BASE/NEUTRALS AND ACIDS (6410)/Introduction 6-65

6410 EXTRACTABLE BASE/NEUTRALS AND ACIDS*

6410 A. Introduction

1. Sources and Significance 2. Selection of Method

The semivolatile compounds covered by this section include Method 6410B is a broad-spectrum gas chromatographic/mass
many classes of compounds, each characterized by different spectrometric (GC/MS) packed- or capillary-column method for
sources. The compounds include polynuclear aromatic hydrocar- detection of these compounds following liquid-liquid extraction.
bons, often as by-products of petroleum processing or combus- Although this method can be used to determine all the listed
tion; phthalates, used as plasticizers; phenolics, found most often compounds, it is not the most sensitive method for individual
in wood preservatives; organochlorine pesticides, found most classes of compounds, which are detected at lower concentra-
often in agricultural runoff or in wastewaters draining such tions by GC methods such as those listed in 6420C (phenols),
areas; and PCBs (also see Section 6431A). Many of the listed 6440B and C (polynuclear aromatic hydrocarbons), and 6630C
compounds are toxic or carcinogenic. However, they generally and D (organochlorine pesticides and PCBs). In some cases,
are relatively insoluble in water so they do not occur frequently
notably the pesticides, the GC method is substantially more
in potable waters or most wastewaters.
sensitive than the GC/MS method. In other cases, such as the
phenols, there is less difference between the methods.
* Approved by Standard Methods Committee, 2000.
6-66 INDIVIDUAL ORGANIC COMPOUNDS (6000)

6410 B. Liquid-Liquid Extraction Gas Chromatographic/Mass Spectrometric Method

This method1 is applicable to the determination of organic Matrix interferences may be caused by coextracted contami-
compounds that are partitioned into an organic solvent and are nants. The extent of matrix interferences will vary considerably
amenable to gas chromatography,* in municipal and industrial depending on the sample.
discharges. 2) Special precautions—Benzidine can be lost by oxidation
during solvent concentration. Under the alkaline conditions of
the extraction step, ␣-BHC, ␥-BHC, endosulfan I and II, and
1. General Discussion endrin are subject to decomposition. Hexachlorocyclopentadiene
is subject to thermal decomposition in the inlet of the gas
a. Principle: A measured volume of sample is extracted seri- chromatograph, chemical reaction in acetone solution, and pho-
ally with methylene chloride at a pH above 11 and again at a pH tochemical decomposition. N-nitrosodimethylamine is difficult
below 2. The extract is dried, concentrated, and analyzed by to separate from the solvent under the chromatographic condi-
GC/MS.2,3 Qualitative compound identification is based on re- tions described. N-nitrosodiphenylamine decomposes in the gas
tention time and relative abundance of three characteristic chromatographic inlet and cannot be separated from diphe-
masses (m/z). Quantitative analysis uses internal-standard tech- nylamine. Other methods may be preferred for these com-
niques with a single characteristic m/z. pounds.1
b. Interferences: The base-neutral extraction may cause significantly reduced
1) General precautions—See Section 6010C. Method interfer- recovery of phenol, 2-methylphenol, and 2,4-dimethylphenol.
ences may be caused by contaminants in solvents, reagents, Results obtained under these conditions are minimum concen-
glassware, and other sample-processing hardware that lead to trations.
discrete artifacts and/or elevated base lines in detector output. The packed gas chromatographic columns recommended for
Routinely demonstrate that all materials are free from interfer- the basic fraction may not be able to resolve certain isomeric
ences under the conditions of the analysis by running laboratory pairs including the following: anthracene and phenanthrene;
reagent blanks as described in Section 6200A.5c3). chrysene and benzo(a)anthracene; and benzo(b)fluoranthene and
Clean all glassware thoroughly4 as soon as possible after use benzo(k)fluoranthene because retention time and mass spectra
by rinsing with the last solvent used in it, followed by detergent for these pairs are not sufficiently different to make unambigu-
washing with hot water and rinsing with tap water and distilled ous identification possible. Use alternative techniques, such as
the method for polynuclear aromatic hydrocarbons (Section
water. Drain glassware dry and heat in a muffle furnace at 400°C
6440B), to identify and quantify these compounds.
for 15 to 30 min. Some thermally stable materials, such as
In samples containing many interferences, use chemical ion-
PCBs, may not be eliminated by this treatment. Solvent rinses
ization (CI) mass spectrometry to make identification easier.
with acetone and pesticide-quality hexane may be substituted
Tables 6410:I and II give characteristic CI ions for most com-
for the baking. Thorough rinsing with such solvents usually
pounds covered by this method. Use of CI mass spectrometry to
eliminates PCB interference. Do not heat volumetric ware in
support electron ionization (EI) mass spectrometry is encouraged
a muffle furnace. After drying and cooling, seal and store but not required.
glassware in a clean environment to prevent accumulation of c. Detection levels: The method detection level (MDL) is the
dust or other contaminants. Store inverted or capped with minimum concentration of a substance that can be measured and
aluminum foil. reported with 99% confidence that the value is above zero.5 The
Use high-purity reagents and solvents to minimize interfer- MDL concentrations listed in Tables 6410:I and II were obtained
ence. Purification of solvents by distillation in all-glass systems with reagent water.6 The MDL actually obtained in a given
may be required. analysis will vary, depending on instrument sensitivity and ma-
trix effects.
d. Safety: The toxicity or carcinogenicity of each reagent has
* Base/neutral extractables: acenaphthene, acenaphthylene, anthracene, aldrin, not been defined precisely. Benzo(a)anthracene, benzidine, 3,3⬘-
benzo(a)anthracene, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo-
(a)pyrene, benzo(ghi)perylene, benzyl butyl phthalate, ␤-BHC, ␦-BHC, bis(2- dichlorobenzidine, benzo(a)pyrene, ␣-BHC, ␤-BHC, ␦-BHC,
chloroethyl) ether, bis(2-chloroethoxy) methane, bis(2-ethylhexyl) phthalate, ␥-BHC, dibenzo(a,h)anthracene, n-nitrosodimethylamine, 4,4⬘-
bis(2-chloroisopropyl) ether more correctly known as 2,2-oxybis (1-chloropro- DDT, and polychlorinated biphenyls (PCBs) have been tenta-
pane), 4-bromophenyl phenyl ether, chlordane, 2-chloronaphthalene, 4-chlorophe-
nyl phenyl ether, chrysene, 4,4⬘-DDD, 4,4⬘-DDE, 4,4⬘-DDT, dibenzo(a,h)anthra- tively classified as known or suspected, human or mammalian
cene, di-n-butylphthalate, 1,3-dichlorobenzene, 1,2-dichlorobenzene, 1,4-dichlo- carcinogens. Prepare primary standards of these compounds in a
robenzene, 3,3⬘-dichlorobenzidine, dieldrin, diethyl phthalate, dimethyl phthalate,
2,4-dinitrotoluene, 2,6-dinitrotoluene, di-n-octylphthalate, endosulfan sulfate, en-
hood and wear a NIOSH/MESA-approved toxic gas respirator
drin aldehyde, fluoranthene, fluorene, heptachlor, heptachlor epoxide, hexachlo- when handling high concentrations.
robenzene, hexachlorobutadiene, hexachloroethane, indeno(1,2,3-cd)pyrene, iso-
phorone, naphthalene, nitrobenzene, N-nitrosodi-n-propylamine, PCB-1016,
PCB-1221, PCB-1232, PCB-1242, PCB-1248, PCB-1254, PCB-1260, phenan- 2. Sampling and Storage
threne, pyrene, toxaphene, 1,2,4-trichlorobenzene.
Acid extractables: 4-chloro-3-methylphenol, 2-chlorophenol, 2,4-dichlorophe-
nol, 2,4-dimethylphenol, 2,4-dinitrophenol, 2-methyl-4,6-dinitrophenol, 2-nitro- Collect grab samples in 1-L amber glass bottles fitted with a
phenol, 4-nitrophenol, pentachlorophenol, phenol, 2,4,6-trichlorophenol. screw cap lined with TFE. Foil may be substituted for TFE if the
The method may be extended to include the following compounds: benzidine,
␣-BHC, ␥-BHC, endosulfan I, endosulfan II, endrin, hexachlorocyclopentadiene, sample is not corrosive. If amber bottles are not available,
N-nitrosodimethylamine, N-nitrosodiphenylamine. protect samples from light. Wash and rinse bottle and cap liner
BASE/NEUTRALS AND ACIDS (6410)/Liquid-Liquid Extraction GC/MS Method 6-67

