<<

SUBCHAPTER D— PROGRAMS (CONTINUED)

PART 136—GUIDELINES ESTAB- to perform the measurements required LISHING TEST PROCEDURES FOR for an application submitted to the Ad- THE ANALYSIS OF POLLUTANTS ministrator or to a State for a sludge permit under section 405(f) of Sec. the Clean Water Act and for record- 136.1 Applicability. keeping and reporting requirements 136.2 Definitions. under part 503 of title 40. 136.3 Identification of test procedures. (c) For the purposes of the NPDES 136.4 Application for and approval of alter- program, when more than one test pro- nate test procedures for nationwide use. 136.5 Approval of alternate test procedures cedure is approved under this part for for limited use. the analysis of a pollutant or pollutant 136.6 Method modifications and analytical parameter, the test procedure must be requirements. sufficiently sensitive as defined at 40 136.7 Quality assurance and quality control. CFR 122.21(e)(3) and 122.44(i)(1)(iv). APPENDIX A TO PART 136—METHODS FOR OR- GANIC CHEMICAL ANALYSIS OF MUNICIPAL [72 FR 14224, Mar. 26, 2007, as amended at 77 AND INDUSTRIAL WASTEWATER FR 29771, May 18, 2012; 79 FR 49013, Aug. 19, APPENDIX B TO PART 136—DEFINITION AND 2014; 82 FR 40846, Aug. 28, 2017] PROCEDURE FOR THE DETERMINATION OF THE METHOD DETECTION LIMIT—REVISION § 136.2 Definitions. 1.11 APPENDIX C TO PART 136—DETERMINATION OF As used in this part, the term: METALS AND TRACE ELEMENTS IN WATER (a) Act means the Clean Water Act of AND WASTES BY INDUCTIVELY COUPLED 1977, Pub. L. 95–217, 91 Stat. 1566, et seq. PLASMA-ATOMIC EMISSION SPECTROMETRY (33 U.S.C. 1251 et seq.) (The Federal METHOD 200.7 Water Control Act Amend- APPENDIX D TO PART 136—PRECISION AND RE- COVERY STATEMENTS FOR METHODS FOR ments of 1972 as amended by the Clean MEASURING METALS Water Act of 1977). (b) Administrator means the Adminis- AUTHORITY: Secs. 301, 304(h), 307 and 501(a), Pub. L. 95–217, 91 Stat. 1566, et seq. (33 U.S.C. trator of the U.S. Environmental Pro- 1251, et seq.) (the Federal Water Pollution tection Agency. Control Act Amendments of 1972 as amended (c) Regional Administrator means one by the Clean Water Act of 1977). of the EPA Regional Administrators. § 136.1 Applicability. (d) Director means the director as de- fined in 40 CFR 122.2. (a) The procedures prescribed herein (e) National Pollutant Discharge Elimi- shall, except as noted in §§ 136.4, 136.5, nation System (NPDES) means the na- and 136.6, be used to perform the meas- tional system for the issuance of per- urements indicated whenever the waste constituent specified is required to be mits under section 402 of the Act and measured for: includes any State or interstate pro- (1) An application submitted to the gram which has been approved by the Director and/or reports required to be Administrator, in whole or in part, submitted under NPDES permits or pursuant to section 402 of the Act. other requests for quantitative or qual- (f) Detection limit means the minimum itative effluent data under parts 122 concentration of an analyte (sub- through 125 of this chapter; and stance) that can be measured and re- (2) Reports required to be submitted ported with a 99% confidence that the by dischargers under the NPDES estab- analyte concentration is distinguish- lished by parts 124 and 125 of this chap- able from the method blank results as ter; and determined by the procedure set forth (3) Certifications issued by States at appendix B of this part. pursuant to section 401 of the Clean Water Act (CWA), as amended. [38 FR 28758, Oct. 16, 1973, as amended at 49 (b) The procedure prescribed herein FR 43250, Oct. 26, 1984; 82 FR 40846, Aug. 28, and in part 503 of title 40 shall be used 2017]

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§ 136.3 Identification of test proce- the methods listed in these tables, the dures. provisions of 40 CFR parts 122 and 125 (a) Parameters or pollutants, for are controlling and will determine a which methods are approved, are listed permittee’s reporting requirements. together with test procedure descrip- The full texts of the referenced test tions and references in Tables IA, IB, procedures are incorporated by ref- IC, ID, IE, IF, IG, and IH of this sec- erence into Tables IA, IB, IC, ID, IE, tion. The methods listed in Tables IA, IF, IG, and IH. The year after the IB, IC, ID, IE, IF, IG, and IH are incor- method number indicates the latest porated by reference, see paragraph (b) editorial change of the method. The of this section, with the exception of discharge parameter values for which EPA Methods 200.7, 601–613, 624.1, 625.1, reports are required must be deter- 1613, 1624, and 1625. The full texts of mined by one of the standard analyt- Methods 601–613, 624.1, 625.1, 1613, 1624, ical test procedures incorporated by and 1625 are printed in appendix A of reference and described in Tables IA, this part, and the full text of Method IB, IC, ID, IE, IF, IG, and IH or by any 200.7 is printed in appendix C of this alternate test procedure which has part. The full text for determining the been approved by the Administrator method detection limit when using the under the provisions of paragraph (d) of test procedures is given in appendix B this section and §§ 136.4 and 136.5. Under of this part. In the event of a conflict certain circumstances (paragraph (c) of between the reporting requirements of this section, in § 136.5(a) through (d) or 40 CFR parts 122 and 125 and any re- 40 CFR 401.13) other additional or alter- porting requirements associated with nate test procedures may be used.

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. . 19 . ® 13 17 18 13 24 13 18 21 29 ® ® ® 13 18 Other ® Colilert-18

...... Enterolert . . . 4 4 4 9 ...... Colilert LUDGE USGS S 10 AOAC, ASTM, EWAGE S ...... B–0050–85 ...... 991.15 . 12 14 30 30 13 ASTEWATER AND 9221F–2006 W 9221 C E–2006.

11 15 . ETHODS FOR 1680, ...... mColiBlue-24 ...... 9230 C–2007...... 9222 D–2006 ...... 9221 C E–2006...... 9222 D–2006 ...... 9221 B–2006...... 9222 B–2006 ...... B–0025–85 9221 B–2006...... 9222 B–2006...... 9230 B–2007...... 9230 C–2007 ...... B–0055–85 9230 B–2007. . . 11 20 M 3 3 3 3 3 3 3 3 3 3 3 3 3 22 25 1681 ...... 9230 D–2007 ...... D6503–99 ...... Colilert-18 ...... 9223 B–2004 ...... 9221B.2–2006/ 1600 p. 132, p. 132 p. 114 p. 114 p. 139 p. 139 p. 124 p. 108 p. 111 IOLOGICAL .... p. 124 B 5 EPA Standard methods Standard EPA 1 multiple single step 1603 single step . , single step. , multiple 5 2 PPROVED 6816 68 with enrich- A , single step , single step or , or ...... p. 136 2 2 2 2678 2 2678 or. ber (MPN), 5 tube, 3 dilution, or. tion, or. tion, or. tion, or. tion, or. tube/multiple well, or. well, or. (MF) well, or. ment tube, or. tion, or. two step. Most Probable Num- MPN, 5 tube, 3 dilu- MPN, 5 tube, 3 dilu- Multiple tube/multiple Membrane filter MF MF MF multiple tube/multiple MF MF Plate count ...... p. 143 MPN MF Plate count ...... p. 143 IST OF ...... MPN, 5 tube, 3 dilu- IA—L 21 ...... MPN ABLE 21 T Parameter and units Method number per 100 mL E. coli, number per gram dry weight. number per 100 mL. number per 100 mL. 1. Coliform (fecal), number per 100 mL or 2. Coliform (fecal) in presence of , 3. Coliform (total), number per 100 mL ...... MPN, 5 tube, 3 dilu- 4. Coliform (total), in presence of chlorine, 5. 6. Fecal streptococci, number per 100 mL MPN, 5 tube, 3 dilu- 7. Enterococci, number per 100 mL

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Sep- b E. coli C, and a water bath incubator is used. ° 0.2 ± may be obtained from IDEXX Laboratories, Inc. ® for the determination of total coliforms and ® test and is recommended for marine water samples. ® , and Quanti-Tray ® ) in Water by Membrane Filtration Using Modified membrane-Thermotolerant test, is available from Hach Company. ® E. coli test may be obtained from IDEXX Laboratories Inc. ( ® , Colilert-18 in Sewage Sludge (Biosolids) by Modified Semisolid Rappaport-Vassiliadis (MSRV) Medium, EPA–821–R–14–012. September ® to assay for fecal coliforms, the incubation temperature is 44.5 ® 3 h of incubation shall be submitted to 9221F–2006. Commercially available EC–MUG media or EC supplemented in the laborat and the MPN calculated from table provided by manufacturer. Salmonella Escherichia coli ± ® is an optimized formulation of the Colilert ® E. coli. g/mL of MUG may be used. μ Method 1600: Enterococci in Water by Membrane Filtration Using membrane-Enterococcus Indoxyl- Official Methods of Analysis AOAC International. 16th Edition, 4th Revision, 1998. Approved for enumeration of target organism in sewage sludge. The multiple-tube fermentation test is used in 9221B.2–2006. Lactose broth may be lieu of lauryl tryptose (LTB), These tests are collectively known as defined substrate tests, where, for example, a is used to detect the enz After prior enrichment in a presumptive medium for total coliform using 9221B.2–2006, all tubes or bottles showing Samples shall be enumerated by the multiple-tube or multiple-well procedure. Using procedures, employ an appropri Colilert-18 Descriptions of the Colilert A description of the mColiBlue24 Method 1681: Fecal Coliforms in Sewage Sludge (Biosolids) by Multiple-Tube Fermentation using A–1 Medium, EPA–821–R–06–013. Jul Approved for enumeration of target organism in wastewater effluent. Method 1603: Method 1682: A description of the Enterolert Methods for Measuring the Acute Toxicity of Effluents and Receiving to Freshwater Marine Organisms, EPA–821–R–02–012 Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, EPA–821–R–02– Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Marine Estuarine Organisms, EPA To use Colilert-18 On a monthly basis, at least ten colonies from the medium must be verified using Lauryl Tryptose Broth and EC broth, follo Method 1680: Fecal Coliforms in Sewage Sludge (Biosolids) by Multiple-Tube Fermentation Using Lauryl-Tryptose Broth (LTB) and E Tests must be conducted to provide organism enumeration (density). Select the appropriate configuration of tubes/filtrations an When the MF method has been used previously to test waters with high turbidity, large numbers of noncoliform bacteria, or sampl To assess the comparability of results obtained with individual methods, it is suggested that side-by-side tests be conducted a Annual Book of ASTM Standards-Water and Environmental Technology, Section 11.02. 2000, 1999, 1996. International. 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 C rather than the 24 h required for Colilert R–14–009. September 2014. U.S. EPA. for the quality, character, consistency, and anticipated organism density of water sample. acidity within 48 h conducted between this broth and LTB using the water samples normally tested, comparison demonstrates that false-p rate for total coliform using lactose broth is less than 10 percent. No requirement exists to run the completed phase on per tubes on a seasonal basis. figuration of the sample as needed and report Most Probable Number (MPN). Samples tested with Colilert ber 2002. U.S. EPA. October 2002. U.S. EPA. Edition, October 2002. U.S. EPA. based on these results; and representative non-blue colonies should be verified using Lauryl Tryptose Broth. Where possible, ve randomized sample sources. duced by tember 2014. U.S. EPA. dures, Quanti-Tray ° procedure (ATP) guidelines. the water samples routinely tested in accordance with most current Standard Methods for Examination of Water and Wastew nisms stressed by chlorine, a parallel test should be conducted with multiple-tube technique to demonstrate applicability and with 50 2014. U.S. EPA. EPA–821–R–14–010. September 2014. U.S. EPA.

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TABLE IB—LIST OF APPROVED INORGANIC TEST PROCEDURES

58 52 Standard meth- USGS/AOAC/ Parameter Methodology EPA ods ASTM other

1. Acidity, as Electrometric ...... 2310 B–2011 ... D1067–11 ...... I–1020–85.2 CaCO3, mg/L. endpoint or - phthalein endpoint. 2. Alkalinity, as Electrometric or ...... 2320 B–2011 ... D1067–11 ...... 973.43,3 I– 2 CaCO3, mg/L. Colorimetric 1030–85. titration to pH 4.5, Manual. Automatic ...... 310.2 (Rev...... I–2030–85.2 1974) 1. 3. Aluminum— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 D–2011 or ...... I–3051–85.2 ration 36. 3111 E–2011. AA furnace ...... 3113 B–2010. STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES 36 ...... 200.5, Rev 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4471–97.50 Direct Current ...... D4190–08 ...... See footnote.34 Plasma (DCP) 36. Colorimetric ...... 3500-Al B–2011. (Eriochrome cyanine R). 3 4. (as Manual distilla- 350.1, Rev. 2.0 4500–NH3 B– ...... 973.49. N), mg/L. tion 6 or gas (1993). 2011. diffusion (pH > 11), fol- lowed by any of the fol- lowing:. Nesslerization ...... D1426–08 (A) .. 973.49,3 I– 3520–85.2

Titration ...... 4500–NH3 C– 2011.

Electrode ...... 4500–NH3 D– D1426–08 (B). 2011 or E– 2011. 60 Manual ...... 4500–NH3 F- ...... See footnote. phenate, sa- 2011. licylate, or other sub- stituted phe- nols in Berthelot re- action based methods.

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TABLE IB—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued

58 52 Standard meth- USGS/AOAC/ Parameter Methodology EPA ods ASTM other

30 2 Automated 350.1, Rev. 4500–NH3 G- ...... I–4523–85. phenate, sa- 2.0 (1993). 2011, 4500– licylate, or NH3 H-2011. other sub- stituted phe- nols in Berthelot re- action based methods. Automated elec- ...... See footnote.7 trode. Chroma- ...... D6919–09. tography. Automated gas ...... Timberline Am- diffusion, fol- monia-001.74 lowed by con- ductivity cell analysis. 5. Antimony— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011. ration 36. AA furnace ...... 3113 B–2010. STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES 36 ...... 200.5, Rev 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4471–97.50 6. - Digestion,4 fol- 206.5 (Issued Total,4 mg/L. lowed by any 1978) 1. of the fol- lowing:. AA gaseous hy- ...... 3114 B–2011 or D2972–08 (B) .. I–3062–85.2 dride. 3114 C–2011. AA furnace ...... 3113 B–2010 ... D2972–08 (C) .. I–4063–98.49 STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES 36 ...... 200.5, Rev 4.2 3120 B–2011 ... D1976–12. (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4020–05.70 Colorimetric ...... 3500-As B– D2972–08 (A) .. I–3060–85.2 (SDDC). 2011. 7. Barium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 D–2011 ...... I–3084–85.2 ration 36. AA furnace ...... 3113 B–2010 ... D4382–12. ICP/AES 36 ...... 200.5, Rev 4.2 3120 B–2011 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994).

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TABLE IB—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued

58 52 Standard meth- USGS/AOAC/ Parameter Methodology EPA ods ASTM other

ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4471–97.50 DCP 36 ...... See footnote.34 8. Beryllium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing:. AA direct aspi- ...... 3111 D–2011 or D3645–08 (A) .. I–3095–85.2 ration. 3111 E–2011. AA furnace ...... 3113 B–2010 ... D3645–08 (B). STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES ...... 200.5, Rev 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4471–97.50 DCP ...... D4190–08 ...... See footnote.34 Colorimetric ...... See footnote.61 (aluminon). 9. Biochemical Dissolved Oxy- ...... 5210 B–2011 ...... 973.44,3 p. 17,9 de- gen Depletion. I–1578–78,8 mand (BOD5), See foot- mg/L. note.10 63 10. Boron— Colorimetric ...... 4500–B B–2011 ...... I–3112–85.2 Total,37 mg/L. (curcumin). ICP/AES ...... 200.5, Rev 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4471–97.50 DCP ...... D4190–08 ...... See footnote.34 11. Bromide, Electrode ...... D1246–10 ...... I–1125–85.2 mg/L. Ion Chroma- 300.0, Rev 2.1 4110 B–2011, D4327–03 ...... 993.30.3 tography. (1993) and C–2011, D– 300.1, Rev 2011. 1.0 (1997). CIE/UV ...... 4140 B–2011 ... D6508–10 ...... D6508, Rev. 2.54 12. Cadmium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 or D3557–12 (A or 974.27,3 p. 37,9 ration 36. 3111 C–2011. B). I–3135–852 or I–3136– 85.2 AA furnace ...... 3113 B–2010 ... D3557–12 (D) .. I–4138–89.51 STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES 36 ...... 200.5, Rev 4.2 3120 B–2011 ... D1976–12 ...... I–1472–85 2 or (2003); 68 I–4471–97.50 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4471–97.50 DCP 36 ...... D4190–08 ...... See footnote.34 Voltametry 11 ...... D3557–12 (C).

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TABLE IB—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued

58 52 Standard meth- USGS/AOAC/ Parameter Methodology EPA ods ASTM other

Colorimetric (Di- ...... 3500-Cd-D- thizone). 1990. 13. Calcium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 ... D511–09(B) ..... I–3152–85.2 ration. ICP/AES ...... 200.5, Rev 4.2 3120 B–2011 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14.3 (1994). DCP ...... See footnote.34 Titrimetric ...... 3500-Ca B– D511–09 (A). (EDTA). 2011. Ion Chroma- ...... D6919–09. tography. 14. Carbo- Dissolved Oxy- ...... 5210 B–2011 ...... See foot- naceous bio- gen Depletion note.35 63 chemical oxy- with nitrifica- gen demand tion inhibitor. (CBOD5), mg/ L 12. 15. Chemical ox- Titrimetric ...... 410.3 (Rev. 5220 B–2011 or D1252–06 (A) .. 973.46,3 p. 17,9 ygen demand 1978) 1. C–2011. I–3560–85.2 (COD), mg/L. Spectrophotom- 410.4, Rev. 2.0 5220 D–2011 ... D1252–06 (B) .. See foot- etric, manual (1993). notes.13 14, I– or automatic. 3561–85.2 16. , Titrimetric: (sil- ...... 4500–Cl¥ B– D512–04 (B) .... I–1183–85.2 mg/L. ver nitrate). 2011. (Mercuric ni- ...... 4500–Cl¥ C– D512–04 (A) .... 973.51,3 I– trate). 2011. 1184–85.2 Colorimetric: ...... I–1187–85.2 Manual. Automated (fer- ...... 4500–Cl¥ E– ...... I–2187–85.2 ricyanide). 2011. Potentiometric ...... 4500–Cl¥ D– Titration. 2011. Ion Selective ...... D512–04 (C). Electrode. Ion Chroma- 300.0, Rev 2.1 4110 B–2011 or D4327–03 ...... 993.30,3 I– tography. (1993) and 4110 C–2011. 2057–90.51 300.1, Rev 1.0 (1997). CIE/UV ...... 4140 B–2011 ... D6508–10 ...... D6508, Rev. 2.54 17. Chlorine- Amperometric ...... 4500–Cl D– D1253–08. Total residual, direct. 2011. mg/L. Amperometric ...... 4500–Cl E– direct (low 2011. level). Iodometric di- ...... 4500–Cl B– rect. 2011. Back titration ...... 4500–Cl C– ether end- 2011. point 15. DPD–FAS ...... 4500–Cl F-2011.

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TABLE IB—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued

58 52 Standard meth- USGS/AOAC/ Parameter Methodology EPA ods ASTM other

Spectrophotom- ...... 4500–Cl G- etric, DPD. 2011. Electrode ...... See footnote.16 17A. Chlorine- Amperometric ...... 4500–Cl D– D1253–08. Free Avail- direct. 2011. able, mg/L. Amperometric ...... 4500–Cl E– direct (low 2011. level). DPD–FAS ...... 4500–Cl F-2011. Spectrophotom- ...... 4500–Cl G- etric, DPD. 2011. 18. Chromium VI 0.45-micron fil- dissolved, mg/ tration fol- L. lowed by any of the fol- lowing: AA chelation- ...... 3111 C–2011 ...... I–1232–85.2 extraction. Ion Chroma- 218.6, Rev. 3.3 3500-Cr C– D5257–11 ...... 993.23.3 tography. (1994). 2011. Colorimetric (di- ...... 3500-Cr B– D1687–12 (A) .. I–1230–85.2 phenyl- 2011. carbazide). 19. Chromium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 ... D1687–12 (B) .. 974.27,3 I– ration 36. 3236–85.2 AA chelation- ...... 3111 C–2011. extraction. AA furnace ...... 3113 B–2010 ... D1687–12 (C) .. I–3233–93.46 STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES 36 ...... 200.5, Rev 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003),68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4020–05.70 DCP 36 ...... D4190–08 ...... See footnote.34 Colorimetric (di- ...... 3500-Cr B– phenyl- 2011. carbazide). 20. Cobalt— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 or D3558–08 (A or p. 37,9 I–3239– ration. 3111 C–2011. B). 85.2 AA furnace ...... 3113 B–2010 ... D3558–08 (C) .. I–4243–89.51 STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES 36 ...... 200.7, Rev. 4.4 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4020–05.70 DCP ...... D4190–08 ...... See footnote.34

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21. Color, plat- Colorimetric ...... 2120 F-2011 78. inum cobalt (ADMI). units or domi- nant wave- length, hue, luminance pu- rity. cobalt ...... 2120 B–2011 ...... I–1250–85.2 visual com- parison. Spectrophotom- ...... See footnote.18 etric. 22. Copper— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 or D1688–12 (A or 974.27,3 p. 37,9 ration 36. 3111 C–2011. B). I–3270–852 or I–3271– 85.2 AA furnace ...... 3113 B–2010 ... D1688–12 (C) .. I–4274–89.51 STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES 36 ...... 200.5, Rev 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4020–05.70 DCP 36 ...... D4190–08 ...... See footnote.34 Colorimetric ...... 3500-Cu B– (Neocuproine). 2011. Colorimetric ...... 3500-Cu C– ...... See footnote.19 (Bathocuproi- 2011. ne). 23. — Automated UV ...... Kelada-01.55 Total, mg/L. digestion/dis- tillation and Colorimetry. Segmented ...... D7511–12. Flow Injec- tion, In-Line Di- gestion, fol- lowed by gas diffusion am- perometry. Manual distilla- 335.4, Rev. 1.0 4500–CN¥ B– D2036–09(A), 10–204–00–1– tion with (1993) 57. 2011 and C– D7284–13. X.56 MgCl2, fol- 2011. lowed by any of the fol- lowing:. Flow Injection, ...... D2036–09(A) gas diffusion D7284–13. amperometry. Titrimetric ...... 4500–CN¥ D– D2036–09(A) ... p. 22.9 2011. Spectrophotom- ...... 4500–CN¥ E– D2036–09(A) ... I–3300–85.2 etric, manual. 2011.

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58 52 Standard meth- USGS/AOAC/ Parameter Methodology EPA ods ASTM other

Semi-Auto- 335.4, Rev. 1.0 ...... 10–204–00–1– mated 20. (1993) 57. X,56 I–4302– 85.2 Ion Chroma- ...... D2036–09(A). tography. Ion Selective ...... 4500–CN¥ F- D2036–09(A). Electrode. 2011. 24. Cyanide— Cyanide Ame- ...... 4500–CN¥ G- D2036–09(B). Available, mg/ nable to 2011. L. Chlorination (CATC); Man- ual distillation with MgCl2, followed by Titrimetric or Spectrophoto- metric. Flow injection ...... D6888–09 ...... OIA–1677–09.44 and exchange, fol- lowed by gas diffusion am- perometry 59. Automated Dis- ...... Kelada-01.55 tillation and Colorimetry (no UV diges- tion). 24.A Cyanide— Flow Injection, ...... D7237–10 ...... OIA–1677–09.44 Free, mg/L. followed by gas diffusion amperometry. Manual micro- ...... D4282–02. diffusion and colorimetry. 25. Fluoride— Manual distilla- ...... 4500–F¥ B– Total, mg/L. tion,6 followed 2011. by any of the following: Electrode, man- ...... 4500–F¥ C– D1179–10 (B). ual. 2011. Electrode, auto- ...... I–4327–85.2 mated. Colorimetric, ...... 4500–F¥ D– D1179–10 (A). (SPADNS). 2011. Automated ...... 4500–F¥ E– complexone. 2011. Ion Chroma- 300.0, Rev 2.1 4110 B–2011 or D4327–03 ...... 993.30.3 tography. (1993) and C–2011. 300.1, Rev 1.0 (1997). CIE/UV ...... 4140 B–2011 ... D6508–10 ...... D6508, Rev. 2.54 26. — Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011. ration. AA furnace ...... 231.2 (Issued 3113 B–2010. 1978) 1.

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TABLE IB—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued

58 52 Standard meth- USGS/AOAC/ Parameter Methodology EPA ods ASTM other

ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14.3 (1994). DCP ...... See footnote.34 27. Hardness— Automated col- 130.1 (Issued Total, as orimetric. 1971) 1. CaCO3, mg/L. Titrimetric ...... 2340 C–2011 ... D1126–12 ...... 973.52B,3 I– (EDTA). 1338–85.2 Ca plus Mg as ...... 2340 B–2011. their - ates, by any approved method for Ca and Mg (See Param- eters 13 and 33), provided that the sum of the lowest point of quan- titation for Ca and Mg is below the NPDES per- mit require- ment for Hardness.. 28. Hydrogen Electrometric ...... 4500–H+ B– D1293–99 (A or 973.41,3 I– ion (pH), pH measurement. 2011. B). 1586–85.2 units. Automated elec- 150.2 (Dec...... See footnote,21 trode. 1982) 1. I–2587–85.2 29. Iridium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011. ration. AA furnace ...... 235.2 (Issued 1978) 1. ICP/MS ...... 3125 B–2011. 30. —Total,4 Digestion,4 fol- mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 or D1068–10 (A) .. 974.27,3 I– ration 36. 3111 C–2011. 3381–85.2 AA furnace ...... 3113 B–2010 ... D1068–10 (B). STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES 36 ...... 200.5, Rev. 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14.3 (1994). DCP 36 ...... D4190–08 ...... See footnote.34 Colorimetric ...... 3500-Fe B– D1068–10 (C) .. See footnote.22 (Phenan- 2011. throline).

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45 31. Kjeldahl Ni- Manual diges- ...... 4500–Norg B– D3590–11 (A) .. I–4515–91. trogen 5— tion 20 and 2011 or C– Total, (as N), distillation or 2011 and mg/L. gas diffusion, 4500–NH3 B– followed by 2011. any of the fol- lowing:. 3 Titration ...... 4500–NH3 C– ...... 973.48. 2011. Nesslerization ...... D1426–08 (A). Electrode ...... 4500–NH3 D– D1426–08 (B). 2011 or E– 2011. Semi-automated 350.1, Rev. 2.0 4500–NH3 G- phenate. (1993). 2011 4500– NH3 H-2011. 60 Manual ...... 4500–NH3 F- ...... See footnote. phenate, sa- 2011. licylate, or other sub- stituted phe- nols in Berthelot re- action based methods. Automated gas ...... Timberline Am- diffusion, fol- monia-001.74 lowed by con- ductivity cell analysis.

Automated Methods for TKN that do not require manual distillation.

