2.5 If the sample is being prepared for the
furnace AA analysis of Sb, or ICP analysis
of Al, Sb, Ba, Be, Ca, Cd, Cr, Co, Cu,
Fe, Pb, Mg, Mn, Ni, K, Ag, Na, Tl, V, and
Zn, add 5 mL of 1:1 HCl and 10 mL of
192
-------
deionized distilled water, return the
covered beaker to the hot plate, and heat
for an additional 10 minutes. After
cooling, filter through Whatman No. 42
filter paper (or equivalent) and dilute to
100 ml with deionized distilled water.
(NOTE: In place of filtering the sample,
after dilution and mixing the sample may
be centrifuged or allowed to settle by
gravity overnight to remove insoluble
material.) The diluted sample has an
approximate acid concentration of 2.5%
(v/v) HCl and 5% (v/v) HNO,. Dilute the
digestate 1:1 (200 ml final volume) with
acidified water to maintain constant acid
strength. The sample is now ready for
analysis.
10.1.2.6 if the sample is being prepared for the
furnace analysis of As, Be, Cd, Cr, Co,
Cu, Fe, Pb, Mn, Ni, Se, Ag, Tl, V, and Zn,
continue heating the acid-peroxide
digestate until the volume has been
reduced to approximately 2 ml, add 10 ml
of deionized distilled water, and warm the
mixture. After cooling, filter through
Whatman No. 42 filter paper (or
equivalent) and dilute to 100 ml with
deionized distilled water. (NOTE: In
place of filtering the sample, after
dilution and mixing the sample may be
centrifuged or allowed to settle by
gravity overnight to remove insoluble
material.) The diluted digestate solution
contains approximately 2% (v/v) HNOj.
Dilute the digestate 1:1 (200 ml final
volume) with acidified water to maintain
constant acid strength. For analysis,
withdraw aliquots of appropriate volume,
and add any required reagent or matrix
modifier. The sample is now ready for
analysis.
10.1.3 Sample Analysis
10.1.3.1
10.1.3.2
Initiate the appropriate
configuration of the computer.
operating
Profile and calibrate the instrument
according to instrument manufacturer's
recommended procedures, using mixed
calibration standard solutions such as
those described in Section 6.1.4. Flush
the system with the calibration blank
(Section 6.1.8.1) between each standard.
NOTE: For boron concentrations greater
than 500 ug/L, extended flush times of 1 -
2 minutes may be required.
10.1.3.3 Begin the sample run, flushing the system
with the calibration blank solution
(Section 6.1.8.1) between each sample.
(See NOTE in Section 10.1.3.2.) Analyze
the CCV standard (Section 6.1.6) and the
calibration blank (Section 6.1.8.1)
following each 10 analytical samples.
10.1.3.4 A minimum of two replicate exposures are
required for standardization and for all
QC and sample analyses. The average
result of the multiple exposures for the
standardization and all QC and sample
analyses shall be used.
10.2 Semiquantitative Screen of 42 Elements by
ICP
10.2.1 All element files should be set up with
the narrowest possible survey and peak
windows. Wherever possible, automatic or
manual background correction for each
element should be employed to compensate
for interferences.
10.2.2 Wavelength calibration standards should be
run as many times as needed to bring all
analytes within the specified survey
window. This may require as many as five
replicate readings on the wavelength
standard. The lower threshold limit (LTD
for each element is established by
analyzing each analyte at a level of twice
the expected LTL in seven replicates. The
LTL is the value obtained by multiplying
three times the standard deviation of the
replicate readings.
10.2.3 Following wavelength calibration,
instrument calibration standards and
blanks are run. The system should be
flushed with the calibration blank
solution between readings.
10.2.4 Analysis of solutions following
calibration can be performed using single
readings. Wavelength profiles should be
stored on a magnetic device for future
reference.
10.3 Analysis of Mercury in Water by Manual
Cold Vapor Technique
10.3.1 Transfer 100 ml of sample, or a sample
aliquot diluted to 100 ml, containing not
more than 1.0 ug of mercury, to a 300 ml
BOD bottle. Add 5 ml of sulfuric acid
(Section 6.4.1) and 2.5 ml of cone, nitric
acid (Section 6.4.2), mixing after each
addition. Add 15 ml of potassium
193
-------
permanganate solution (Section 6.4.5) to
each sample bottle. The same amount of
KMnO, added to the samples should be
present in standards and blanks. (NOTE:
For sewage samples additional permanganate
may be required.) Shake and add
additional portions of potassium
permanganate solution, if necessary, until
the purple color persists for at least 15
minutes. Add 8 ml of potassium persulfate
(Section 6.4.6) to each bottle and heat
for 2 hours in a water bath at 95 °C
10.3.2 Cool and add 6 ml of sodium chloride-
hydroxylamine sulfate (Section 6.4.4) to
reduce the excess permanganate (NOTE: Add
reductant in 6 ml increments until KMnO,
is completely reduced.) Purge the head
space in the BOD bottle for at least 1
minute, add 5 ml of stannous sulfate
(Section 6.4.3), and immediately attach
the bottle to the aeration apparatus.
Continue as described under Section 7.2.1.
10.3.3 Sludge samples having less than 1% solids
should be treated by the above method.
Whereas, sludge samples having between 1
to 30% solids should be diluted to less
than 1% solids and then treated by the
above method.
10.4 Analysis of Mercury in Water by Automated
Cold Vapor Technique
10.4.1 Set up manifold as shown in Figure 3.
10.4.2 Feeding all the reagents through the
system, with acid wash solution (Section
6.5.2.1) through the sample line, adjust
heating bath to 105 "C.
10.4.3 Turn on atomic absorption spectrophoto-
meter, adjust instrument settings as
recommended by the manufacturer, align
absorption cell in light path for maximum
transmittance, and place heat lamp
directly over absorption cell.
10.4.4 Arrange working mercury standards from 0.2
to 20.0 ug Hg/L in sampler and start
sampling. Complete loading of sample tray
with unknown samples.
10.4.5 Prepare standard curve by plotting peak
height of processed standards against
concentration values. Determine
concentration of samples by comparing
sample peak height with standard curve.
10.4.6 After the analysis is complete put all
lines except the HjSO, line in distilled
water to wash out system. After flushing
the system, wash out the H-SO, line. Also
flush the coils in the high temperature
heating bath by pumping stannous sulfate
(Section 6.5.3) through the sample lines,
followed by deionized distilled water.
This Mill prevent build-up of oxides of
manganese. Because of the toxic nature of
mercury vapor, precaution must be taken to
avoid its inhalation. Venting the mercury
vapor into an exhaust hood or passing the
vapor through some absorbing media such
as, equal volumes of 0.1 N KMnO, (Section
6.5.6) and 10% HgSO, (Section 6.3.1.2), or
0.25% iodine in a 3% KI solution, is
recommended. A specially treated charcoal
that will absorb mercury vapor is also
available.
10.4.7 For treatment of sludge samples, see
Section 10.3.3.
10.5 Analysis of Mercury in Soil/Sediment by
Manual Cold Vapor Technique
10.5.1 Weigh a representative 0.2 g portion of
wet sample and place in the bottom of a
BOD bottle. Add 5 ml of sulfuric acid
(Section 6.6.1) and 2.5 ml of cone, nitric
acid (6.6.2), mixing after each addition.
Heat two minutes in a water bath at 95 °C.
Cool, add 50 ml distilled water, 15 ml
potassium permanganate solution (Section
6.6.5), and 8 ml of potassium persulfate
solution (Section 6.6.6) to each sample
bottle. Mix thoroughly and place in the
water bath for 30 minutes at 95 °C. Cool
and add 6 ml of sodium chloride-
hydroxylamine sulfate (Section 6.6.4) to
reduce the excess permanganate. Add 55 ml
of distilled Hater. Treating each bottle
individually, purge the head space of the
sample bottle for at least one minute, add
5 ml of stannous sulfate (Section 6.6.3),
and immediately attach the bottle to the
aeration apparatus. Continue as described
under Section 7.3.1.
10.5.2 An alternate digestion procedure employing
an autoclave may also be used. In this
method, add 5 ml cone. HpSO, and 2 ml
cone. HNO, to the 0.2 g of sample. Then
add 5 ml saturated KMnO, solution and 8 ml
potassium persulfate solution and cover
the bottle with a piece of aluminum foil.
Autoclave the sample at 121 °C and 15 psi
for 15 minutes. Cool, make up to a volume
of 100 ml with distilled water, and
194
-------
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195
-------
add 6 mL of sodium chloride-hydroxylamine
sulfate solution (Section 6.6.4) to reduce
the excess permanganate. Purge the
headspace of the sample bottle for at
least 1 minute and continue as described
under Section 7.3.1.