with acetone or methylene chloride, and dry before use. Follow 3. Apparatus
conventional sampling practices7 but do not rinse bottle with
sample. Collect composite samples in refrigerated glass contain- a. Separatory funnel, 2-L, with TFE stopcock.
ers. Optionally, use automatic sampling equipment as free as b. Drying column, chromatographic, 400 mm long ⫻ 19 mm
possible of plastic tubing and other potential sources of contam- ID, with coarse frit filter disk.
ination; incorporate glass sample containers for collecting a c. Concentrator tube, Kuderna-Danish, 10-mL, graduated.†
minimum of 250 mL. Refrigerate sample containers at 4°C and Check calibration at volumes used. Use ground-glass stopper to
protect from light during compositing. If the sampler includes a prevent evaporation.
peristaltic pump, use a minimum length of compressible silicone d. Evaporative flask, Kuderna-Danish, 500-mL.‡ Attach to
rubber tubing, but before use, thoroughly rinse it with methanol concentrator tube with springs.
and rinse repeatedly with distilled water to minimize contami- e. Snyder column, Kuderna-Danish, three-ball macro.§
nation. Use an integrating flow meter to collect flow-proportional
f. Snyder column, Kuderna-Danish, two-ball micro.㛳
composites.
g. Vials, 10- to 15-mL, amber glass, with TFE-lined screw cap.
Fill sample bottles and, if residual chlorine is present, add 80
mg sodium thiosulfate per liter of sample and mix well. Ice all
samples or refrigerate at 4°C from time of collection until
extraction. † Kontes K-570050-1025 or equivalent.
‡ Kontes K-570001-0500 or equivalent.
Extract samples within 7 d of collection and analyze com- § Kontes K-503000-0121 or equivalent.
pletely within 40 d of extraction. 㛳 Kontes K-569001-0219 or equivalent.

TABLE 6410:I. CHROMATOGRAPHIC CONDITIONS, METHOD DETECTION LEVELS, AND CHARACTERISTIC MASSES FOR BASE/NEUTRAL EXTRACTABLES

Method Characteristic Masses

Retention Detection
Electron Impact Chemical Ionization
Time Level
Compound min ␮g/L Primary Secondary Secondary Methane Methane Methane

1,3-Dichlorobenzene 7.4 1.9 146 148 113 146 148 150

1,4-Dichlorobenzene 7.8 4.4 146 148 113 146 148 150
Hexachloroethane 8.4 1.6 117 201 199 199 201 203
bis(2-Chloroethyl) ether 8.4 5.7 93 63 95 63 107 109
1,2-Dichlorobenzene 8.4 1.9 146 148 113 146 148 150
bis(2-Chloroisopropyl) ether* 9.3 5.7 45 77 79 77 135 137
N-Nitrosodi-n-propylamine 130 42 101
Nitrobenzene 11.1 1.9 77 123 65 124 152 164
Hexachlorobutadiene 11.4 0.9 225 223 227 223 225 227
1,2,4-Trichlorobenzene 11.6 1.9 180 182 145 181 183 209
Isophorone 11.9 2.2 82 95 138 139 167 178
Naphthalene 12.1 1.6 128 129 127 129 157 169
bis(2-Chloroethoxy) methane 12.2 5.3 93 95 123 65 107 137
Hexachlorocyclopentadiene† 13.9 237 235 272 235 237 239
2-Chloronaphthalene 15.9 1.9 162 164 127 163 191 203
Acenaphthylene 17.4 3.5 152 151 153 152 153 181
Acenaphthene 17.8 1.9 154 153 152 154 155 183
Dimethyl phthalate 18.3 1.6 163 194 164 151 163 164
2,6-Dinitrotoluene 18.7 1.9 165 89 121 183 211 223
Fluorene 19.5 1.9 166 165 167 166 167 195
4-Chlorophenyl phenyl ether 19.5 4.2 204 206 141
2,4-Dinitrotoluene 19.8 5.7 165 63 182 183 211 223
Diethyl phthalate 20.1 1.9 149 177 150 177 223 251
N-Nitrosodiphenylamine† 20.5 1.9 169 168 167 169 170 198
Hexachlorobenzene 21.0 1.9 284 142 249 284 286 288
␣-BHC† 21.1 183 181 109
4-Bromophenyl phenyl ether 21.2 1.9 248 250 141 249 251 277
␥-BHC† 22.4 183 181 109
Phenanthrene 22.8 5.4 178 179 176 178 179 207
Anthracene 22.8 1.9 178 179 176 178 179 207
␤-BHC 23.4 4.2 181 183 109
Heptachlor 23.4 1.9 100 272 274
␦-BHC 23.7 3.1 183 109 181
Aldrin 24.0 1.9 66 263 220
Dibutyl phthalate 24.7 2.5 149 150 104 149 205 279
6-68 INDIVIDUAL ORGANIC COMPOUNDS (6000)

TABLE 6410:I. CONT.

Method Characteristic Masses
Retention Detection
Electron Impact Chemical Ionization
Time Level
Compound min ␮g/L Primary Secondary Secondary Methane Methane Methane