Automated 351.1 (Rev...... I–4551–78.8 phenate, sa- 1978) 1. licylate, or other sub- stituted phe- nols in Berthelot re- action based methods col- orimetric (auto diges- tion and dis- tillation). 45 Semi-automated 351.2, Rev. 2.0 4500–Norg D– D3590–11 (B) .. I–4515–91. block digestor (1993). 2011. colorimetric (distillation not required). Block digester, ...... See footnote.39 followed by Auto distilla- tion and Titra- tion. Block digester, ...... See footnote.40 followed by Auto distilla- tion and Nesslerization.

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Block Digester, ...... See footnote.41 followed by Flow injection gas diffusion (distillation not required). Digestion with ...... Hach 10242.76 peroxdisulfat- e, followed by Spectrophoto- metric (2,6-di- methyl phe- nol). Digestion with ...... NCASI TNTP persulfate, W10900.77 followed by Colorimetric. 32. Lead— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 or D3559–08 (A or 974.27,3 I– ration 36. 3111 C–2011. B). 3399–85.2 AA furnace ...... 3113 B–2010 ... D3559–08 (D) .. I–4403–89.51 STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES 36 ...... 200.5, Rev. 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4471–97.50 DCP 36 ...... D4190–08 ...... See footnote.34 Voltametry 11 ...... D3559–08 (C). Colorimetric (Di- ...... 3500-Pb B– thizone). 2011. 33. Magne- Digestion,4 fol- sium—Total,4 lowed by any mg/L. of the fol- lowing: AA direct aspi- ...... 3111 B–2011 ... D511–09 (B) .... 974.27,3 I– ration. 3447–85.2 ICP/AES ...... 200.5, Rev. 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14.3 (1994). DCP ...... See footnote.34 ...... Ion Chroma- ...... D6919–09. tography. 34. Man- Digestion,4 fol- ganese— lowed by any Total,4 mg/L. of the fol- lowing: AA direct aspi- ...... 3111 B–2011 ... D858–12 (A or 974.27,3 I– ration 36. B). 3454–85.2 AA furnace ...... 3113 B–2010 ... D858–12 (C). STGFAA ...... 200.9, Rev. 2.2 (1994).

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ICP/AES 36 ...... 200.5, Rev. 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4471–97.50 DCP 36 ...... D4190–08 ...... See footnote.34 Colorimetric ...... 3500-Mn B– ...... 920.203.3 (Persulfate). 2011. Colorimetric ...... See footnote.23 (Periodate). 35. Mercury— Cold vapor, 245.1, Rev. 3.0 3112 B–2011 ... D3223–12 ...... 977.22,3 I– Total,4 mg/L. Manual. (1994). 3462–85.2 Cold vapor, 245.2 (Issued Automated. 1974) 1. Cold vapor 245.7 Rev. 2.0 ...... I–4464–01.71 atomic fluo- (2005) 17. rescence spectrometry (CVAFS). Purge and Trap 1631E 43. CVAFS. 36. Molyb- Digestion,4 fol- denum— lowed by any Total,4 mg/L. of the fol- lowing: AA direct aspi- ...... 3111 D–2011 ...... I–3490–85.2 ration. AA furnace ...... 3113 B–2010 ...... I–3492–96.47 ICP/AES 36 ...... 200.7, Rev. 4.4 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4471–97.50 DCP ...... See footnote.34 37. Nickel— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 or D1886–08 (A or I–3499–85.2 ration 36. 3111 C–2011. B). AA furnace ...... 3113 B–2010 ... D1886–08 (C) .. I–4503–89.51 STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES 36 ...... 200.5, Rev. 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4020–05.70 DCP 36 ...... D4190–08 ...... See footnote.34 38. Nitrate (as Ion Chroma- 300.0, Rev. 2.1 4110 B–2011 or D4327–03 ...... 993.30.3 N), mg/L. tography. (1993) and C–2011. 300.1, Rev. 1.0 (1997). CIE/UV ...... 4140 B–2011 ... D6508–10 ...... D6508, Rev. 2.54 ¥ Ion Selective ...... 4500–NO3 D– Electrode. 2011. Colorimetric 352.1 (Issued ...... 973.50,3 (Brucine sul- 1971) 1. 419D17 p. fate). 28.9

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Spectrophotom- ...... Hach 10206.75 etric (2,6- dimethylphen- ol). Nitrate- N minus Nitrite N (See pa- rameters 39 and 40). ¥ 39. Nitrate-nitrite Cadmium re- ...... 4500–NO3 E– D3867–04 (B). (as N), mg/L. duction, Man- 2011. ual. ¥ 51 Cadmium re- 353.2, Rev. 2.0 4500–NO3 F- D3867–04 (A) .. I–2545–90. duction, Auto- (1993). 2011. mated. ¥ Automated hy- ...... 4500–NO3 H- drazine. 2011. Reduction/Col- ...... See footnote.62 orimetric. Ion Chroma- 300.0, Rev. 2.1 4110 B–2011 or D4327–03 ...... 993.30.3 tography. (1993) and C–2011. 300.1, Rev. 1.0 (1997). CIE/UV ...... 4140 B–2011 ... D6508–10 ...... D6508, Rev. 2.54 Enzymatic re- ...... I–2547–11,72 I– duction, fol- 2548–11,72 lowed by N07–0003.73 automated colorimetric determination. Spectrophotom- ...... Hach 10206.75 etric (2,6- dimethylphen- ol). ¥ 25 40. Nitrite (as Spectrophotom- ...... 4500–NO2 B– ...... See footnote. N), mg/L. etric: Manual. 2011. Automated ...... I–4540–85,2 (Diazotizatio- See foot- n). note.62 ¥ 2 Automated 353.2, Rev. 2.0 4500–NO3 F– D3867–04 (A) .. I–4545–85. (*bypass cad- (1993). 2011. mium reduc- tion). ¥ Manual ...... 4500–NO3 E– D3867–04 (B). (*bypass cad- 2011. mium reduc- tion). Ion Chroma- 300.0, Rev. 2.1 4110 B–2011 or D4327–03 ...... 993.30.3 tography. (1993) and C–2011. 300.1, Rev. 1.0 (1997). CIE/UV ...... 4140 B–2011 ... D6508–10 ...... D6508, Rev. 2.54 Automated ...... I–2547–11,72 I– (*bypass En- 2548–11,72 zymatic re- N07–0003.73 duction).

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41. Oil and Hexane extract- 1664 Rev. A; 5520 B–2011 38. grease—Total able material 1664 Rev. recoverable, (HEM): n- B 42. mg/L. Hexane ex- traction and gravimetry. Silica gel treat- 1664 Rev. A; 5520 B–2011 38 ed HEM 1664 Rev. and 5520 F- (SGT–HEM): B 42. 201138. Silica gel treatment and gravimetry. 42. Organic car- Combustion ...... 5310 B–2011 ... D7573–09 ...... 973.47,3 p. bon—Total 14.24 (TOC), mg/L. Heated ...... 5310 C–2011, D4839–03 ...... 973.47,3 p. persulfate or 5310 D–2011. 14.24 UV persulfate oxidation. 43. Organic ni- Total Kjeldahl N trogen (as N), (Parameter mg/L. 31) minus ammonia N (Parameter 4). 44. Ortho-phos- Ascorbic acid phate (as P), method: mg/L. Automated ...... 365.1, Rev. 2.0 4500–P F–2011 ...... 973.56,3 I– (1993). or G–2011. 4601–85.2 Manual single ...... 4500–P E–2011 D515–88 (A) .... 973.55.3 reagent. Manual two rea- 365.3 (Issued gent. 1978) 1. Ion Chroma- 300.0, Rev. 2.1 4110 B–2011 or D4327–03 ...... 993.30.3 tography. (1993) and C–2011. 300.1, Rev. 1.0 (1997). CIE/UV ...... 4140 B–2011 ... D6508–10 ...... D6508, Rev. 2.54 45. Osmium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 D–2011. ration. AA furnace ...... 252.2 (Issued 1978) 1. 46. Oxygen, dis- Winkler ( ...... 4500–O (B–F)– D888–09 (A) .... 973.45B,3 I– solved, mg/L. modification). 2011. 1575–78.8 Electrode ...... 4500–O G– D888–09 (B) .... I–1576–78.8 2011. Luminescence ...... D888–09 (C) .... See footnote.63 Based Sensor. See foot- note.64 47. Palladium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011. ration.

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AA furnace ...... 253.2 (Issued 1978) 1. ICP/MS ...... 3125 B–2011. DCP ...... See footnote.34 48. , mg/ Manual distilla- 420.1 (Rev. 5530 B–2010 ... D1783–01. L. tion,26 fol- 1978) 1. lowed by any of the fol- lowing: Colorimetric 420.1 (Rev. 5530 D–2010 27 D1783–01 (A or (4AAP) man- 1978) 1. B). ual. Automated col- 420.4 Rev. 1.0 orimetric (1993). (4AAP). 49. Phosphorus Gas-liquid chro- ...... See footnote.28 (elemental), matography. mg/L. 50. Phos- Digestion,20 fol- ...... 4500–P B(5)– ...... 973.55.3 phorus—Total, lowed by any 2011. mg/L. of the fol- lowing: Manual ...... 365.3 (Issued 4500–P E–2011 D515–88 (A). 1978) 1. Automated 365.1 Rev. 2.0 4500–P (F-H)– ...... 973.56,3 I– ascorbic acid (1993). 2011. 4600–85.2 reduction. ICP/AES 436 ..... 200.7, Rev. 4.4 3120 B–2011 ...... I–4471–97.50 (1994). Semi-automated 365.4 (Issued ...... D515–88 (B) .... I–4610–91.48 block digestor 1974) 1. (TKP diges- tion). Digestion with ...... NCASI TNTP persulfate, W10900.77 followed by Colorimetric. 51. Platinum— Digestion,4 fol- Total 4, mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011. ration. AA furnace ...... 255.2 (Issued 1978) 1. ICP/MS ...... 3125 B–2011. DCP ...... See footnote.34 52. Potassium— Digestion,4 fol- Total 4, mg/L. lowed by any of the fol- lowing:. AA direct aspi- ...... 3111 B–2011 ...... 973.53,3 I– ration. 3630–85.2 ICP/AES ...... 200.7, Rev. 4.4 3120 B–2011. (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14.3 (1994). Flame photo- ...... 3500–K B–2011. metric. Electrode ...... 3500–K C–2011. Ion Chroma- ...... D6919–09. tography.

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TABLE IB—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued

58 52 Standard meth- USGS/AOAC/ Parameter Methodology EPA ods ASTM other

53. Residue— Gravimetric, ...... 2540 B–2011 ...... I–3750–85.2 Total, mg/L. 103–105°. 54. Residue—fil- Gravimetric, ...... 2540 C–2011 ... D5907–13 ...... I–1750–85.2 terable, mg/L. 180°. 55. Residue— Gravimetric, ...... 2540 D–2011 ... D5907–13 ...... I–3765–85.2 non-filterable 103–105° (TSS), mg/L. post washing of residue. 56. Residue— Volumetric, ...... 2540 F–2011. settleable, mg/ (Imhoff cone), L. or gravimetric. 57. Residue— Gravimetric, 160.4 (Issued 2540–E–2011 ...... I–3753–85.2 Volatile, mg/L. 550°. 1971) 1. 58. Rhodium— Digestion,4 fol- Total 4, mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011. ration, or. AA furnace ...... 265.2 (Issued 1978) 1. ICP/MS ...... 3125 B–2011. 59. Ruthenium— Digestion,4 fol- Total 4, mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011. ration, or. AA furnace ...... 267.2 1. ICP/MS ...... 3125 B–2011. 60. — Digestion,4 fol- Total 4, mg/L. lowed by any of the fol- lowing: AA furnace ...... 3113 B–2010 ... D3859–08 (B) .. I–4668–98.49 STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES 36 ...... 200.5, Rev 4.2 3120 B–2011 ... D1976–12. (2003) 68; 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4020–05.70 AA gaseous hy- ...... 3114 B–2011, D3859–08 (A) .. I–3667–85.2 dride. or 3114 C– 2011. 61. Silica—Dis- 0.45-micron fil- solved,37 mg/L. tration fol- lowed by any of the fol- lowing: 2 Colorimetric, ...... 4500–SiO2 C– D859–10 ...... I–1700–85. Manual. 2011. 2 Automated ...... 4500–SiO2 E– ...... I–2700–85. (Molybdosilic- 2011 or F– ate). 2011. ICP/AES ...... 200.5, Rev. 4.2 3120 B–2011 ...... I–4471–97.50 (2003) 68; 200.7, Rev. 4.4 (1994).

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TABLE IB—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued

58 52 Standard meth- USGS/AOAC/ Parameter Methodology EPA ods ASTM other

ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14.3 (1994). 62. — Digestion,429 Total,431 mg/L. followed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 or ...... 974.27,3 p. 37,9 ration. 3111 C–2011. I–3720–85.2 AA furnace ...... 3113 B–2010 ...... I–4724–89.51 STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES ...... 200.5, Rev. 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003) 68; 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4471–97.50 DCP ...... See footnote.34 63. Sodium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 ...... 973.54,3 I– ration. 3735–85.2 ICP/AES ...... 200.5, Rev. 4.2 3120 B–2011 ...... I–4471–97.50 (2003) 68; 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14.3 (1994). DCP ...... See footnote.34 Flame photo- ...... 3500–Na B– metric. 2011. Ion Chroma- ...... D6919–09. tography. 64. Specific con- Wheatstone 120.1 (Rev. 2510 B–2011 ... D1125–95(99) 973.40,3 I– ductance, bridge. 1982) 1. (A). 2781–85.2 micromhos/cm at 25 °C. 2¥ 65. Sulfate (as Automated col- 375.2, Rev. 2.0 4500–SO4 SO4), mg/L. orimetric. (1993). F–2011 or G- 2011. 2¥ 3 Gravimetric ...... 4500–SO4 ...... 925.54. C–2011 or D– 2011. 2¥ ...... Turbidimetric ...... 4500–SO4 D516–11. E–2011. Ion Chroma- 300.0, Rev. 2.1 4110 B–2011 or D4327–03 ...... 993.30,3 I– tography. (1993) and C–2011. 4020–05.70 300.1, Rev. 1.0 (1997). CIE/UV ...... 4140 B–2011 ... D6508–1010 .... D6508, Rev. 2.54 66. Sulfide (as Sample ...... 4500–S2¥ B, S), mg/L. Pretreatment. C–2011. Titrimetric (io- ...... 4500–S2¥ F– ...... I–3840–85.2 dine). 2011. Colorimetric ...... 4500–S2¥ D– (methylene 2011. blue). Ion Selective ...... 4500–S2¥ G- D4658–09. Electrode. 2011.

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TABLE IB—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued

58 52 Standard meth- USGS/AOAC/ Parameter Methodology EPA ods ASTM other

2¥ 67. Sulfite (as Titrimetric (io- ...... 4500–SO3 SO3), mg/L. dine-iodate). B–2011. 68. Surfactants, Colorimetric ...... 5540 C–2011 ... D2330–02. mg/L. (methylene blue). 69. Tempera- Thermometric ...... 2550 B–2010 ...... See footnote.32 ture, °C. 70. Thallium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011. ration. AA furnace ...... 279.2 (Issued 3113 B–2010. 1978) 1. STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES ...... 200.7, Rev. 4.4 3120 B–2011 ... D1976–12. (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4471–97.50 71. —Total,4 Digestion,4 fol- mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 ...... I–3850–78.8 ration. AA furnace ...... 3113 B–2010. STGFAA ...... 200.9, Rev. 2.2 (1994). ICP/AES ...... 200.5, Rev. 4.2 (2003) 68; 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14.3 (1994). 72. Titanium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 D–2011. ration. AA furnace ...... 283.2 (Issued 1978) 1. ICP/AES ...... 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14.3 (1994). DCP ...... See footnote.34 73. Turbidity, Nephelometric .. 180.1, Rev. 2.0 2130 B–2011 ... D1889–00 ...... I–3860–85.2 NTU 53. (1993). See foot- note.65 See footnote.66 See foot- note.67 74. Vanadium— Digestion,4 fol- Total,4 mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 D–2011. ration. AA furnace ...... 3113 B–2010 ... D3373–12.

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TABLE IB—LIST OF APPROVED INORGANIC TEST PROCEDURES—Continued

58 52 Standard meth- USGS/AOAC/ Parameter Methodology EPA ods ASTM other

ICP/AES ...... 200.5, Rev. 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003); 68 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4020–05.70 DCP ...... D4190–08 ...... See footnote.34 Colorimetric ...... 3500–V B–2011. (Gallic Acid). 75. —Total,4 Digestion,4 fol- mg/L. lowed by any of the fol- lowing: AA direct aspi- ...... 3111 B–2011 or D1691–12 (A or 974.27,3 p. 37,9 ration 36. 3111 C–2011. B). I–3900–85.2 AA furnace ...... 289.2 (Issued 1978) 1. ICP/AES 36 ...... 200.5, Rev. 4.2 3120 B–2011 ... D1976–12 ...... I–4471–97.50 (2003) 68; 200.7, Rev. 4.4 (1994). ICP/MS ...... 200.8, Rev. 5.4 3125 B–2011 ... D5673–10 ...... 993.14,3 I– (1994). 4020–05.70 DCP 36 ...... D4190–08 ...... See footnote.34 Colorimetric ...... 3500 Zn B– ...... See footnote.33 (Zincon). 2011. 76. Acid Mine ...... 1627 69. Drainage. Table IB Notes: 1 Methods for Chemical Analysis of Water and Wastes, EPA–600/4–79–020. Revised March 1983 and 1979, where applicable. U.S. EPA. 2 Methods for Analysis of Inorganic Substances in Water and Fluvial Sediments, Techniques of Water-Re- source Investigations of the U.S. Geological Survey, Book 5, Chapter A1., unless otherwise stated. 1989. USGS. 3 Official Methods of Analysis of the Association of Official Analytical Chemists, Methods Manual, Sixteenth Edition, 4th Revision, 1998. AOAC International. 4 For the determination of total metals (which are equivalent to total recoverable metals) the sample is not filtered before processing. A digestion procedure is required to solubilize analytes in suspended material and to break down organic-metal complexes (to convert the analyte to a detectable form for colorimetric analysis). For non-platform graphite furnace atomic absorption determinations, a digestion using (as specified in Section 4.1.3 of Methods for the Chemical Analysis of Water and Wastes) is required prior to analysis. The procedure used should subject the sample to gentle, acid refluxing and at no time should the sample be taken to dryness. For direct aspiration flame atomic absorption determinations (FLAA) a combination acid (nitric and hydrochloric acids) digestion is preferred prior to analysis. The approved total recoverable digestion is de- scribed as Method 200.2 in Supplement I of ‘‘Methods for the Determination of Metals in Environmental Sam- ples’’ EPA/600R–94/111, May, 1994, and is reproduced in EPA Methods 200.7, 200.8, and 200.9 from the same Supplement. However, when using the gaseous hydride technique or for the determination of certain elements such as antimony, arsenic, selenium, silver, and tin by non-EPA graphite furnace atomic absorption methods, mercury by cold vapor atomic absorption, the noble metals and titanium by FLAA, a specific or modi- fied sample digestion procedure may be required and in all cases the referenced method write-up should be consulted for specific instruction and/or cautions. For analyses using inductively coupled plasma-atomic emis- sion spectrometry (ICP–AES), the direct current plasma (DCP) technique or EPA spectrochemical techniques (platform furnace AA, ICP–AES, and ICP–MS) use EPA Method 200.2 or an approved alternate procedure (e.g., CEM microwave digestion, which may be used with certain analytes as indicated in Table IB); the total recoverable digestion procedures in EPA Methods 200.7, 200.8, and 200.9 may be used for those respective methods. Regardless of the digestion procedure, the results of the analysis after digestion procedure are re- ported as ‘‘total’’ metals. 5 Copper sulfate or other catalysts that have been found suitable may be used in place of mercuric sulfate.

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6 Manual distillation is not required if comparability data on representative effluent samples are on file to show that this preliminary distillation step is not necessary: However, manual distillation will be required to re- solve any controversies. In general, the analytical method should be consulted regarding the need for distilla- tion. If the method is not clear, the laboratory may compare a minimum of 9 different sample matrices to evaluate the need for distillation. For each matrix, a matrix spike and matrix spike duplicate are analyzed both with and without the distillation step. (A total of 36 samples, assuming 9 matrices). If results are comparable, the laboratory may dispense with the distillation step for future analysis. Comparable is defined as <20% RPD for all tested matrices). Alternatively the two populations of spike recovery percentages may be compared using a recognized statistical test. 7 Industrial Method Number 379–75 WE Ammonia, Automated Electrode Method, Technicon Auto Analyzer II. February 19, 1976. Bran & Luebbe Analyzing Technologies Inc. 8 The approved method is that cited in Methods for Determination of Inorganic Substances in Water and Flu- vial Sediments, Techniques of Water-Resources Investigations of the U.S. Geological Survey, Book 5, Chapter A1. 1979. USGS. 9 American National Standard on Effluents. April 2, 1975. American National Standards Institute. 10 In-Situ Method 1003–8–2009, Biochemical Oxygen Demand (BOD) Measurement by Optical Probe. 2009. In-Situ Incorporated. 11 The use of normal and differential pulse voltage ramps to increase sensitivity and resolution is acceptable. 12 Carbonaceous biochemical oxygen demand (CBOD5) must not be confused with the traditional BOD5 test method which measures ‘‘total 5-day BOD.’’ The addition of the nitrification inhibitor is not a procedural option, but must be included to report the CBOD5 parameter. A discharger whose permit requires reporting the tradi- tional BOD5 may not use a nitrification inhibitor in the procedure for reporting the results. Only when a dis- charger’s permit specifically states CBOD5 is required can the permittee report data using a nitrification inhib- itor. 13 OIC Chemical Oxygen Demand Method. 1978. Oceanography International Corporation. 14 Method 8000, Chemical Oxygen Demand, Hach Handbook of Water Analysis, 1979. Hach Company. 15 The back titration method will be used to resolve controversy. 16 Orion Research Instruction Manual, Residual Chlorine Electrode Model 97–70. 1977. Orion Research In- corporated. The calibration graph for the Orion residual chlorine method must be derived using a reagent blank and three standard solutions, containing 0.2, 1.0, and 5.0 mL 0.00281 N potassium iodate/100 mL solu- tion, respectively. 17 Method 245.7, Mercury in Water by Cold Vapor Atomic Fluorescence Spectrometry, EPA–821–R–05–001. Revision 2.0, February 2005. US EPA. 18 National Council of the Paper Industry for Air and Stream Improvement (NCASI) Technical Bulletin 253 (1971) and Technical Bulletin 803, May 2000. 19 Method 8506, Bicinchoninate Method for Copper, Hach Handbook of Water Analysis. 1979. Hach Com- pany. 20 When using a method with block digestion, this treatment is not required. 21 Industrial Method Number 378–75WA, Hydrogen ion (pH) Automated Electrode Method, Bran & Luebbe (Technicon) Autoanalyzer II. October 1976. Bran & Luebbe Analyzing Technologies. 22 Method 8008, 1,10-Phenanthroline Method using FerroVer Iron Reagent for Water. 1980. Hach Company. 23 Method 8034, Periodate Oxidation Method for Manganese, Hach Handbook of Wastewater Analysis. 1979. Hach Company. 24 Methods for Analysis of Organic Substances in Water and Fluvial Sediments, Techniques of Water-Re- sources Investigations of the U.S. Geological Survey, Book 5, Chapter A3, (1972 Revised 1987). 1987. USGS. 25 Method 8507, , Nitrite-Low Range, Diazotization Method for Water and Wastewater. 1979. Hach Company. 26 Just prior to distillation, adjust the sulfuric-acid-preserved sample to pH 4 with 1 + 9 NaOH. 27 The colorimetric reaction must be conducted at a pH of 10.0 ± 0.2. 28 Addison, R.F., and R.G. Ackman. 1970. Direct Determination of Elemental Phosphorus by Gas-Liquid Chromatography, Journal of Chromatography, 47(3):421–426. 29 Approved methods for the analysis of silver in industrial wastewaters at concentrations of 1 mg/L and above are inadequate where silver exists as an inorganic halide. Silver halides such as the bromide and chlo- ride are relatively insoluble in reagents such as nitric acid but are readily soluble in an aqueous buffer of so- dium and to pH of 12. Therefore, for levels of silver above 1 mg/L, 20 mL of sample should be diluted to 100 mL by adding 40 mL each of 2 M Na2S2O3and NaOH. Standards should be prepared in the same manner. For levels of silver below 1 mg/L the approved method is satisfactory. 30 The use of EDTA decreases method sensitivity. Analysts may omit EDTA or replace with another suitable complexing reagent provided that all method specified quality control acceptance criteria are met. 31 For samples known or suspected to contain high levels of silver (e.g., in excess of 4 mg/L), io- dide should be used to keep the silver in solution for analysis. Prepare a cyanogen solution by adding 4.0 mL of concentrated NH4OH, 6.5 g of KCN, and 5.0 mL of a 1.0 N solution of I2 to 50 mL of reagent water in a volumetric flask and dilute to 100.0 mL. After digestion of the sample, adjust the pH of the digestate to >7 to prevent the formation of HCN under acidic conditions. Add 1 mL of the solution to the sample digestate and adjust the volume to 100 mL with reagent water (NOT acid). If cyanogen iodide is added to sample digestates, then silver standards must be prepared that contain cyanogen iodide as well. Prepare working standards by diluting a small volume of a silver stock solution with water and adjusting the pH>7 with NH4OH. Add 1 mL of the cyanogen iodide solution and let stand 1 hour. Transfer to a 100-mL volumetric flask and dilute to volume with water. 32 ‘‘Water Temperature-Influential Factors, Field Measurement and Data Presentation,’’ Techniques of Water-Resources Investigations of the U.S. Geological Survey, Book 1, Chapter D1. 1975. USGS. 33 Method 8009, Zincon Method for Zinc, Hach Handbook of Water Analysis, 1979. Hach Company. 34 Method AES0029, Direct Current Plasma (DCP) Optical Emission Spectrometric Method for Trace Ele- mental Analysis of Water and Wastes. 1986-Revised 1991. Thermo Jarrell Ash Corporation.