10.5.3 Sludge samples having more than 30% solids
11 QUANT I TAT I ON DETERMINATION
11.1 ICP and GFAA Spectroscopic Techniques
11.1.1 Analytical results for water samples are
expressed in ug/L; for soil samples,
analytical results are expressed as mg/kg
on a dry weight basis. Therefore, a
determination of percent solids is
required for soils, sediments, and sludge
samples containing greater than 30%
solids, as follows.
11.1.1.1 Immediately following the weighing of the
sample to be processed for analysis (see
Section 10), add 5-10 g of sample to a
tared weighing dish. Weigh and record the
weight to the nearest 0.01 g.
11.1.1.2 Place weighing dish plus sample, with the
cover tipped to allow for moisture escape,
in a drying oven maintained at 103-105 "C.
NOTE: Sample handling and drying should
be conducted in a well-ventilated area.
11.1.1.3 Dry the sample overnight (12-24 hours),
but no longer than 24 hours. If dried
less than 12 hours, it must be documented
that constant weight was attained. Remove
the sample from the oven and cool in a
dessicator with the weighing dish cover in
place before weighing. Weigh and record
weight to nearest 0.01 g. Do not analyze
the dried sample.
NOTE: Drying time is defined as the
elapsed time in the oven. Therefore, time
in and out of the oven should be recorded
to document the 12-hour drying time
minimum. In the event it is necessary to
demonstrate the attainment of constant
weight, data must be recorded for a
minimum of two repetitive weigh/dry/
dessicate/ weigh cycles with a minimum of
one- hour drying time in each cycle.
Constant weight is defined as a loss in
weight of no greater than 0.01 g between
the start weight and final weight of the
last cycle.
11.1.1.4 Calculate percent solids by the formula
below. This value will be used for
calculating analyte concentration on a dry
weight basis.
11.1.2
11.1.2.1
11.1.3
X Solids
Sample Dry Weight
Sample Wet Weight
x 100
The concentrations determined in the
digest are to be reported on the basis of
the dry weight of the sample for
soil/sediment samples and sludge samples
containing greater than 30% solids.
Concentration (dry wt) (mg/kg)
_ C x V
W x S
Where,
C
V
Concentration (mg/L)
Final volume in liters after sample
preparation
Weight in kg of wet sample
X Solids/100
For aqueous samples and sludge samples
containing less than 30% solids, the
concentration of the elements in the
digest can determined as follows:
Concentration (ug/L) =
C x Vr
Where,
C =
Concentration (ug/L)
Final volume in liters after
sample preparation
Volume in liters of the sample
digested.
Preparation (reagent) blanks should be
treated as specified in Section 10.
11.1.4 If dilutions were performed, the
appropriate factor must be applied to
sample values.
11.1.5 Report results for semiquantitative ICP
screen of 42 elements in ug/L or mg/kg,
depending on the matrix. Samples are
semiquantified by comparing each analyte
result to the established LTL for that
analyte. All "peak offsets" or similar
designations reported by ICP should be
searched through stored spectrum files or
the data confirmed through sample spikes
before reporting.
196
-------
11.2
11.2.1
11.2.2
11.2.3
11.3
11.3.1
11.3.2
Analysis of Mercury in Water by Manual and
Automated Cold Vapor Technique
Determine the peak height of the unknown
from the chart and read the mercury value
from the standard curve.
Calculate the mercury concentration in the
sample by the formula:
ug Hg/L
ug Hg in aliquot
volume of aliquot in ml'
x 1000
Report mercury concentrations as follows:
below 0.20 ug/L,to 0.20 U; between 0.20
and 10.0 ug/L, to two significant figures;
equal to or above 10.0 ug/L, to three
significant figures.
Analysis of Mercury in Soil/Sediments by
Manual Cold Vapor Technique
Measure the height of the unknown peak
from the chart and read the mercury value
from the standard curve.
Calculate the mercury concentration in the
sample by the formula:
ug Hg/g =
ug Hg in the aliquot
wt of the aliquot in gms
(based upon dry weight of the sample)
11.3.3 Report mercury concentrations for
soil/sediment samples converted to units
of mg/kg. The sample result or the
detection limit for each sample must be
corrected for sample weight and percent
solids before reporting.
12 ANALYSIS OF COMPLEX SAMPLES
12.1 Some samples may contain high levels
(>1500 mg/L) of the compounds of interest,
interfering compounds, and/or polymeric
materials. These may lead to inaccuracies
in the determination of trace elements.
12.2. Physical, chemical, and/or spectral
interference effects may arise. These
interferences can be overcome by dilution
of the sample, matrix matching, varying
the temperature or by employing the Method
of Standard Addition. These effects are
described in Section 3.
12.3 The acceptable range for recovery of the
predigested spike is 75-125 percent for
all analytes. If any analyte falls
outside the QC limits, proceed as
described in Section 8.8.
13 METHOD PERFORMANCE
13.1 In an EPA round robin study, seven
laboratories applied the ICP technique to
acid-distilled water matrices that had
been dosed with various metal
concentrations. Table 12 lists the true
values, the mean reported values, and the
mean percent relative standard deviations
from this study.
13.2 The precision data obtainable for
electrothermal atomization method is given
in Table 13.
13.3 The precision data for CVAA technique for
analysis of mercury is given in Table 14.
14 GLOSSARY OF TERMS
14.1 Calibration blank -- A volume of deionized
distilled water acidified with HNOj and
HCl used in establishing the analytical
curve.
14.2 Calibration standards -- A series of known
standard solutions used by the analyst for
calibration of the instrument (i.e.,
preparation of the analytical curve).
14.3 Continuing calibration verification (CCV)
standard solutions -- A multi-element
standard of known concentrations prepared
by the laboratory to monitor and verify
instrument performance on a daily basis.
14.4 Dissolved elements -- Those elements which
will pass through a 0.45 urn membrane
filter.
14.5 Initial calibration verification (ICV)
standard solutions -- A solution obtained
from an outside source having known
concentration values, used to verify the
calibration standards.
14.6 Instrumental detection limits (IDL) --
Determined by multiplying by three the
standard deviation obtained for the
analysis of a standard solution (each
analyte in reagent water) at a
concentration of 3-5x IDL on three
nonconsecutive days, with seven
consecutive measurements per day.
14.7 Interference check sample (ICS) -- A
solution containing both interfering and
analyte elements of known concentration,
197
-------
used to verify background and inter-
el ement correction factors.
14.8 Laboratory control sample -- A control
sample of known composition. Aqueous and
solid laboratory control samples are
analyzed using the same sample
preparation, reagents, and analysis
methods employed for the analytical
samples.
14.9 Linear range -- The concentration range
over which the analytical curve remains
linear.
14.10 Lower threshold limit (LTD " Based on
signal-to-noise ration of 2:1 for each
element, expressed as mg/L. Levels lower
than LTL are considered "not detected."
The LTL for each element is highly
dependent on sample matrix.
14.1 Method of Standard Addition (MSA) -- The
standard addition technique involves the
use of the unknown and the unknown-plus-a-
known amount of standard by adding known
amounts of standard to one or more
aliquots of the processed sample solution
The MSA procedure is described in Section
8.15.
14.12 Minimum level (ML) -- The minimum level is
defined as the minimum concentration of a
substance that can be measured and
reported with 99% confidence that the
value is above zero. The laboratory is
required to achieve the ML listed for each
element in Table 11.
14.13 Preparation (reagent) blank -- A volume of
deionized distilled water containing the
same acid matrix as the calibration
standards, that is carried through the
entire analytical scheme.
14.14 Sensitivity -- The slope of the analytical
curve, i.e., functional relationship
between emission intensity or absorption
and concentration.
14.15 Serial dilution analysis -- A five-fold
dilution analysis used to establish a
chemical or physical interference effect.
14.16 Soil samples
sludge samples
solids.
Soils, sediments, and
containing more than 30%
14.17 Suspended elements -- Those elements which
are retained by a 0.45 urn membrane filter.
14.18 Total elements -- The concentration
determined on an unfiltered sample
following vigorous digestion.
14.19 Water samples -- Aqueous samples and
sludge samples containing 30% or less
solids which are diluted and treated as
water samples.
15 BIBLIOGRAPHY
15.1 Annual Book of ASTH Standards, Part 31,
"Water," Standard 03223-73 (1976).
15.2 "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.
15.3 Handbook for Analytical Quality Control in
Water and Wastewater Laboratories, EPA-
600/4-79-019.