Heptachlor epoxide 25.6 2.2 353 355 351

Endosulfan I† 26.4 237 338 341
Fluoranthene 26.5 2.2 202 101 100 203 231 243
Dieldrin 27.2 2.5 79 263 279
4,4⬘-DDE 27.2 5.6 246 248 176
Pyrene 27.3 1.9 202 101 100 203 231 243
Endrin† 27.9 81 263 82
Endosulfan II† 28.6 237 339 341
4,4⬘-DDD 28.6 2.8 235 237 165
Benzidine† 28.8 44 184 92 185 185 213 225
4,4⬘-DDT 29.3 4.7 235 237 165
Endosulfan sulfate 29.8 5.6 272 387 422
Endrin aldehyde 67 345 250
Butyl benzyl phthalate 29.9 2.5 149 91 206 149 299 327
bis(2-Ethylhexyl) phthalate 30.6 2.5 149 167 279 149
Chrysene 31.5 2.5 228 226 229 228 229 257
Benzo(a)anthracene 31.5 7.8 228 229 226 228 229 257
3,3⬘-Dichlorobenzidine 32.2 16.5 252 254 126
Di-n-octyl phthalate 32.5 2.5 149
Benzo(b)fluoranthene 34.9 4.8 252 253 125 252 253 281
Benzo(k)fluoranthene 34.9 2.5 252 253 125 252 253 281
Benzo(a)pyrene 36.4 2.5 252 253 125 252 253 281
Indeno(1,2,3-cd)pyrene 42.7 3.7 276 138 277 276 277 305
Dibenzo(a,h)anthracene 43.2 2.5 278 139 279 278 279 307
Benzo(ghi)perylene 45.1 4.1 276 138 277 276 277 305
N-Nitrosodimethylamine† 42 74 44
Chlordane‡ 19–30 373 375 377
Toxaphene‡ 25–34 159 231 233
PCB 1016‡ 18–30 224 260 294
PCB 1221‡ 15–30 30 190 224 260
PCB 1232‡ 15–32 190 224 260
PCB 1242‡ 15–32 224 260 294
PCB 1248‡ 12–34 294 330 262
PCB 1254‡ 22–34 36 294 330 362
PCB 1260‡ 23–32 330 362 394
*The proper chemical name is 2,2⬘-oxybis(1-chloropropane).
†See introductory section of text.
‡These compounds are mixtures of various isomers. (See Figures 6410:2 through 12.)
Column conditions: Supelcoport (100/120 mesh) coated with 3% SP-2250 packed in a 1.8 m long ⫻ 2 mm ID glass column with helium carrier gas at 30 mL/min flow
rate. Column temperature held isothermal at 50°C for 4 min, then programmed at 8°C/min to 270°C and held for 30 min.

h. Continuous liquid-liquid extractor, equipped with TFE or accessories including syringes, analytical columns, and gases.
glass connecting joints and stopcocks requiring no lubrication.# Use chromatograph with the injection port designed for on-
i. Boiling chips, approximately 10/40 mesh. Heat to 400°C for column injection when packed columns are used and for splitless
30 min or extract in a Soxhlet extractor with methylene chloride. injection when capillary columns are used.
j. Water bath, heated, with concentric ring cover and temper- 1) Column for base/neutrals, 1.8 m long ⫻ 2-mm ID glass,
ature control to ⫾2°C. Use bath in a hood. packed with 3% SP-2250 on Supelcoport (100/200 mesh) or
k. Balance, analytical, capable of accurately weighing 0.0001 g. equivalent. This column was used to develop the detection level
l. Gas chromatograph:** An analytical system complete with and precision and bias data presented herein. Guidelines for the
a temperature-programmable gas chromatograph and all required use of alternate columns (e.g., DB-5 fused silica capillary) are
provided in ¶ 5b.
# Hershberg-Wolf Extractor, Ace Glass Co., Vineland, NJ, P/N 6841-10, or
2) Column for acids, 1.8 m long ⫻ 2-mm ID glass, packed
equivalent. with 1% SP-1240DA on Supelcoport (100/120 mesh) or equiv-
** Gas chromatographic methods are extremely sensitive to the materials used. alent. The detection level and precision and bias data presented
Mention of trade names by Standard Methods does not preclude the use of other
existing or as-yet-undeveloped products that give demonstrably equivalent results. herein were developed with this column. For guidelines for the
BASE/NEUTRALS AND ACIDS (6410)/Liquid-Liquid Extraction GC/MS Method 6-69

TABLE 6410:II. CHROMATOGRAPHIC CONDITIONS, METHOD DETECTION LEVELS, AND CHARACTERISTIC MASSES FOR ACID EXTRACTABLES
Method Characteristic Masses
Detection
Electron Impact Chemical Ionization
Retention Time Level
Compound min ␮g/L Primary Secondary Secondary Methane Methane Methane

2-Chlorophenol 5.9 3.3 128 64 130 129 131 157

2-Nitrophenol 6.5 3.6 139 65 109 140 168 122
Phenol 8.0 1.5 94 65 66 95 123 135
2,4-Dimethylphenol 9.4 2.7 122 107 121 123 151 163
2,4-Dichlorophenol 9.8 2.7 162 164 98 163 165 167
2,4,6-Trichlorophenol 11.8 2.7 196 198 200 197 199 201
4-Chloro-3-methylphenol 13.2 3.0 142 107 144 143 171 183
2,4-Dinitrophenol 15.9 42 184 63 154 185 213 225
2-Methyl-4,6-dinitrophenol 16.2 24 198 182 77 199 227 239
Pentachlorophenol 17.5 3.6 266 264 268 267 265 269
4-Nitrophenol 20.3 2.4 65 139 109 140 168 122
Column conditions: Supelcoport (100/120 mesh) coated with 1% SP-1240DA packed in a 1.8-m-long ⫻ 2-mm-ID glass column with helium carrier gas at 30 mL/min
flow rate. Column temperature held isothermal at 70°C for 2 min then programmed at 8°C/min to 200°C.

use of alternate columns (e.g., DB-5 fused silica capillary) see d. Sodium thiosulfate, Na2S2O3 䡠 5H2O, granular.
¶ 5b. e. Sulfuric acid, H2SO4, 1 ⫹ 1: Slowly add 50 mL conc H2SO4
m. Mass spectrometer, capable of scanning from 35 to 450 to 50 mL reagent water.
amu every 7 s or less, utilizing 70-V (nominal) electron energy f. Acetone, methanol, methylene chloride, pesticide quality or
in the electron impact ionization mode, and producing a mass equivalent.
spectrum that meets all the criteria in Table 6410:III when 50 ng g. Stock standard solutions: Prepare from pure standard ma-
of decafluorotriphenyl phosphine [DFTPP; bis(perfluorophenyl) terials or purchase as certified solutions. Prepare by accurately
phenyl phosphine] is injected through the GC inlet. weighing about 0.0100 g of pure material, dissolve in pesticide-
n. GC/MS interface: Any GC to MS interface that gives quality acetone or other suitable solvent, and dilute to volume in
acceptable calibration points at 50 ng or less per injection for a 10-mL volumetric flask; 1 ␮L ⫽ 1.00 ␮g compound. When
each of the compounds of interest and achieves all acceptable compound purity is assayed to be 96% or greater, use the weight
performance criteria may be used. GC to MS interfaces con- without correction to calculate concentration of the stock stand-
structed of all glass or glass-lined materials are recommended. ard. Use commercially prepared stock standards at any concen-
Glass can be deactivated by silanizing with dichlorodimethylsilane. tration if certified by the manufacturer or by an independent
o. Data system: See Section 6200B.2f. source.
Transfer stock standard solutions into TFE-sealed screw-cap
4. Reagents bottles. Store at 4°C and protect from light. Check stock standard
solutions frequently for signs of degradation or evaporation,
a. Reagent water: See Section 6200B.3a. especially just before preparing calibration standards. Replace
b. Sodium hydroxide solution, NaOH, 10N: Dissolve 40 g stock standard solutions after 6 months, or sooner if comparison
NaOH in reagent water and dilute to 100 mL. with check standards indicates a problem.
c. Sodium sulfate, Na2SO4, granular, anhydrous. Purify by h. Surrogate standard known-addition solution: Select a min-
heating at 400°C for 4 h in a shallow tray. imum of three surrogate compounds from Table 6410:IV. Pre-
pare a surrogate standard solution containing each selected sur-
rogate compound at a concentration of 100 ␮g/mL in acetone.
TABLE 6410:III. DFTPP KEY MASSES AND ABUNDANCE CRITERIA
Adding 1.00 mL to 1000 mL sample is equivalent to a concen-
Mass m/z Abundance Criteria tration of 100 ␮g/L of each surrogate standard. Store at 4°C in
TFE-sealed glass container. Check solution frequently for sta-
51 30–60% of mass 198 bility. Replace solution after 6 months, or sooner if comparison
68 Less than 2% of mass 69
with quality-control check standards indicates a problem.
70 Less than 2% of mass 69
127 40–60% of mass 198 i. DFTPP standard: Prepare a 25-␮g/mL solution of DFTPP
197 Less than 1% of mass 198 in acetone.
198 Base peak, 100% relative abundance j. Calibration standards: Prepare calibration standards at a
199 5–9% of mass 198 minimum of three concentration levels for each compound by
275 10–30% of mass 198 adding appropriate volumes of one or more stock standards to a
365 Greater than 1% of mass 198 volumetric flask. To each calibration standard or standard mix-
441 Present but less than mass 443 ture, add a known constant amount of one or more internal
442 Greater than 40% of mass 198 standards (such as those listed in Table 6410:IV), and dilute to
443 17–23% of mass 442
volume with acetone. Prepare one calibration standard at a con-
6-70 INDIVIDUAL ORGANIC COMPOUNDS (6000)