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35 In-Situ Method 1004–8–2009, Carbonaceous Biochemical Oxygen Demand (CBOD) Measurement by Op- tical Probe. 2009. In-Situ Incorporated. 36 Microwave-assisted digestion may be employed for this metal, when analyzed by this methodology. Closed Vessel Microwave Digestion of Wastewater Samples for Determination of Metals. April 16, 1992. CEM Corporation. 37 When determining boron and silica, only , PTFE, or quartz laboratory ware may be used from start until completion of analysis. 38 Only use n-hexane (n-Hexane—85% minimum purity, 99.0% min. saturated C6 isomers, residue less than 1 mg/L) extraction when determining Oil and Grease parameters—Hexane Extractable Material (HEM), or Silica Gel Treated HEM (analogous to EPA Methods 1664 Rev. A and 1664 Rev. B). Use of other extrac- tion is prohibited. 39 Method PAI–DK01, Nitrogen, Total Kjeldahl, Block Digestion, Steam Distillation, Titrimetric Detection. Re- vised December 22, 1994. OI Analytical. 40 Method PAI–DK02, Nitrogen, Total Kjeldahl, Block Digestion, Steam Distillation, Colorimetric Detection. Revised December 22, 1994. OI Analytical. 41 Method PAI–DK03, Nitrogen, Total Kjeldahl, Block Digestion, Automated FIA Gas Diffusion. Revised De- cember 22, 1994. OI Analytical. 42 Method 1664 Rev. B is the revised version of EPA Method 1664 Rev. A. U.S. EPA. February 1999, Revi- sion A. Method 1664, n-Hexane Extractable Material (HEM; Oil and Grease) and Silica Gel Treated n-Hexane Extractable Material (SGT–HEM; Non-polar Material) by Extraction and Gravimetry. EPA–821–R–98–002. U.S. EPA. February 2010, Revision B. Method 1664, n-Hexane Extractable Material (HEM; Oil and Grease) and Silica Gel Treated n-Hexane Extractable Material (SGT–HEM; Non-polar Material) by Extraction and Gravim- etry. EPA–821–R–10–001. 43 Method 1631, Revision E, Mercury in Water by Oxidation, Purge and Trap, and Cold Vapor Atomic Fluo- rescence Spectrometry, EPA–821–R–02–019. Revision E. August 2002, U.S. EPA. The application of clean techniques described in EPA’s Method 1669: Sampling Ambient Water for Trace Metals at EPA Water Quality Criteria Levels, EPA–821–R–96–011, are recommended to preclude contamination at low-level, trace metal determinations. 44 Method OIA–1677–09, Available Cyanide by Ligand Exchange and Flow Injection Analysis (FIA). 2010. OI Analytical. 45 Open File Report 00–170, Methods of Analysis by the U.S. Geological Survey National Water Quality Laboratory—Determination of Ammonium Plus Organic Nitrogen by a Kjeldahl Digestion Method and an Auto- mated Photometric Finish that Includes Digest Cleanup by Gas Diffusion. 2000. USGS. 46 Open File Report 93–449, Methods of Analysis by the U.S. Geological Survey National Water Quality Laboratory—Determination of Chromium in Water by Graphite Furnace Atomic Absorption Spectrophotometry. 1993. USGS. 47 Open File Report 97–198, Methods of Analysis by the U.S. Geological Survey National Water Quality Laboratory—Determination of Molybdenum by Graphite Furnace Atomic Absorption Spectrophotometry. 1997. USGS. 48 Open File Report 92–146, Methods of Analysis by the U.S. Geological Survey National Water Quality Laboratory—Determination of Total Phosphorus by Kjeldahl Digestion Method and an Automated Colorimetric Finish That Includes Dialysis. 1992. USGS. 49 Open File Report 98–639, Methods of Analysis by the U.S. Geological Survey National Water Quality Laboratory—Determination of Arsenic and Selenium in Water and Sediment by Graphite Furnace-Atomic Ab- sorption Spectrometry. 1999. USGS. 50 Open File Report 98–165, Methods of Analysis by the U.S. Geological Survey National Water Quality Laboratory—Determination of Elements in Whole-water Digests Using Inductively Coupled Plasma-Optical Emission Spectrometry and Inductively Coupled Plasma-Mass Spectrometry. 1998. USGS. 51 Open File Report 93–125, Methods of Analysis by the U.S. Geological Survey National Water Quality Laboratory—Determination of Inorganic and Organic Constituents in Water and Fluvial Sediments. 1993. USGS. 52 Unless otherwise indicated, all EPA methods, excluding EPA Method 300.1, are published in U.S. EPA. May 1994. Methods for the Determination of Metals in Environmental Samples, Supplement I, EPA/600/R–94/ 111; or U.S. EPA. August 1993. Methods for the Determination of Inorganic Substances in Environmental Samples, EPA/600/R–93/100. EPA Method 300.1 is US EPA. Revision 1.0, 1997, including errata cover sheet April 27, 1999. Determination of Inorganic in Drinking Water by Ion Chromatography. 53 Styrene divinyl beads (e.g., AMCO–AEPA–1 or equivalent) and stabilized formazin (e.g., Hach StablCalTM or equivalent) are acceptable substitutes for formazin. 54 Method D6508–10, Test Method for Determination of Dissolved Inorganic Anions in Aqueous Matrices Using Capillary Ion Electrophoresis and Chromate Electrolyte. 2010. ASTM. 55 Kelada-01, Kelada Automated Test Methods for Total Cyanide, Acid Dissociable Cyanide, and , EPA 821–B–01–009, Revision 1.2, August 2001. US EPA. Note: A 450–W UV lamp may be used in this method instead of the 550–W lamp specified if it provides performance within the quality control (QC) acceptance criteria of the method in a given instrument. Similarly, modified flow cell configurations and flow conditions may be used in the method, provided that the QC acceptance criteria are met. 56 QuikChem Method 10–204–00–1–X, Digestion and Distillation of Total Cyanide in Drinking and Wastewaters using MICRO DIST and Determination of Cyanide by Flow Injection Analysis. Revision 2.2, March 2005. Lachat Instruments. 57 When using sulfide removal test procedures described in EPA Method 335.4–1, reconstitute particulate that is filtered with the sample prior to distillation. 58 Unless otherwise stated, if the language of this table specifies a sample digestion and/or distillation ‘‘fol- lowed by’’ analysis with a method, approved digestion and/or distillation are required prior to analysis.

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59 Samples analyzed for available cyanide using OI Analytical method OIA–1677–09 or ASTM method D6888–09 that contain particulate matter may be filtered only after the ligand exchange reagents have been added to the samples, because the ligand exchange process converts complexes containing available cyanide to free cyanide, which is not removed by filtration. Analysts are further cautioned to limit the time between the addition of the ligand exchange reagents and sample filtration to no more than 30 minutes to preclude settling of materials in samples. 60 Analysts should be aware that pH optima and chromophore absorption maxima might differ when phenol is replaced by a substituted phenol as the color reagent in Berthelot Reaction (‘‘phenol- reaction’’) colorimetric ammonium determination methods. For example when phenol is used as the color reagent, pH optimum and wavelength of maximum absorbance are about 11.5 and 635 nm, respectively—see, Patton, C.J. and S.R. Crouch. March 1977. Anal. Chem. 49:464–469. These reaction parameters increase to pH > 12.6 and 665 nm when salicylate is used as the color reagent—see, Krom, M.D. April 1980. The Analyst 105:305– 316. 61 If atomic absorption or ICP instrumentation is not available, the aluminon colorimetric method detailed in the 19th Edition of Standard Methods may be used. This method has poorer precision and bias than the meth- ods of choice. 62 Easy (1-Reagent) Nitrate Method, Revision November 12, 2011. Craig Chinchilla. 63 Hach Method 10360, Luminescence Measurement of Dissolved Oxygen in Water and Wastewater and for Use in the Determination of BOD5 and cBOD5. Revision 1.2, October 2011. Hach Company. This method may be used to measure dissolved oxygen when performing the methods approved in Table IB for measurement of biochemical oxygen demand (BOD) and carbonaceous biochemical oxygen demand (CBOD). 64 In-Situ Method 1002–8–2009, Dissolved Oxygen (DO) Measurement by Optical Probe. 2009. In-Situ In- corporated. 65 Mitchell Method M5331, Determination of Turbidity by Nephelometry. Revision 1.0, July 31, 2008. Leck Mitchell. 66 Mitchell Method M5271, Determination of Turbidity by Nephelometry. Revision 1.0, July 31, 2008. Leck Mitchell. 67 Orion Method AQ4500, Determination of Turbidity by Nephelometry. Revision 5, March 12, 2009. Thermo Scientific. 68 EPA Method 200.5, Determination of Trace Elements in Drinking Water by Axially Viewed Inductively Coupled Plasma-Atomic Emission Spectrometry, EPA/600/R–06/115. Revision 4.2, October 2003. US EPA. 69 Method 1627, Kinetic Test Method for the Prediction of Mine Drainage Quality, EPA–821–R–09–002. De- cember 2011. US EPA. 70 Techniques and Methods Book 5–B1, Determination of Elements in Natural-Water, Biota, Sediment and Soil Samples Using Collision/Reaction Cell Inductively Coupled Plasma-Mass Spectrometry, Chapter 1, Sec- tion B, Methods of the National Water Quality Laboratory, Book 5, Laboratory Analysis, 2006. USGS. 71 Water-Resources Investigations Report 01–4132, Methods of Analysis by the U.S. Geological Survey Na- tional Water Quality Laboratory—Determination of Organic Plus Inorganic Mercury in Filtered and Unfiltered Natural Water with Cold Vapor-Atomic Fluorescence Spectrometry, 2001. USGS. 72 USGS Techniques and Methods 5–B8, Chapter 8, Section B, Methods of the National Water Quality Lab- oratory Book 5, Laboratory Analysis, 2011 USGS. 73 NECi Method N07–0003, ’’Nitrate Reductase Nitrate-Nitrogen Analysis,’’ Revision 9.0, March 2014, The Nitrate Elimination Co., Inc. 74 Timberline Instruments, LLC Method Ammonia-001, ‘‘Determination of Inorganic Ammonia by Continuous Flow Gas Diffusion and Conductivity Cell Analysis,’’ June 2011, Timberline Instruments, LLC. 75 Hach Company Method 10206, ‘‘Spectrophotometric Measurement of Nitrate in Water and Wastewater,’’ Revision 2.1, January 2013, Hach Company. 76 Hach Company Method 10242, ‘‘Simplified Spectrophotometric Measurement of Total Kjeldahl Nitrogen in Water and Wastewater,’’ Revision 1.1, January 2013, Hach Company. 77 National Council for Air and Stream Improvement (NCASI) Method TNTP–W10900, ‘‘Total (Kjeldahl) Nitro- gen and Total Phosphorus in Pulp and Paper Biologically Treated Effluent by Alkaline Persulfate Digestion,’’ June 2011, National Council for Air and Stream Improvement, Inc. 78 The pH adjusted sample is to be adjusted to 7.6 for NPDES reporting purposes.

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...... See footnote, OMPOUNDS C RGANIC O ...... See footnote, ESTICIDE ...... See footnote, -P ON N Standard methods ASTM Standard Other ROCEDURES FOR 27 P 1624B ...... 1624B ...... , 1625B ...... 6410 B–2000. 4 4 EST 5 T PPROVED A IST OF IC—L GC/MS ...... HPLC 625.1, 1625B ...... 610 6410 B–2000 ...... GC/MS ...... 6440 B–2005 ...... HPLC ...... 625.1, 1625B ...... See footnote, D4657–92 (98). 610 6410 B–2000 ...... GC/MS ...... 6440 B–2005 ...... 624.1, See footnote, GC/MS D4657–92 (98)...... 624.1, GC/MS ...... HPLC 625.1, 1625B ...... 610 6410 B–2000 ...... GC/MS ...... 6440 B–2005 ...... 624.1, 1624B ...... See footnote, GC/MS D4657–92 (98)...... 6200 B–2011. HPLC 625.1 ...... 605 ...... GC/MS ...... HPLC 625.1, 1625B ...... 610 6410 B–2000 ...... GC/MS ...... 6440 B–2005 ...... HPLC ...... 625.1, 1625B ...... See footnote, D4657–92 (98). 610 6410 B–2000 ...... GC/MS ...... 6440 B–2005 ...... HPLC ...... 625.1, 1625B ...... See footnote, D4657–92 (98). 610 6410 B–2000 ...... GC/MS ...... 6440 B–2005 ...... HPLC ...... 625.1, 1625B ...... See footnote, D4657–92 (98). 610 6410 B–2000 ...... GC/MS ...... 6440 B–2005 ...... HPLC ...... 625.1, 1625B ...... See footnote, D4657–92 (98). 610 6410 B–2000 ...... GC/MS ...... 6440 B–2005 ...... See footnote, GC/MS D4657–92 (98)...... 625.1, 1625B ...... GC/MS ...... 6410 B–2000 ...... 625.1, 1625B ...... 6410 B–2000 See footnote, ...... See footnote, ABLE T Method EPA Method 1 Parameter 1. Acenaphthene ...... GC ...... 6102. Acenaphthylene ...... GC ...... 6103. Acrolein ...... GC ...... 4. ...... 603 ...... GC ...... 5. Anthracene ...... 603 ...... GC ...... 6106. Benzene ...... GC ...... 7. ...... 602 ...... Spectro-photometric 6200 C–2011. 8. Benzo(a)anthracene ...... GC ...... 6109. Benzo(a)pyrene ...... GC ...... 61010. Benzo(b)fluoranthene ...... GC ...... 61011. Benzo(g,h,i)perylene ...... GC ...... 61012. Benzo(k)fluoranthene ...... GC ...... 61013. Benzyl chloride ...... GC ...... 14. Butyl benzyl phthalate ...... GC ...... 15. bis(2-Chloroethoxy) GC 606 ...... 61116. bis(2-Chloroethyl) ether ...... GC ...... 611 ......

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8 8 8 8 8 8 footnote. footnote. footnote. footnote. footnote. footnote. —Continued OMPOUNDS C ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, RGANIC O ESTICIDE -P ON N Standard methods ASTM Standard Other 27 ...... 1625B ...... 6410 B–2000 ...... 1625B ...... 6410 B–2000 See footnote, ...... 1625B ...... 6410 B–2000 See footnote, ...... See footnote, 10 10 5 5 5 ROCEDURES FOR P EST T PPROVED A IST OF GC ...... 611 ...... GC/MS ...... HPLC 625.1, 1625B ...... 610 6410 B–2000 ...... GC/MS ...... 6440 B–2005. HPLC ...... 625.1, 1625B ...... See footnote, ...... 6410 B–2000 ...... GC/MS ...... D4657–92 (98). 625.1, 1625B ...... See footnote, GC/MS ...... 6410 B–2000 ...... 625.1, 1625B ...... GC/MS ...... 6410 B–2000 See footnote, ...... 625.1, ...... GC/MS ...... See footnote, 625.1, GC/MS ...... 625.1, GC/MS ...... 625.1, 1625B ...... 6410 B–2000 ...... GC/MS ...... 625.1 ...... See footnote, 6410 B–2000. GC/MS ...... 625.1 ...... 6410 B–2000. GC/MS ...... 625.1 ...... 6410 B–2000. GC/MS ...... 625.1 ...... 6410 B–2000. GC/MS ...... 625.1 ...... 6410 B–2000. IC—L -dioxin GC/MS ...... 1613B ABLE p T Method EPA Method 1 [also known 12 Parameter as bis(2-Chloro-1-methylethyl) ether]. chloropropane) 78. Naphthalene ...... GC ...... 61079. Nitrobenzene ...... GC ...... 60980. 2- ...... GC ...... 81. 4-Nitrophenol ...... 604 ...... GC ...... 82. N-Nitrosodimethylamine 6420 B–2000...... 604 GC ...... 83. N-Nitrosodi-n-propylamine 6420 B–2000. ... 607 ...... GC ...... 84. N-Nitrosodiphenylamine ...... 607 ...... GC ...... 85. Octachlorodibenzofuran ...... 60786. Octachlorodibenzo- ...... GC/MS ...... 87. 2,2’-oxybis(1- . 1613B 88. PCB–1016 ...... GC ...... 608.389. PCB–1221 ...... GC ...... 608.390. PCB–1232 ...... GC ...... 608.391. PCB–1242 ...... GC ...... 608.392. PCB–1248 ...... GC ...... 608.393. PCB–1254 ...... GC ...... 608.3 ......

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8 footnote...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, .

5a ...... 10 1613B. GC/MS ...... 1613B ...... GC/MS ...... 1613B ...... GC/MS ...... 1613B ...... GC/MS ...... 1613B GC/MS ...... 613, 625.1, GC/MS ...... 625.1 ...... 6410 B–2000. GC/MS ...... 625.1 ...... 6410 B–2000. GC/MS ...... 625.1, 1625B ...... GC/MS ...... 6410 B–2000 ...... HPLC 625.1, 1625B ...... 610 6410 B–2000 ...... See footnote, ...... GC/MS ...... 6440 B–2005 ...... 625.1, 1625B ...... See footnote, GC/MS D4657–92 (98)...... 6410 B–2000 ...... HPLC 625.1, 1625B ...... 610 6410 B–2000 ...... See footnote, ...... 6440 B–2005 ...... See footnote, D4657–92 (98). GC/MS ...... 624.1, 1624B ...... GC/MS ...... 6200 B–2011. 624.1, 1624B ...... GC/MS ...... 6200 B–2011. 624.1, 1624B ...... GC/MS ...... 6200 B–2011. 625.1, 1625B ...... GC/MS ...... 6410 B–2000 ...... 624.1, 1624B ...... GC/MS ...... 6200 B–2011. See footnote, 624.1, 1624B ...... GC/MS ...... 6200 B–2011. 624.1, 1624B ...... GC/MS ...... 6200 B–2011. 624.1 ...... GC/MS ...... 6200 B–2011. 625.1, 1625B ...... GC/MS ...... 6410 B–2000 ...... 624.1, 1624B ...... 6200 B–2011. See footnote, -dioxin. -dioxin. p p dibenzofuran. dibenzofuran. dibenzo- dibenzofuran. dibenzo- 94. PCB–1260 ...... GC ...... 608.395. 1,2,3,7,8-Pentachloro- ...... 96. 2,3,4,7,8-Pentachloro- .. 97. 1,2,3,7,8,-Pentachloro- 98. Pentachlorophenol ...... GC ...... 99. Phenanthrene ...... 604 ...... GC ...... 6420 B–2000 ...... 610100. Phenol ...... GC ...... 101. Pyrene ...... 604 ...... GC ...... 6420 B–2000. 610102. 2,3,7,8-Tetrachloro- ...... 103. 2,3,7,8-Tetrachloro- 104. 1,1,2,2-Tetrachloroethane GC105. Tetrachloroethene ...... 601 GC ...... 106. Toluene ...... 6200 C–2011 601 ...... GC ...... 107. 1,2,4-Trichlorobenzene 6200 C–2011 ...... 602 ...... GC ...... 108. 1,1,1-Trichloroethane 6200 C–2011...... 612 ...... GC ...... 109. 1,1,2-Trichloroethane ...... 601 ...... GC ...... 110. Trichloroethene 6200 C–2011...... 601 ...... GC ...... 111. Trichlorofluoromethane 6200 C–2011 ...... 601 ...... GC ...... 112. 2,4,6-Trichlorophenol 6200 C–2011...... 601 ...... GC ...... 113. Vinyl chloride 6200 C–2011...... 604 ...... GC ...... 6420 B–2000. 601 ...... 6200 C–2011.

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f this part, Definition and Proce- f sample introduction techniques e criteria in the pertinent method, nated Dioxins and Furans by Iso- —Continued as individual PDF files. OMPOUNDS C 65–11. RGANIC O . D7065–11. ESTICIDE https://www.epa.gov/cwa-methods -P ON N three standard deviations around the mean of a minimum five replicate meas- ± Disk. Revised October 28, 1994. 3M Corporation. Standard methods ASTM Standard Other TM 27 ...... ROCEDURES FOR 11 11 P g/L) except for Method 1613B, in which the parameters are expressed picograms per liter (pg/L). 1650 1653 EST μ T . 1981. American Public Health Association (APHA). PPROVED A Coulometric Titra- tion. and GC/MS. IST OF GC/MS ...... GC/MS ...... D7065–11...... and ...... D7065–11. IC—L ABLE T Method EPA Method 1 Parameter -Octylphenol (OP) ...... GC/MS ...... D7065–11. Method 625.1 may be extended to include benzidine, hexachlorocyclopentadiene, N-nitrosodimethylamine, N-nitrosodi-n-propylamine p-tert Analysts may use Fluid Management Systems, Inc. Power-Prep system in place of manual cleanup provided the analyst meets req Method 625.1 screening only. The full text of Methods 601–613, 1613B, 1624B, and 1625B are provided at appendix A, Test Procedures for Analysis Organic P Each analyst must make an initial, one-time demonstration of their ability to generate acceptable precision and accuracy with M Method 624.1 may be used for quantitative determination of acrolein and acrylonitrile, provided that the laboratory has documen Selected Analytical Methods Approved and Cited by the United States Environmental Protection Agency, Supplement to 15th Edi Method O–3116–87 is in Open File Report 93–125, Methods of Analysis by U.S. Geological Survey National Water Quality Laboratory All parameters are expressed in micrograms per liter ( Methods for Benzidine: Chlorinated Organic Compounds, Pentachlorophenol and in Water Wastewater. September 1978. Organochlorine Pesticides and PCBs in Wastewater Using Empore Monoethoxylate (NP1EO). (NP2EO). Halides (AOX). Table IC notes: 1 2 3 4 5 5a 6 7 8 9 10 114. ...... 115. (BPA) GC/MS ...... 116. GC/MS ...... 117. Nonylphenol ...... 118. Nonylphenol Diethoxylate D70 119. Adsorbable Organic 120. Chlorinated Phenolics ...... In Situ Acetylation for the Examination of Water and Wastewater. 1613B (as specified in Section 9 of the method) and permitting authorities. Method 1613, Revision B, Tetra- through Octa-Chlori tope Dilution HRGC/HRMS. Revision B, 1994. U.S. EPA. The full text of this method is provided in appendix A to part and at ods/approved-cwa-methods-organic-compounds ity to detect and quantify these analytes at levels necessary comply with any associated regulations. In addition, the use o other than simple purge-and-trap may be required. QC acceptance criteria from Method 603 should used when analyzing samples in the absence of such criteria Method 624.1. ganic and Organic Constituents in Water Fluvial Sediments. 1993. USGS. 1625B in accordance with procedures each Section 8.2 of these Methods. Additionally, laboratory, on an on-going 10% (5% for Methods 624.1 and 625.1 100% methods 1624B 1625B) of all samples to monitor evaluate laboratory dat with Sections 8.3 and 8.4 of these methods. When the recovery any parameter falls outside quality control (QC) acceptanc analytical results for that parameter in the unspiked sample are suspect. The should be reported but cannot used to ance. If the method does not contain QC acceptance criteria, control limits of urements must be used. These quality control requirements also apply to the Standard Methods, ASTM and other methods c nitrosodiphenylamine. However, when they are known to be present, Methods 605, 607, and 612, or Method 1625B, preferred met ized test procedure to be used determine the method detection limit (MDL) for these procedures is given at appendix B o dure for the Determination of Method Detection Limit. These methods are available at:

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8 4 8 9 6 6 6 6 p. 6 O–3106– 9 nated Phenolics in p. 104; See foot- O–1121–91. O–2060–01. O–1126–95. O–1126–95. O–1126–95. p. 7; See footnote, p. 7; See footnote, p. 83; See footnote, p. 94; See footnote, p. 83; See footnote, p. 83; See footnote, p. 25; See footnote, 3M0222. 3M0222. 3 3 3 3 3 3 3 3 14 12 11 11 11 8 8 of Methods 611, and 1625 p. S64. tnote, ppendix A in part 430 of this 6 footnote, O–3104–83; See footnote, 3M0222. note, 93. O–3106–93; See footnote, p. S60. p. S68. p. S68; See footnote, p. S51. S68. 3M0222. See footnote, See footnote, See footnote, See footnote, .. See footnote, ..... See footnote,

1 ...... See footnote, ESTICIDES P ...... See footnote, ...... See footnote, (02). 96(02). 96(02). 96(02)...... See footnote, ...... See footnote, ...... See footnote, ROCEDURES FOR P EST T PPROVED A Standard methods ASTM Standard Other 2710 IST OF ...... 6410 B–2000 ...... See footnote, 5 ID—L ABLE T GC/MS ...... 625.1 ...... 6410 B–2000. GC/MS ...... 525.2, 625.1 ...... HPLC ...... 632. See GC/MS ...... 625.1. HPLC/MS ...... GC/MS ...... 525.1, 525.2, 625.1 ...... GC/MS ...... 625.1 ...... See foo ...... HPLC ...... GC/MS 632...... 625.1. GC/MS ...... 625.1 GC/MS ...... 625.1 ...... 6410 B–2000. Parameter Method EPA Parameter Method Method 1650, Adsorbable Organic Halides by Adsorption and Coulometric Titration. Revision C, 1997 U.S. EPA. 1653, Chlori -BHC ...... GC ...... 617, 608.3 ...... 6630 B–2007 & C–2007 D3086–90, D5812– The compound was formerly inaccurately labeled as 2,2’-oxybis(2-chloropropane) and bis(2-chloroisopropyl) ether. Some versions -BHC ...... GC ...... 617, 608.3 ...... 6630 B–2007 & C–2007 D3086–90, D5812– d -BHC ...... GC ...... 617, 608.3 ...... 6630 B–2007 & C–2007 D3086–90, D5812– a b 11 12 1. ...... GC ...... 617, 608.3 ...... 2. Ametryn 6630 B–2007 & C–2007 ...... D3086–90, D5812–96 GC ...... 507, 619 ...... 3. ...... TLC ...... 4. Atraton ...... GC ...... 619 ...... 5. ...... GC ...... 507, 619, 608.3 ...... 6. Azinphos methyl ...... GC ...... 614, 622, 16577. Barban ...... TLC ...... 8. 9. 10. Wastewater by In Situ Acetylation and GCMS. Revision A, 1997 U.S. EPA. The full text for both of these methods is provided at a chapter, The Pulp, Paper, and Paperboard Point Source Category. inaccurately list the analyte as ‘‘bis(2-chloroisopropyl)ether,’’ but use correct CAS number of 108–60–1.

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8 8 8 8 8 4 4 4 4 4 6 6 p. 104; See foot- p. 115; See foot- page 27; See foot- O–1126–95. O–2060–01. O–1126–95. O–2060–01. O–1126–95. p. 7; See footnote, p. 7; See footnote, p. 7; See footnote, p. 7; See footnote, p. 7; See footnote, p. 94, See footnote, p. 25; See footnote, p. 7. 3 3 4 3 3 3 3 3 3 3 11 3 12 11 12 11 te, p. S73. p. S64. O–3105–83. 6 6 4 O–3104–83; See footnote, O–3104–83; See footnote, O–3105–83; See footnote, O–3104–83; See footnote, O–3104–83; See footnote, note, note, p. S60. p. S51. note, 3M0222. 3M0222. 3M0222. 3M0222. 3M0222. See footnote, See footnote, See footnote, See footnote, See footnote, See footnote, . See footnote, .. See footnote, ..... See footnote, ...... See footnote, —Continued 1 ...... See footnote, ...... See footnote, ...... See footnote, 96(02). 96(02). 96(02). 96(02). 96(02). 96(02). ESTICIDES P ...... See footnote, ROCEDURES FOR P EST T Standard methods ASTM Standard Other PPROVED A 2710 ...... 6410 B–2000 ...... See footnote, 5 IST OF ID—L ABLE T GC/MS ...... 625.1 ...... 6410 B–2000. GC/MS ...... 625.1 HPLC ...... HPLC/MS 531.1, 632...... GC/MS 553 ...... 625.1 ...... GC/MS ...... 625.1. GC/MS ...... 625.1 ...... 6410 B–2000. HPLC ...... GC/MS 632...... 625.1. HPLC/MS ...... GC/MS ...... 625.1 ...... 6410 B–2000. GC/MS ...... 625.1 ...... 6410 B–2000 ...... GC/MS ...... See footno 625.1 ...... 6410 B–2000. -DDD ...... GC ...... 617, 608.3 ...... 6630 B–2007 & C–2007 -DDE ...... D3086–90, D5812– GC ...... 617, 608.3 ...... -DDT 6630 B–2007 & C–2007 ...... D3086–90, D5812– GC ...... 617, 608.3 ...... 6630 B–2007 & C–2007 D3086–90, D5812– Parameter Method EPA Parameter Method ′ ′ ′ -BHC () ...... GC ...... 617, 608.3 ...... 6630 B–2007 & C–2007 D3086–90, D5812– g 11. 12. Captan ...... GC ...... 13. ...... 617, 608.3 ...... TLC ...... 6630 B–2007 ...... D3086–90, D5812– ...... 14. ..... GC ...... 617, 608.315. ...... 6630 B–2007 GC ...... 617, 608.3 ...... 16. Chloropropham 6630 B–2007 & C–2007 ...... D3086–90, D5812– TLC ...... 17. 2,4-D ...... GC ...... 615 ...... 18. 4,4 6640 B–2006 ...... 19. 4,4 20. 4,4 21. Demeton-O ...... GC ...... 614, 622 ......