15.4 "Inductively Coupled Plasma-Atomic
Emission Spectrometric Method of Trace
Elements Analysis of Water and Waste",
Method 200.7 modified by CLP Inorganic
Data/Protocol Review Committee; original
method by Theodore D. Martin,
EMSL/Cincinnati.
15.5 "Interim Methods for the Sampling and
Analysis of Priority Pollutants in
Sediments and Fish . Tissue," USEPA
Environmental Monitoring and Support
Laboratory/Cincinnati, Ohio, August 1977,
revised October 1930.
15.6 Methods for Chemical Analysis of Water and
Wastes, EPA-600/4-79-020.
15.7 "OSHA Safety and Health Standards, General
Industry," (29 CFR 1910), Occupational
Safety and Health Administration, OSHA
2206, (Revised, January 1976).
15.8 "Safety in Academic Chemistry
Laboratories," American Chemical Society
Publications, Committee on Chemical
Safety, 3rd Edition, 1979.
15.9 Standard Methods for the Examination of
Water and Wastewater, 14th Edition, p. 156
(1975).
15.10 Statement of Work for Inorganics Analysis,
Multi-Media, Multi-Concentration, SOW No.
788, USEPA Contract Laboratory Program
(July, 1988).
198
-------
15.11 Bishop, J. N., "Mercury in Sediments,"
Ontario Water Resources Com., Toronto,
Ontario, Canada, 1971.
15.12 Brandenberger, H. and Bader, H., "The
Determination of Nanogram Levels of
Mercury in Solution by a FlameIess Atomic
Absorption Technique," Atomic Absorption
Newsletter 6, 101 (1967).
15.13 Brandenberger, H. and Bader, H., "The
Determination of Mercury by FlameIess
Atomic Absorption II, A Static Vapor
Method," Atomic Absorption Newsletter 7:53
(1963).
15.14 Garbarino, J.R. and Taylor, H.E., "An
Inductively-Coupled Plasma Atomic Emission
Spectrometric Method for Routine Water
Quality Testing," Applied Spectroscopy 33,
No. 3 (1979).
15.15 Goulden, P.O. and Afghan, B.K. "An
Automated Method for Determining Mercury
in Water," Technicon, Adv. in Auto. Analy.
2, p. 317 (1970).
15.16 Hatch, W.R. and Ott, W.L., "Determination
of Sub-Microgram Quantities of Mercury by
Atomic Absorption Specrophotometry," Anal.
Chem. 40, 2085 (1968).
15.17 Kopp, J.F., Longbottom, M.C. and Lobring,
L.B., "Cold Vapor Method for Determining
Mercury," AWWA, vol. 64, p. 20, Jan. 1972.
15.18 Salma, M., personal communication, EPA
Cal/Nev. Basin Office, Almeda, California.
15.19 Wallace R.A., Fulkerson, W., Shults, W.D.,
and Lyon, W.S., "Mercury in the
Environment-The Hunan Element," Oak Ridge
National Laboratory, ORNL/NSF-EP-1, p. 31,
(January, 1971).
15.20 Winefordner, J.D., "Trace Analysis:
Spectroscopic Methods for Elements,"
Chemical Analysis, Vol. 46, pp. 41-42.
15.21 Winge, R.K., V.J. Peterson, and V.A.
Fassel, "Inductively Coupled Plasma-Atomic
Emission Spectroscopy Prominent Lines,"
EPA-600/4-79-017.
199
-------
Table 1
RECOMMENDED WAVELENGTHS AND ESTIHATiD
INSTRUMENTAL DETECTION LIMITS ANALYZED BY ICP
Table 2
RECOMMENDED WAVELENGTHS, ESTIMATED INSTRUMENTAL
DETECTION LIMITS, AND OPTIMUM CONCENTRATION RANGE FOR
ELEMENTS ANALYZED BY GFAA SPECTROSCOPY (1)
Element
Aluminum
Antimony
Arsenic
Barium
Beryl lium
Boron
Cadmium
Calcium
Chromium
Cobalt
Copper
Iron
Lead
Magnesium
Manganese
Molybdenum
Nickel
Potassium
Selenium
Silica (SiO,)
Silver
Sodium
Thallium
Vanadium
Zinc
Wavelength (1)
nm
308.215
206.833
193.696
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 (2)
ug/L
45
32
531
2
0.3
5
4
10
7
7
6
7
42
30
2
8
15
(3)
75
58
7
29
40
8
2
Estimated Optimum
Detection Concentration
Element Wavelength Limit (2) Range (2)
(nm) (ug/L) (ug/L)
Antimony 217.6 3 20-300
Arsenic 193.7 1 5-100
Beryllium 234.9 0.2 1-30
Cadmium 228.8 0.1 0.5-10
Chromium 357.9 1 5-100
Lead 283.3 1 5-100
Selenium 196.0 2 5-100
Thallium 276.8 1 5-100
(1) Values are taken from Methods 204.2 (Sb), 206.2
(As), 210.2 (Be), 213.2 (Cd), 218.2 (Cr), 239.2
(Pb), 270.2 (Se), 272.2 (Ag), 279.2 (Tl),
"Methods for Chemical Analysis of Water and
Wastes" (EPA-600/4-79-020), Metals-4.
(2) Concentration values and instrument conditions
given are for a Perkin-Elmer HGA-2100, based on
the use of a 20 uL injection, continous flow
purge gas, and non-pyrolytic graphite, and are
to be used as guidelines only. Smaller size
furnace devices or those employing faster rates
of atomization can be operated using lower
atomization temperatures for shorter time
periods than these recommended settings.
(1) These wavelengths are reeomnended 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 Section 3.1.1). The use of
alternate wavelengths should be reported (in nm)
with the sample data.
(2) Estimated detection limits are taken from
"Inductively Coupled Plasma-Atomic Emission
Spectroseopy-Proininent 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.
(3) Highly dependent on operating conditions and
plasma position.
-------
Table 3
RECOMMENDED INSTRUMENTAL PARAMETERS FOR ANALYSIS OF TRACE ELEMENTS BY GFAA SPECTROSCOPY (1)
Element
Antimony
Arsenic
Beryllium
Cadmium
Chromium
Lead
Selenium
Thallium
Drying
Time and Temperature
(sec) (°C)
30
30
30
30
30
30
30
30
125
125
125
125
125
125
125
125
Ashing
Time and Temperature
(sec) <°C)
30
30
30
30
30
30
30
30
800
1100
1000
500
1000
500
1200
400
Atomizing
Time and Temperature
(sec) (°C)
10
10
10
10
10
10
10
10
2700
2700
2800
1900
2700
2700
2700
2400
Purge Gas
Atmosphere
Argon (2)
Argon
Argon
Argon
Argon
Argon
Argon
Argon (2)
(1) Other operating parameters should be set as specified by the particular instrument manufacturer.
(2) Nitrogen may be substituted as the purge gas (see Section 3.2.2).
201
-------
Table 4
ICP SCREEN ELEMENTS, WAVELENGTHS, AND LOWER THRESHOLD
LIMITS
Element
Bismuth
Cerium
Dysprosium
Erbium
Europium
Gadolinium
Gallium
Germanium
Gold
Hafnium
Holmium
Indium
Iodine
Indium
Lanthanum
Lithium
Lutetium
Neodymium
Niobium
Osmium
Palladium
Phosphorus
Platinum
Potassium
Praseobymium
Rhenium
Rhodium
Ruthenium
Samarium
Scandium
Si I icon
Strontium
Sulfur
Tantalium
Tellurium
Terbium
Thorium
Thulium
Tungsten
Uranium
Ytterbium
Zirconium
Symbol
Bi-SS
Ce-SS
Dy-SS
Er-SS
Eu-SS
Gd-SS
Ga-SS
Ge-SS
Au-SS
Hf-SS
Ho-SS
In-SS
I-SS
Ir-SS
La-SS
Li-SS
Lu-SS
Nd-SS
Nb-SS
Os-SS
Pd-SS
P-SS
Pt-SS
K-SS
Pr-SS
Re-SS
Rh-SS
RU-SS
Sm-SS
Sc-SS
Si-SS
Sr-SS
S-SS
Ta-SS
Te-SS
Tb-SS
Th-SS
Tm-SS
W-SS
U-SS
Yb-SS
Zr-SS
Wavelength (1)
396.152
413.765
353.170
349.910
381 .967
342.247
294.364
265.118
242.765
277.336
345.600
230.606
183.038
224.268
379.478
670.781
261.542
309.418
401.225
228.226
340.458
213.618
214.423
766.490
390.844
221.426
233.477
240.272
359.260
361 .384
251.611
407.771
180.731
226.230
214.281
350.917
283.730
313.126
207.911
385.958
328.937
343.823
LTL (2)
(mg/L)
0.1
1
0.1
0.1
0.1
0.5
0.5
0.5
1
1
0.5
1
1
1
0.1
0.1
0.1
0.5
1
0.1
0.5
1
1
1
1
1
1
1
0.5
0.1
0.1
0.1
1
0.5
1
0.5
1
0.5
1
1
0.1
0.1
(1) Wavelength: Most sensitive line for analysis.