TABLE 6410:IV. SUGGESTED INTERNAL AND SURROGATE STANDARDS centration near, but above, the MDL and others corresponding to
the expected range of sample concentrations or defining the
Base/Neutral Fraction Acid Fraction
working range of the GC/MS system.
Aniline-d5 2-Fluorophenol k. Quality control (QC) check sample concentrate: Obtain a
Anthracene-d10 Pentafluorophenol check sample concentrate containing each compound at a con-
Benzo(a)anthracene-d12 Phenol-d5 centration of 100 ␮g/mL in acetone. Multiple solutions may be
4,4⬘-Dibromobiphenyl 2-Perfluoromethyl phenol required. PCBs and multicomponent pesticides may be omitted.
4,4⬘-Dibromooctafluorobiphenyl If such a sample is not available from an external source, prepare
Decafluorobiphenyl
using stock standards prepared independently from those used
2,2⬘-Difluorobiphenyl
4-Fluoroaniline for calibration.
1-Fluoronaphthylene
2-Fluoronaphthylene 5. Procedure
Naphthalene-d8
Nitrobenzene-d5 a. Extraction: Extraction by means of a separatory funnel,
2,3,4,5,6-Pentafluorobiphenyl ¶ 1), is most common, but if emulsions will prevent acceptable
Phenanthrene-d10 solvent recovery, use continuous extraction, ¶ 2).
Pyridine-d5 1) Separatory funnel extraction—Normally use a sample vol-
ume of 1 L. For sample volumes of 2 L, use 250-, 100-, and
100-mL volumes of methylene chloride for the serial extraction
of the base/neutrals and 200-, 100-, and 100-mL volumes of
methylene chloride for the acids.

Figure 6410:1. Gas chromatogram of base/neutral fraction. Column: 3% SP-2250 on Supelcoport; program: 50°C for 4 min, 8°C/min to 270°C; detector: mass
spectrometer.
BASE/NEUTRALS AND ACIDS (6410)/Liquid-Liquid Extraction GC/MS Method 6-71

Figure 6410:2. Gas chromatogram of acid fraction. Column: 1% SP-1240DA on Supelcoport; program: 70°C for 2 min, 8°C/min to 200°C; detector: mass
spectrometer.

Mark water meniscus on side of sample bottle for later deter- ratory funnel and extract sample by shaking for 2 min with
mination of sample volume. Pour entire sample into a 2-L periodic venting to release excess pressure. Let organic layer
separatory funnel. Pipet 1.00 mL surrogate standard solution into separate from water phase for a minimum of 10 min. If emulsion
separatory funnel and mix well. Check pH with wide-range pH interface between layers is more than one-third the volume of the
paper and adjust to pH ⬎ 11 with NaOH solution.
Add 60 mL methylene chloride to sample bottle, seal, and
shake for 30 s to rinse inner surface. Transfer solvent to sepa-

Figure 6410:3. Gas chromatogram of pesticide fraction. Column: 3% Figure 6410:4. Gas chromatogram of chlordane. Column: 3% SP-2250 on
SP-2250 on Supelcoport; program: 50°C for 4 min, 8°C/min Supelcoport; program: 50°C for 4 min, 8°C/min to 270°C;
to 270°C; detector: mass spectrometer. detector: mass spectrometer.
6-72 INDIVIDUAL ORGANIC COMPOUNDS (6000)

Figure 6410:7. Gas chromatogram of PCB-1221. Column: 3% SP-2250 on

Supelcoport; program: 50°C for 4 min, 8°C/min to 270°C;
detector: mass spectrometer.

Figure 6410:5. Gas chromatogram of toxaphene. Column: 3% SP-2250 on

Supelcoport; program: 50°C for 4 min, 8°C/min to 270°C;
detector: mass spectrometer.

Figure 6410:6. Gas chromatogram of PCB-1016. Column: 3% SP-2250 on Figure 6410:8. Gas chromatogram of PCB-1232. Column: 3% SP-2250 on
Supelcoport; program: 50°C for 4 min, 8°C/min to 270°C; Supelcoport; program: 50°C for 4 min, 8°C/min to 270°C;
detector: mass spectrometer. detector: mass spectrometer.
BASE/NEUTRALS AND ACIDS (6410)/Liquid-Liquid Extraction GC/MS Method 6-73

Figure 6410:11. Gas chromatogram of PCB-1254. Column: 3% SP-2250

on Supelcoport; program: 50°C for 4 min, 8°C/min to
270°C; detector: mass spectrometer.

Figure 6410:9. Gas chromatogram of PCB-1242. Column: 3% SP-2250 on

Supelcoport; program: 50°C for 4 min, 8°C/min to 270°C;
detector: mass spectrometer.

Figure 6410:10. Gas chromatogram of PCB-1248. Column: 3% SP-2250 Figure 6410:12. Gas chromatogram of PCB-1260. Column: 3% SP-2250
on Supelcoport; program: 50°C for 4 min, 8°C/min to on Supelcoport; program: 50°C for 4 min, 8°C/min to
270°C; detector: mass spectrometer. 270°C; detector: mass spectrometer.
6-74 INDIVIDUAL ORGANIC COMPOUNDS (6000)