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8 8 8 4 4 8 4 6 4 6 p. 6 p. 104; See foot- page 27; See foot- page 27; See foot- page 27; See foot- O–1126–95. O–2060–01. O–1126–95. O–1126–95. O–2060–01. O–2002–01. O–2002–01. p. 7; See footnote, p. 7; See footnote, p. 7; See footnote, p. 7; See footnote, p. 25; See footnote, p. 25; See footnote, p. 25; See footnote, p. 115. p. 7. O–3104–83. 3M0222. 3M0222. 3 4 4 4 3 3 3 3 3 3 3 11 3 12 3 4 11 11 12 13 13 8 8 te, p. S73. p. S73. p. S64. p. S73. 6 6 6 6 footnote, footnote, O–3104–83; See footnote, O–3104–83; See footnote, 3M0222. O–3104–83; See footnote, note, p. S51. O–3104–83; See footnote, S51. p. S51. note, note, note, 3M0222. 3M0222). 3M0222. See footnote, See footnote, See footnote, See footnote, . See footnote, ..... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, 96(02). 96(02). 96(02). 96(02)...... See footnote, ...... See footnote, 6630 B–2007 & C–2007 D3086–90, D5812– ..... 6410 B–2000. 5 ...... 6410 B–2000 ...... See footnote, 6410 B–2000 ...... See footnote, 5 5 608.3. GC/MS ...... 625.1. GC/MS ...... 625.1. GC/MS ...... 525.2, 625.1 ...... HPLC/MS ...... See ...... GC/MS ...... 625.1 ...... 6410 B–2000 ...... GC/MS See footno ...... 525.2, 625.1 ...... HPLC ...... HPLC/MS 632...... See 553 ...... GC/MS ...... 625.1 GC/MS ...... 625.1 GC/MS ...... 625.1 ...... 6410 B–2000. GC/MS ...... 525.1, 525.2, 625.1 GC/MS ...... 625.1. 22. Demeton-S ...... GC ...... 614, 62223. ...... GC ...... 507, 614, 622, 1657 ...... 24. ...... GC ...... 25. Dichlofenthion ...... 615 ...... GC ...... 26. Dichloran ...... 622.1 ...... 27. ...... GC ...... 28. GC ...... 608.2, 617, 608.3 ...... GC 617, 608.3 ...... 6630 B–2007 ...... 617, 608.3 ...... 29. .... 6630 B–2007 & C–2007 ...... D3086–90, D5812– GC ...... 30. ...... 614.1, 1657 ...... GC ...... 507, 614, 622, 1657 .....31. Diuron ...... TLC ...... 32. I ...... GC ...... 617, 608.3 ...... 33. Endosulfan II 6630 B–2007 & C–2007 ...... D3086–90, D5812– GC ...... 617, 608.334. Endosulfan Sulfate ...... 6630 B–2007 & C–2007 GC ...... D3086–90, D5812– 35. ...... 617, 608.3 ...... GC ...... 6630 C–2007 ...... 505, 508, 617, 1656, ...... 36. Endrin ...... GC ...... 37. ...... 617, 608.3 ...... GC ...... 6630 C–2007 ...... 614, 614.1, 1657 ......

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8 8 4 4 4 6 6 6 p. 6 3M0222. 8 O–3104–83; See p. 104; See foot- p. 104; See foot- p. 104; See foot- O–2002–01. O–2060–01. O–2060–01. O–1126–95. O–1126–95. O–2060–01. O–1126–95. p. 7; See footnote, p. 7; See footnote, p. 7; See footnote, p. 25; See footnote, p. 94; See footnote, p. 94; See footnote, 4 3 3 3 3 3 3 3 3 3 13 12 12 11 11 12 11 p. S73. 6 p. S64. p. S64. p. S64. tnote, 6 6 6 O–3104–83; See footnote, O–3104–83; See footnote, O–3104–83; See footnote, note, note, note, p. S51. p. S60. p. S60. S73; See footnote, footnote, 3M0222. 3M0222. See footnote, See footnote, See footnote, . See footnote, .. See footnote, .. See footnote, ..... See footnote, ..... See footnote, ...... See footnote, ...... See footnote, —Continued ...... See footnote, 1 96(02). 96(02). 96(02)...... See footnote, ...... See footnote, ESTICIDES ...... See footnote, P ...... See footnote, ...... See footnote, ROCEDURES FOR P 6630 B–2007 & C–2007 D3086–90, D5812– 6630 B–2007 & C–2007 D3086–90, D5812– EST T Standard methods ASTM Standard Other PPROVED A 2710 IST OF 608.3. 1656, 608.3. ID—L ABLE T GC/MS ...... 625.1 ...... HPLC ...... HPLC/MS 632...... HPLC ...... 632. GC/MS ...... 525.1, 525.2, 625.1 ...... 6410 B–2000. GC/MS ...... 625.1 ...... 6410 B–2000. GC/MS ...... 625.1. HPLC ...... HPLC/MS 632...... GC/MS 553 ...... GC/MS ...... 625.1 ...... HPLC ...... HPLC/MS 632...... GC/MS ...... 525.1, 525.2, 625.1 ...... HPLC ...... GC/MS 632...... See foo 625.1. Parameter Method EPA Parameter Method 38. Fenuron ...... TLC ...... 39. Fenuron-TCA ...... TLC ...... 40. ...... GC ...... 505, 508, 617, 1656, 41. Heptachlor epoxide GC ...... 617, 608.3 ...... 42. Isodrin 6630 B–2007 & C–2007 ...... D3086–90, D5812– GC ...... 617, 608.343. ...... 6630 B–2007 & C–2007 GC ...... 44. ...... GC ...... 614, 165745. ...... 6630 B–2007 TLC ...... 46. ...... GC ...... 505, 508, 608.2, 617, 47. Mexacarbate ...... TLC ......

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4 6 6 6 6 6 O–3106– O–3106– O–3106– 9 9 9 p. 104; See foot- p. 104; See foot- p. 104; See foot- p. 104; See foot- page 27; See foot- page 27; See foot- O–2060–01. O–1126–95. O–1126–95. O–1126–95. O–2002–01. O–2060–01. p. 7; See footnote, p. 83; See footnote, p. 83; See footnote, p. 83; See footnote, p. 94; See footnote, p. 83; See footnote, p. 7. O–3104–83. 3 3 3 3 4 4 3 3 3 3 3 3 12 11 11 3 4 11 13 12 p. S64. p. S64. p. S64. p. 25. p. 25. p. S64. tnote, 6 6 6 3 3 6 footnote, O–3104–83. note, note, note, note, 93. 93. 93. p. S68; See footnote, p. S68; See footnote, p. S68; See footnote, p. S60. p. S68. note, note, See footnote, See footnote, See footnote, .. See footnote, ..... See footnote, ..... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, 0, D5812– ...... See footnote, ...... See footnote, ...... See footnote, 96(02). 96(02). 96(02)...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, ...... See footnote, GC/MS ...... 625.1. HPLC ...... 632. HPLC ...... 632. HPLC ...... HPLC/MS 632...... GC/MS ...... 625.1 ...... GC/MS ...... GC/MS ...... 525.2, 625.1 ...... GC/MS ...... See 525.1, 525.2, 625.1 ...... GC/MS ...... See foo 525.1, 525.2, 625.1. HPLC ...... HPLC/MS 632...... HPLC ...... 632. 48. ...... GC ...... 617, 608.349. Monuron ...... 6630 B–2007 & C–2007 TLC D3086–90, D5812– ...... 50. Monuron-TCA ...... TLC ...... 51. Neburon ...... TLC ...... 52. methyl ..... GC ...... 614, 622, 165753. Parathion ethyl ...... 6630 B–2007 GC ...... 61454. PCNB ...... 6630 B–2007 GC ...... 55. Perthane ...... 608.1, 617, 608.3 ...... GC ...... 6630 B–2007 & C–2007 56. Prometon ...... D3086–90, D5812– 617, 608.3 ...... GC ...... 507, 619 D3086–9 ...... 57. Prometryn ...... GC ...... 507, 619 ...... 58. Propazine ...... GC ...... 507, 619, 1656, 608.3 ..59. Propham ...... TLC ...... 60. ...... TLC ...... 61. Secbumeton ...... TLC ......

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9 6 6 8 O–3106– 9 Standard Methods O–3105–83. p. 104; See foot- p. 104; See foot- p. 115; See foot- p. 115; See foot- O–2060–01. O–1126–95. O–2002–01. O–1126–95. p. 7; See footnote, p. 7; See footnote; 4 p. 83; See footnote, p. 83; See footnote, U.S. EPA. This EPA p. 7. 3 3 3 3 3 3 3 3 12 11 3 13 11 s of the U.S. Geological Table IC of this section, tion of p. S64. p. S64. O–3105–83. O–3105–83. tnote, 6 6 4 4 appendix B of this part, Defini- footnote, O–3106–93. note, note, note, note, 93. p. S68; See footnote, p. S68. See footnote, See footnote, ..... See footnote, ...... See footnote, ...... See footnote, —Continued 1 ...... See footnote, ...... See footnote, ...... See footnote, 96(02). ESTICIDES P ...... See footnote, ...... See footnote, ROCEDURES FOR P ...... See footnote, 6630 B–2007 & C–2007 D3086–90, D5812– 6630 B–2007 ...... See footnote, EST T Standard methods ASTM Standard Other PPROVED A 2710 -BHC, endosulfan I, II, and endrin. However, when they are known to exist, Method 608.3 is the g IST OF 608.3. 608.3. 608.3. 1981. American Public Health Association (APHA). -BHC, a ID—L ABLE T GC ...... 619. HPLC ...... HPLC/MS 632...... GC/MS ...... 525.1, 525.2, 625.1 ...... HPLC ...... See foo 632. GC/MS ...... GC/MS ...... 525.1, 525.2, 625.1 ...... 6410 B–2000. GC/MS ...... 525.2, 625.1 ...... See Parameter Method EPA Parameter Method Pesticides are listed in this table by common name for the convenience of reader. Additional pesticides may be found under The standardized test procedure to be used determine the method detection limit (MDL) for these procedures is given at Methods for Benzidine, Chlorinated Organic Compounds, Pentachlorophenol and Pesticides in Water Wastewater. September 1978. Methods for the Determination of Organic Substances in Water and Fluvial Sediments, Techniques Water-Resources Investigation The method may be extended to include Selected Analytical Methods Approved and Cited by the United States Environmental Protection Agency, Supplement to 15th Edi Table ID notes: 1 2 3 4 5 6 62. Siduron ...... TLC ...... 63. ...... GC ...... 505, 507, 619, 1656, 64. Strobane ...... 65. Swep GC ...... TLC 617, 608.3 ...... 6630 B–2007 & C–2007 66. 2,4,5-T ...... GC ...... 67. 2,4,5-TP (Silvex) ...... 615 ...... GC ...... 6640 B–200668. ...... 615 ...... GC 6640 B–2006 ...... 619, 1656, 608.3 ...... 69. ...... GC ...... 505, 508, 617, 1656, 70. ...... GC ...... 508, 617, 627, 1656, Survey, Book 5, Chapter A3. 1987. USGS. for the Examination of Water and Wastewater. where entries are listed by chemical name. preferred method. tion and Procedure for the Determination of Method Detection Limit. publication includes thin-layer chromatography (TLC) methods.

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2 3 3 l Water Quality Lab- l Water Quality Lab- ory—Determination of tory—Determination of tory—Determination of conventional Pesticides are in Methods for the ectrometry, Revision 2.0, omatography/Mass Spec- ndustrial Wastewater, Vol- A. EPA Method 525.2 is in ity in accordance with Sec- ected-ion monitoring. 1992. y/mass spectrometry. 2001...... p. 81 rol requirements also apply to st spike and analyze 5% of all monitoring. 1995. USGS. Methods 608.3 and 625.1 in ac- for that parameter in the unspiked ASTM USGS

erefore, the two results must be added to obtain n Agency, August 1980. l Survey, Open-File Report 76–177 (1976)...... D1890–90, 96 ...... p. 79 ROCEDURES line P B–00 ...... D1890–90, 96 ...... pp. 75 and 78 EST T Standard Methods On- Reference (method number or page) EST T https://www.epa.gov/cwa-methods/approved-cwa-test-methods-organic- ADIOLOGIC R Standard Methods 18th, 19th, 20th Ed. Disk. Revised October 28, 1994. 3M Corporation. TM

PPROVED 1 A EPA IST OF 900.0 ...... 7110 B ...... Appendix B ...... 7110 B–00 ...... 7110 B ...... D1943–90, 96 ...... 7110 B–00 ...... pp. 75 and 78 D1943–90, 96 ...... p. 79 IE—L ABLE T counter. counter. Proportional or scintillation Proportional counter ...... 903.0 ...... 7500-Ra B ...... 7500-Ra B–01 ...... D2460–90, 97. Scintillation counter ...... 903.1 ...... 7500-Ra C ...... 7500-Ra C–01 ...... D3454–91, 97 .. . Parameter and units Method EPA Methods 608.1, 608.2, 614, 614.1, 615, 617, 619, 622, 622.1, 627, and 632 are found in for the Determination of Non Method O–1126–95 is in Open-File Report 95–181, Methods of Analysis by the U.S. Geological Survey National Water Quality Labora Method O–2060–01 is in Water-Resources Investigations Report 01–4134, Methods of Analysis by the U.S. Geological Survey Nationa Method O–2002–01 is in Water-Resources Investigations Report 01–4098, Methods of Analysis by the U.S. Geological Survey Nationa Method O–1121–91 is in Open-File Report 91–519, Methods of Analysis by the U.S. Geological Survey National Water Quality Labora Each analyst must make an initial, one-time, demonstration of their ability to generate acceptable precision and accuracy with Organochlorine Pesticides and PCBs in Wastewater Using Empore Method O–3106–93 is in Open File Report 94–37, Methods of Analysis by the U.S. Geological Survey National Water Quality Laborat Prescribed Procedures for Measurement of Radioactivity in Drinking Water, EPA–600/4–80–032 (1980), U.S. Environmental Protectio Fishman, M. J. and Brown, Eugene, ‘‘Selected Methods of the U.S. Geological Survey Analysis Wastewaters,’’ Geologica The method found on p. 75 measures only the dissolved portion while 78 suspended portion. Th 7 8 9 10 11 12 13 14 per liter. liter. 1 2 3 1. Alpha-Total, pCi per liter .... Proportional or scintillation 2. Alpha-Counting error, pCi 3. Beta-Total, pCi per liter ...... 4. Beta-Counting error, pCi Proportional counter ...... 5. (a) Radium Total pCi per Proportional counter 900.0 ...... Appendix B(b) Ra, pCi per liter 7110 B ...... 7110 B 7110 ...... 7110 B–00 .. and Other Nitrogen-Containing Compounds by Gas Chromatography With Nitrogen Phosphorus Detectors. 1994. USGS. pesticides in water by C–18 solid-phase extraction and capillary-column gas chromatography/mass spectrometry with selected-ion organonitrogen in water by solid-phase extraction and capillary-column gas chromatography/mass spectrometry with sel USGS. in Municipal and Industrial Wastewater, EPA 821–R–92–002, April 1992, U.S. EPA. Methods 505, 507, 508, 525.1, 531.1 553 Determination of Nonconventional Pesticides in Municipal and Industrial Wastewater, Volume II, EPA 821–R–93–010B, 1993, U.S. EP Determination of Organic Compounds in Drinking Water by Liquid-Solid Extraction and Capillary Column Gas Chromatography/Mass Sp ume I, EPA 821–R–93–010A, 1993, U.S. EPA. Methods 608.3 and 625.1 are available at compounds 1995, U.S. EPA. EPA methods 1656 and 1657 are in Methods for the Determination of Nonconventional Pesticides Municipal I trometry. 2001. USGS. oratory—Determination of Pesticides in Water by Graphitized Carbon-Based Solid-Phase Extraction and High-Performance Liquid Chr oratory—Determination of moderate-use pesticides in water by C–18 solid-phase extraction and capillary-column gas chromatograph USGS. ‘‘total.’’ cordance with procedures given in Section 8.2 of each these methods. Additionally, laboratory, on an on-going basis, mu samples analyzed with Method 608.3 or 5% of all 625.1 to monitor and evaluate laboratory data qual tions 8.3 and 8.4 of these methods. When the recovery any parameter falls outside warning limits, analytical results sample are suspect. The results should be reported, but cannot used to demonstrate regulatory compliance. These quality cont the Standard Methods, ASTM and other methods cited.

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TABLE IF—LIST OF APPROVED METHODS FOR PHARMACEUTICAL POLLUTANTS

CAS registry Pharmaceuticals pollutants No. Analytical method number

Acetonitrile ...... 75–05–8 1666/1671/D3371/D3695/624.1 n-Amyl acetate ...... 628–63–7 1666/D3695 n-Amyl alcohol ...... 71–41–0 1666/D3695 Benzene ...... 71–43–2 D4763/D3695/502.2/524.2/624.1 n-Butyl-acetate ...... 123–86–4 1666/D3695 tert-Butyl alcohol ...... 75–65–0 1666/624.1 Chlorobenzene ...... 108–90–7 502.2/524.2/624.1 Chloroform ...... 67–66–3 502.2/524.2/551/624.1 o-Dichlorobenzene ...... 95–50–1 1625C/502.2/524.2/624.1 1,2-Dichloroethane ...... 107–06–2 D3695/502.2/524.2/624.1 Diethylamine ...... 109–89–7 1666/1671 ...... 67–68–5 1666/1671 ...... 64–17–5 1666/1671/D3695/624.1 Ethyl acetate ...... 141–78–6 1666/D3695/624.1 n- ...... 142–82–5 1666/D3695 n-Hexane ...... 110–54–3 1666/D3695 Isobutyraldehyde ...... 78–84–2 1666/1667 Isopropanol ...... 67–63–0 1666/D3695 Isopropyl acetate ...... 108–21–4 1666/D3695 Isopropyl ether ...... 108–20–3 1666/D3695 Methanol ...... 67–56–1 1666/1671/D3695/624.1 Methyl Cellosolve® (2-Methoxy ethanol) ...... 109–86–4 1666/1671 Methylene chloride ...... 75–09–2 502.2/524.2/624.1 Methyl formate ...... 107–31–3 1666 4-Methyl-2-pentanone (MIBK) ...... 108–10–1 1624C/1666/D3695/D4763/524.2/624.1 Phenol ...... 108–95–2 D4763 n-Propanol ...... 71–23–8 1666/1671/D3695/624.1 2-Propanone () ...... 67–64–1 D3695/D4763/524.2/624.1 Tetrahydrofuran ...... 109–99–9 1666/524.2/624.1 Toluene ...... 108–88–3 D3695/D4763/502.2/524.2/624.1 Triethlyamine ...... 121–44–8 1666/1671 Xylenes ...... (Note 1) 1624C/1666/624.1 Table IF note: 1 1624C: m-xylene 108–38–3, o,p-xylene, E–14095 (Not a CAS number; this is the number provided in the Environmental Monitoring Methods Index [EMMI] database.); 1666: m,p-xylene 136777–61–2, o-xylene 95–47–6.

TABLE IG—TEST METHODS FOR ACTIVE INGREDIENTS [40 CFR part 455]

EPA survey code Pesticide name CAS No. EPA analytical method No.(s) 3

8 ...... Triadimefon ...... 43121–43–3 507/633/525.1/525.2/1656/625.1. 12 ...... ...... 62–73–7 1657/507/622/525.1/525.2/625.1. 16 ...... 2,4-D; 2,4-D and Esters [2,4- 94–75–7 1658/515.1/615/515.2/555. Dichloro-phenoxyacetic acid]. 17 ...... 2,4-DB; 2,4-DB Salts and Esters [2,4- 94–82–6 1658/515.1/615/515.2/555. Dichlorophenoxybutyric acid]. 22 ...... ...... 7786–34–7 1657/507/622/525.1/525.2/625.1. 25 ...... ...... 21725–46–2 629/507/608.3/625.1. 26 ...... ...... 1918–16–7 1656/508/608.1/525.1/525.2/608.3/625.1. 27 ...... MCPA; MCPA Salts and Esters ...... 94–74–6 1658/615/555. [2-Methyl-4-chlorophenoxyacetic acid] ...... 30 ...... ; Dichlorprop Salts and Esters 120–36–5 1658/515.1/615/515.2/555. [2-(2,4-Dichlorophenoxy) propionic acid]. 31 ...... MCPP; MCPP Salts and Esters [2-(2-Meth- 93–65–2 1658/615/555. yl-4-chlorophenoxy) propionic acid]. 35 ...... TCMTB [2-(Thiocyanomethylthio) benzo- 21564–17–0 637. thiazole]. 39 ...... Pronamide ...... 23950–58–5 525.1/525.2/507/633.1/625.1. 41 ...... ...... 709–98–8 632.1/1656/608.3. 45 ...... ...... 21087–64–9 507/633/525.1/525.2/1656/608.3/625.1. 52 ...... ...... 30560–19–1 1656/1657/608.3. 53 ...... ...... 50594–66–6 515.1/515.2/555. 54 ...... ...... 15972–60–8 505/507/645/525.1/525.2/1656/608.3/ 625.1. 55 ...... ...... 116–06–3 531.1. 58 ...... Ametryn ...... 834–12–8 507/619/525.2/625.1. 60 ...... Atrazine ...... 1912–24–9 505/507/619/525.1/525.2/1656/ 608.3/ 625.1. 62 ...... Benomyl ...... 17804–35–2 631.

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TABLE IG—TEST METHODS FOR PESTICIDE ACTIVE INGREDIENTS—Continued [40 CFR part 455]

EPA survey code Pesticide name CAS No. EPA analytical method No.(s) 3

68 ...... Bromacil; Bromacil Salts and Esters ...... 314–40–9 507/633/525.1/525.2/1656/608.3/625.1. 69 ...... ...... 1689–84–5 1625/1661/625.1. 69 ...... Bromoxynil Octanoate ...... 1689–99–2 1656/608.3. 70 ...... ...... 23184–66–9 507/645/525.1/525.2/1656/608.3/625.1. 73 ...... Captafol ...... 2425–06–1 1656/608.3/625.1. 75 ...... Carbaryl [Sevin] ...... 63–25–2 531.1/632/553/625.1. 76 ...... ...... 1563–66–2 531.1/632/625.1. 80 ...... Chloroneb ...... 2675–77–6 1656/508/608.1/525.1/525.2/608.3/625.1. 82 ...... Chlorothalonil ...... 1897–45–6 508/608.2/525.1/525.2/1656/608.3/625.1. 84 ...... Stirofos ...... 961–11–5 1657/507/622/525.1/525.2/625.1. 86 ...... ...... 2921–88–2 1657/508/622/625.1. 90 ...... ...... 51630–58–1 1660. 103 ...... Diazinon ...... 333–41–5 1657/507/614/622/525.2/625.1. 107 ...... ...... 298–00–0 1657/614/622/625.1. 110 ...... DCPA [Dimethyl 2,3,5,6-tetrachloro- 1861–32–1 508/608.2/525.1/525.2/515.1 2/515.2 2/ terephthalate]. 1656/608.3/625.1. 112 ...... ...... 88–85–7 1658/515.1/615/515.2/555/625.1. 113 ...... Dioxathion ...... 78–34–2 1657/614.1. 118 ...... Nabonate [Disodium cyanodithio- 138–93–2 630.1. imidocarbonate]. 119 ...... Diuron ...... 330–54–1 632/553. 123 ...... ...... 145–73–3 548/548.1. 124 ...... Endrin ...... 72–20–8 1656/505/508/617/525.1/525.2/608.3/ 625.1. 125 ...... Ethalfluralin ...... 55283–68–6 1656/627/608.3 See footnote 1. 126 ...... Ethion ...... 563–12–2 1657/614/614.1/625.1. 127 ...... Ethoprop ...... 13194–48–4 1657/507/622/525.1/525.2/625.1. 132 ...... Fenarimol ...... 60168–88–9 507/633.1/525.1/525.2/1656/608.3/625.1. 133 ...... ...... 55–38–9 1657/622/625.1. 138 ...... [N-(Phosphonomethyl) glycine] 1071–83–6 547. 140 ...... Heptachlor ...... 76–44–8 1656/505/508/617/525.1/525.2/608.3/ 625.1. 144 ...... Isopropalin ...... 33820–53–0 1656/627/608.3. 148 ...... Linuron ...... 330–55–2 553/632. 150 ...... Malathion ...... 121–75–5 1657/614/625.1. 154 ...... ...... 10265–92–6 1657. 156 ...... ...... 16752–77–5 531.1/632. 158 ...... Methoxychlor ...... 72–43–5 1656/505/508/608.2/617/525.1/525.2/ 608.3/625.1. 172 ...... Nabam ...... 142–59–6 630/630.1. 173 ...... ...... 300–76–5 1657/622/625.1. 175 ...... Norflurazon ...... 27314–13–2 507/645/525.1/525.2/1656/608.3/625.1. 178 ...... ...... 1861–40–1 1656/627/608.3 See footnote 1. 182 ...... Fensulfothion ...... 115–90–2 1657/622/625.1. 183 ...... Disulfoton ...... 298–04–4 1657/507/614/622/525.2/625.1. 185 ...... ...... 732–11–6 1657/622.1/625.1. 186 ...... Azinphos Methyl ...... 86–50–0 1657/614/622/625.1. 192 ...... Organo-tin pesticides ...... 12379–54–3 Ind-01/200.7/200.9. 197 ...... Bolstar ...... 35400–43–2 1657/622. 203 ...... Parathion ...... 56–38–2 1657/614/625.1. 204 ...... ...... 40487–42–1 1656. 205 ...... Pentachloronitrobenzene ...... 82–68–8 1656/608.1/617/608.3/625.1. 206 ...... Pentachlorophenol ...... 87–86–5 1625/515.2/555/515.1/525.1/525.2/625.1. 208 ...... ...... 52645–53–1 608.2/508/525.1/525.2/1656/1660/608.3 4/ 625.1 4. 212 ...... ...... 298–02–2 1657/622/625.1. 218 ...... Busan 85 [Potassium 128–03–0 630/630.1. dimethyldithiocarbamate]. 219 ...... Busan 40 [Potassium N-hydroxymethyl-N- 51026–28–9 630/630.1. methyldithiocarbamate]. 220 ...... KN Methyl [Potassium N-methyl- 137–41–7 630/630.1. dithiocarbamate]. 223 ...... Prometon ...... 1610–18–0 507/619/525.2/625.1. 224 ...... Prometryn ...... 7287–19–6 507/619/525.1/525.2/625.1. 226 ...... Propazine ...... 139–40–2 507/619/525.1/525.2/1656/608.3/625.1. 230 ...... I ...... 121–21–1 1660. 232 ...... Pyrethrin II ...... 121–29–9 1660. 236 ...... DEF [S,S,S-Tributyl phosphorotrithioate] .... 78–48–8 1657. 239 ...... Simazine ...... 122–34–9 505/507/619/525.1/525.2/1656/608.3/ 625.1. 241 ...... Carbam-S [Sodium dimethyldithio-carba- 128–04–1 630/630.1. mate].