Line choice is dependent on sample matrix. Use
of secondary lines is necessary for some elements
for spectral interference confirmation.
(2) LTL: Lower Threshold Limit. Based upon signal-
to-noise ratio for each element; expressed as
mg/L. Lower levels would be recorded as ND. The
LTL for each analyte is highly dependent upon
sample matrix.
202
-------
Table 5
EXAMPLE OF ANALYTE CONCENTRATION EQUIVALENTS (MG/L) ARISING FROM INTERFERENTS AT THE 100 MG/L LEVEL
Element
Aluminum
Antimony
Arsenic
Barium
Beryllium
Boron
Cadmium
Calcium
Chromium
Cobalt
Copper
Iron
Lead
Magnesium
Manganese
Molybdenum
Nickel
Selenium
Silicon
Sodium
Thallium
Vanadium
Zinc
Wavelength
(nm)
308.215
206.833
193.696
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
196.026
288.158
588.995
190.864
292.402
213.856
Al Ca
..
0.47 --
1.3
..
..
0.04
..
..
..
..
..
--
0.17 --
0.02
0.005 --
0.05
..
0.23 --
..
..
0.30 --
..
..
Cr
--
2.9
0.44
--
--
--
--
0.08
--
0.03
--
--
--
0.11
0.01
--
--
--
0.07
--
--
0.05
--
Cu Fe
..
0.08
--
..
..
0.32
0.03
0.01
0.003
0.005
0.003
..
..
0.13
0.002
0.03
..
0.09
..
..
..
0.005
0.14
Mg Hn
0.21
..
-.
..
-.
..
..
0.01 0.04
0.04
..
..
0.12
..
0.25
0.002 --
--
..
..
..
..
..
..
--
Ni Ti
..
.25
..
..
0.04
..
0.02
0.03
..
0.03 0.15
0.05
--
..
0.07
--
--
..
..
..
0.08
..
0.02
0.29
V
1.4
0.45
1.1
--
0.05
--
--
0.03
0.04
--
0.02
--
--
0.12
--
--
--
--
0.01
--
--
--
--
203
-------
Table 6
ANALYTE AND INTERFERENT ELEMENTAL CONCENTRATIONS USED
FOR INTERFERENCE MEASUREMENTS IN TABLE 5
Table 7
WORKING STANDARD CONCENTRATIONS
Analytes
Al
As
A1
AS
B
Ba
Be
Ca
Cd
Co
Cr
Cu
Fe
Mg
Mn
Mo
Na
Ni
Pb
Sb
Se
Si
T I
V
Zn
mg/L
10
10
10
10
10
1
1
1
10
1
1
1
1
1
1
10
10
10
10
10
10
1
10
1
10
Interferents mg/L
Al 1000
Ca 1000
Al 1000 Blsmuth
Ca 1000 Cerlun
Cr 200 Dysprosium
Cu 200 Erbiun
Fe 1000 Europium
Mg 1000 Gadoliniun
Mn 200 Galllun
„, 200 Germanium
Ti 200 Gold
V 200 Hafnium
Holmium
Indium
Iodine
Iridium
Lanthanum
Lithium
Lutetian
Neodymium
Niobium
Osmium
Palladium
Phosphorus
Platinum
Pnl-assium
Praseobymium
Rhenium
Rhodium
Ruthenium
Samarium
Scandium
Silicon
Strontium
Sulfur
Tantalium
Tellurium
Terbium
Thorium
Thulium
Tungsten
Uranium
Ytterbium
Zirconium
Symbol
Bi
Ce
Dy
Er
Eu
Gd
Ga
Ge
Au
Hf
Ho
In
I
Ir
La
Li
Lu
Nd
Nb
Os
Pd
P
Pt
K
Pr
Re
Rh
Ru
Sm
Sc
Si
Sr
S
Ta
Te
Tb
Th
Tm
U
U
Yb
Zr
Working
Standard (1)
(mg/L)
1.0
1.0
1.0
1.0
1.0
1.0
1.0
10.0
10.0
10.0
10.0
10.0
10.0
10.0
1.0
1.0
1.0
1.0
10.0
10.0
1.0
10.0
10.0
150.0
10.0
10.0
10.0
10.0
1.0
1.0
1.0
1.0
10.0
1.0
10.0
1.0
10.0
1.0
1.0
10.0
1.0
1.0
(1) Working Standard: For each 1 mg/L of final
concentration needed, pipette 1 mL of stock
solution and dilute to 1 L final volume. For
example, for a 10 mg/L final concentration,
pipette 10.0 mL of stock solution.
204
-------
Table 8
QC SPECIFICATIONS FOR ANALYSIS OF PRECISION AND
ACCURACY STANDARDS (1)
Table 10
INITIAL AND CONTINUING CALIBRATION VERIFICATION
CONTROL LIMITS
ICP
Element
Be
Mn
V
As
Cr
Cu
Fe
Al
Cd
Co
Ni
Pb
Zn
Se
Spectroscopy
Mean
(2) % RSD (3)
70.7
4.23
1.93
15.83
8.37
17.67
8
17.2
14.67
11.7
10.27
20.67
20
24.07
GFAA Spectroscopy
Mean
Element (2) % RSD (4)
Pb 2.73
Cr 5.97
As (5) 12.83
Se (5) 9.7
Analytical
Method
ICP (1)/AA
Cold Vapor AA
Inorganic
Species
Metals
Mercury
% of True Value (EPA Set)
Low Limit
90
80
High Limit
110
120
(1) Limits apply to quantitative ICP and semiquanti-
tative ICP screen of 42 elements.
Table 11
ANALYTE AND INTERFERENT ELEMENTAL CONCENTRATIONS USED
FOR ICP INTERFERENCE CHECK SAMPLE
(1) Acceptable range of percent recovery for all
elements is 75-125%. As more data becomes
available, these limits Mill be re-evaluated.
(2) Other elements Mill be added as data becomes
available to EPA.
(3) Values derived from 21 determinations.
(4) Values derived from 30 determinations, except
for Pb. A total of 36 determinations Mere made
for Pb.
(5) Automated sample injection.
Table 9
MINIMUM LEVELS (ML) OF DETECTION
Analytes
ICP
Analytes
Calcium
Magnesium
Sodium
Aluminum
Manganese
Lead
Vanadium
Boron
Barium
Beryllium
Cadmium
Molybdenum
Tin
Yttrium
Cobalt
Chromium
Copper
Iron
Nickel
Titanium
Zinc
ML
(ug/L)
5000
5000
5000
200
15
50
50
10
200
5
5
10
30
5
50
10
25
100
40
5
20
Furnace AA
Analytes
Selenium
Thallium
Silver
Arsenic
Antimony
Mercury
ML
(ug/L)
5
10
10
10
20
0.2
Ag
8a
Be
Cd
Co
Cr
Cu
Mn
Ni
Pb
V
Zn
mg/L
1.0
0.5
0.5
1.0
0.5
0.5
1.0
1.0
0.5
1.0
Interferents
mg/L
Al
Ca
Fe
Mg
500
500
200
500
205
-------
Sample
Table 12
ICP PRECISION AND ACCURACY DATA (1)
Sample #2
Sample #3
Element
Be
Nn
V
As
Cr
Cu
Fe
Al
Cd
Co
Mi
Pb
Zn
Se
True
Value
(ug/L)
750
350
750
200
150
250
600
700
50
500
250
250
200
40
Mean
Reported
Value
(ug/L)
733
345
749
208
149
235
594
696
48
512
245
236
201
32
Mean
Percent
RSD
6.2
2.7
1.8
7.5
3.8
5.1
3.0
5.6
12
10
5.8
16
5.6
21.9
True
Value
(ug/L)
20
15
70
22
10
11
20
60
2.5
20
30
24
16
6
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
(ug/L)
180
100
170
60
50
70
180
160
14
120
60
80
80
10
Mean
Reported
Value
(ug/L)
176
99
169
63
50
67
178
161
13
108
55
80
82
8.5
Mean
Percent
RSD
5.2
3.3
1.1
17
3.3
7.9
6.0
13
16
21
14
14
9.4
8.3
(1) Not all elements were analyzed by all laboratories.