solvent layer, use mechanical techniques to complete phase

separation. The optimum technique depends on the sample, but
may include stirring, filtering emulsion through glass wool,
centrifuging, or other physical methods. Collect methylene chlo-
ride extract in a 250-mL erlenmeyer flask.
If the emulsion cannot be broken (recovery of less than 80%
of the methylene chloride, corrected for water solubility of
methylene chloride) in the first extraction, transfer sample, sol-
vent, and emulsion into extraction chamber of a continuous
extractor and proceed as described in ¶ 2) below.
Add a second 60-mL volume of methylene chloride to sample
bottle and repeat extraction procedure, combining extracts in the
erlenmeyer flask. Perform a third extraction in the same manner.
After the third extraction, adjust pH of aqueous phase to ⬍2
using H2SO4. Serially extract acidified aqueous phase three
times with 60-mL portions of methylene chloride. Collect and
combine extracts in a 250-mL erlenmeyer flask and label com-
bined extracts as the acid fraction.
For each fraction, assemble a Kuderna-Danish (K-D) concen-
trator by attaching a 10-mL concentrator tube to a 500-mL
evaporative flask. Other concentration devices or techniques may
be used if the requirements of ¶ 7 are met.
Pour combined extract through a solvent-rinsed drying column
containing at least 10 cm anhydrous Na2SO4 or more and collect
extract in concentrator. Rinse erlenmeyer flask and column with
20 to 30 mL methylene chloride to complete transfer.
Add one or two clean boiling chips to the evaporative flask
and attach a three-ball Snyder column. Prewet Snyder column by
adding about 1 mL methylene chloride to the top. Place K-D
apparatus on a hot water bath (60 to 65°C) in a hood so that
concentrator tube is partially immersed in the hot water and
entire lower rounded surface of flask is bathed with hot vapor.
Adjust vertical position of apparatus and water temperature as
required to complete concentration in 15 to 20 min. At proper
Figure 6410:13. Tailing factor calculation. Example calculation: rate of distillation the column balls actively chatter but the
Peak height ⫽ DE ⫽ 100 mm 10% peak height ⫽ BD ⫽ 10 mm chambers are not flooded with condensed solvent. When the
Peak width at 10% peak height ⫽ AC ⫽ 23 mm apparent volume of liquid reaches 1 mL, remove K-D apparatus
AB ⫽ 11 mm BC ⫽ 12 mm and let drain and cool for at least 10 min.
12 Remove Snyder column and rinse flask and its lower joint into
Tailing factor ⫽ ⫽ 1.1
11 the concentrator tube with 1 to 2 mL methylene chloride, pref-
erably using a 5-mL syringe.
Add another one or two clean boiling chips to concentrator
tube for each fraction and attach a two-ball micro-Snyder col-

TABLE 6410:V. QC ACCEPTANCE CRITERIA*

Test Limits Range Range
Concentration for s for X̄ for P, Ps
Compound ␮g/L ␮g/L ␮g/L %

Acenaphthene 100 27.6 60.1–132.3 47–145

Acenaphthylene 100 40.2 53.5–126.0 33–145
Aldrin 100 39.0 7.2–152.2 D–166
Anthracene 100 32.0 43.4–118.0 27–133
Benzo(a)anthracene 100 27.6 41.8–133.0 33–143
Benzo(b)fluoranthene 100 38.8 42.0–140.4 24–159
Benzo(k)fluoranthene 100 32.3 25.2–145.7 11–162
Benzo(a)pyrene 100 39.0 31.7–148.0 17–163
Benzo(ghi)perylene 100 58.9 D–195.0 D–219
Benzyl butyl phthalate 100 23.4 D–139.9 D–152
BASE/NEUTRALS AND ACIDS (6410)/Liquid-Liquid Extraction GC/MS Method 6-75

TABLE 6410:V. CONT.

Test Limits Range Range
Concentration for s ៮
for X for P, Ps
Compound ␮g/L ␮g/L ␮g/L %

␦-BHC 100 31.5 41.5–130.6 24–149

␤-BHC 100 21.6 D–100.0 D–110
bis(2-Chloroethyl) ether 100 55.0 42.9–126.0 12–158
bis(2-Chloroethoxy) methane 100 34.5 49.2–164.7 33–184
bis(2-Chloroisopropyl) ether† 100 46.3 62.8–138.6 36–166
bis(2-Ethylhexyl) phthalate 100 41.1 28.9–136.8 8–158
4-Bromophenyl phenyl ether 100 23.0 64.9–114.4 53–127
2-Chloronaphthalene 100 13.0 64.5–113.5 60–118
4-Chlorophenyl phenyl ether 100 33.4 38.4–144.7 25–158
Chrysene 100 48.3 44.1–139.9 17–168
4,4⬘-DDD 100 31.0 D–134.5 D–145
4,4⬘-DDE 100 32.0 19.2–119.7 4–136
4,4⬘-DDT 100 61.6 D–170.6 D–203
Dibenzo(a,h)anthracene 100 70.0 D–199.7 D–227
Di-n-butyl phthalate 100 16.7 8.4–111.0 1–118
1,2-Dichlorobenzene 100 30.9 48.6–112.0 32–129
1,3-Dichlorobenzene 100 41.7 16.7–153.9 D–172
1,4-Dichlorobenzene 100 32.1 37.3–105.7 20–124
3,3⬘-Dichlorobenzidine 100 71.4 8.2–212.5 D–262
Dieldrin 100 30.7 44.3–119.3 29–136
Diethyl phthalate 100 26.5 D–100.0 D–114
Dimethyl phthalate 100 23.2 D–100.0 D–112
2,4-Dinitrotoluene 100 21.8 47.5–126.9 39–139
2,6-Dinitrotoluene 100 29.6 68.1–136.7 50–158
Di-n-octylphthalate 100 31.4 18.6–131.8 4–146
Endosulfan sulfate 100 16.7 D–103.5 D–107
Endrin aldehyde 100 32.5 D–188.8 D–209
Fluoranthene 100 32.8 42.9–121.3 26–137
Fluorene 100 20.7 71.6–108.4 59–121
Heptachlor 100 37.2 D–172.2 D–192
Heptachlor epoxide 100 54.7 70.9–109.4 26–155
Hexachlorobenzene 100 24.9 7.8–141.5 D–152
Hexachlorobutadiene 100 26.3 37.8–102.2 24–116
Hexachloroethane 100 24.5 55.2–100.0 40–113
Indeno(1,2,3-cd)pyrene 100 44.6 D–150.9 D–171
Isophorone 100 63.3 46.6–180.2 21–196
Naphthalene 100 30.1 35.6–119.6 21–133
Nitrobenzene 100 39.3 54.3–157.6 35–180
N-Nitrosodi-n-propylamine 100 55.4 13.6–197.9 D–230
PCB-1260 100 54.2 19.3–121.0 D–164
Phenanthrene 100 20.6 65.2–108.7 54–120
Pyrene 100 25.2 69.6–100.0 52–115
1,2,4-Trichlorobenzene 100 28.1 57.3–129.2 44–142
4-Chloro-3-methylphenol 100 37.2 40.8–127.9 22–147
2-Chlorophenol 100 28.7 36.2–120.4 23–134
2,4-Dichlorophenol 100 26.4 52.5–121.7 39–135
2,4-Dimethylphenol 100 26.1 41.8–109.0 32–119
2,4-Dinitrophenol 100 49.8 D–172.9 D–191
2-Methyl-4,6-dinitrophenol 100 93.2 53.0–100.0 D–181
2-Nitrophenol 100 35.2 45.0–166.7 29–182
4-Nitrophenol 100 47.2 13.0–106.5 D–132
Pentachlorophenol 100 48.9 38.1–151.8 14–176
Phenol 100 22.6 16.6–100.0 5–112
2,4,6-Trichlorophenol 100 31.7 52.4–129.2 37–144