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TABLE IG—TEST METHODS FOR PESTICIDE ACTIVE INGREDIENTS—Continued [40 CFR part 455]

EPA survey code Pesticide name CAS No. EPA analytical method No.(s) 3

243 ...... Vapam [Sodium methyldithiocarbamate] .... 137–42–8 630/630.1. 252 ...... ...... 34014–18–1 507/525.1/525.2/625.1. 254 ...... Terbacil ...... 5902–51–2 507/633/525.1/525.2/1656/608.3/625.1. 255 ...... ...... 13071–79–9 1657/507/614.1/525.1/525.2/625.1. 256 ...... Terbuthylazine ...... 5915–41–3 619/1656/608.3. 257 ...... Terbutryn ...... 886–50–0 507/619/525.1/525.2/625.1. 259 ...... Dazomet ...... 533–74–4 630/630.1/1659. 262 ...... Toxaphene ...... 8001–35–2 1656/505/508/617/525.1/525.2/608.3/ 625.1. 263 ...... Merphos [Tributyl phosphorotrithioate] ...... 150–50–5 1657/507/525.1/525.2/622/625.1. 264 ...... Trifluralin 1 ...... 1582–09–8 1656/508/617/627/525.2/608.3/625.1. 268 ...... Ziram [Zinc dimethyldithiocarbamate] ...... 137–30–4 630/630.1. Table IG notes: 1 Monitor and report as total Trifluralin. 2 Applicable to the analysis of DCPA degradates. 3 EPA Methods 608.1 through 645, 1645 through 1661, and Ind-01 are available in Methods for the Determination of Non- conventional Pesticides in Municipal and Industrial Wastewater, Volume I, EPA 821–R–93–010A, Revision I, August 1993, U.S. EPA. EPA Methods 200.9 and 505 through 555 are available in Methods for the Determination of Nonconventional Pesticides in Municipal and Industrial Wastewater, Volume II, EPA 821–R–93–010B, August 1993, U.S. EPA. The full text of Methods 608.3, 625.1, and 1625 are provided at appendix A of this part. The full text of Method 200.7 is provided at appendix C of this part. Methods 608.3 and 625.1 are available at https://www.epa.gov/cwa-methods/approved-cwa-test-methods-organic-compounds. 4 Permethrin is not listed within methods 608.3 and 625.1; however, cis-permethrin and trans-permethrin are listed. Permethrin can be calculated by adding the results of cis- and trans-permethrin.

TABLE IH—LIST OF APPROVED MICROBIOLOGICAL METHODS FOR AMBIENT WATER

Parameter and Standard meth- AOAC, Method 1 EPA ASTM, Other units ods USGS

Bacteria

1. Coliform Most Probable p. 132 3 ...... 9221 C E–2006. (fecal), number Number per 100 mL or (MPN), 5 number per tube, 3 dilu- gram dry tion, or. weight. Membrane filter p. 124 3 ...... 9222 D–2006 27 B–0050– (MF),2 single 85. 4 step. 2. Coliform MPN, 5 tube, 3 p. 132 3 ...... 9221 C E–2006. (fecal) in pres- dilution, or. ence of chlo- rine, number per 100 mL. MF,2 single p. 124 3 ...... 9222 D– step 5. 2006. 27 3. Coliform MPN, 5 tube, 3 p. 114 3 ...... 9221 B–2006. (total), number dilution, or. per 100 mL. MF,2 single step p. 108 3 ...... 9222 B–2006 ... B–0025– or two step. 85. 4 4. Coliform MPN, 5 tube, 3 p. 114 3 ...... 9221 B–2006. (total), in pres- dilution, or. ence of chlo- rine, number per 100 mL. MF 2 with en- p. 111 3 ...... 9222 B–2006. richment. 5. E. coli, num- MPN,6814 mul- ...... 9221 B.2–2006/ ber per 100 tiple tube, or. 9221 F– mL. 2006 11 13 Multiple tube/ ...... 9223 B–2004 12 991.15 10 Colilert®,12 16 multiple well, Colilert- or. 18®.12 15 16

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TABLE IH—LIST OF APPROVED MICROBIOLOGICAL METHODS FOR AMBIENT WATER—Continued

Parameter and Standard meth- AOAC, Method 1 EPA ASTM, Other units ods USGS

MF,25678 two 1103.1 19 ...... 9222 B–2006/ D–5392– step, or. 9222 G– 93. 9 2006,18 9213 D–2007. Single step ...... 1603,20 1604 21 ...... mColiBlue- 24®.17 6. Fecal MPN, 5 tube, 3 p. 139 3 ...... 9230 B–2007. streptococci, dilution, or. number per 100 mL. MF 2, or ...... p. 136 3 ...... 9230 C–2007 ... B–0055– 85 4. Plate count ...... p. 143. 3 7. Enterococci, MPN,68 multiple ...... 9230 D–2007 ... D6503– Enterolert®.12 22 number per tube/multiple 99 9. 100 mL. well, or. MF 25678 two 1106.1 23 ...... 9230 C–2007 ... D5259– step, or. 92. 9 Single step, or 1600 24 ...... 9230 C–2007. Plate count ...... p. 143. 3

Protozoa

8. Filtration/IMS/ 1622, 25 1623. 26 Cryptosporidiu- FA. m. 9. Giardia ...... Filtration/IMS/ 1623. 26 FA. Table IH notes: 1 The method must be specified when results are reported. 2 A 0.45-μm membrane filter (MF) or other pore size certified by the manufacturer to fully retain organisms to be cultivated and to be free of extractables which could interfere with their growth. 3 Microbiological Methods for Monitoring the Environment, Water, and Wastes. EPA/600/8–78/017. 1978. U.S. EPA. 4 U.S. Geological Survey Techniques of Water-Resource Investigations, Book 5, Laboratory Analysis, Chap- ter A4, Methods for Collection and Analysis of Aquatic Biological and Microbiological Samples. 1989. USGS. 5 Because the MF technique usually yields low and variable recovery from chlorinated wastewaters, the Most Probable Number method will be required to resolve any controversies. 6 Tests must be conducted to provide organism enumeration (density). Select the appropriate configuration of tubes/filtrations and dilutions/volumes to account for the quality, character, consistency, and anticipated or- ganism density of the water sample. 7 When the MF method has not been used previously to test waters with high turbidity, large numbers of noncoliform bacteria, or samples that may contain organisms stressed by chlorine, a parallel test should be conducted with a multiple-tube technique to demonstrate applicability and comparability of results. 8 To assess the comparability of results obtained with individual methods, it is suggested that side-by-side tests be conducted across seasons of the year with the water samples routinely tested in accordance with the most current Standard Methods for the Examination of Water and Wastewater or EPA alternate test procedure (ATP) guidelines. 9 Annual Book of ASTM Standards—Water and Environmental Technology. Section 11.02. 2000, 1999, 1996. ASTM International. 10 Official Methods of Analysis of AOAC International, 16th Edition, Volume I, Chapter 17. 1995. AOAC International. 11 The multiple-tube fermentation test is used in 9221B.2–2006. Lactose broth may be used in lieu of lauryl tryptose broth (LTB), if at least 25 parallel tests are conducted between this broth and LTB using the water samples normally tested, and this comparison demonstrates that the false-positive rate and false-negative rate for total coliform using lactose broth is less than 10 percent. No requirement exists to run the completed phase on 10 percent of all total coliform-positive tubes on a seasonal basis. 12 These tests are collectively known as defined enzyme substrate tests, where, for example, a substrate is used to detect the enzyme b-glucuronidase produced by E. coli. 13 After prior enrichment in a presumptive medium for total coliform using 9221B.2–2006, all presumptive tubes or bottles showing any amount of gas, growth or acidity within 48 h ± 3 h of incubation shall be sub- mitted to 9221F–2006. Commercially available EC–MUG media or EC media supplemented in the laboratory with 50 μg/mL of MUG may be used.

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14 Samples shall be enumerated by the multiple-tube or multiple-well procedure. Using multiple-tube proce- dures, employ an appropriate tube and dilution configuration of the sample as needed and report the Most Probable Number (MPN). Samples tested with Colilert® may be enumerated with the multiple-well procedures, Quanti-Tray® or Quanti-Tray®/2000, and the MPN calculated from the table provided by the manufacturer. 15 Colilert-18® is an optimized formulation of the Colilert® for the determination of total coliforms and E. coli that provides results within 18 h of incubation at 35 °C, rather than the 24 h required for the Colilert® test, and is recommended for marine water samples. 16 Descriptions of the Colilert®, Colilert-18®, and Quanti-Tray® may be obtained from IDEXX Laboratories Inc. 17 A description of the mColiBlue24® test may be obtained from Hach Company. 18 Subject total coliform positive samples determined by 9222B–2006 or other membrane filter procedure to 9222G–2006 using NA–MUG media. 19 Method 1103.1: Escherichia coli (E. coli) in Water by Membrane Filtration Using membrane- Thermotolerant Escherichia coli Agar (mTEC), EPA–821–R–10–002. March 2010. U.S. EPA. 20 Method 1603: Escherichia coli (E. coli) in Water by Membrane Filtration Using Modified membrane- Thermotolerant Escherichia coli Agar (Modified mTEC), EPA–821–R–14–010. September 2014. U.S. EPA. 21 Preparation and use of MI agar with a standard membrane filter procedure is set forth in the article, Bren- ner et al. 1993. New Medium for the Simultaneous Detection of Total Coliform and Escherichia coli in Water. Appl. Environ. Microbiol. 59:3534–3544 and in Method 1604: Total Coliforms and Escherichia coli (E. coli) in Water by Membrane Filtration by Using a Simultaneous Detection Technique (MI Medium), EPA 821–R–02– 024, September 2002, U.S. EPA. 22 A description of the Enterolert® test may be obtained from IDEXX Laboratories Inc. 23 Method 1106.1: Enterococci in Water by Membrane Filtration Using membrane-Enterococcus-Esculin Iron Agar (mE-EIA), EPA–821–R–09–015. December 2009. U.S. EPA. 24 Method 1600: Enterococci in Water by Membrane Filtration Using membrane-Enterococcus Indoxyl-b-D- Glucoside Agar (mEI), EPA–821–R–14–011. September 2014. U.S. EPA. 25 Method 1622 uses a filtration, concentration, immunomagnetic separation of oocysts from captured mate- rial, immunofluorescence assay to determine concentrations, and confirmation through vital dye staining and differential interference contrast microscopy for the detection of Cryptosporidium. Method 1622: Cryptosporidium in Water by Filtration/IMS/FA, EPA–821–R–05–001. December 2005. U.S. EPA. 26 Method 1623 uses a filtration, concentration, immunomagnetic separation of oocysts and cysts from cap- tured material, immunofluorescence assay to determine concentrations, and confirmation through vital dye staining and differential interference contrast microscopy for the simultaneous detection of Cryptosporidium and Giardia oocysts and cysts. Method 1623: Cryptosporidium and Giardia in Water by Filtration/IMS/FA. EPA– 821–R–05–002. December 2005. U.S. EPA. 27 On a monthly basis, at least ten blue colonies from the medium must be verified using Lauryl Tryptose Broth and EC broth, followed by count adjustment based on these results; and representative non-blue colo- nies should be verified using Lauryl Tryptose Broth. Where possible, verifications should be done from ran- domized sample sources.

(b) Certain material is incorporated (i) Microbiological Methods for Moni- by reference into this part with the ap- toring the Environment, Water, and proval of the Director of the Federal Wastes. 1978. EPA/600/8–78/017, Pub. No. Register under 5 U.S.C. 552(a) and 1 PB–290329/A.S. CFR part 51. All approved material is (A) Part III Analytical Methodology, available for inspection at EPA’s Water Section B Total Coliform Methods, Docket, EPA West, 1301 Constitution page 108. Table IA, Note 3; Table IH, Avenue NW., Room 3334, Washington, Note 3. (B) Part III Analytical Methodology, DC 20004, Telephone: 202–566–2426, and is Section B Total Coliform Methods, 2.6.2 available from the sources listed below. Two-Step Enrichment Procedure, page It is also available for inspection at the 111. Table IA, Note 3; Table IH, Note 3. National Archives and Records Admin- (C) Part III Analytical Methodology, istration (NARA). For information on Section B Total Coliform Methods, 4 the availability of this material at Most Probable Number (MPN) Method, NARA, call 202–741–6030, or go to: page 114. Table IA, Note 3; Table IH, https://www.archives.gov/federal-register/ Note 3. cfr/ibr-locations.html. (D) Part III Analytical Methodology, (1) Environmental Monitoring and Section C Fecal Coliform Methods, 2 Support Laboratory, U.S. Environ- Direct Membrane Filter (MF) Method, mental Protection Agency, Cincinnati page 124. Table IA, Note 3; Table IH, OH (US EPA). Available at http:// Note 3. water.epa.gov/scitech/methods/cwa/ (E) Part III, Analytical Methodology, index.cfm or from: National Technical Section C Fecal Coliform Methods, 5 Information Service, 5285 Port Royal Most Probable Number (MPN) Method, page 132. Table IA, Note 3; Table IH, Road, Springfield, Virginia 22161 Note 3.

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(F) Part III Analytical Methodology, (E) Method 351.2, Determination of Section D Fecal Streptococci, 2 Mem- Total Kjeldahl Nitrogen by Semi-Auto- brane Filter (MF) Method, page 136. mated Colorimetry. Revision 2.0. Table Table IA, Note 3; Table IH, Note 3. IB, Note 52. (G) Part III Analytical Methodology, (F) Method 353.2, Determination of Section D Fecal Streptococci, 4 Most Nitrate-Nitrite Automated Colorim- Probable Number Method, page 139. etry. Revision 2.0. Table IB, Note 52. Table IA, Note 3; Table IH, Note 3. (G) Method 365.1, Determination of (H) Part III Analytical Methodology, Phosphorus by Automated Colorim- Section D Fecal Streptococci, 5 Pour etry. Revision 2.0. Table IB, Note 52. Plate Method, page 143. Table IA, Note (H) Method 375.2, Determination of 3; Table IH, Note 3. Sulfate by Automated Colorimetry. Re- (ii) [Reserved] vision 2.0. Table IB, Note 52. (2) Environmental Monitoring and (I) Method 410.4, Determination of Support Laboratory, U.S. Environ- Chemical Oxygen Demand by Semi- mental Protection Agency, Cincinnati Automated Colorimetry. Revision 2.0. OH (US EPA). Available at http:// Table IB, Note 52. water.epa.gov/scitech/methods/cwa/ (ii) Methods for the Determination of index.cfm. Metals in Environmental Samples, (i) Method 300.1 (including Errata Supplement I. May 1994. EPA/600/R–94/ Cover Sheet, April 27, 1999), Determina- 111, Pub. No. PB 95125472. Table IB, tion of Inorganic Ions in Drinking Note 52. Water by Ion Chromatography, Revi- (A) Method 200.7, Determination of sion 1.0, 1997. Table IB, Note 52. Metals and Trace Elements in Water (ii) Method 551, Determination of and Wastes by Inductively Coupled Chlorination Disinfection Byproducts Plasma-Atomic Emission Spectrom- and Chlorinated Solvents in Drinking etry. Revision 4.4. Table IB, Note 52. Water by Liquid-Liquid Extraction and Gas Chromatography With Electron- (B) Method 200.8, Determination of Capture Detection. 1990. Table IF. Trace Elements in Water and Wastes (3) National Exposure Risk Labora- by Inductively Coupled Plasma Mass tory-Cincinnati, U.S. Environmental Spectrometry. Revision 5.3. Table IB, Protection Agency, Cincinnati OH (US Note 52. EPA). Available from http:// (C) Method 200.9, Determination of water.epa.gov/scitech/methods/cwa/ Trace Elements by Stabilized Tempera- index.cfm or from the National Tech- ture Graphite Furnace Atomic Absorp- nical Information Service (NTIS), 5285 tion Spectrometry. Revision 2.2. Table Port Royal Road, Springfield, VA 22161. IB, Note 52. Telephone: 800–553–6847. (D) Method 218.6, Determination of (i) Methods for the Determination of Dissolved Hexavalent Chromium in Inorganic Substances in Environ- Drinking Water, Groundwater, and In- mental Samples. August 1993. EPA/600/ dustrial Wastewater Effluents by Ion R–93/100, Pub. No. PB 94120821. Table Chromatography. Revision 3.3. Table IB, Note 52. IB, Note 52. (A) Method 180.1, Determination of (E) Method 245.1, Determination of Turbidity by Nephelometry. Revision Mercury in Water by Cold Vapor Atom- 2.0. Table IB, Note 52. ic Absorption Spectrometry. Revision (B) Method 300.0, Determination of 3.0. Table IB, Note 52. Inorganic Anions by Ion Chroma- (4) National Exposure Risk Labora- tography. Revision 2.1. Table IB, Note tory-Cincinnati, U.S. Environmental 52. Protection Agency, Cincinnati OH (US (C) Method 335.4, Determination of EPA). Available at http://water.epa.gov/ Total Cyanide by Semi-Automated Col- scitech/methods/cwa/index.cfm. orimetry. Revision 1.0. Table IB, Notes (i) EPA Method 200.5, Determination 52 and 57. of Trace Elements in Drinking Water (D) Method 350.1, Determination of by Axially Viewed Inductively Coupled Ammonium Nitrogen by Semi-Auto- Plasma-Atomic Emission Spectrom- mated Colorimetry. Revision 2.0. Table etry. Revision 4.2, October 2003. EPA/ IB, Notes 30 and 52. 600/R–06/115. Table IB, Note 68.

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(ii) EPA Method 525.2, Determination (K) Method 265.2, Rhodium, Atomic of Organic Compounds in Drinking Absorption, Furnace Technique. Issued Water by Liquid-Solid Extraction and 1978. Table IB, Note 1. Capillary Column Gas Chroma- (L) Method 279.2, Thallium, Atomic tography/Mass Spectrometry. Revision Absorption, Furnace Technique. Issued 2.0, 1995. Table ID, Note 10. 1978. Table IB, Note 1. (5) Office of Research and Develop- (M) Method 283.2, Titanium, Atomic ment, Cincinnati OH. U.S. Environ- Absorption, Furnace Technique. Issued mental Protection Agency, Cincinnati 1978. Table IB, Note 1. OH (US EPA). Available at http:// (N) Method 289.2, Zinc, Atomic Ab- water.epa.gov/scitech/methods/cwa/ sorption, Furnace Technique. Issued index.cfm or from ORD Publications, 1978. Table IB, Note 1. CERI, U.S. Environmental Protection (O) Method 310.2, Alkalinity, Colori- Agency, Cincinnati OH 45268. metric, Automated, Methyl Orange. (i) Methods for Benzidine, Revision 1974. Table IB, Note 1. Chlorinated Organic Compounds, (P) Method 351.1, Nitrogen, Kjeldahl, Pentachlorophenol, and Pesticides in Total, Colorimetric, Automated Water and Wastewater. 1978. Table IC, Phenate. Revision 1978. Table IB, Note Note 3; Table ID, Note 3. 1. (ii) Methods for Chemical Analysis of (Q) Method 352.1, Nitrogen, Nitrate, Water and Wastes. March 1979. EPA– Colorimetric, Brucine. Issued 1971. 600/4–79–020. Table IB, Note 1. Table IB, Note 1. (iii) Methods for Chemical Analysis (R) Method 365.3, Phosphorus, All of Water and Wastes. Revised March Forms, Colorimetric, Ascorbic Acid, 1983. EPA–600/4–79–020. Table IB, Note 1. Two Reagent. Issued 1978. Table IB, (A) Method 120.1, Conductance, Spe- Note 1. cific Conductance, μmhos at 25 °C. Re- (S) Method 365.4, Phosphorus, Total, vision 1982. Table IB, Note 1. Colorimetric, Automated, Block (B) Method 130.1, Hardness, Total Digestor AA II. Issued 1974. Table IB, (mg/L as CaCO3), Colorimetric, Auto- Note 1. mated EDTA. Issued 1971. Table IB, (T) Method 410.3, Chemical Oxygen Note 1. Demand, Titrimetric, High Level for (C) Method 150.2, pH, Continuous Saline Waters. Revision 1978. Table IB, Monitoring (Electrometric). December Note 1. 1982. Table IB, Note 1. (U) Method 420.1, Phenolics, Total (D) Method 160.4, Residue, Volatile, Recoverable, Spectrophotometric, Gravimetric, Ignition at 550 °C. Issued Manual 4–AAP With Distillation. Revi- 1971. Table IB, Note 1. sion 1978. Table IB, Note 1. (E) Method 206.5, Arsenic, Sample Di- (iv) Prescribed Procedures for Meas- gestion Prior to Total Arsenic Analysis urement of Radioactivity in Drinking by Silver Diethyldithiocarbamate or Water. 1980. EPA–600/4–80–032. Table IE. Hydride Procedures. Issued 1978. Table (A) Method 900.0, Gross Alpha and IB, Note 1. Gross Beta Radioactivity. Table IE. (F) Method 231.2, Gold, Atomic Ab- (B) Method 903.0, Alpha-Emitting sorption, Furnace Technique. Issued iRadio Isotopes. Table IE. 1978. Table IB, Note 1. (C) Method 903.1, Radium-226, Radon (G) Method 245.2, Mercury, Auto- Emanation Technique. Table IE. mated Cold Vapor Technique. Issued (D) Appendix B, Error and Statistical 1974. Table IB, Note 1. Calculations. Table IE. (H) Method 252.2, Osmium, Atomic (6) Office of Science and Technology, Absorption, Furnace Technique. Issued U.S. Environmental Protection Agen- 1978. Table IB, Note 1. cy, Washington DC (US EPA). Avail- (I) Method 253.2, Palladium, Atomic able at http://water.epa.gov/scitech/meth- Absorption, Furnace Technique. Issued ods/cwa/index.cfm. 1978. Table IB, Note 1. (i) Method 1625C, Semivolatile Or- (J) Method 255.2, Platinum, Atomic ganic Compounds by Isotope Dilution Absorption, Furnace Technique. Issued GCMS. 1989. Table IF. 1978. Table IB, Note 1. (ii) [Reserved]

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(7) Office of Water, U.S. Environ- (F) Method 617, Organohalide Pes- mental Protection Agency, Washington ticides and PCBs. Table ID, Note 10; DC (US EPA). Available at http:// Table IG, Note 3. water.epa.gov/scitech/methods/cwa/ (G) Method 619, Triazine Pesticides. index.cfm or from National Technical Table ID, Note 10; Table IG, Note 3. Information Service, 5285 Port Royal (H) Method 622, Organophosphorus Road, Springfield, Virginia 22161. Pesticides. Table ID, Note 10; Table IG, (i) Method 1631, Mercury in Water by Note 3. Oxidation, Purge and Trap, and Cold (I) Method 622.1, Thiophosphate Pes- Vapor Atomic Fluorescence Spectrom- ticides. Table ID, Note 10; Table IG, etry. Revision E, August 2002. EPA–821– Note 3. R–02–019, Pub. No. PB2002–108220. Table IB, Note 43. (J) Method 627, Dinitroaniline Pes- (ii) Kelada-01, Kelada Automated ticides. Table ID, Note 10; Table IG, Test Methods for Total Cyanide, Acid Notes 1 and 3. Dissociable Cyanide, and Thiocyanate. (K) Method 629, Cyanazine. Table IG, Revision 1.2, August 2001. EPA 821–B– Note 3. 01–009, Pub. No. PB 2001–108275. Table (L) Method 630, Dithiocarbamate Pes- IB, Note 55. ticides. Table IG, Note 3. (iii) In the compendium Analytical (M) Method 630.1, Dithiocarbamate Methods for the Determination of Pollut- Pesticides. Table IG, Note 3. ants in Pharmaceutical Manufacturing (N) Method 631, Benomyl and Industry Wastewaters. July 1998. EPA Carbendazim. Table IG, Note 3. 821–B–98–016, Pub. No. PB95201679. Table (O) Method 632, and IF, Note 1. Pesticides. Table ID, Note 10; Table IG, (A) EPA Method 1666, Volatile Or- Note 3. ganic Compounds Specific to the Phar- (P) Method 632.1, Carbamate and maceutical Industry by Isotope Dilu- Amide Pesticides. Table IG, Note 3. tion GC/MS. Table IF, Note 1. (Q) Method 633, Organonitrogen Pes- (B) EPA Method 1667, , ticides. Table IG, Note 3. Isobutyraldehyde, and Furfural by Derivatization Followed by High Per- (R) Method 633.1, Neutral Nitrogen- formance Liquid Chromatography. Containing Pesticides. Table IG, Note Table IF. 3. (C) Method 1671, Volatile Organic (S) Method 637, MBTS and TCMTB. Compounds Specific to the Pharma- Table IG, Note 3. ceutical Manufacturing Industry by (T) Method 644, . Table IG, GC/FID. Table IF. Note 3. (iv) Methods For The Determination (U) Method 645, Certain Amine Pes- of Nonconventional Pesticides In Mu- ticides and Lethane. Table IG, Note 3. nicipal and Industrial Wastewater, Vol- (V) Method 1656, Organohalide Pes- ume I. Revision I, August 1993. EPA ticides. Table ID, Note 10; Table IG, 821–R–93–010A, Pub. No. PB 94121654. Notes 1 and 3. Tables ID, IG. (W) Method 1657, Organophosphorus (A) Method 608.1, Organochlorine Pes- Pesticides. Table ID, Note 10; Table IG, ticides. Table ID, Note 10; Table IG, Note 3. Note 3. (X) Method 1658, Phenoxy-Acid Herbi- (B) Method 608.2, Certain cides. Table IG, Note 3. Organochlorine Pesticides. Table ID, Note 10; Table IG, Note 3. (Y) Method 1659, Dazomet. Table IG, (C) Method 614, Organophosphorus Note 3. Pesticides. Table ID, Note 10; Table IG, (Z) Method 1660, and Note 3. . Table IG, Note 3. (D) Method 614.1, Organophosphorus (AA) Method 1661, Bromoxynil. Table Pesticides. Table ID, Note 10; Table IG, IG, Note 3. Note 3. (BB) Ind-01. Methods EV–024 and EV– (E) Method 615, Chlorinated Herbi- 025, Analytical Procedures for Deter- cides. Table ID, Note 10; Table IG, Note Total Tin and Triorganotin in 3. Wastewater. Table IG, Note 3.