Table 13
PRECISION DATA FOR ELECTROTHERMAL ATOMIZATION METHODS (1)
Metal Wavelength
(rai)
Pb 217.0
Cr 357.9
As (2) 193.7
Se (2) 196.0
Sample
Size
(UL>
25
25
25
20
20
20
50
50
50
50
50
50
No. of
Replicate
Determinations
12
12
12
10
10
10
10
10
10
10
10
10
Mean
Concentration
(ug/L)
36.6
103
161
10.5
55.2
76.6
12.5
28.4
58.4
12.5
29.6
55.8
Relative
Standard
Deviation
3.8
2.9
1.5
9.3
4.3
4.3
17.6
13.7
7.2
17.6
5.6
5.9
(1) Values taken from "Standard Methods for the Examination of Water and Wastewater," 16th edition, p 179
(1985).
(2) Automated sample injection.
-------
Table 14
PRECISION DATA FOR CVAA TECHNIQUE FOR ANALYSIS OF MERCURY (1)
Metal
(Dissolved)
Inorganic
Organic
Netal
Concentration
(ug/L)
0.34
4.2
4.2
Relative
Standard
Deviation
(%)
0.077
0.56
0.36
Relative
Error
(%>
21
14.4
8.4
Number of
Participants
23
21
21
(1) Data from Kopp, J.F., M.C. Longbottom, and L. 8. Lobring, 1972, "Cold Vapour Method for Determinig
Mercury," J. Amer. Water Works Ass. 64:20, for distilled water samples.
207
-------
208
-------
EPA METHOD 160.3
RESIDUE, TOTAL
GRAVIMETRIC, DRIED AT 103-105
209
-------
Modification to Method 160.3 for analysis of solids:
Accurately weigh approximately 50 grams of soil, sediment, or sludge sample to
the nearest 0.1 mg. Proceed with drying the sample at 103-105 °C per Section 7.3.
210
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RESIDUE, TOTAL
Method 160.3 (Gravimetric, Dried at 103-105°C)
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
211
-------
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:
_,,,., ,, (A - B)xl,000
Total residue, mg/1 = v •£—'-
where:
A = weight of sample + dish in mg
B — weight of dish in mg
C = volume of sample in ml
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).
212
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EPA METHOD
CYANIDE, TOTAL
TITRIMETRIC,
213
-------
Modification to Method 335.2 for analysis of solids:
Accurately weigh approximately 5 grams of soil, sediment, or sludge sample to the
nearest 0.1 mg. Transfer the sample quantitatively into the CN distillation flask.
Add deionized distilled water to bring the sample to the required 500 mL volume.
Proceed with analysis starting with Section 8.2.1.
214
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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 of ascorbic
acid for each liter of sample volume.
Approved for NPDES
Issued 1974
Editorial revision 1974 and 1978
Technical Revision 1980
215
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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 other 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.
5.4 High results may be obtained for samples that contain nitrate and/or nitrite. During
the distillation nitrate and nitrite will form nitrous acid which will react with some
organic compounds to form oximes. These compounds formed will decompose under
test conditions to generate HCN. The interference of nitrate and nitrite is eliminated
by pretreatment 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.
6.4 Reflux distillation apparatus for sulfide removal as shown in Figure 3. The boiling
flask same as 6.1. The sulfide scrubber may be a Wheaton Rubber #709682 with 29/42
joints, size 100 ml. The air inlet tube should not be fritted. The cyanide absorption
vessel should be the same as the sulfide scrubber. The air inlet tube should be fritted.
6.5 Flow meter, such as Lab Crest with stainless steel float (Fisher 11-164-50).
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.
216
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7.2 Lead acetate: Dissolve 30 g of Pb (CzHaOz^S^O 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 H2SO4 to 500 ml of distilled
water.
7.6 Sodium dihydrogenphosphate, 1 M: Dissolve 138 g of NaH2PO4ttH2O 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 ml of distilled
water. Standardize with 0.0192 N AgNO3. Dilute to appropriate concentration so that
1 ml = 1 mg CN.
7.8 Standard cyanide solution, intermediate: Dilute 100. Oml of stock(l ml = l mgCN)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-Methyl-l-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'-diphenyl-[4,4'-bi-2 pyrazoline]-5,5'dione (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 MgCl2«6H2O into a 1000 ml flask, dissolve
and dilute to 1 liter with distilled water.
7.15 Sulfamic acid.i
217
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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. Adj ust 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 1.5 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 HaSO4.
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 100 ml 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-pyrazolene method: Add 0.5 ml of chloramine T (7.12) and mix. See
Note 1 and 2. After 1 to 2 minutes add 5 ml of pyridine-pyrazolone solution
218
-------
(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 Kl-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 fjg CN) 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 fJg/\. 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 that at least 3
standards be distilled.
8.9.2 Prepare a standard curve by plotting absorbance of standard vs. cyanide con-
centrations.
219
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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
220
-------
9.2 Using the titrimetric procedure, calculate concentration of CN as follows:
CN, mg/1 = v" ~ "'*•"«" -- 25°
ml orig. 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 nig/1 CN^ the standard deviations were
±0.005, ±0.007, ±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 Colorimetric 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.
221
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ALLIHN CONDENSER-—
AIR INLET TUBE
CONNECTING TUBING
ONE LITER-
BOILING FLASK
SUCTION
1
CYANIDE DISTILLATION
222
-------
COOLING WATER
INLET
SCREW CLAMP
I
HEATER-
TO LOW VACUUM
SOURCE
- ABSORBER
DISTILLING FLASK
O
FIGURE 2
CYANIDE DISTILLATION APPARATUS
223
-------
224
-------
EPA METHOD 340.2
FLUORIDE
POTENTIOMETRIC, ION SELECTIVE ELECTRODE
225
-------
Modification to Method 340.2 for analysis of solids:
For determination of total fluoride in solids, Bellack distillation (Section 1.4) is
necessary. Accurately weigh 5 grams of soil, sediment, or sludge sample to the
nearest 0.1 mg. Quantitatively transfer the sample into the distillation flask. Add
deionized distilled water to bring sample to 50 mL volume. Perform Bellack
distillation per EPA Method 340.1, Section 6.1, using a stirring heating mantle as
the heat source. To prevent bumping, place a stirring bar into the flask and stir the
contents during the heating process. After distillation is complete, proceed with
analysis by Method 340.2.
NOTE: Method 340.1 is included as part of this modification.
226
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FLUORIDE, TOTAL
Method 340.1 (Colorimetric, SPADNS with Bellack Distillation)
STORET 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 The method covers the range from 0.1 to about 1.4 mg/1 F. This range may be extended
to 1000 mg/1 using the Fluoride Ion Selective Electrode Method (340.2) after
distillation.
2. Summary of Method
2.1 Following distillation to remove interferences, the sample is treated with the SPADNS
reagent. The loss of color resulting from the reaction of fluoride with the zirconyl-
SPADNS dye is a function of the fluoride concentration.
3. Comments
3.1 The SPADNS reagent is more tolerant of interfering materials than other accepted
fluoride reagents. Reference to Table 414:1, p 388, Standard Methods for the
Examination of Waters and Wastewaters, 14th Edition, will help the analyst decide if
distillation is required. The addition of the highly colored SPADNS reagent must be
done with utmost accuracy because the fluoride concentration is measured as a difference
of absorbance in the blank and the sample. A small error in reagent additon is the most
prominent source of error in this test.
3.2 Care must be taken to avoid overheating the flask above the level of the solution. This is
done by maintaining an even flame entirely under the boiling flask.
4. Apparatus
4.1 Distillation apparatus: A 1-liter round-bottom, long-necked pyrex boiling flask,
connecting tube, efficient condenser, thermometer adapter and thermometer reading to
200°C. All connections should be ground glass. Any apparatus equivalent to that shown
in Figure 1 is acceptable.
4.2 Colorimeter: One of the folio wing
4.2.1 Spectrophotometer for use at 570 nm providing a light path of at least 1 cm.
4.2.2 Filter photometer equipped with a greenish yellow filter having maximum
transmittance at 550 to 580 nm and a light path of at least 1 cm.
5. Reagents
5.1 Sulfuric acid, H2SO4, cone.
Approved for NPDES and SDWA
Issued 1971
Editorial revision 1974 and 1978
227
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5.2 Silver sulfate, Ag2SO4 crystals.
5.3 Stock fluoride solution: Dissolve 0.221 g anhydrous sodium fluoride, NaF, in distilled
water in a 1-liter volumetric flask and dilute to the mark with distilled water; 1.00 ml =
0.1 mg F.