*s ⫽ standard deviation for four recovery measurements,

X̄ ⫽ average recovery for four recovery measurements,
P, Ps ⫽ percent recovery measured, and
D ⫽ detected; results must be greater than zero.
† The proper chemical name is 2,2⬘-oxybis(1-chloropropane).
NOTE: These criteria are based directly upon the method performance data in Table 6410:VI. Where necessary, the limits for recovery were broadened to assure applicability
of the limits to concentrations below those used to develop Table 6410:VI.
6-76 INDIVIDUAL ORGANIC COMPOUNDS (6000)

umn. Prewet Snyder column by adding about 0.5 mL of meth- spectrometer and repeat test until all criteria are achieved. Meet
ylene chloride to the top. Place K-D apparatus on a hot-water performance criteria before any samples, blanks, or standards are
bath (60 to 65°C) so that concentrator tube is partially immersed analyzed. The tailing factor tests in ¶s 2) and 3) may be per-
in hot water and continue concentrating as directed above with- formed simultaneously with the DFTPP test.
out further solvent addition until apparent volume of liquid 2) Column performance test for base/neutrals—At beginning
reaches about 0.5 mL. After cooling, remove Snyder column and of each day that base/neutral fraction is to be analyzed for
rinse flask and its lower joint into the concentrator tube with benzidine, calculate benzidine tailing factor. Inject 100 ng ben-
approximately 0.2 mL acetone or methylene chloride. Adjust zidine either separately or as a part of a standard mixture that
final volume to 1.0 mL with solvent. Stopper concentrator tube may contain DFTPP, and calculate tailing factor, which must be
and store refrigerated if further processing will not be done less than 3.0. Calculation of the tailing factor is illustrated in
immediately. If extract is to be stored longer than 2 d, transfer to Figure 6410:13.9 Replace column packing if tailing factor crite-
a TFE-sealed screw-cap vial and label base/neutral or acid frac- rion cannot be met.
tion as appropriate. 3) Column performance test for acids—At beginning of each
Determine original sample volume by refilling sample bottle day that acids are to be determined, inject 50 ng pentachloro-
to mark and transferring liquid to a 1000-mL graduated cylinder. phenol either separately or as a part of a standard mix that may
Record sample volume to nearest 5 mL. contain DFTPP. The tailing factor for pentachlorophenol must be
2) Continuous extraction—Mark water meniscus on side of less than 5. Calculation of the tailing factor is illustrated in
sample bottle, and determine sample volume later as described in Figure 6410:13.9 Replace column packing if tailing factor crite-
¶ 1). Check pH with wide-range pH paper and adjust to pH ⬎ 11 rion cannot be met.
with NaOH solution. Transfer sample to continuous extractor d. Calibration of GC/MS system: Calibrate system daily after
and, using a pipet, add 1.00 mL surrogate standard solution and performance tests.
mix well. Add 60 mL methylene chloride to sample bottle, seal, Select three or more internal standards similar in analytical
and shake for 30 s to rinse inner surface. Transfer solvent to behavior to the compounds of interest. Demonstrate that the
extractor. Repeat rinse with an additional 50- to 100-mL portion measurement of the internal standards is not affected by method
methylene chloride and add rinse to extractor. or matrix interferences. Some recommended internal standards
Add 200 to 500 mL methylene chloride to distilling flask, add are listed in Table 6410:IV. Use base peak m/z as the primary m/z
sufficient reagent water to ensure proper operation, and extract for quantification. If interferences are noted, use one of the next
for 24 h. Let cool and detach distilling flask. Dry, concentrate, two most intense m/z quantities for quantification. Using injec-
and seal extract as in ¶ 1) above. tions of 2 to 5 ␮L, analyze each calibration standard according
Charge a clean distilling flask with 500 mL methylene chlo- to ¶ e below and tabulate area of primary characteristic m/z
ride and attach it to continuous extractor. Carefully, while stir- (Tables 6410:I and II) against concentration for each compound
ring, adjust pH of aqueous phase to less than 2 with H2SO4. and internal standard. Calculate response factors (RF) for each
Extract for 24 h. Dry, concentrate, and seal extract as in ¶ 1) compound by the equation given in Section 6200B.4c2). If the
above. RF value over the working range is a constant (⬍35% RSD), it
b. GC/MS operating conditions: Table 6410:I summarizes the can be assumed to be invariant; use the average RF for calcula-
recommended gas chromatographic operating conditions for the tions. Alternatively, use the results to plot a calibration curve of
base/neutral fraction and Table 6410:II for the acid fraction. response ratios, As /Ais vs. RF.
Included in these tables are retention times and MDLs that can Verify working calibration curve or RF on each working day
be achieved under these conditions. Examples of the separations by measuring one or more calibration standards. If the response
obtained with these columns are shown in Figures 6410:1 for any compound varies from the predicted response by more
through 12. Other packed or capillary (open-tubular) columns or than 20%, repeat test using a fresh calibration standard. Alter-
chromatographic conditions may be used if the requirements of natively, prepare a new calibration curve for that compound.
¶ 7 are met. e. Sample analysis: Add internal standard to sample extract,
c. GC/MS performance tests: At the beginning of each day on mix thoroughly, and immediately inject 2 to 5 ␮L of sample
which analyses are to be performed, check GC/MS system to see extract or standard into GC/MS system using solvent-flush tech-
if acceptable performance criteria are achieved for DFTPP.8 nique10 to minimize losses due to adsorption, chemical reaction,
Each day that benzidine is to be determined, the tailing factor or evaporation. Smaller (1.0-␮L) volumes may be injected if
criterion described in ¶ 2) must be achieved. Each day that the automatic devices are used. Record volume injected to nearest
acids are to be determined, the tailing factor criterion described 0.05 ␮L. If response for any m/z exceeds the working range of
in ¶ 3) must be achieved. the GC/MS system, dilute extract and reanalyze. Make all qual-
These performance tests have the requirements given in Sec- itative and quantitative measurements as described below and in
tion 6200B.4b, but use following conditions: ¶ 6. When extract is not being used, store at 4°C, protected from
Electron energy: 70 V (nominal) light, in screw-cap vial equipped with unpierced TFE-lined sep-
Mass range: 35 to 450 amu tum.
Scan time: To give at least 5 scans per peak but not to exceed Obtain EICPs for the primary m/z and the two other masses
7 s per scan. listed in Tables 6410:I and II. See ¶ d for masses to be used with
1) DFTPP performance test—At beginning of each day, inject internal and surrogate standards. Use the following criteria to
2 ␮L (50 ng) DFTPP standard solution. Obtain a background- make a qualitative identification:
corrected mass spectrum of DFTPP and confirm that all the key • The characteristic masses of each compound maximize in
m/z criteria in Table 6410:III are achieved. If not, retune mass the same or within one scan of each other.
BASE/NEUTRALS AND ACIDS (6410)/Liquid-Liquid Extraction GC/MS Method 6-77

TABLE 6410:VI. METHOD BIAS AND PRECISION AS FUNCTIONS OF CONCENTRATION*

Bias as Single-Analyst Overall
Recovery, X⬘ Precision, sr⬘ Precision, S⬘
Compound ␮g/L ␮g/L ␮g/L