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(v) Methods For The Determination (K) Method 548.1, Determination of of Nonconventional Pesticides In Mu- Endothall in Drinking Water by Ion- nicipal and Industrial Wastewater, Vol- Exchange Extraction, Acidic Methanol ume II. August 1993. EPA 821–R–93– Methylation and Gas Chromatography/ 010B, Pub. No. PB 94166311. Table IG. Mass Spectrometry. Table IG, Note 3. (A) Method 200.9, Determination of (L) Method 553, Determination of Trace Elements by Stabilized Tempera- and Nitrogen-Containing ture Graphite Furnace Atomic Absorp- Pesticides in Water by Liquid-Liquid tion Spectrometry. Table IG, Note 3. Extraction or Liquid-Solid Extraction (B) Method 505, Analysis of and Reverse Phase High Performance Organohalide Pesticides and Commer- Liquid Chromatography/Particle Beam/ cial Polychlorinated Biphenyl (PCB) Mass Spectrometry Table ID, Note 10; Products in Water by Microextraction Table IG, Note 3. and Gas Chromatography. Table ID, (M) Method 555, Determination of Note 10; Table IG, Note 3. Chlorinated Acids in Water by High (C) Method 507, The Determination of Performance Liquid Chromatography Nitrogen- and Phosphorus-Containing With a Photodiode Array Ultraviolet Pesticides in Water by Gas Chroma- Detector. Table IG, Note 3. tography with a Nitrogen-Phosphorus (vi) In the compendium Methods for Detector. Table ID, Note 10; Table IG, the Determination of Organic Compounds Note 3. in Drinking Water. Revised July 1991, December 1998. EPA–600/4–88–039, Pub. (D) Method 508, Determination of No. PB92–207703. Table IF. Chlorinated Pesticides in Water by Gas (A) EPA Method 502.2, Volatile Or- Chromatography with an Electron Cap- ganic Compounds in Water by Purge ture Detector. Table ID, Note 10; Table and Trap Capillary Column Gas Chro- IG, Note 3. matography with Photoionization and (E) Method 515.1, Determination of Electrolytic Conductivity Detectors in Chlorinated Acids in Water by Gas Series. Table IF. Chromatography with an Electron Cap- (B) [Reserved] ture Detector. Table IG, Notes 2 and 3. (vii) In the compendium Methods for (F) Method 515.2, Determination of the Determination of Organic Compounds Chlorinated Acids in Water Using Liq- in Drinking Water-Supplement II. August uid-Solid Extraction and Gas Chroma- 1992. EPA–600/R–92–129, Pub. No. PB92– tography with an Electron Capture De- 207703. Table IF. tector. Table IG, Notes 2 and 3. (A) EPA Method 524.2, Measurement (G) Method 525.1, Determination of of Purgeable Organic Compounds in Organic Compounds in Drinking Water Water by Capillary Column Gas Chro- by Liquids-Solid Extraction and Cap- matography/Mass Spectrometry. Table illary Column Gas Chromatography/ IF. Mass Spectrometry. Table ID, Note 10; (B) [Reserved] Table IG, Note 3. (viii) Methods for Measuring the (H) Method 531.1, Measurement of N- Acute Toxicity of Effluents and Re- Methylcarbamoyloximes and N- ceiving Waters to Freshwater and Ma- Methylcarbamates in Water by Direct rine Organisms, Fifth Edition. October Aqueous Injection HPLC with Post- 2002. EPA 821–R–02–012, Pub. No. Column Derivatization. Table ID, Note PB2002–108488. Table IA, Note 26. 10; Table IG, Note 3. (ix) Short-Term Methods for Meas- (I) Method 547, Determination of uring the Chronic Toxicity of Effluents Glyphosate in Drinking Water by Di- and Receiving Waters to Freshwater rect-Aqueous-Injection HPLC, Post- Organisms, Fourth Edition. October Column Derivatization, and Fluores- 2002. EPA 821–R–02–013, Pub. No. cence Detection. Table IG, Note 3. PB2002–108489. Table IA, Note 27. (J) Method 548, Determination of (x) Short-Term Methods for Meas- Endothall in Drinking Water by Aque- uring the Chronic Toxicity of Effluents ous Derivatization, Liquid-Solid Ex- and Receiving Waters to Marine and traction, and Gas Chromatography Estuarine Organisms, Third Edition. with Electron-Capture Detector. Table October 2002. EPA 821–R–02–014, Pub. IG, Note 3. No. PB2002–108490. Table IA, Note 28.

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(8) Office of Water, U.S. Environ- (xi) Method 1664, n-Hexane Extract- mental Protection Agency, Washington able Material (HEM; Oil and Grease) DC (US EPA). Available at http:// and Silica Gel Treated n-Hexane Ex- water.epa.gov/scitech/methods/cwa/ tractable Material (SGT-HEM; Non- index.cfm. polar Material) by Extraction and Gra- (i) Method 245.7, Mercury in Water by vimetry. Revision B, February 2010. Cold Vapor Atomic Fluorescence Spec- EPA–821–R–10–001. Table IB, Notes 38 trometry. Revision 2.0, February 2005. and 42. EPA–821–R–05–001. Table IB, Note 17. (xii) Method 1669, Sampling Ambient (ii) Method 1103.1: Escherichia coli (E. Water for Trace Metals at EPA Water coli) in Water by Membrane Filtration Quality Criteria Levels. July 1996. Using membrane-Thermotolerant Esch- Table IB, Note 43. erichia coli Agar (mTEC). March 2010. (xiii) Method 1680: Fecal Coliforms in EPA–621–R–10–002. Table IH, Note 19. Sewage Sludge (Biosolids) by Multiple- (iii) Method 1106.1: Enterococci in Tube Fermentation using Lauryl Water by Membrane Filtration Using Tryptose Broth (LTB) and EC Medium. membrane-Enterococcus-Esculin Iron September 2014. EPA–821–R–14–009. Agar (mE–EIA). December 2009. EPA– Table IA, Note 15. 621–R–09–015. Table IH, Note 23. (xiv) Method 1681: Fecal Coliforms in (iv) Method 1600: Enterococci in Sewage Sludge (Biosolids) by Multiple- Water by Membrane Filtration Using Tube Fermentation using A–1 Medium. membrane-Enterococcus Indoxyl-b-D- July 2006. EPA 821–R–06–013. Table IA, Glucoside Agar (mEI). September 2014. Note 20. EPA–821–R–14–011. Table IA, Note 25; (xv) Method 1682: Salmonella in Sew- Table IH, Note 24. age Sludge (Biosolids) by Modified (v) Method 1603: Escherichia coli (E. Semisolid Rappaport-Vassiliadis coli) in Water by Membrane Filtration (MSRV) Medium. September 2014. EPA Using Modified membrane- 821–R–14–012. Table IA, Note 23. Thermotolerant Escherichia coli Agar (9) American National Standards In- (Modified mTEC). September 2014. stitute, 1430 Broadway, New York NY EPA–821–R–14–010. Table IA, Note 22; 10018. Table IH, Note 20. (i) ANSI. American National Stand- (vi) Method 1604: Total Coliforms and ard on Photographic Processing Escherichia coli (E. coli) in Water by Effluents. April 2, 1975. Table IB, Note Membrane Filtration Using a Simulta- 9. neous Detection Technique (MI Me- (ii) [Reserved] dium). September 2002. EPA–821–R–02– (10) American Public Health Associa- 024. Table IH, Note 21. tion, 1015 15th Street NW., Washington, (vii) Method 1622: Cryptosporidium in DC 20005. Standard Methods Online is Water by Filtration/IMS/FA. December available through the Standard Meth- 2005. EPA–821–R–05–001. Table IH, Note ods Web site (http:// 25. www.standardmethods.org). (viii) Method 1623: Cryptosporidium (i) Standard Methods for the Exam- and Giardia in Water by Filtration/IMS/ ination of Water and Wastewater. 14th FA. December 2005. EPA–821–R–05–002. Edition, 1975. Table IB, Notes 17 and 27. Table IH, Note 26. (ii) Standard Methods for the Exam- (ix) Method 1627, Kinetic Test Method ination of Water and Wastewater. 15th for the Prediction of Mine Drainage Edition, 1980, Table IB, Note 30; Table Quality. December 2011. EPA–821–R–09– ID. 002. Table IB, Note 69. (iii) Selected Analytical Methods Ap- (x) Method 1664, n-Hexane Extract- proved and Cited by the United States able Material (HEM; Oil and Grease) Environmental Protection Agency, and Silica Gel Treated n-Hexane Ex- Supplement to the 15th Edition of tractable Material (SGT-HEM; Non- Standard Methods for the Examination polar Material) by Extraction and Gra- of Water and Wastewater. 1981. Table vimetry. Revision A, February 1999. IC, Note 6; Table ID, Note 6. EPA–821–R–98–002. Table IB, Notes 38 (iv) Standard Methods for the Exam- and 42. ination of Water and Wastewater. 18th

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Edition, 1992. Tables IA, IB, IC, ID, IE, (xxxiii) 3500-Zn, Zinc. 2011. Table IB. and IH. (xxxiv) 4110, Determination of Anions (v) Standard Methods for the Exam- by Ion Chromatography. 2011. Table IB. ination of Water and Wastewater. 19th (xxxv) 4140, Inorganic Anions by Cap- Edition, 1995. Tables IA, IB, IC, ID, IE, illary Ion Electrophoresis. 2011. Table and IH. IB. (vi) Standard Methods for the Exam- (xxxvi) 4500-B, Boron. 2011. Table IB. ination of Water and Wastewater. 20th (xxxvii) 4500-Cl¥, Chloride. 2011. Edition, 1998. Tables IA, IB, IC, ID, IE, Table IB. and IH. (xxxviii) 4500-Cl, Chlorine (Residual). (vii) Standard Methods for the Exam- 2011. Table IB. ination of Water and Wastewater. 21st (xxxix) 4500-CN¥, Cyanide. 2011. Table Edition, 2005. Table IB, Notes 17 and 27. IB. ¥ (viii) 2120, Color. 2011. Table IB. (xl) 4500-F , Fluoride. 2011. Table IB. ∂ (ix) 2130, Turbidity. 2011. Table IB. (xli) 4500-H , pH Value. 2011. Table (x) 2310, Acidity. 2011. Table IB. IB. (xi) 2320, Alkalinity. 2011. Table IB. (xlii) 4500-NH3, Nitrogen (Ammonia). 2011. Table IB. (xii) 2340, Hardness. 2011. Table IB. ¥ (xiii) 2510, Conductivity. 2011. Table (xliii) 4500-NO2 , Nitrogen (Nitrite). 2011. Table IB. IB. ¥ (xiv) 2540, Solids. 2011. Table IB. (xliv) 4500–NO3 , Nitrogen (Nitrate). (xv) 2550, Temperature. 2010. Table 2011. Table IB. IB. (xlv) 4500–Norg, Nitrogen (Organic). (xvi) 3111, Metals by Flame Atomic 2011. Table IB. Absorption Spectrometry. 2011. Table (xlvi) 4500–O, Oxygen (Dissolved). IB. 2011. Table IB. (xlvii) 4500–P, Phosphorus. 2011. Table (xvii) 3112, Metals by Cold-Vapor IB. Atomic Absorption Spectrometry. 2011. (xlviii) 4500–SiO , Silica. 2011. Table Table IB. 2 IB. (xviii) 3113, Metals by Electrothermal (xlix) 4500–S2¥, Sulfide. 2011. Table Atomic Absorption Spectrometry. 2010. IB. Table IB. 2¥ (l) 4500–SO3 , Sulfite. 2011. Table IB. (xix) 3114, Arsenic and Selenium by 2¥ (li) 4500–SO4 , Sulfate. 2011. Table Hydride Generation/Atomic Absorption IB. Spectrometry. 2011. Table IB. (lii) 5210, Biochemical Oxygen De- (xx) 3120, Metals by Plasma Emission mand (BOD). 2011. Table IB. Spectroscopy. 2011. Table IB. (liii) 5220, Chemical Oxygen Demand (xxi) 3125, Metals by Inductively Cou- (COD). 2011. Table IB. pled Plasma-Mass Spectrometry. 2011. (liv) 5310, Total Organic Carbon Table IB. (TOC). 2011. Table IB. (xxii) 3500-Al, Aluminum. 2011. Table (lv) 5520, Oil and Grease. 2011. Table IB. IB. (xxiii) 3500-As, Arsenic. 2011. Table (lvi) 5530, Phenols. 2010. Table IB. IB. (lvii) 5540, Surfactants. 2011. Table (xxiv) 3500-Ca, Calcium. 2011. Table IB. IB. (lviii) 6200, Volatile Organic Com- (xxv) 3500-Cr, Chromium. 2011. Table pounds. 2011. Table IC. IB. (lix) 6410, Extractable Base/Neutrals (xxvi) 3500-Cu, Copper. 2011. Table IB. and Acids. 2000. Tables IC, ID. (xxvii) 3500-Fe, Iron. 2011. Table IB. (lx) 6420, Phenols. 2000. Table IC. (xxviii) 3500-Pb, Lead. 2011. Table IB. (lxi) 6440, Polynuclear Aromatic Hy- (xxix) 3500-Mn, Manganese. 2011. drocarbons. 2005. Table IC. Table IB. (lxii) 6630, Organochlorine Pesticides. (xxx) 3500-K, Potassium. 2011. Table 2007. Table ID. IB. (lxiii) 6640, Acidic Com- (xxxi) 3500-Na, Sodium. 2011. Table pounds. 2006. Table ID. IB. (lxiv) 7110, Gross Alpha and Gross (xxxii) 3500-V, Vanadium. 2011. Table Beta Radioactivity (Total, Suspended, IB. and Dissolved). 2000. Table IE.

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(lxv) 7500, Radium. 2001. Table IE. (G) 973.45, Oxygen (Dissolved) in (lxvi) 9213, Recreational Waters. 2007. Water, Titrimetric Methods. Table IB, Table IH. Note 3. (lxvii) 9221, Multiple-Tube Fermenta- (H) 973.46, Chemical Oxygen Demand tion Technique for Members of the (COD) of Water, Titrimetric Methods. Coliform Group. 2006. Table IA, Notes Table IB, Note 3. 12 and 14; Table IH, Notes 11 and 13. (I) 973.47, Organic Carbon in Water, (lxviii) 9222, Membrane Filter Tech- Infrared Analyzer Method. Table IB, nique for Members of the Coliform Note 3. Group. 2006. Table IA; Table IH, Note (J) 973.48, Nitrogen (Total) in Water, 18. Kjeldahl Method. Table IB, Note 3. (lxix) 9223, Enzyme Substrate Coli- (K) 973.49, Nitrogen (Ammonia) in form Test. 2004. Table IA; Table IH. Water, Colorimetric Method. Table IB, (lxx) 9230, Fecal Enterococcus/Strepto- Note 3. coccus Groups. 2007. Table IA; Table IH. (L) 973.50, Nitrogen (Nitrate) in (11) The Analyst, The Royal Society Water, Brucine Colorimetric Method. of Chemistry, RSC Publishing, Royal Table IB, Note 3. Society of Chemistry, Thomas Graham (M) 973.51, Chloride in Water, Mer- House, Science Park, Milton Road, curic Nitrate Method. Table IB, Note 3. Cambridge CB4 0WF, United Kingdom. (N) 973.52, Hardness of Water. Table (Also available from most public li- IB, Note 3. braries.) (O) 973.53, Potassium in Water, Atom- (i) Spectrophotometric Determina- ic Absorption Spectrophotometric tion of Ammonia: A Study of a Modi- Method. Table IB, Note 3. fied Berthelot Reaction Using Salicy- (P) 973.54, Sodium in Water, Atomic late and Dichloroisocyanurate. Krom, Absorption Spectrophotometric Meth- M.D. 105:305–316, April 1980. Table IB, od. Table IB, Note 3. Note 60. (Q) 973.55, Phosphorus in Water, Pho- tometric Method. Table IB, Note 3. (ii) [Reserved] (R) 973.56, Phosphorus in Water, (12) Analytical Chemistry, ACS Pub- Automated Method. Table IB, Note 3. lications, 1155 Sixteenth St. NW., (S) 974.27, Cadmium, Chromium, Cop- Washington DC 20036. (Also available per, Iron, Lead, Magnesium, Man- from most public libraries.) ganese, Silver, Zinc in Water, Atomic (i) Spectrophotometric and Kinetics Absorption Spectrophotometric Meth- Investigation of the Berthelot Reaction od. Table IB, Note 3. for the Determination of Ammonia. (T) 977.22, Mercury in Water, Patton, C.J. and S.R. Crouch. 49(3):464– Flameless Atomic Absorption 469, March 1977. Table IB, Note 60. Spectrophotometric Method. Table IB, (ii) [Reserved] Note 3. (13) AOAC International, 481 North (U) 991.15. Total Coliforms and Esch- Frederick Avenue, Suite 500, Gaithers- erichia coli in Water Defined Substrate burg, MD 20877–2417. Technology (Colilert) Method. Table (i) Official Methods of Analysis of IA, Note 10; Table IH, Note 10. AOAC International. 16th Edition, 4th (V) 993.14, Trace Elements in Waters Revision, 1998. and Wastewaters, Inductively Coupled (A) 920.203, Manganese in Water, Plasma-Mass Spectrometric Method. Persulfate Method. Table IB, Note 3. Table IB, Note 3. (B) 925.54, Sulfate in Water, (W) 993.23, Dissolved Hexavalent Gravimetric Method. Table IB, Note 3. Chromium in Drinking Water, Ground (C) 973.40, Specific Conductance of Water, and Industrial Wastewater Water. Table IB, Note 3. Effluents, Ion Chromatographic Meth- (D) 973.41, pH of Water. Table IB, od. Table IB, Note 3. Note 3. (X) 993.30, Inorganic Anions in Water, (E) 973.43, Alkalinity of Water, Ion Chromatographic Method. Table Titrimetric Method. Table IB, Note 3. IB, Note 3. (F) 973.44, Biochemical Oxygen De- (ii) [Reserved] mand (BOD) of Water, Incubation (14) Applied and Environmental Method. Table IB, Note 3. Microbiology, American Society for

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Microbiology, 1752 N Street NW., Wash- (xiv) ASTM D1125–95 (Reapproved ington DC 20036. (Also available from 1999), Standard Test Methods for Elec- most public libraries.) trical Conductivity and Resistivity of (i) New Medium for the Simultaneous Water. December 1995. Table IB. Detection of Total Coliforms and Esch- (xv) ASTM D1126–12, Standard Test erichia coli in Water. Brenner, K.P., C.C. Method for Hardness in Water. March Rankin, Y.R. Roybal, G.N. Stelma, Jr., 2012. Table IB. P.V. Scarpino, and A.P. Dufour. 59:3534– (xvi) ASTM D1179–10, Standard Test 3544, November 1993. Table IH, Note 21. Methods for Fluoride Ion in Water. (ii) [Reserved] July 2010. Table IB. (15) ASTM International, 100 Barr (xvii) ASTM D1246–10, Standard Test Harbor Drive, P.O. Box C700, West Method for Bromide Ion in Water. July Conshohocken, PA 19428–2959, or online 2010. Table IB. at http://www.astm.org. (xviii) ASTM D1252–06, Standard Test (i) Annual Book of ASTM Standards, Water, and Environmental Technology, Methods for Chemical Oxygen Demand Section 11, Volumes 11.01 and 11.02. (Dichromate Oxygen Demand) of 1994. Tables IA, IB, IC, ID, IE, and IH. Water. February 2006. Table IB. (ii) Annual Book of ASTM Standards, (xix) ASTM D1253–08, Standard Test Water, and Environmental Technology, Method for Residual Chlorine in Water. Section 11, Volumes 11.01 and 11.02. October 2008. Table IB. 1996. Tables IA, IB, IC, ID, IE, and IH. (xx) ASTM D1293–99, Standard Test (iii) Annual Book of ASTM Stand- Methods for pH of Water. March 2000. ards, Water, and Environmental Tech- Table IB. nology, Section 11, Volumes 11.01 and (xxi) ASTM D1426–08, Standard Test 11.02. 1999. Tables IA, IB, IC, ID, IE, and Methods for Ammonia Nitrogen in IH. Water. September 2008. Table IB. (iv) Annual Book of ASTM Stand- (xxii) ASTM D1687–12 (Approved Sep- ards, Water, and Environmental Tech- tember 1, 2012), Standard Test Methods nology, Section 11, Volumes 11.01 and for Chromium in Water. August 2007. 11.02. 2000. Tables IA, IB, IC, ID, IE, and Table IB. IH. (xxiii) ASTM D1688–12, Standard Test (v) ASTM D511–09, Standard Test Methods for Copper in Water. Sep- Methods for Calcium and Magnesium tember 2012. Table IB. in Water. May 2009. Table IB. (xxiv) ASTM D1691–12, Standard Test (vi) ASTM D512–04, Standard Test Methods for Zinc in Water. September Methods for Chloride Ion in Water. 2012. Table IB. July 2004. Table IB. (xxv) ASTM D1783–01 (Reapproved (vii) ASTM D515–88, Test Methods for 2007), Standard Test Methods for Phe- Phosphorus in Water, March 1989. nolic Compounds in Water. January Table IB. 2008). Table IB. (viii) ASTM D516–11, Standard Test (xxvi) ASTM D1886–08, Standard Test Method for Sulfate Ion in Water, Sep- Methods for Nickel in Water. October tember 2011. Table IB. 2008. Table IB. (ix) ASTM D858–12, Standard Test Methods for Manganese in Water. Sep- (xxvii) ASTM D1889–00, Standard Test tember 2012. Table IB. Method for Turbidity of Water. October (x) ASTM D859–10, Standard Test 2000. Table IB. Method for Silica in Water. July 2010. (xxviii) ASTM D1890–96, Standard Table IB. Test Method for Beta Particle Radioac- (xi) ASTM D888–09, Standard Test tivity of Water. April 1996. Table IE. Methods for Dissolved Oxygen in (xxix) ASTM D1943–96, Standard Test Water. December 2009. Table IB. Method for Alpha Particle Radioac- (xii) ASTM D1067–11, Standard Test tivity of Water. April 1996. Table IE. Methods for Acidity or Alkalinity of (xxx) ASTM D1976–12, Standard Test Water. April 2011. Table IB. Method for Elements in Water by In- (xiii) ASTM D1068–10, Standard Test ductively-Coupled Argon Plasma Methods for Iron in Water. October Atomic Emission Spectroscopy. March 2010. Table IB. 2012. Table IB.

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(xxxi) ASTM D2036–09, Standard Test (xlix) ASTM D4327–03, Standard Test Methods for in Water. Octo- Method for Anions in Water by Chemi- ber 2009. Table IB. cally Suppressed Ion Chromatography. (xxxii) ASTM D2330–02, Standard Test January 2003. Table IB. Method for Methylene Blue Active (l) ASTM D4382–12, Standard Test Substances. August 2002. Table IB. Method for Barium in Water, Atomic (xxxiii) ASTM D2460–97, Standard Absorption Spectrophotometry, Graph- Test Method for Alpha-Particle-Emit- ite Furnace. September 2012. Table IB. ting Isotopes of Radium in Water. Oc- (li) ASTM D4657–92 (Reapproved 1998), tober 1997. Table IE. Standard Test Method for Polynuclear (xxxiv) ASTM D2972–08, Standard Aromatic Hydrocarbons in Water. Jan- Tests Method for Arsenic in Water. Oc- uary 1993. Table IC. tober 2008. Table IB. (lii) ASTM D4658–09, Standard Test (xxxv) ASTM D3223–12, Standard Test Method for Sulfide Ion in Water. May Method for Total Mercury in Water. 2009. Table IB. September 2012. Table IB. (liii) ASTM D4763–88 (Reapproved (xxxvi) ASTM D3371–95, Standard 2001), Standard Practice for Identifica- Test Method for in Aqueous tion of Chemicals in Water by Fluores- Solution by Gas-Liquid Chroma- cence Spectroscopy. September 1988. tography, February 1996. Table IF. Table IF. (xxxvii) ASTM D3373–12, Standard (liv) ASTM D4839–03, Standard Test Test Method for Vanadium in Water. Method for Total Carbon and Organic September 2012. Table IB. Carbon in Water by Ultraviolet, or (xxxviii) ASTM D3454–97, Standard Persulfate Oxidation, or Both, and In- Test Method for Radium-226 in Water. frared Detection. January 2003. Table February 1998. Table IE. IB. (xxxix) ASTM D3557–12, Standard (lv) ASTM D5257–11, Standard Test Test Method for Cadmium in Water. Method for Dissolved Hexavalent Chro- September 2012. Table IB. mium in Water by Ion Chroma- (xl) ASTM D3558–08, Standard Test tography. April 2011. Table IB. Method for Cobalt in Water. November (lvi) ASTM D5259–92, Standard Test 2008. Table IB. Method for Isolation and Enumeration (xli) ASTM D3559–08, Standard Test of Enterococci from Water by the Methods for Lead in Water. October Membrane Filter Procedure. October 2008. Table IB. 1992. Table IH, Note 9. (xlii) ASTM D3590–11, Standard Test (lvii) ASTM D5392–93, Standard Test Methods for Total Kjeldahl Nitrogen in Method for Isolation and Enumeration Water. April 2011. Table IB. of Escherichia coli in Water by the Two- (xliii) ASTM D3645–08, Standard Test Step Membrane Filter Procedure. Sep- Methods for Beryllium in Water. Octo- tember 1993. Table IH, Note 9. ber 2008. Table IB. (lviii) ASTM D5673–10, Standard Test (xliv) ASTM D3695–95, Standard Test Method for Elements in Water by In- Method for Volatile Alcohols in Water ductively Coupled Plasma—Mass Spec- by Direct Aqueous-Injection Gas Chro- trometry. September 2010. Table IB. matography. April 1995. Table IF. (lix) ASTM D5(19)907–13, Standard (xlv) ASTM D3859–08, Standard Test Test Method for Filterable Matter Methods for Selenium in Water. Octo- (Total Dissolved Solids) and Nonfilter- ber 2008. Table IB. able Matter (Total Suspended Solids) (xlvi) ASTM D3867–04, Standard Test in Water. July 2013. Table IB. Method for Nitrite-Nitrate in Water. (lx) ASTM D6503–99, Standard Test July 2004. Table IB. Method for Enterococci in Water Using (xlvii) ASTM D4190–08, Standard Test Enterolert. April 2000. Table IA Note 9, Method for Elements in Water by Di- Table IH, Note 9. rect-Current Plasma Atomic Emission (lxi) ASTM. D6508–10, Standard Test Spectroscopy. October 2008. Table IB. Method for Determination of Dissolved (xlviii) ASTM D4282–02, Standard Inorganic Anions in Aqueous Matrices Test Method for Determination of Free Using Capillary Ion Electrophoresis Cyanide in Water and Wastewater by and Chromate Electrolyte. October Microdiffusion. August 2002. Table IB. 2010. Table IB, Note 54.

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(lxii) ASTM. D6888–09, Standard Test mination of Metals. April 16, 1992. Method for Available Cyanide with Table IB, Note 36. Ligand Displacement and Flow Injec- (ii) [Reserved] tion Analysis (FIA) Utilizing Gas Diffu- (18) Craig R. Chinchilla, 900 Jorie sion Separation and Amperometric De- Blvd., Suite 35, Oak Brook IL 60523. tection. October 2009. Table IB, Note 59. Telephone: 630–645–0600. (lxiii) ASTM. D6919–09, Standard Test (i) Nitrate by Discrete Analysis Easy Method for Determination of Dissolved (1-Reagent) Nitrate Method, (Colori- Alkali and Alkaline Earth Cations and metric, Automated, 1 Reagent). Revi- Ammonium in Water and Wastewater sion 1, November 12, 2011. Table IB, by Ion Chromatography. May 2009. Note 62. Table IB. (ii) [Reserved] (lxiv) ASTM. D7065–11, Standard Test (19) Hach Company, P.O. Box 389, Method for Determination of Loveland CO 80537. Nonylphenol, Bisphenol A, p-tert- (i) Method 8000, Chemical Oxygen De- Octylphenol, Nonylphenol mand. Hach Handbook of Water Anal- Monoethoxylate and Nonylphenol ysis. 1979. Table IB, Note 14. Diethoxylate in Environmental Waters (ii) Method 8008, 1,10-Phenanthroline by Gas Chromatography Mass Spec- Method using FerroVer Iron Reagent trometry. July 2011. Table IB. for Water. 1980. Table IB, Note 22. (lxv) ASTM. D7237–10, Standard Test (iii) Method 8009, Zincon Method for Method for Free Cyanide with Flow In- Zinc. Hach Handbook for Water Anal- jection Analysis (FIA) Utilizing Gas ysis. 1979. Table IB, Note 33. Diffusion Separation and Ampero- metric Detection. June 2010. Table IB. (iv) Method 8034, Periodate Oxidation (lxvi) ASTM. D7284–13, Standard Test Method for Manganese. Hach Handbook Method for Total Cyanide in Water by for Water Analysis. 1979. Table IB, Note Micro Distillation followed by Flow In- 23. jection Analysis with Gas Diffusion (v) Method 8506, Bicinchoninate Separation and Amperometric Detec- Method for Copper. Hach Handbook of tion. July 2013. Table IB. Water Analysis. 1979. Table IB, Note 19. (lxvii) ASTM. D7365–09a, Standard (vi) Method 8507, Nitrogen, Nitrite— Practice for Sampling, Preservation, Low Range, Diazotization Method for and Mitigating Interferences in Water Water and Wastewater. 1979. Table IB, Samples for Analysis of Cyanide. Octo- Note 25. ber 2009. Table II, Notes 5 and 6. (vii) Method 10206, Hach Company (lxviii) ASTM. D7511–12, Standard TNTplus 835/836 Nitrate Method 10206, Test Method for Total Cyanide by Seg- Spectrophotometric Measurement of mented Flow Injection Analysis, In- Nitrate in Water and Wastewater. Re- Line Ultraviolet Digestion and Amper- vision 2.1, January 10, 2013. Table IB, ometric Detection. January 2012. Table Note 75. IB. (viii) Method 10242, Hach Company (lxix) ASTM. D7573–09, Standard Test TNTplus 880 Total Kjeldahl Nitrogen Method for Total Carbon and Organic Method 10242, Simplified Carbon in Water by High Temperature Spectrophotometric Measurement of Catalytic Combustion and Infrared De- Total Kjeldahl Nitrogen in Water and tection. November 2009. Table IB. Wastewater. Revision 1.1, January 10, (16) Bran & Luebbe Analyzing Tech- 2013. Table IB, Note 76. nologies, Inc., Elmsford NY 10523. (ix) Hach Method 10360, Lumines- (i) Industrial Method Number 378– cence Measurement of Dissolved Oxy- 75WA, Hydrogen Ion (pH) Automated gen in Water and Wastewater and for Electrode Method, Bran & Luebbe Use in the Determination of BOD5 and (Technicon) Auto Analyzer II. October cBOD5. Revision 1.2, October 2011. 1976. Table IB, Note 21. Table IB, Note 63. (ii) [Reserved] (x) m-ColiBlue24® Method, for total (17) CEM Corporation, P.O. Box 200, Coliforms and E. coli. Revision 2, 1999. Matthews NC 28106–0200. Table IA, Note 18; Table IH, Note 17. (i) Closed Vessel Microwave Diges- (20) IDEXX Laboratories Inc., One tion of Wastewater Samples for Deter- Idexx Drive, Westbrook ME 04092.