5.4 Standard fluoride solution: Place 100 ml stock fluoride solution (5.3) in a 1 liter
volumetric flask and dilute to the mark with distilled water; 1.00 ml = 0.010 mg F.
5.5 SPADNS solution: Dissolve 0.958 g SPADNS, sodium 2-(parasulfophenylazo)-l,8-
dihydroxy-3,6-naphthalene disulfonate, in distilled water in a 500 ml volumetric flask
and dilute to the mark. Stable indefinitely if protected from direct sunlight.
5.6 Zirconyl-acid reagent: Dissolve 0.133 g zirconyl chloride octahydrate, ZrOCl2«8H2O in
approximately 25 ml distilled water in a 500 ml volumetric flask. Add 350 ml cone HC1
and dilute to the mark with distilled water.
5.7 Acid-zirconyl-SPADNS reagent: Mix equal volumes of SPADNS solution (5.5) and
zirconyl-acid reagent (5.6). The combined reagent is stable for at least 2 years.
5.8 Reference solution: Add 10 ml SPADNS solution (5.5) to 100 ml distilled water. Dilute 7
ml cone HC1 to 10 ml and add to the dilute SPADNS solution. This solution is used for
zeroing the spectrophotometer or photometer. It is stable and may be used indefinitely.
5.9 Sodium arsenite solution: Dissolve 5.0 g NaAsO2 in distilled water in a 1-liter volumetric
flask and dilute to the mark with distilled water (CAUTION: Toxic-avoid ingestion).
6. Procedure
6.1 Preliminary distillation
6.1.1 Place 400 ml distilled water in the distilling flask.
6.1.2 Carefully add 200 ml cone. H2SO4 and swirl until contents are homogeneous.
6.1.3 Add 25 to 35 glass beads, connect the apparatus (Figure 1) making sure all joints
are tight:
6.1.4 Heat slowly at first, then as rapidly as the efficiency of the condenser will permit
(distillate must be cool) until the temperature of the flask contents reaches exactly
180°C. Discard the distillate. This process removes fluoride contamination and
adjusts the acid-water ratio for subsequent distillations.
6.1.5 Cool to 120°C or below.
6.1.6 Add 300 ml sample, mix thoroughly, distill as in 6.1.4 until temperature reaches
180°C. Do not heat above 180°C to prevent sulfate carryover.
6.1.7 Add Ag2SO4 (5.2) at a rate of 5 mg/mg Cl when high chloride samples are distilled.
6.1.8 Use the sulfuric acid solution in the flask repeatedly until the contaminants from
the samples accumulate to such an extent that recovery is affected or interferences
appear in the distillate. Check periodically by distilling standard fluoride samples.
6.1.9 High fluoride samples may require that the still be flushed by using distilled water
and combining distillates.
6.2 Colorimetric Determination:
6.2.1 Prepare fluoride standards in the range 0 to 1.40 mg/1 by diluting appropriate
quantities of standard fluoride solution (5.4) to 50 ml with distilled water.
228
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CONNECTING TUBE
THERMOMETER
THERMOMETER ADAPTER
1-liter
ROUND BOTTOM
FLASK
X
124/40
JOINTS
ADAPTER
24/40
JOINT
BURNER
CONDENSER
300-ml
O VOLUMETRIC
FLASK
FIGURE 1 DIRECT DISTILLATION APPARATUS
FOR FLUORIDE.
229
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6.2.2 Pipet 5.00 ml each of SPADNS solution (5.5) and zirconyl-acid reagent (5.6) or
10.00 ml of the mixed acid-zirconyl-SPADNS reagent (5.7) to each standard and
mix well.
6.2.3 Set photometer to zero with reference solution (5.8) and immediately obtain
absorbance readings of standards.
6.2.4 Plot absorbance versus concentration. Prepare a new standard curve whenever
fresh reagent is made.
6.2.5 If residual chlorine is present pretreat the sample with 1 drop (0.05 ml) NaAsO2
solution (5.9) per 0.1 mg residual chlorine and mix. Sodium arsenite
concentrations of 1300 mg/1 produce an error of 0. 1 mg/1 at 1.0 mg/1 F.
6.2.6 Use a 50 ml sample or a portion diluted to 50 ml. Adjust the temperature of the
sample to that used for the standard curve.
6.2.7 Perform step 6.2.2 and 6.2.3.
7. Calculations
7. 1 Read the concentration in the 50 ml sample using the standard curve (6.2.4)
7.2 Calculate as follows:
mg/lF= x l'
mg/
ml sample
7.3 When a sample (ml sample) is diluted to a volume (B) and then a portion (C) is analyzed,
use:
,.„ mgF x 1,000 B
mg/1F= ml sample x "C
8. Precision and Accuracy
8.1 On a sample containing 0.83 mg/1 F with no interferences, 53 analysts using the Bellack
distillation and the SPADNS reagent obtained a mean of 0.81 mg/1 F with a standard
deviation of ±0.089 mg/1.
8.2 On a sample containing 0.57 mg/1 F (with 200 mg/1 SO4 and 10 mg/1 Al as
interferences) 53 analysts using the Bellack distillation obtained a mean of 0.60 mg/lF
with a standard deviation of ±0. 103 mg/ 1 .
8.3 On a sample containing 0.68 mg/1 F (with 200 mg/1 SO4, 2 mg/1 Al and 2.5 mg/1
[Na(PO3)6] as interferences), 53 analysts using the Bellack distillation obtained a mean of
0.72 mg/1 F with a standard deviation of ±0.092 mg/1. (Analytical Reference Service,
Sample 1 1 1-B water, Fluoride, August, 1961.)
Bibliography
1. Standard Methods for the Examination of Water and Wastewater, p. 389-390 (Method No.
414A, Preliminary Distillation Step) and p. 393-394 (Method 414C SPADNS) 14th Edition,
(1975).
2. Annual Book of ASTM Standards, Part 31, "Water", Standard D 1 179-72, Method A, p. 310
(1976).
230
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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 nig/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),
F(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"1"4, Fe"1"3 and Al+3 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. 94-090>
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
231
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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 Nad 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.
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
232
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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).
233
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234
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EPA METHOD 351.3
NITROGEN, KJELDAHL, TOTAL
COLORIMETRIC; TITRIMETRIC; POTENTIOMETRIC
235
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Modification to Method 351.3 for analysis of solids:
Accurately weigh approximately 10 grams of soil, sediment, or sludge sample to
the nearest 0.1 mg. Quantitatively transfer the sample to an 800 mL flask. Add
deionized distilled water to bring the sample to 500 mL volume. Proceed with
analysis.
236
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NITROGEN, KJELDAHL, TOTAL
Method 351.3 (Colorimetric; Titrimetric; Potentiometric)
STORET NO. 00625
1. Scope and Application
1.1 This method covers the determination of total Kjeldahl nitrogen in drinking, surface and
saline 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 nitrogenous compounds of some industrial wastes such as
amines, nitro compounds, hydrazones, oximes, semicarbazones and some refractory
tertiary amines.
1.2 Three alternatives are listed for the determination of ammonia after distillation: the
titrimetric method which is applicable to concentrations above 1 mg N/liter; the
Nesslerization method which is applicable to concentrations below 1 mg N/liter; and the
potentiometric method applicable to the range 0.05 to 1400 mg/1.
1.3 This method is described for macro and micro glassware systems.
2. Definitions
2.1 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.
2.2 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. This may be determined directly by removal of ammonia before
digestion.
3. Summary of Method
3.1 The sample is heated in the presence of cone, sulfuric acid, K2SO4 and HgSO4 and
evaporated until SO3 fumes are obtained and the solution becomes colorless or pale
yellow. The residue is cooled, diluted, and is treated and made alkaline with a hydroxide-
thiosulfate solution. The ammonia is distilled and determined after distillation by
Nesslerization, titration or potentiometry.
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. Preserved samples should be analyzed as soon as possible.
5. Interference
5.1 High nitrate concentrations (10X or more than the TKN level) result in low TKN
values. The reaction between nitrate and ammonia can be prevented by the use of an
anion exchange resin (chloride form) to remove the nitrate prior to the TKN analysis.
Approved for NPDES
Issued 1971
Editorial revision 1974 and 1978
237
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6. Apparatus
6.1 Digestion apparatus: A Kjeldahl digestion apparatus with 800 or 100 ml flasks and
suction takeoff to remove SO3 fumes and water.
6.2 Distillation apparatus: The macro Kjeldahl flask is connected to a condenser and an
adaptor so that the distillate can be collected. Micro Kjeldahl steam distillation
apparatus is commercially available.