Acenaphthene 0.96C ⫹ 0.19 0.15X ⫺ 0.12 0.21X ⫺ 0.67



Acenaphthylene 0.89C ⫹ 0.74 0.24X ⫺ 1.06 0.26X ⫺ 0.54



Aldrin 0.78C ⫹ 1.66 0.27X ⫺ 1.28 0.43X ⫹ 1.13



Anthracene 0.80C ⫹ 0.68 0.21X ⫺ 0.32 0.27X ⫺ 0.64



Benzo(a)anthracene 0.88C ⫺ 0.60 0.15X ⫹ 0.93 0.26X ⫺ 0.28



Benzo(b)fluoranthene 0.93C ⫺ 1.80 0.22X ⫹ 0.43 0.29X ⫹ 0.96



Benzo(k)fluoranthene 0.87C ⫺ 1.56 0.19X ⫹ 1.03 0.35X ⫹ 0.40



Benzo(a)pyrene 0.90C ⫺ 0.13 0.22X ⫹ 0.48 0.32X ⫹ 1.35



Benzo(ghi)perylene 0.98C ⫺ 0.86 0.29X ⫹ 2.40 0.51X ⫺ 0.44



Benzyl butyl phthalate 0.66C ⫺ 1.68 0.18X ⫹ 0.94 0.53X ⫹ 0.92



␤-BHC 0.87C ⫺ 0.94 0.20X ⫺ 0.58 0.30X ⫺ 1.94



␦-BHC 0.29C ⫺ 1.09 0.34X ⫹ 0.86 0.93X ⫺ 0.17



bis(2-Chloroethyl) ether 0.86C ⫺ 1.54 0.35X ⫺ 0.99 0.35X ⫹ 0.10



bis(2-Chloroethoxy) methane 1.12C ⫺ 5.04 0.16X ⫹ 1.34 0.26X ⫹ 2.01



bis(2-Chloroisopropyl) ether† 1.03C ⫺ 2.31 0.24X ⫹ 0.28 0.25X ⫹ 1.04



bis(2-Ethylhexyl) phthalate 0.84C ⫺ 1.18 0.26X ⫹ 0.73 0.36X ⫹ 0.67



4-Bromophenyl phenyl ether 0.91C ⫺ 1.34 0.13X ⫹ 0.66 0.16X ⫹ 0.66



2-Chloronaphthalene 0.89C ⫹ 0.01 0.07X ⫹ 0.52 0.13X ⫹ 0.34



4-Chlorophenyl phenyl ether 0.91C ⫹ 0.53 0.20X ⫺ 0.94 0.30X ⫺ 0.46



Chrysene 0.93C ⫺ 1.00 0.28X ⫹ 0.13 0.33X ⫺ 0.09



4,4⬘-DDD 0.56C ⫺ 0.40 0.29X ⫺ 0.32 0.66X ⫺ 0.96



4,4⬘-DDE 0.70C ⫺ 0.54 0.26X ⫺ 1.17 0.39X ⫺ 1.04



4,4⬘-DDT 0.79C ⫺ 3.28 0.42X ⫹ 0.19 0.65X ⫺ 0.58



Dibenzo(a,h)anthracene 0.88C ⫹ 4.72 0.30X ⫹ 8.51 0.59X ⫹ 0.25



Di-n-butyl phthalate 0.59C ⫹ 0.71 0.13X ⫹ 1.16 0.39X ⫹ 0.60



1,2-Dichlorobenzene 0.80C ⫹ 0.28 0.20X ⫹ 0.47 0.24X ⫹ 0.39



1,3-Dichlorobenzene 0.86C ⫺ 0.70 0.25X ⫹ 0.68 0.41X ⫹ 0.11



1,4-Dichlorobenzene 0.73C ⫺ 1.47 0.24X ⫹ 0.23 0.29X ⫹ 0.36



3,3⬘-Dichlorobenzidine 1.23C ⫺ 12.65 0.28X ⫹ 7.33 0.47X ⫹ 3.45



Dieldrin 0.82C ⫺ 0.16 0.20X ⫺ 0.16 0.26X ⫺ 0.07



Diethyl phthalate 0.43C ⫹ 1.00 0.28X ⫹ 1.44 0.52X ⫹ 0.22



Dimethyl phthalate 0.20C ⫹ 1.03 0.54X ⫹ 0.19 1.05X ⫺ 0.92



2,4-Dinitrotoluene 0.92C ⫺ 4.81 0.12X ⫹ 1.06 0.21X ⫹ 1.50



2,6-Dinitrotoluene 1.06C ⫺ 3.60 0.14X ⫹ 1.26 0.19X ⫹ 0.35



Di-n-octylphthalate 0.76C ⫺ 0.79 0.21X ⫹ 1.19 0.37X ⫹ 1.19



Endosulfan sulfate 0.39C ⫹ 0.41 0.12X ⫹ 2.47 0.63X ⫺ 1.03



Endrin aldehyde 0.76C ⫺ 3.86 0.18X ⫹ 3.91 0.73X ⫺ 0.62



Fluoranthene 0.81C ⫹ 1.10 0.22X ⫺ 0.73 0.28X ⫺ 0.60



Fluorene 0.90C ⫺ 0.00 0.12X ⫹ 0.26 0.13X ⫹ 0.61



Heptachlor 0.87C ⫺ 2.97 0.24X ⫺ 0.56 0.50X ⫺ 0.23



Heptachlor epoxide 0.92C ⫺ 1.87 0.33X ⫺ 0.46 0.28X ⫹ 0.64



Hexachlorobenzene 0.74C ⫹ 0.66 0.18X ⫺ 0.10 0.43X ⫺ 0.52



Hexachlorobutadiene 0.71C ⫺ 1.01 0.19X ⫹ 0.92 0.26X ⫹ 0.49



Hexachloroethane 0.73C ⫺ 0.83 0.17X ⫹ 0.67 0.17X ⫹ 0.80



Indeno(1,2,3-cd)pyrene 0.78C ⫺ 3.10 0.29X ⫹ 1.46 0.50X ⫹ 0.44



Isophorone 1.12C ⫹ 1.41 0.27X ⫹ 0.77 0.33X ⫹ 0.26



Naphthalene 0.76C ⫹ 1.58 0.21X ⫺ 0.41 0.30X ⫺ 0.68



Nitrobenzene 1.09C ⫺ 3.05 0.19X ⫹ 0.92 0.27X ⫹ 0.21



N-Nitrosodi-n-propylamine 1.12C ⫺ 6.22 0.27X ⫹ 0.68 0.44X ⫹ 0.47



PCB-1260 0.81C ⫺ 10.86 0.35X ⫹ 3.61 0.43X ⫹ 1.82



Phenanthrene 0.87C ⫺ 0.06 0.12X ⫹ 0.57 0.15X ⫹ 0.25



Pyrene 0.84C ⫺ 0.16 0.16X ⫹ 0.06 0.15X ⫹ 0.31



1,2,4-Trichlorobenzene 0.94C ⫺ 0.79 0.15X ⫹ 0.85 0.21X ⫹ 0.39



4-Chloro-3-methylphenol 0.84C ⫹ 0.35 0.23X ⫹ 0.75 0.29X ⫹ 1.31



2-Chlorophenol 0.78C ⫹ 0.29 0.18X ⫹ 1.46 0.28X ⫹ 0.97



2,4-Dichlorophenol 0.87C ⫹ 0.13 0.15X ⫹ 1.25 0.21X ⫹ 1.28



2,4-Dimethylphenol 0.71C ⫹ 4.41 0.16X ⫹ 1.21 0.22X ⫹ 1.31



2,4-Dinitrophenol 0.81C ⫺ 18.04 0.38X ⫹ 2.36 0.42X ⫹ 26.29



2-Methyl-4,6-dinitrophenol 1.04C ⫺ 28.04 0.10X ⫹ 42.29 0.26X ⫹ 23.10
6-78 INDIVIDUAL ORGANIC COMPOUNDS (6000)

TABLE 6410:VI. CONT.