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(i) Colilert. 2013. Table IA, Notes 17 ments, Inc. (NCASI), 260 Madison Ave- and 18; Table IH, Notes 14, 15 and 16. nue, New York NY 10016. (ii) Colilert-18. 2013. Table IA, Notes (i) NCASI Method TNTP–W10900, 17 and 18; Table IH, Notes 14, 15 and 16. Total Nitrogen and Total Phophorus in (iii) Enterolert. 2013. Table IA, Note Pulp and Paper Biologically Treated 24; Table IH, Note 12. Effluent by Alkaline Persulfate Diges- (iv) Quanti-Tray Insert and Most tion. June 2011. Table IB, Note 77. Probable Number (MPN) Table. 2013. (ii) NCASI Technical Bulletin No. 253, Table IA, Note 18; Table IH, Notes 14 An Investigation of Improved Proce- and 16. dures for Measurement of Mill Effluent (21) In-Situ Incorporated, 221 E. Lin- and Receiving Water Color. December coln Ave., Ft. Collins CO 80524. Tele- 1971. Table IB, Note 18. phone: 970–498–1500. (iii) NCASI Technical Bulletin No. (i) In-Situ Inc. Method 1002–8–2009, 803, An Update of Procedures for the Dissolved Oxygen Measurement by Op- Measurement of Color in Pulp Mill tical Probe. 2009. Table IB, Note 64. Wastewaters. May 2000. Table IB, Note (ii) In-Situ Inc. Method 1003–8–2009, 18. Biochemical Oxygen Demand (BOD) (26) The Nitrate Elimination Co., Inc. Measurement by Optical Probe. 2009. (NECi), 334 Hecla St., Lake Linden NI Table IB, Note 10. 49945. (iii) In-Situ Inc. Method 1004–8–2009, (i) NECi Method N07–0003, Method for Carbonaceous Biochemical Oxygen De- Nitrate Reductase Nitrate-Nitrogen mand (CBOD) Measurement by Optical Analysis. Revision 9.0. March 2014. Probe. 2009. Table IB, Note 35. Table IB, Note 73. (22) Journal of Chromatography, (ii) [Reserved] Elsevier/North-Holland, Inc., Journal (27) Oceanography International Cor- Information Centre, 52 Vanderbilt Ave- poration, 512 West Loop, P.O. Box 2980, nue, New York NY 10164. (Also avail- College Station TX 77840. able from most public libraries. (i) OIC Chemical Oxygen Demand (i) Direct Determination of Ele- Method. 1978. Table IB, Note 13. mental Phosphorus by Gas-Liquid Chromatography. Addison, R.F. and (ii) [Reserved] R.G. Ackman. 47(3): 421–426, 1970. Table (28) OI Analytical, Box 9010, College IB, Note 28. Station TX 77820–9010. (ii) [Reserved] (i) Method OIA–1677–09, Available Cy- (23) Lachat Instruments, 6645 W. Mill anide by Ligand Exchange and Flow In- Road, Milwaukee WI 53218, Telephone: jection Analysis (FIA). Copyright 2010. 414–358–4200. Table IB, Note 59. (i) QuikChem Method 10–204–00–1–X, (ii) Method PAI–DK01, Nitrogen, Digestion and Distillation of Total Cy- Total Kjeldahl, Block Digestion, Steam anide in Drinking and Wastewaters Distillation, Titrimetric Detection. Re- using MICRO DIST and Determination vised December 22, 1994. Table IB, Note of Cyanide by Flow Injection Analysis. 39. Revision 2.2, March 2005. Table IB, Note (iii) Method PAI–DK02, Nitrogen, 56. Total Kjeldahl, Block Digestion, Steam (ii) [Reserved] Distillation, Colorimetric Detection. (24) Leck Mitchell, Ph.D., P.E., 656 Revised December 22, 1994. Table IB, Independence Valley Dr., Grand Junc- Note 40. tion CO 81507. Telephone: 970–244–8661. (iv) Method PAI–DK03, Nitrogen, (i) Mitchell Method M5271, Deter- Total Kjeldahl, Block Digestion, Auto- mination of Turbidity by mated FIA Gas Diffusion. Revised De- Nephelometry. Revision 1.0, July 31, cember 22, 1994. Table IB, Note 41. 2008. Table IB, Note 66. (29) ORION Research Corporation, 840 (ii) Mitchell Method M5331, Deter- Memorial Drive, Cambridge, Massachu- mination of Turbidity by setts 02138. Nephelometry. Revision 1.0, July 31, (i) ORION Research Instruction Man- 2008. Table IB, Note 65. ual, Residual Chlorine Electrode Model (25) National Council of the Paper In- 97–70. 1977. Table IB, Note 16. dustry for Air and Stream Improve- (ii) [Reserved]

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(30) Technicon Industrial Systems, U.S. Geological Survey, Book 5, Chap- Tarrytown NY 10591. ter A1. 1979. Table IB, Note 8. (i) Industrial Method Number 379– (iii) Methods for Determination of In- 75WE Ammonia, Automated Electrode organic Substances in Water and Flu- Method, Technicon Auto Analyzer II. vial Sediments, Techniques of Water- February 19, 1976. Table IB, Note 7. Resources Investigations of the U.S. (ii) [Reserved] Geological Survey, Book 5, Chapter A1. (31) Thermo Jarrell Ash Corporation, 1989. Table IB, Note 2. 27 Forge Parkway, Franklin MA 02038. (iv) Methods for the Determination of (i) Method AES0029. Direct Current Organic Substances in Water and Flu- Plasma (DCP) Optical Emission Spec- vial Sediments. Techniques of Water- trometric Method for Trace Elemental Resources Investigations of the U.S. Analysis of Water and Wastes. 1986, Re- Geological Survey, Book 5, Chapter A3. vised 1991. Table IB, Note 34. 1987. Table IB, Note 24; Table ID, Note (ii) [Reserved] 4. (32) Thermo Scientific, 166 Cummings (v) OFR 76–177, Selected Methods of Center, Beverly MA 01915. Telephone: the U.S. Geological Survey of Analysis 1–800–225–1480. www.thermoscientific.com. of Wastewaters. 1976. Table IE, Note 2. (i) Thermo Scientific Orion Method (vi) OFR 91–519, Methods of Analysis AQ4500, Determination of Turbidity by by the U.S. Geological Survey National Nephelometry. Revision 5, March 12, Water Quality Laboratory—Determina- 2009. Table IB, Note 67. tion of Organonitrogen Herbicides in (ii) [Reserved] Water by Solid-Phase Extraction and (33) 3M Corporation, 3M Center Build- Capillary-Column Gas Chroma- ing 220–9E–10, St. Paul MN 55144–1000. tography/Mass Spectrometry With Se- (i) Organochlorine Pesticides and lected-Ion Monitoring. 1992. Table ID, PCBs in Wastewater Using Empore TM Note 14. Disk’’ Test Method 3M 0222. Revised (vii) OFR 92–146, Methods of Analysis October 28, 1994. Table IC, Note 8; Table by the U.S. Geological Survey National ID, Note 8. Water Quality Laboratory—Determina- (ii) [Reserved] tion of Total Phosphorus by a Kjeldahl (34) Timberline Instruments, LLC, Digestion Method and an Automated 1880 South Flatiron Ct., Unit I, Boulder Colorimetric Finish That Includes Di- CO 80301. alysis. 1992. Table IB, Note 48. (i) Timberline Amonia-001, Deter- (viii) OFR 93–125, Methods of Anal- mination of Inorganic Ammonia by ysis by the U.S. Geological Survey Na- Continuous Flow Gas Diffusion and tional Water Quality Laboratory—De- Conductivity Cell Analysis. June 24, termination of Inorganic and Organic 2011. Table IB, Note 74. Constituents in Water and Fluvial (ii) [Reserved] Sediments. 1993. Table IB, Note 51; (35) U.S. Geological Survey (USGS), Table IC, Note 9. U.S. Department of the Interior, Res- (ix) OFR 93–449, Methods of Analysis ton, Virginia. Available from USGS by the U.S. Geological Survey National Books and Open-File Reports (OFR) Water Quality Laboratory—Determina- Section, Federal Center, Box 25425, tion of Chromium in Water by Graphite Denver, CO 80225. Furnace Atomic Absorption (i) Colorimetric determination of ni- Spectrophotometry. 1993. Table IB, trate plus nitrite in water by enzy- Note 46. matic reduction, automated discrete (x) OFR 94–37, Methods of Analysis by analyzer methods. U.S. Geological Sur- the U.S. Geological Survey National vey Techniques and Methods, Book 5— Water Quality Laboratory—Determina- Laboratory Analysis, Section B—Meth- tion of Triazine and Other Nitrogen- ods of the National Water Quality Lab- containing Compounds by Gas Chroma- oratory, Chapter 8. 2011. Table IB, Note tography With Nitrogen Phosphorus 72. Detectors. 1994. Table ID, Note 9. (ii) Methods for Determination of In- (xi) OFR 95–181, Methods of Analysis organic Substances in Water and Flu- by the U.S. Geological Survey National vial Sediments, editors, Techniques of Water Quality Laboratory—Determina- Water-Resources Investigations of the tion of Pesticides in Water by C–18

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Solid-Phase Extraction and Capillary- matography/Mass Spectrometry. 2001. Column Gas Chromatography/Mass Table ID, Note 13. Spectrometry With Selected-Ion Moni- (xix) Water-Resources Investigations toring. 1995. Table ID, Note 11. Report 01–4132, Methods of Analysis by (xii) OFR 97–198, Methods of Analysis the U.S. Geological Survey National by the U.S. Geological Survey National Water Quality Laboratory—Determina- Water Quality Laboratory—Determina- tion of Organic Plus Inorganic Mercury tion of Molybdenum in Water by in Filtered and Unfiltered Natural Graphite Furnace Atomic Absorption Water With Cold Vapor-Atomic Fluo- Spectrophotometry. 1997. Table IB, rescence Spectrometry. 2001. Table IB, Note 47. Note 71. (xiii) OFR 98–165, Methods of Anal- (xx) Water-Resources Investigation ysis by the U.S. Geological Survey Na- Report 01–4134, Methods of Analysis by tional Water Quality Laboratory—De- the U.S. Geological Survey National termination of Elements in Whole- Water Quality Laboratory—Determina- Water Digests Using Inductively Cou- tion of Pesticides in Water by pled Plasma-Optical Emission Spec- Graphitized Carbon-Based Solid-Phase trometry and Inductively Coupled Extraction and High-Performance Liq- Plasma-Mass Spectrometry. 1998. Table uid Chormatography/Mass Spectrom- IB, Note 50. etry. 2001. Table ID, Note 12. (xiv) OFR 98–639, Methods of Analysis (xxi) Water Temperature—Influential by the U.S. Geological Survey National Factors, Field Measurement and Data Water Quality Laboratory—Determina- Presentation, Techniques of Water-Re- tion of Arsenic and Selenium in Water sources Investigations of the U.S. Geo- and Sediment by Graphite Furnace— logical Survey, Book 1, Chapter D1. Atomic Absorption Spectrometry. 1999. 1975. Table IB, Note 32. Table IB, Note 49. (36) Waters Corporation, 34 Maple (xv) OFR 00–170, Methods of Analysis Street, Milford MA 01757, Telephone: by the U.S. Geological Survey National 508–482–2131, Fax: 508–482–3625. Water Quality Laboratory—Determina- (i) Method D6508, Test Method for De- tion of Ammonium Plus Organic Nitro- termination of Dissolved Inorganic gen by a Kjeldahl Digestion Method Anions in Aqueous Matrices Using Cap- and an Automated Photometric Finish illary Ion Electrophoresis and Chro- that Includes Digest Cleanup by Gas mate Electrolyte. Revision 2, Decem- Diffusion. 2000. Table IB, Note 45. ber 2000. Table IB, Note 54. (xvi) Techniques and Methods Book (ii) [Reserved] 5–B1, Determination of Elements in (c) Under certain circumstances, the Natural-Water, Biota, Sediment and Director may establish limitations on Soil Samples Using Collision/Reaction the discharge of a parameter for which Cell Inductively Coupled Plasma-Mass there is no test procedure in this part Spectrometry. Chapter 1, Section B, or in 40 CFR parts 405 through 499. In Methods of the National Water Quality these instances the test procedure shall Laboratory, Book 5, Laboratory Anal- be specified by the Director. ysis. 2006. Table IB, Note 70. (d) Under certain circumstances, the (xvii) U.S. Geological Survey Tech- Administrator may approve additional niques of Water-Resources Investiga- alternate test procedures for nation- tions, Book 5, Laboratory Analysis, wide use, upon recommendation by the Chapter A4, Methods for Collection and Alternate Test Procedure Program Co- Analysis of Aquatic Biological and ordinator, Washington, DC. Microbiological Samples. 1989. Table (e) Sample preservation procedures, IA, Note 4; Table IH, Note 4. container materials, and maximum al- (xviii) Water-Resources Investigation lowable holding times for parameters Report 01–4098, Methods of Analysis by are cited in Tables IA, IB, IC, ID, IE, the U.S. Geological Survey National IF, IG, and IH are prescribed in Table Water Quality Laboratory—Determina- II. Information in the table takes prec- tion of Moderate-Use Pesticides and edence over information in specific Selected Degradates in Water by C–18 methods or elsewhere. Any person may Solid-Phase Extraction and Gas Chro- apply for a change from the prescribed

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preservation techniques, container ma- will review the application and then terials, and maximum holding times notify the applicant and the appro- applicable to samples taken from a spe- priate State agency of approval or re- cific discharge. Applications for such jection of the use of the alternate test limited use changes may be made by procedure. A decision to approve or letters to the Regional Alternative deny any request on deviations from Test Procedure (ATP) Program Coordi- the prescribed Table II requirements nator or the permitting authority in will be made within 90 days of receipt the Region in which the discharge will of the application by the Regional Ad- occur. Sufficient data should be pro- ministrator. An analyst may not mod- vided to assure such changes in sample ify any sample preservation and/or preservation, containers or holding times do not adversely affect the integ- holding time requirements of an ap- rity of the sample. The Regional ATP proved method unless the requirements Coordinator or permitting authority of this section are met.

TABLE II—REQUIRED CONTAINERS, PRESERVATION TECHNIQUES, AND HOLDING TIMES

Parameter number/name Container 1 Preservation 23 Maximum holding time 4

Table IA—Bacterial Tests

1–5. Coliform, total, fecal, and PA, G ...... Cool, <10 °C, 0.008% 8 hours.22 23 5 E. coli. Na2S2O3 . 6. Fecal streptococci ...... PA, G ...... Cool, <10 °C, 0.008% 8 hours.22 5 Na2S2O3 . 7. Enterococci ...... PA, G ...... Cool, <10 °C, 0.008% 8 hours.22 5 Na2S2O3 . 8. Salmonella ...... PA, G ...... Cool, <10 °C, 0.008% 8 hours.22 5 Na2S2O3 .

Table IA—Aquatic Toxicity Tests

9–12. Toxicity, acute and P, FP, G ...... Cool, ≤6 °C 16 ...... 36 hours. chronic.

Table IB—Inorganic Tests

1. Acidity ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 14 days. 2. Alkalinity ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 14 days. 18 4. Ammonia ...... P, FP, G ...... Cool, ≤6 °C, H2SO4 to pH 28 days. <2. 9. Biochemical oxygen de- P, FP, G ...... Cool, ≤6 °C 18 ...... 48 hours. mand.

10. Boron ...... P, FP, or Quartz ...... HNO3 to pH <2 ...... 6 months. 11. Bromide ...... P, FP, G ...... None required ...... 28 days. 14. Biochemical oxygen de- P, FP G ...... Cool, ≤6 °C 18 ...... 48 hours. mand, carbonaceous. 18 15. Chemical oxygen demand P, FP, G ...... Cool, ≤6 °C, H2SO4 to pH 28 days. <2. 16. Chloride ...... P, FP, G ...... None required ...... 28 days. 17. Chlorine, total residual ...... P, G ...... None required ...... Analyze within 15 minutes. 21. Color ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 48 hours. 23–24. Cyanide, total or avail- P, FP, G ...... Cool, ≤6 °C,18 NaOH to pH 14 days. able (or CATC) and free. >10,56 if oxidizer present. 25. Fluoride ...... P ...... None required ...... 28 days.

27. Hardness ...... P, FP, G ...... HNO3 or H2SO4 to pH <2 ...... 6 months. 28. Hydrogen ion (pH) ...... P, FP, G ...... None required ...... Analyze within 15 minutes. 18 31, 43. Kjeldahl and organic N P, FP, G ...... Cool, ≤6 °C, H2SO4 to pH 28 days. <2.

Table IB—Metals 7

18. Chromium VI ...... P, FP, G ...... Cool, ≤6 °C,18 pH = 9.3– 28 days. 9.7 20.

35. Mercury (CVAA) ...... P, FP, G ...... HNO3 to pH <2 ...... 28 days. 35. Mercury (CVAFS) ...... FP, G; and FP-lined cap 17 .... 5 mL/L 12N HCl or 5 mL/L 90 days.17 BrCl 17.

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TABLE II—REQUIRED CONTAINERS, PRESERVATION TECHNIQUES, AND HOLDING TIMES—Continued

Parameter number/name Container 1 Preservation 23 Maximum holding time 4

3, 5–8, 12, 13, 19, 20, 22, 26, P, FP, G ...... HNO3 to pH <2, or at least 24 6 months. 29, 30, 32–34, 36, 37, 45, hours prior to analysis 19. 47, 51, 52, 58–60, 62, 63, 70–72, 74, 75. Metals, ex- cept boron, chromium VI, and mercury. 38. Nitrate ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 48 hours. 18 39. Nitrate-nitrite ...... P, FP, G ...... Cool, ≤6 °C, H2SO4 to pH 28 days. <2. 40. Nitrite ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 48 hours. 41. Oil and grease ...... G ...... Cool to ≤6 °C,18 HCl or 28 days. H2SO4 to pH <2. 18 42. Organic Carbon ...... P, FP, G ...... Cool to ≤6 °C, HCl, H2SO4, 28 days. or H3PO4 to pH <2. 44. Orthophosphate ...... P, FP, G ...... Cool, to ≤6 °C 18 24 ...... Filter within 15 minutes; Ana- lyze within 48 hours. 46. Oxygen, Dissolved Probe .. G, Bottle and top ...... None required ...... Analyze within 15 minutes. 47. Winkler ...... G, Bottle and top ...... Fix on site and store in dark .. 8 hours. 18 48. Phenols ...... G ...... Cool, ≤6 °C, H2SO4 to pH 28 days. <2. 49. Phosphorous (elemental) .. G ...... Cool, ≤6 °C 18 ...... 48 hours. 18 50. Phosphorous, total ...... P, FP, G ...... Cool, ≤6 °C, H2SO4 to pH 28 days. <2. 53. Residue, total ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 7 days. 54. Residue, Filterable ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 7 days. 55. Residue, Nonfilterable P, FP, G ...... Cool, ≤6 °C 18 ...... 7 days. (TSS). 56. Residue, Settleable ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 48 hours. 57. Residue, Volatile ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 7 days. 61. Silica ...... P or Quartz ...... Cool, ≤6 °C 18 ...... 28 days. 64. Specific conductance ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 28 days. 65. Sulfate ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 28 days. 66. Sulfide ...... P, FP, G ...... Cool, ≤6 °C,18 add zinc ace- 7 days. tate plus sodium hydroxide to pH >9. 67. Sulfite ...... P, FP, G ...... None required ...... Analyze within 15 minutes. 68. Surfactants ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 48 hours. 69. Temperature ...... P, FP, G ...... None required ...... Analyze within 15 minutes. 73. Turbidity ...... P, FP, G ...... Cool, ≤6 °C 18 ...... 48 hours.

Table IC—Organic Tests 8

13, 18–20, 22, 24, 25, 27,28, G, FP-lined septum ...... Cool, ≤6 °C,18 0.008% 14 days. 5 34–37, 39–43, 45–47, 56, Na2S2O3, HCl to pH 2. 76, 104, 105, 108–111, 113. Purgeable Halocarbons. 26. 2-Chloroethylvinyl ether ..... G, FP-lined septum ...... Cool, ≤6 °C,18 0.008% 14 days. 5 Na2S2O3 . 6, 57, 106. Purgeable aromatic G, FP-lined septum ...... Cool, ≤6 °C,18 0.008% 14 days.9 5 9 hydrocarbons. Na2S2O3, HCl to pH 2 . 3, 4. Acrolein and acrylonitrile G, FP-lined septum ...... Cool, ≤6 °C,18 0.008% 14 days.10 10 Na2S2O3, pH to 4–5 . 23, 30, 44, 49, 53, 77, 80, 81, G, FP-lined cap ...... Cool, ≤6 °C,18 0.008% 7 days until extraction, 40 11 98, 100, 112. Phenols . Na2S2O3. days after extraction. 7, 38. Benzidines 11 12 ...... G, FP-lined cap ...... Cool, ≤6 °C,18 0.008% 7 days until extraction.13 5 Na2S2O3 . 14, 17, 48, 50–52. Phthalate G, FP-lined cap ...... Cool, ≤6 °C 18 ...... 7 days until extraction, 40 esters 11. days after extraction. 82–84. Nitrosamines 11 14 ...... G, FP-lined cap ...... Cool, ≤6 °C,18 store in dark, 7 days until extraction, 40 5 0.008% Na2S2O3 . days after extraction. 88–94. PCBs 11 ...... G, FP-lined cap ...... Cool, ≤6 °C 18 ...... 1 year until extraction, 1 year after extraction. 54, 55, 75, 79. Nitroaromatics G, FP-lined cap ...... Cool, ≤6 °C,18 store in dark, 7 days until extraction, 40 11 5 and isophorone . 0.008% Na2S2O3 . days after extraction. 1, 2, 5, 8–12, 32, 33, 58, 59, G, FP-lined cap ...... Cool, ≤6 °C,18 store in dark, 7 days until extraction, 40 5 74, 78, 99, 101. Polynuclear 0.008% Na2S2O3 . days after extraction. aromatic hydrocarbons 11. 15, 16, 21, 31, 87. G, FP-lined cap ...... Cool, ≤6 °C,18 0.008% 7 days until extraction, 40 11 5 Haloethers . Na2S2O3 . days after extraction. 29, 35–37, 63–65, 107. G, FP-lined cap ...... Cool, ≤6 °C 18 ...... 7 days until extraction, 40 Chlorinated hydrocarbons 11. days after extraction.

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TABLE II—REQUIRED CONTAINERS, PRESERVATION TECHNIQUES, AND HOLDING TIMES—Continued

Parameter number/name Container 1 Preservation 23 Maximum holding time 4

60–62, 66–72, 85, 86, 95–97, G ...... See footnote 11 ...... See footnote 11. 102, 103. CDDs/CDFs 11. Aqueous Samples: Field G ...... Cool, ≤6 °C,18 0.008% 1 year. 5 and Lab Preservation. Na2S2O3, pH <9. Solids and Mixed-Phase G ...... Cool, ≤6 °C 18 ...... 7 days. Samples: Field Preser- vation. Tissue Samples: Field G ...... Cool, ≤6 °C 18 ...... 24 hours. Preservation. Solids, Mixed-Phase, and G ...... Freeze, ≤¥10 °C ...... 1 year. Tissue Samples: Lab Preservation.

114–118. Alkylated phenols .... G ...... Cool, <6 °C, H2SO4 to pH <2 28 days until extraction, 40 days after extraction. 119. Adsorbable Organic G ...... Cool, <6 °C, 0.008% Hold at least 3 days, but not Halides (AOX). Na2S2O3, HNO3 to pH <2. more than 6 months. 120. Chlorinated Phenolics ..... G, FP-lined cap ...... Cool, <6 °C, 0.008% 30 days until acetylation, 30 Na2S2O3, H2SO4 to pH <2. days after acetylation.

Table ID—Pesticides Tests

1–70. Pesticides 11 ...... G, FP-lined cap ...... Cool, ≤6 °C,18 pH 5–9 15 ...... 7 days until extraction, 40 days after extraction.

Table IE—Radiological Tests

1–5. Alpha, beta, and radium .. P, FP, G ...... HNO3 to pH <2 ...... 6 months.

Table IH—Bacterial Tests

1–4. Coliform, total, fecal ...... PA, G ...... Cool, <10 °C, 0.008% 8 hours.22 23 5 Na2S2O3 . 5. E. coli ...... PA, G ...... Cool, <10 °C, 0. 008% 8 hours.22 5 Na2S2O3 . 6. Fecal streptococci ...... PA, G ...... Cool, <10 °C, 0.008% 8 hours.22 5 Na2S2O3 . 7. Enterococci ...... PA, G ...... Cool, <10 °C, 0. 008% 8 hours.22 5 Na2S2O3 .