6.3 Spectrophotometer for use at 400 to 425 nm with a light path of 1 cm or longer.
7. Reagents
7.1 Distilled water should be free of ammonia. Such water is best prepared by the passage of
distilled water through an ion exchange column containing a strongly acidic cation
exchange resin mixed with a strongly basic anion exchange resin. Regeneration of the
column should be carried out according to the manufacturer's instructions.
NOTE 1: All solutions must be made with ammonia-free water.
7.2 Mercuric sulfate solution: Dissolve 8 g red mercuric oxide (HgO) in 50 ml of 1:4 sulfuric
acid (10.0 ml cone. H2SO4 : 40 ml distilled water) and dilute to 100 ml with distilled
water.
7.3 Sulfuric acid-mercuric sulfate-potassium sulfate solution: Dissolve 267 g K2SO4 in 1300
ml distilled water and 400 ml cone. H2SO4. Add 50 ml mercuric sulfate solution (7.2) and
dilute to 2 liters with distilled water.
7.4 Sodium hydroxide-sodium thiosulfate solution: Dissolve 500 g NaOH and 25 g
Na2S2O3®5H2O in distilled water and dilute to 1 liter.
7.5 Mixed indicator: Mix 2 volumes of 0.2% methyl red in 95% ethanol with 1 volume of
0.2% methylene blue in ethanol. Prepare fresh every 30 days.
7.6 Boric acid solution: Dissolve 20 g boric acid, H3BO3, in water and dilute to 1 liter with
distilled water.
7.7 Sulfuric acid, standard solution: (0.02 N) 1 ml = 0.28 mg NH3-N. Prepare a stock
solution of approximately 0.1 N acid by diluting 3 ml of cone. H2SO4 (sp. gr. 1.84) to 1
liter with CO2-free distilled water. Dilute 200 ml of this solution to 1 liter with CO2-free
distilled water. Standardize the approximately 0.02 N acid so prepared against 0.0200 N
Na2CO3 solution. This last solution is prepared by dissolving 1.060 g anhydrous Na2CO3,
oven-dried at 140°C, and diluting to 1 liter with CO2-free distilled water.
NOTE 2: An alternate and perhaps preferable method is to standardize the
approximately 0.1 N H2SO4 solution against a 0.100 N Na2CO3 solution. By proper
dilution the 0.02 N acid can the be prepared.
7.8 Ammonium chloride, stock solution: 1.0 ml = 1.0 mg NH3-N. Dissolve 3.819 g NH4C1
in water and make up to 1 liter in a volumetric flask with distilled water.
7.9 Ammonium chloride, standard solution: 1.0 ml = 0.01 mg NH3-N. Dilute 10.0 ml of the
stock solution (7.8) with distilled water to 1 liter in a volumetric flask.
7.10 Nessler reagent: Dissolve 100 g of mercuric iodide and 70 g potassium iodide in a small
volume of distilled water. Add this mixture slowly, with stirring, to a cooled solution of
160 g of NaOH in 500 ml of distilled water. Dilute the mixture to 1 liter. The solution is
stable for at least one year if stored in a pyrex bottle out of direct sunlight.
238
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NOTE 3: Reagents 7.7, 7.8, 7.9, and 7.10 are identical to reagents 6.8, 6.2, 6.3, and 6.6
described under Nitrogen, Ammonia (Colorimetric; Titrimetric; Potentiometric-
Distillation Procedure, Method 350.2).
8. Procedure
8.1 The distillation apparatus should be pre-steamed before use by distilling a 1:1 mixture of
distilled water and sodium hydroxide-sodium thiosulfate solution (7.4) until the distillate
is ammonia-free. This operation should be repeated each time the apparatus is out of
service long enough to accumulate ammonia (usually 4 hours or more).
8.2 Macro Kjeldahl system
8.2.1 Place a measured sample or the residue from the distillation in the ammonia
determination (for Organic Kjeldahl only) into an 800 ml Kjeldahl flask. The
sample size can be determined from the following table:
Kjeldahl Nitrogen Sample Size
in Sample, mg/1 ml
0-5 500
5-10 250
10-20 100
20-50 50.0
50-500 25.0
Dilute the sample, if required, to 500 ml with distilled water, and add 100 ml
sulfuric acid-mercuric sulfate-potassium sulfate solution (7.3). Evaporate the
mixture in the Kjeldahl apparatus until SO3 fumes are given off and the solution
turns colorless or pale yellow. Continue heating for 30 additional minutes. Cool the
residue and add 300 ml distilled water.
8.2.2 Make the digestate alkaline by careful addition of 100 ml of sodium hydroxide -
thiosulfate solution (7.4) without mixing.
NOTE Si Slow addition of the heavy caustic solution down the tilted neck of the
digestion flask will cause heavier solution to underlay the aqueous sulfuric acid
solution without loss of free-ammonia. Do not mix until the digestion flask has
been connected to the distillation apparatus.
8.2.3 Connect the Kjeldahl flask to the condenser with the tip of condenser or an
extension of the condenser tip below the level of the boric acid solution (7.6) in the
receiving flask.
8.2.4 Distill 300 ml at the rate of 6-10 ml/min., into 50 ml of 2% boric acid (7.6)
contained in a 500 ml Erlenmeyer flask.
8.2.5 Dilute the distillate to 500 ml in the flask. These flasks should be marked at the 350
and the 500 ml volumes. With such marking, it is not necessary to transfer the
distillate to volumetric flasks. For concentrations above 1 mg/1, the ammonia can
be determined titrimetrically. For concentrations below this value, it is determined
colorimetrically. The potentiometric method is applicable to the range 0.05 to 1400
mg/1.
239
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8.3 Micro Kjeldahl system
8.3.1 Place 50.0 ml of sample or an aliquot diluted to 50 ml in a 100 ml Kjeldahl flask
and add 10 ml sulfuric acid-mercuric sulfate-potassium sulfate solution (7.3).
Evaporate the mixture in the Kjeldahl apparatus until SO3 fumes are given off and
the solution turns colorless or pale yellow. Then digest for an additional 30
minutes. Cool the residue and add 30 ml distilled water.
8.3.2 Make the digestate alkaline by careful addition of 10 ml of sodium hydroxide-
thiosulfate solution (7.4) without mixing. Do not mix until the digestion flask has
been connected to the distillation apparatus.
8.3.3 Connect the Kjeldahl flask to the condenser with the tip of condenser or an
extension of the condenser tip below the level of the boric acid solution (7.6) in the
receiving flask or 50 ml short-form Nessler tube.
8.3.4 Steam distill 30 ml at the rate of 6-10 ml/min., into 5 ml of 2% boric acid (7.6).
8.3.5 Dilute the distillate to 50 ml. For concentrations above 1 mg/1 the ammonia can be
determined titrimetrically. For concentrations below this value, it is determined
colorimetrically. The potentiometric method is applicable to the range 0.05 to 1400
mg/1.
8.4 Determination of ammonia in distillate: Determine the ammonia content of the distillate
titrimetrically, colorimetrically, or potentiometrically, as described below.
8.4.1 Titrimetric determination: Add 3 drops of the mixed indicator (7.5) to the distillate
and titrate the ammonia with the 0.02 N H2SO4 (7.7), matching the endpoint
against a blank containing the same volume of distilled water and H3BO3 (7.6)
solution.
8.4.2 Colorimetric determination: Prepare a series of Nessler tube standards as follows:
ml of Standard
1.0 ml = 0.01 mg NH3-N mg NH3-N/50.0 ml
0.0 0.0
0.5 0.005
1.0 0.010
2.0 0.020
4.0 0.040
5.0 0.050
8.0 0.080
10.0 0.10
Dilute each tube to 50 ml with ammonia free water, add 1 ml of Nessler Reagent
(7.10) and mix. After 20 minutes read the absorbance at 425 nm against the blank.
From the values obtained for the standards plot absorbance vs. mg NH3-N for the
standard curve. Develop color in the 50 ml diluted distillate in exactly the same
manner and read mg NH3-N from the standard curve.
8.4.3 Potentiometric determination: Consult the method entitled Nitrogen, Ammonia:
Potentiometric, Ion Selective Electrode Method, (Method 350.3) in this manual.
8.4.4 It is not imperative that all standards be treated in the same manner as the samples.
It is recommended that at least 2 standards (a high and low) be digested, distilled,
240
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and compared to similar values on the curve to insure that the digestion-distillation
technique is reliable. If treated standards do not agree with untreated standards the
operator should find the cause of the apparent error before proceeding.