Bias as Single-Analyst Overall
Recovery, X⬘ Precision, sr⬘ Precision, S⬘
Compound ␮g/L ␮g/L ␮g/L



2-Nitrophenol 1.07C ⫺ 1.15 0.16X ⫹ 1.94 0.27X ⫹ 2.60



4-Nitrophenol 0.61C ⫺ 1.22 0.38X ⫹ 2.57 0.44X ⫹ 3.24



Pentachlorophenol 0.93C ⫹ 1.99 0.24X ⫹ 3.03 0.30X ⫹ 4.33



Phenol 0.43C ⫹ 1.26 0.26X ⫹ 0.73 0.35X ⫹ 0.58



2,4,6-Trichlorophenol 0.91C ⫺ 0.18 0.16X ⫹ 2.22 0.22X ⫹ 1.81
*X⬘ ⫽ expected recovery for one or more measurements of a sample containing a concentration of C,


sr⬘ ⫽ expected single-analyst standard deviation of measurements at an average concentration found of X,


S⬘ ⫽ expected interlaboratory standard deviation of measurements at an average concentration found of X,
C ⫽ true value for the concentration, and


X ⫽ average recovery found for measurements of samples containing a concentration of C.
† The proper chemical name is 2,2⬘-oxybis(1-chloropropane).

• The retention time falls within ⫾30 s of the retention time ence for the primary m/z, use a secondary characteristic m/z to
of the authentic compound. quantitate.
• The relative peak heights of the three characteristic masses Calculate sample concentration using the response factor (RF)
in the EICPs fall within ⫾20% of the relative intensities of these determined in ¶ 5d and the equation:
masses in a reference mass spectrum obtained from a standard
analyzed in the GC/MS system or from a reference library. (A s) (I s)
Structural isomers that have very similar mass spectra and less Concentration, ␮g/L ⫽
(A is) (RF) (V e)
than 30 s difference in retention time can be identified explicitly
only if the resolution between authentic isomers in a standard
mix is acceptable. Acceptable resolution is achieved if the base- where:
line to valley height between the isomers is less than 25% of the As ⫽ area of characteristic m/z for compound or surrogate
standard to be measured,
sum of the two peak heights. Otherwise, structural isomers are
Ais ⫽ area of characteristic m/z for internal standard,
identified as isomeric pairs.
Is ⫽ amount of internal standard added to each extract, ␮g, and
f. Screening procedure for 2,3,7,8-tetrachlorodibenzo-p-di- Ve ⫽ volume of water extracted, L.
oxin (2,3,7,8-TCDD): CAUTION: In screening a sample for
2,3,7,8-TCDD, do not handle reference material without taking Report results in ␮g/L without correction for recovery. Report
extensive safety precautions. It is sufficient to analyze the base/ all QC data with sample results.
neutral extract by selected ion monitoring (SIM) GC/MS tech-
niques, as follows: 7. Quality Control
Concentrate base/neutral extract to a final volume of 0.2 mL.
Adjust temperature of base/neutral column to 220°C. Operate a. Quality control program: See Section 6200A.5.
mass spectrometer to acquire data in the SIM mode using the b. Initial quality control: Proceed according to Section
ions at m/z 257, 320, and 322 and a dwell time no greater than 6200A.5a1) and 2). Use Table 6410:V for acceptance criteria.
333 ms/mass. Inject 5 to 7 ␮L of base/neutral extract. Collect c. Analyses of samples with known additions: Use quality
SIM data for a total of 10 min. The possible presence of 2,3,7,8- acceptance criteria given in Table 6410:V.
TCDD is indicated if all three masses exhibit simultaneous peaks d. Quality-control check standard analysis: Proceed as in
at any point in the selected ion current profiles. For each occur- Section 6200A.5a3); prepare QC check standard with 1.0 mL
rence where the possible presence of 2,3,7,8-TCDD is indicated, QC check standard concentrate and 1 L reagent water.
calculate and retain the relative abundances of each of the three e. Bias assessment and records: Assess method bias and
masses. False positives may be caused by the presence of single maintain records. For example, after the analysis of five waste-


or coeluting combinations of compounds whose mass spectra water samples, calculate the average percent recovery (P) and
contain all of these masses. Conclusive results of the presence the standard deviation of the percent recovery (sp). Express bias



and concentration level of 2,3,7,8-TCDD can be obtained only assessment as a percent recovery interval from P ⫺ 2sp to P ⫹


from a properly equipped laboratory using a specialized test 2sp. If P ⫽ 90% and sp ⫽ 10%, the recovery interval is expressed
method.11 as 70 –110%. Update bias assessment for each compound regu-
larly, (e.g., after each five to ten new accuracy measurements).
6. Calculation f. Use of surrogate compounds: As a quality control check,
make known additions to all samples of surrogate standard
When a compound has been identified, base quantitation on solution as described in ¶ 5a1), and calculate percent recovery of
the integrated abundance from the EICP of the primary charac- each surrogate compound.
teristic m/z given in Tables 6410:I and II. Use base peak m/z for g. Additional quality-assurance practices: Other desirable
internal and surrogate standards. If sample produces an interfer- practices depend on the needs of the laboratory and the nature of
PHENOLS (6420)/Liquid-Liquid Extraction GC Method 6-79

the samples. Analyze field duplicates to assess precision of 3. U.S. ENVIRONMENTAL PROTECTION AGENCY. 1984. EPA Method Study
environmental measurements. Whenever possible, analyze stan- 30, Method 625—Base/Neutrals, Acids, and Pesticides. EPA-600/
dard reference materials and participate in relevant performance 4-84-053, National Technical Information Serv., PB84-206572,
evaluation studies. Certain compounds, such as phthalates, are Springfield, Va.
common laboratory contaminants. When these are measured 4. AMERICAN SOCIETY FOR TESTING AND MATERIALS. 1978. Standard
above the detection limits in sample blanks, locate their source practices for preparation of sample containers and for preservation
and repeat the analysis after taking corrective action. of organic constituents. ASTM Annual Book of Standards, Part 31,
D3694-78. Philadelphia, Pa.
8. Precision and Bias 5. U.S. ENVIRONMENTAL PROTECTION AGENCY. 1984. Definition and pro-
cedure for the determination of the method detection limit. 40 CFR
Part 136, Appendix B. Federal Register 49, No. 209.
This method was tested by 15 laboratories using reagent water,
6. OLYNYK, P., W.L. BUDDE & J.W. EICHELBERGER. 1980. Method
drinking water, surface water, and industrial wastewaters with Detection Limit for Methods 624 and 625. Unpublished report.
additions at six concentrations over the range 5 to 1300 ␮g/L.3 7. AMERICAN SOCIETY FOR TESTING AND MATERIALS. 1976. Standard
Single-operator precision, overall precision, and method bias practices for sampling water. ASTM Annual Book of Standards,
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