9. Calculation
9.1 If the titrimetric procedure is used, calculate Total Kjeldahl Nitrogen, in mg/1, in the
original sample as follows:
TKN, mg/1 = (A ~ B)N x F x 1.000
where:
A = milliliters of standard 0.020 N H2SO4 solution used in titrating sample.
B = milliliters of standard 0.020 N H2SO4 solution used in titrating blank.
N = normality of sulfuric acid solution.
F = milliequivalent weight of nitrogen (14 mg).
S = milliliters of sample digested.
If the sulfuric acid is exactly 0.02 N the formula is shortened to:
TKN, mg/1 = (A - B> X 28°
9.2 If the Nessler procedure is used, calculate the Total Kjeldahl Nitrogen, in mg/1, in the
original sample as follows:
x .
D ~ C
where:
A = mg NH3-rN read from curve.
B = ml total distillate collected including the H3BO3.
C = ml distillate taken for Nesslerization.
D = ml of original sample taken.
9.3 Calculate Organic Kjeldahl Nitrogen in mg/1, as follows:
Organic Kjeldahl Nitrogen = TKN -
-------
10.
9.4 Potentiometric determination: Calculate Total Kjeldahl Nitrogen, in mg/1, in the
original sample as follows:
TKN, mg/1 = - x A
where:
A = mg NH3-N/1 from electrode method standard curve.
B = volume of diluted distillate in ml.
D = ml of original sample taken.
Precision
10.1 Thirty-one analysts in twenty laboratories analyzed natural water samples containing
exact increments of organic nitrogen, with the following results:
Precision as
Accuracy as
Standard Deviation
mg N/liter
0.197
0.247
1.056
1.191
Bias,
%
+ 15.54
+ 5.45
+ 1.03
- 1.67
Bias,
mg N/liter
+0.03
+0.02
+0.04
-0.08
Increment as
Nitrogen, Kjeldahl
mg N/liter
0.20
0.31
4.10
4.61
(FWPCA Method Study 2, Nutrient Analyses)
Bibliography
1. Standard Methods for the Examination of Water and Wastewater, 14th Edition, p 437,
Method 421(1975).
2. Schlueter, Albert, "Nitrate Interference In Total Kjeldahl Nitrogen Determinations and Its
Removal by Anion Exchange Resins", EPA Report 600/7-77-017.
242
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EPA METHOD 353.2
NITROGEN, NITRATE-NITRITE
COLORIMETRIC, AUTOMATED, CADMIUM REDUCTION
243
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Modification to Method 353.2 for analysis of solids:
Accurately weigh 5 grams of soil, sediment, or sludge sample to the nearest
0.1 mg. Add deionized distilled water to bring the sample to 100 mL volume.
Place the mixture on a shaker for 4 hours, then filter through Whatman #40 (or
equivalent). Proceed with analysis starting with Section 7.1.
244
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NITROGEN, NITRATE-NITRITE
Method 353.2 (Colorimetric, Automated, Cadmium Reduction)
STORET NO. Total 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 to
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 readily
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 stored for
more than 24 hours, they should be preserved with sulfuric acid (2 ml cone. H2SO4 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. Since
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 of 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 Auto Analyzer (A AI or A All) consisting of the following components:
5.1.1 Sampler.
5.1.2 Manifold (AAI) or analytical cartridge (AAll).
5.1.3 Proportioning Pump
5.1.4 Colorimeter equipped with a 15 mm or 50 mm tubular flow cell and 540 nm filters.
5.1.5 Recorder.
Approved for NPDES and SDWA
Issued 1971
Editorial revision 1974 and 1978
245
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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 fromTechnicon Corporation).
246
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INDENTATIONS FOR
SUPPORTING CATALYST
GLASS WOOL
Cd-TURNINGS
TILT COLUMN TO 20° POSTION
FIGURE 1. COPPER CADMIUM REDUCTION COLUMN
(1 1/2 ACTUAL SIZE)
247
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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 KNO2 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= 1.0mgNO2-N.
6.13 Standard nitrate solution: Dilute 10.0 ml of stock nitrate solution (6.11) to 1000 ml.
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.
1.0 ml — 0.01 mgNO2-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/1
CaCl2 - 1.16 g/1
KBr - 0.10 g/1
NaF - 0.003 g/1
MgCl2 - 5.20 g/1
KC1 - 0.70 g/1
H3BO3 - 0.03 g/1
Na2S04 - 4.09 g/1
NaHCO3 - 0.20 g/1
SrCl2 - 0.03 g/1
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.
HC1 or cone. 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 for 20
minutes.
248
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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 NO2 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
+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).
249
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TO SAMPLE WASH
WASTE
ml/min
T
PS-3
HO
C-3* MIXER
DOUBLE MIXER
WASTE
[BO
COLUMN *
BLUE
ff
BLUE
R
BLUE
BLUE
0.42
1.60
H20
0.80 AIR
2.00
JlL
C.42 COLOR REA6ENT
2.00
1.60 SAMPLE
1.20 8.5% NHjCL
1.20 AIR
WASTE
COLORIMETER
50mm TUIULASt f/c
FILTERS
PROPORTIONING PUMP
RECORDER
SAMPLER 2
RATE: 30 PER HR.
* FROM C-3 TO SAMPLE LINE USE
.030 x .048 POLYETHYLENE TUBING.
'* SEE FIGURE 1. FOR DETAIL. COLUMN
SHOULD BE IN 20° INCLINE POSITION
RANBE EXPANDER
FIGURE 2. NITRATE • NITRITE MANIFOLD AA-I
o
m
CM
-------
in
CM
PUMP TUBE ^^
RE
CO
WASTE
RECORDER J[
Q -
DIGITAL
PRINTER
I
Sx,.
f
Hj>
COLORINETE
520 nm FILT
15 mm FLOW
W
T
DUCTIONg,
LUMH
WWPJ
^-WASTF T
PUMP Tl
R
ER
CELL
fASH WATE
0 SAMPLE
A2
")
QUO
•BE
R n
:R
J
mi/mm
BLACK 0.32 AIR
Y Y
BLACK
BLACK
BLACK
W W
GREY
G G
L2 AMMONIUM
CHLORIDE
0.32 SAMPLE ,
0.32 AIR
032 COLOR
REAGENT
0-6 „
2.0 WASH
0
SAMPLER
40/hr
5TE
3TE
PROPORTIONING
PUMP
FIGURE 3 NITRATE-NITRITE MANIFOLD AAII
-------
252
-------
EPA METHOD 365.2
PHOSPHORUS, ALL FORMS
COLORIMETRIC, ASCORBIC ACID, SINGLE REAGENT
253
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Modification to Method 365.2 for analysis of solids:
Accurately weigh approximately 0.5 grams of soil, sediment, or sludge sample to
the nearest 0.1 mg. Transfer the sample quantitatively to a 125 mL Erlenmeyer
flask. Add deionized distilled water to bring the sample to the required 50 mL
volume. Proceed with analysis starting with Section 8.1.
254
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PHOSPHORUS, ALL FORMS
Method 365.2 (Colorimetric, Ascorbic Acid, Single Reagent)
STORE! 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/1 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 sulfuric acid
hydrolysis. Organic phosphorus compounds may be converted to the orthophosphate
form by persulfate digestion™.
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. H2SO4 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 [(PO4)'3] 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 sulfuric acid hydrolysis procedure, and minus pre-determined
orthophosphates. This hydrolyzable phosphorus includes polyphosphorus.
[(PjOy)"1, (PsO^)'5, etc.] plus some organic phosphorus. (00669)
Approved for NPDES
Issued 1971
255
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Total Sample (No Filtration)
\/
Direct
Colorimetry
Hydrolysis f-,
\/ Colorimetrv
Orthophosphate
Hydrolyzable §
Orthophosphate
Filter (through 0.45 p membrane filter)
\
Direct
Colorimetry
/ \
Dissolved
Orthophosphate
H2S04
Hydrolysis §
/ Colorimetry ^
Diss. Hydrolyzable
§ Orthophosphate
Persulfate
Digestion §
/ Colorimetry
Dissolved
Phosphorus
Persulfate
Digestion
\ / Colorimetrv
Phosphorus
FIGURE 1. ANALYTICAL SCHEME FOR DIFFERENTIATION
OF PHOSPHORUS FORMS
VO
IT)
Of
-------
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 distilled water and
257
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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/2H2O,
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, O.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 = 0.05mgP.
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.
-------
8. Procedure
8.1 Phosphorus
8.1.1 Add 1 ml of H2SO4 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 H2SO4 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.
259
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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
+ 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).
260
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