Table IH—Protozoan Tests

8. Cryptosporidium ...... LDPE; field filtration ...... 1–10 °C ...... 96 hours.21 9. Giardia ...... LDPE; field filtration ...... 1–10 °C ...... 96 hours.21

1 ‘‘P’’ is for polyethylene; ‘‘FP’’ is fluoropolymer (polytetrafluoroethylene (PTFE); Teflon®), or other fluoropolymer, unless stated otherwise in this Table II; ‘‘G’’ is glass; ‘‘PA’’ is any plastic that is made of a sterilizable material (polypropylene or other autoclavable plastic); ‘‘LDPE’’ is low density polyethylene. 2 Except where noted in this Table II and the method for the parameter, preserve each grab sample within 15 minutes of col- lection. For a composite sample collected with an automated sample (e.g., using a 24-hour composite sample; see 40 CFR 122.21(g)(7)(i) or 40 CFR part 403, appendix E), refrigerate the sample at ≤6 °C during collection unless specified otherwise in this Table II or in the method(s). For a composite sample to be split into separate aliquots for preservation and/or analysis, main- tain the sample at ≤6 °C, unless specified otherwise in this Table II or in the method(s), until collection, splitting, and preservation is completed. Add the to the sample container prior to sample collection when the preservative will not compromise the integrity of a grab sample, a composite sample, or aliquot split from a composite sample within 15 minutes of collection. If a composite measurement is required but a composite sample would compromise sample integrity, individual grab samples must be collected at prescribed time intervals (e.g., 4 samples over the course of a day, at 6-hour intervals). Grab samples must be analyzed separately and the concentrations averaged. Alternatively, grab samples may be collected in the field and composited in the laboratory if the compositing procedure produces results equivalent to results produced by arithmetic averaging of results of analysis of individual grab samples. For examples of laboratory compositing procedures, see EPA Method 1664 Rev. A (oil and grease) and the procedures at 40 CFR 141.24(f)(14)(iv) and (v) (volatile organics). 3 When any sample is to be shipped by common carrier or sent via the U.S. Postal Service, it must comply with the Depart- ment of Transportation Hazardous Materials Regulations (49 CFR part 172). The person offering such material for transportation is responsible for ensuring such compliance. For the preservation requirement of Table II, the Office of Hazardous Materials, Ma- terials Transportation Bureau, Department of Transportation has determined that the Hazardous Materials Regulations do not apply to the following materials: Hydrochloric acid (HCl) in water solutions at concentrations of 0.04% by weight or less (pH about 1.96 or greater; Nitric acid (HNO3) in water solutions at concentrations of 0.15% by weight or less (pH about 1.62 or great- er); Sulfuric acid (H2SO4) in water solutions at concentrations of 0.35% by weight or less (pH about 1.15 or greater); and Sodium hydroxide (NaOH) in water solutions at concentrations of 0.080% by weight or less (pH about 12.30 or less).

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4 Samples should be analyzed as soon as possible after collection. The times listed are the maximum times that samples may be held before the start of analysis and still be considered valid. Samples may be held for longer periods only if the permittee or monitoring laboratory have data on file to show that, for the specific types of samples under study, the analytes are stable for the longer time, and has received a variance from the Regional ATP Coordinator under § 136.3(e). For a grab sample, the holding time begins at the time of collection. For a composite sample collected with an automated sampler (e.g., using a 24-hour com- posite sampler; see 40 CFR 122.21(g)(7)(i) or 40 CFR part 403, appendix E), the holding time begins at the time of the end of collection of the composite sample. For a set of grab samples composited in the field or laboratory, the holding time begins at the time of collection of the last grab sample in the set. Some samples may not be stable for the maximum time period given in the table. A permittee or monitoring laboratory is obligated to hold the sample for a shorter time if it knows that a shorter time is necessary to maintain sample stability. See § 136.3(e) for details. The date and time of collection of an individual grab sample is the date and time at which the sample is collected. For a set of grab samples to be composited, and that are all collected on the same calendar date, the date of collection is the date on which the samples are collected. For a set of grab samples to be com- posited, and that are collected across two calendar dates, the date of collection is the dates of the two days; e.g., November 14– 15. For a composite sample collected automatically on a given date, the date of collection is the date on which the sample is col- lected. For a composite sample collected automatically, and that is collected across two calendar dates, the date of collection is the dates of the two days; e.g., November 14–15. For static-renewal toxicity tests, each grab or composite sample may also be used to prepare test solutions for renewal at 24 h, 48 h, and/or 72 h after first use, if stored at 0–6 °C, with minimum head space. 5 ASTM D7365–09a specifies treatment options for samples containing oxidants (e.g., chlorine) for cyanide analyses. Also, Section 9060A of Standard Methods for the Examination of Water and Wastewater (20th and 21st editions) addresses dechlorination procedures for microbiological analyses. 6 Sampling, preservation and mitigating interferences in water samples for analysis of cyanide are described in ASTM D7365– 09a. There may be interferences that are not mitigated by the analytical test methods or D7365–09a. Any technique for removal or suppression of interference may be employed, provided the laboratory demonstrates that it more accurately measures cyanide through quality control measures described in the analytical test method. Any removal or suppression technique not described in D7365–09a or the analytical test method must be documented along with supporting data. 7 For dissolved metals, filter grab samples within 15 minutes of collection and before adding . For a composite sample collected with an automated sampler (e.g., using a 24-hour composite sampler; see 40 CFR 122.21(g)(7)(i) or 40 CFR part 403, appendix E), filter the sample within 15 minutes after completion of collection and before adding preservatives. If it is known or suspected that dissolved sample integrity will be compromised during collection of a composite sample collected auto- matically over time (e.g., by interchange of a metal between dissolved and suspended forms), collect and filter grab samples to be composited (footnote 2) in place of a composite sample collected automatically. 8 Guidance applies to samples to be analyzed by GC, LC, or GC/MS for specific compounds. 9 If the sample is not adjusted to pH 2, then the sample must be analyzed within seven days of sampling. 10 The pH adjustment is not required if acrolein will not be measured. Samples for acrolein receiving no pH adjustment must be analyzed within 3 days of sampling. 11 When the extractable analytes of concern fall within a single chemical category, the specified preservative and maximum holding times should be observed for optimum safeguard of sample integrity (i.e., use all necessary preservatives and hold for the shortest time listed). When the analytes of concern fall within two or more chemical categories, the sample may be preserved by cooling to ≤6 °C, reducing residual chlorine with 0.008% , storing in the dark, and adjusting the pH to 6–9; samples preserved in this manner may be held for seven days before extraction and for forty days after extraction. Exceptions to this optional preservation and holding time procedure are noted in footnote 5 (regarding the requirement for thiosulfate reduc- tion), and footnotes 12, 13 (regarding the analysis of benzidine). 12 If 1,2-diphenylhydrazine is likely to be present, adjust the pH of the sample to 4.0 ± 0.2 to prevent rearrangement to benzi- dine. 13 Extracts may be stored up to 30 days at <0 °C. 14 For the analysis of diphenylnitrosamine, add 0.008% Na2S2O3 and adjust pH to 7–10 with NaOH within 24 hours of sam- pling. 15 The pH adjustment may be performed upon receipt at the laboratory and may be omitted if the samples are extracted within 72 hours of collection. For the analysis of aldrin, add 0.008% Na2S2O3. 16 Place sufficient ice with the samples in the shipping container to ensure that ice is still present when the samples arrive at the laboratory. However, even if ice is present when the samples arrive, immediately measure the temperature of the samples and confirm that the preservation temperature maximum has not been exceeded. In the isolated cases where it can be docu- mented that this holding temperature cannot be met, the permittee can be given the option of on-site testing or can request a variance. The request for a variance should include supportive data which show that the toxicity of the effluent samples is not re- duced because of the increased holding temperature. Aqueous samples must not be frozen. Hand-delivered samples used on the day of collection do not need to be cooled to 0 to 6 °C prior to test initiation. 17 Samples collected for the determination of trace level mercury (<100 ng/L) using EPA Method 1631 must be collected in tightly-capped fluoropolymer or glass bottles and preserved with BrCl or HCl solution within 48 hours of sample collection. The time to preservation may be extended to 28 days if a sample is oxidized in the sample bottle. A sample collected for dissolved trace level mercury should be filtered in the laboratory within 24 hours of the time of collection. However, if circumstances pre- clude overnight shipment, the sample should be filtered in a designated clean area in the field in accordance with procedures given in Method 1669. If sample integrity will not be maintained by shipment to and filtration in the laboratory, the sample must be filtered in a designated clean area in the field within the time period necessary to maintain sample integrity. A sample that has been collected for determination of total or dissolved trace level mercury must be analyzed within 90 days of sample collection. 18 Aqueous samples must be preserved at ≤6 °C, and should not be frozen unless data demonstrating that sample freezing does not adversely impact sample integrity is maintained on file and accepted as valid by the regulatory authority. Also, for pur- poses of NPDES monitoring, the specification of ‘‘≤ °C’’ is used in place of the ‘‘4 °C’’ and ‘‘<4 °C’’ sample temperature require- ments listed in some methods. It is not necessary to measure the sample temperature to three significant figures (1/100th of 1 degree); rather, three significant figures are specified so that rounding down to 6 °C may not be used to meet the ≤6 °C require- ment. The preservation temperature does not apply to samples that are analyzed immediately (less than 15 minutes). 19 An aqueous sample may be collected and shipped without acid preservation. However, acid must be added at least 24 hours before analysis to dissolve any metals that adsorb to the container walls. If the sample must be analyzed within 24 hours of collection, add the acid immediately (see footnote 2). Soil and sediment samples do not need to be preserved with acid. The allowances in this footnote supersede the preservation and holding time requirements in the approved metals methods. 20 To achieve the 28-day holding time, use the ammonium sulfate buffer solution specified in EPA Method 218.6. The allow- ance in this footnote supersedes preservation and holding time requirements in the approved hexavalent chromium methods, un- less this supersession would compromise the measurement, in which case requirements in the method must be followed. 21 Holding time is calculated from time of sample collection to elution for samples shipped to the laboratory in bulk and cal- culated from the time of sample filtration to elution for samples filtered in the field. 22 Sample analysis should begin as soon as possible after receipt; sample incubation must be started no later than 8 hours from time of collection. 23 For fecal coliform samples for sewage sludge (biosolids) only, the holding time is extended to 24 hours for the following sample types using either EPA Method 1680 (LTB–EC) or 1681 (A–1): Class A composted, Class B aerobically digested, and Class B anaerobically digested. 24 The immediate filtration requirement in orthophosphate measurement is to assess the dissolved or bio-available form of orthophosphorus (i.e., that which passes through a 0.45-micron filter), hence the requirement to filter the sample immediately upon collection (i.e., within 15 minutes of collection).

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[38 FR 28758, Oct. 16, 1973]

EDITORIAL NOTE: For FEDERAL REGISTER citations affecting § 136.3, see the List of CFR Sec- tions Affected, which appears in the Finding Aids section of the printed volume and at www.govinfo.gov.

§ 136.4 Application for and approval of Coordinator may specify what addi- alternate test procedures for na- tional information might lead to a re- tionwide use. consideration of the application and (a) A written application for review notify the Regional Alternate Test of an alternate test procedure (alter- Procedure Coordinators of the dis- nate method) for nationwide use may approval recommendation. Based on be made by letter via email or by hard the National Coordinator’s rec- copy in triplicate to the National Al- ommended disapproval of a proposed ternate Test Procedure (ATP) Program alternate test procedure and an assess- Coordinator (National Coordinator), ment of any current approvals for lim- Office of Science and Technology ited uses for the unapproved method, (4303T), Office of Water, U.S. Environ- the Regional ATP Coordinator may de- mental Protection Agency, 1200 Penn- cide to withdraw approval of the meth- sylvania Ave. NW., Washington, DC od for limited use in the Region. 20460. Any application for an ATP (2) Where the National Coordinator under this paragraph (a) shall: has recommended approval of an appli- (1) Provide the name and address of the responsible person or firm making cant’s request for nationwide use of an the application. alternate test procedure, the National (2) Identify the pollutant(s) or pa- Coordinator will notify the applicant. rameter(s) for which nationwide ap- The National Coordinator will also no- proval of an alternate test procedure is tify the Regional ATP Coordinators being requested. that they may consider approval of (3) Provide a detailed description of this alternate test procedure for lim- the proposed alternate test procedure, ited use in their Regions based on the together with references to published information and data provided in the or other studies confirming the general application until the alternate test applicability of the alternate test pro- procedure is approved by publication in cedure for the analysis of the pollut- a final rule in the FEDERAL REGISTER. ant(s) or parameter(s) in wastewater (3) EPA will propose to amend this discharges from representative and part to include the alternate test pro- specified industrial or other categories. cedure in § 136.3. EPA shall make avail- (4) Provide comparability data for able for review all the factual bases for the performance of the proposed alter- its proposal, including the method, any native test procedure compared to the performance data submitted by the ap- performance of the reference method. plicant and any available EPA analysis (b) The National Coordinator may re- of those data. quest additional information and anal- yses from the applicant in order to (4) Following public comment, EPA evaluate whether the alternate test shall publish in the FEDERAL REGISTER procedure satisfies the applicable re- a final decision on whether to amend quirements of this part. this part to include the alternate test (c) Approval for nationwide use. (1) procedure as an approved analytical After a review of the application and method for nationwide use. any additional analyses requested from (5) Whenever the National Coordi- the applicant, the National Coordi- nator has recommended approval of an nator will notify the applicant, in writ- applicant’s ATP request for nationwide ing, of whether the National Coordi- use, any person may request an ap- nator will recommend approval or dis- proval of the method for limited use approval of the alternate test proce- under § 136.5 from the EPA Region. dure for nationwide use in CWA pro- grams. If the application is not rec- [77 FR 29809, May 18, 2012, as amended at 82 ommended for approval, the National FR 40874, Aug. 28, 2017]

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§ 136.5 Approval of alternate test pro- (and their associated laboratories) cedures for limited use. specified in the approval for the Re- (a) Any person may request the Re- gion. If the application is not approved, gional ATP Coordinator to approve the the Regional ATP Coordinator shall use of an alternate test procedure in specify what additional information the Region. might lead to a reconsideration of the (b) When the request for the use of an application. alternate test procedure concerns use (2) The Regional ATP Coordinator in a State with an NPDES permit pro- will forward a copy of every approval gram approved pursuant to section 402 and rejection notification to the Na- of the Act, the requestor shall first tional Alternate Test Procedure Coor- submit an application for limited use dinator. to the Director of the State agency [77 FR 29809, May 18, 2012, as amended at 82 having responsibility for issuance of FR 40875, Aug. 28, 2017] NPDES permits within such State (i.e., permitting authority). The Director § 136.6 Method modifications and ana- will forward the application to the Re- lytical requirements. gional ATP Coordinator with a rec- (a) Definitions of terms used in this sec- ommendation for or against approval. tion—(1) Analyst means the person or (c) Any application for approval of an laboratory using a test procedure (ana- alternate test procedure for limited use lytical method) in this part. may be made by letter, email or by (2) Chemistry of the method means the hard copy. The application shall in- reagents and reactions used in a test clude the following: (1) Provide the name and address of procedure that allow determination of the applicant and the applicable ID the analyte(s) of interest in an environ- number of the existing or pending per- mental sample. mit(s) and issuing agency for which use (3) Determinative technique means the of the alternate test procedure is re- way in which an analyte is identified quested, and the discharge serial num- and quantified (e.g., colorimetry, mass ber. spectrometry). (2) Identify the pollutant or param- (4) Equivalent performance means that eter for which approval of an alternate the modified method produces results test procedure is being requested. that meet or exceed the QC acceptance (3) Provide justification for using criteria of the approved method. testing procedures other than those (5) Method-defined analyte means an specified in Tables IA through IH of analyte defined solely by the method § 136.3, or in the NPDES permit. used to determine the analyte. Such an (4) Provide a detailed description of analyte may be a physical parameter, a the proposed alternate test procedure, parameter that is not a specific chem- together with references to published ical, or a parameter that may be com- studies of the applicability of the alter- prised of a number of substances. Ex- nate test procedure to the effluents in amples of such analytes include tem- question. perature, oil and grease, total sus- (5) Provide comparability data for pended solids, total phenolics, tur- the performance of the proposed alter- bidity, chemical oxygen demand, and nate test procedure compared to the biochemical oxygen demand. performance of the reference method. (6) QC means ‘‘quality control.’’ (d) Approval for limited use. (1) The (b) Method modifications. (1) If the un- Regional ATP Coordinator will review derlying chemistry and determinative the application and notify the appli- technique in a modified method are es- cant and the appropriate State agency sentially the same as an approved Part of approval or rejection of the use of 136 method, then the modified method the alternate test procedure. The ap- is an equivalent and acceptable alter- proval may be restricted to use only native to the approved method pro- with respect to a specific discharge or vided the requirements of this section facility (and its laboratory) or, at the are met. However, those who develop or discretion of the Regional ATP Coordi- use a modification to an approved nator, to all dischargers or facilities (Part 136) method must document that

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the performance of the modified meth- ceptance criteria, the modified method od, in the matrix to which the modified must use these QC tests and the modi- method will be applied, is equivalent to fied method must meet the QC accept- the performance of the approved meth- ance criteria with the following condi- od. If such a demonstration cannot be tions: made and documented, then the modi- (A) The analyst may only rely on QC fied method is not an acceptable alter- tests and QC acceptance criteria in a native to the approved method. Sup- method if it includes wastewater ma- porting documentation must, if appli- trix QC tests and QC acceptance cri- cable, include the routine initial dem- teria (e.g., matrix spikes) and both ini- onstration of capability and ongoing tial (start-up) and ongoing QC tests QC including determination of preci- and QC acceptance criteria. sion and accuracy, detection limits, (B) If the approved method does not and matrix spike recoveries. Initial contain QC tests and QC acceptance demonstration of capability typically criteria or if the QC tests and QC ac- includes analysis of four replicates of a ceptance criteria in the method do not mid-level standard and a method detec- meet the requirements of this section, tion limit study. Ongoing quality con- then the analyst must employ QC tests trol typically includes method blanks, published in the ‘‘equivalent’’ of a Part mid-level laboratory control samples, 136 method that has such QC, or the es- and matrix spikes (QC is as specified in sential QC requirements specified at the method). The method is considered 136.7, as applicable. If the approved equivalent if the quality control re- method is from a compendium or VCSB quirements in the reference method are and the QA/QC requirements are pub- achieved. Where the laboratory is using lished in other parts of that organiza- a vendor-supplied method, it is the QC tion’s compendium rather than within criteria in the reference method, not the Part 136 method then that part of the vendor’s method, that must be met the organization’s compendium must to show equivalency. Where a sample be used for the QC tests. preparation step is required (i.e., diges- (C) In addition, the analyst must per- tion, distillation), QC tests are to be form ongoing QC tests, including as- run using standards treated in the sessment of performance of the modi- same way as the samples. The method fied method on the sample matrix (e.g., user’s Standard Operating Procedure analysis of a matrix spike/matrix spike (SOP) must clearly document the duplicate pair for every twenty sam- modifications made to the reference ples), and analysis of an ongoing preci- method. Examples of allowed method sion and recovery sample (e.g., labora- modifications are listed in this section. tory fortified blank or blank spike) and If the method user is uncertain wheth- a blank with each batch of 20 or fewer er a method modification is allowed, samples. the Regional ATP Coordinator or Di- (D) If the performance of the modi- rector should be contacted for approval fied method in the wastewater matrix prior to implementing the modifica- or reagent water does not meet or ex- tion. The method user should also com- ceed the QC acceptance criteria, the plete necessary performance checks to method modification may not be used. verify that acceptable performance is (ii) Requirements for documentation. achieved with the method modification The modified method must be docu- prior to analyses of compliance sam- mented in a method write-up or an ad- ples. dendum that describes the modifica- (2) Requirements. The modified meth- tion(s) to the approved method prior to od must meet or exceed performance of the use of the method for compliance the approved method(s) for the purposes. The write-up or addendum analyte(s) of interest, as documented must include a reference number (e.g., by meeting the initial and ongoing method number), revision number, and quality control requirements in the revision date so that it may be ref- method. erenced accurately. In addition, the or- (i) Requirements for establishing equiv- ganization that uses the modified alent performance. If the approved method must document the results of method contains QC tests and QC ac- QC tests and keep these records, along

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with a copy of the method write-up or the requirements of this section are addendum, for review by an auditor. met include: (3) Restrictions. An analyst may not (i) Changes between manual method, modify an approved Clean Water Act flow analyzer, and discrete instrumen- analytical method for a method-de- tation. fined analyte. In addition, an analyst (ii) Changes in chromatographic col- may not modify an approved method if umns or temperature programs. the modification would result in meas- (iii) Changes between automated and urement of a different form or species manual sample preparation, such as di- of an analyte. Changes in method pro- gestions, distillations, and extractions; cedures are not allowed if such changes in-line sample preparation is an ac- would alter the defined chemistry (i.e., ceptable form of automated sample method principle) of the unmodified preparation for CWA methods. method. For example, phenol method (iv) In general, ICP–MS is a sensitive 420.1 or 420.4 defines phenolics as ferric and selective detector for metal anal- iron oxidized compounds that react ysis; however isobaric interference can with 4-aminoantipyrine (4-AAP) at pH cause problems for quantitative deter- 10 after being distilled from acid solu- mination, as well as identification tion. Because total phenolics rep- based on the isotope pattern. Inter- resents a group of compounds that all ference reduction technologies, such as react at different efficiencies with 4- collision cells or reaction cells, are de- AAP, changing test conditions likely signed to reduce the effect of would change the behavior of these dif- ferent phenolic compounds. An analyst spectroscopic interferences that may may not modify any sample collection, bias results for the element of interest. preservation, or holding time require- The use of interference reduction tech- ments of an approved method. Such nologies is allowed, provided the meth- modifications to sample collection, od performance specifications relevant preservation, and holding time require- to ICP–MS measurements are met. ments do not fall within the scope of (v) The use of EPA Method 200.2 or the flexibility allowed at § 136.6. Meth- the sample preparation steps from EPA od flexibility refers to modifications of Method 1638, including the use of the analytical procedures used for closed-vessel digestion, is allowed for identification and measurement of the EPA Method 200.8, provided the method analyte only and does not apply to performance specifications relevant to sample collection, preservation, or the ICP–MS are met. holding time procedures, which may (vi) Changes in pH adjustment re- only be modified as specified in agents. Changes in compounds used to § 136.3(e). adjust pH are acceptable as long as (4) Allowable changes. Except as noted they do not produce interference. For under paragraph (b)(3) of this section, example, using a different acid to ad- an analyst may modify an approved just pH in colorimetric methods. test procedure (analytical method) pro- (vii) Changes in buffer reagents are vided that the underlying reactions acceptable provided that the changes and principles used in the approved do not produce interferences. method remain essentially the same, (viii) Changes in the order of reagent and provided that the requirements of addition are acceptable provided that this section are met. If equal or better the change does not alter the chem- performance can be obtained with an istry and does not produce an inter- alternative reagent, then it is allowed. ference. For example, using the same A laboratory wishing to use these reagents, but adding them in different modifications must demonstrate ac- order, or preparing them in combined ceptable method performance by per- or separate solutions (so they can be forming and documenting all applica- added separately), is allowed, provided ble initial demonstration of capability reagent stability or method perform- and ongoing QC tests and meeting all ance is equivalent or improved. applicable QC acceptance criteria as (ix) Changes in calibration range described in § 136.7. Some examples of (provided that the modified range cov- the allowed types of changes, provided ers any relevant regulatory limit and

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the method performance specifications tions be extrapolated for instrument for calibration are met). responses that exceed that of the most (x) Changes in calibration model. (A) concentrated calibrator. Examples of Linear calibration models do not ade- methods with nonlinear calibration quately fit calibration data with one or functions include chloride by SM4500– two inflection points. For example, Cl–E–1997, hardness by EPA Method vendor-supplied data acquisition and 130.1, cyanide by ASTM D6888 or processing software on some instru- OIA1677, Kjeldahl nitrogen by PAI– ments may provide quadratic fitting DK03, and anions by EPA Method 300.0. functions to handle such situations. If (B) As an alternative to using the av- the calibration data for a particular erage response factor, the quality of analytical method routinely display the calibration may be evaluated using quadratic character, using quadratic the Relative Standard Error (RSE). fitting functions may be acceptable. In The acceptance criterion for the RSE is such cases, the minimum number of the same as the acceptance criterion calibrators for second order fits should for Relative Standard Deviation (RSD), be six, and in no case should concentra- in the method. RSE is calculated as:

Where: used the value should be ≤20%. Note

x′i = Calculated concentration at level i that the use of the RSE is included as xi = Actual concentration of the calibration an alternative to the use of the correla- level i tion coefficient as a measure of the n = Number of calibration points suitability of a calibration curve. It is p = Number of terms in the fitting equation not necessary to evaluate both the (average = 1, linear = 2, quadratic = 3) RSE and the correlation coefficient. (C) Using the RSE as a metric has (xi) Changes in equipment such as the added advantage of allowing the equipment from a vendor different same numerical standard to be applied from the one specified in the method. to the calibration model, regardless of (xii) The use of micro or midi dis- the form of the model. Thus, if a meth- tillation apparatus in place of macro od states that the RSD should be ≤20% distillation apparatus. for the traditional linear model (xiii) The use of prepackaged re- through the origin, then the RSE ac- agents. ceptance limit can remain ≤20% as (xiv) The use of digital titrators and well. Similarly, if a method provides methods where the underlying chem- an RSD acceptance limit of ≤15%, then istry used for the determination is that same figure can be used as the ac- similar to that used in the approved ceptance limit for the RSE. The RSE method. may be used as an alternative to cor- (xv) Use of selected ion monitoring relation coefficients and coefficients of (SIM) mode for analytes that cannot be determination for evaluating calibra- effectively analyzed in full-scan mode tion curves for any of the methods at and reach the required sensitivity. part 136. If the method includes a nu- False positives are more of a concern merical criterion for the RSD, then the when using SIM analysis, so at a min- same numerical value is used for the imum, one quantitation and two quali- RSE. Some older methods do not in- fying ions must be monitored for each clude any criterion for the calibration analyte (unless fewer than three ions curve—for these methods, if RSE is with intensity greater than 15% of the

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base peak are available). The ratio of time of four minutes is recommended, each of the two qualifying ions to the however a shorter desorb time may be quantitation ion must be evaluated and used, provided that all QC specifica- should agree with the ratio observed in tions in the method are met. an authentic standard within ±20 per- (F) Use of water management tech- cent. Analyst judgment must be ap- niques is allowed. Water is always col- plied to the evaluation of ion ratios be- lected on the trap along with the cause the ratios can be affected by co- analytes and is a significant inter- eluting compounds present in the sam- ference for analytical systems (GC and ple matrix. The signal-to-noise ratio of GC/MS). Modern water management the least sensitive ion should be at techniques (e.g., dry purge or condensa- least 3:1. Retention time in the sample tion points) can remove moisture from should match within 0.05 minute of an the sample stream and improve analyt- authentic standard analyzed under ical performance. identical conditions. Matrix inter- (xvii) If the characteristics of a ferences can cause minor shifts in re- wastewater matrix prevent efficient re- tention time and may be evident as covery of organic pollutants and pre- shifts in the retention times of the in- vent the method from meeting QC re- ternal standards. The total scan time quirements, the analyst may attempt should be such that a minimum of to resolve the issue by adding salts to eight scans are obtained per the sample, provided that such salts do chromatographic peak. not react with or introduce the target (xvi) Changes are allowed in purge- pollutant into the sample (as evidenced and-trap sample volumes or operating by the analysis of method blanks, lab- conditions. Some examples are: oratory control samples, and spiked (A) Changes in purge time and purge- samples that also contain such salts), gas flow rate. A change in purge time and that all requirements of paragraph and purge-gas flow rate is allowed pro- (b)(2) of this section are met. Samples vided that sufficient total purge vol- having residual chlorine or other halo- ume is used to achieve the required gen must be dechlorinated prior to the minimum detectible concentration and addition of such salts. calibration range for all compounds. In (xviii) If the characteristics of a general, a purge rate in the range 20– wastewater matrix result in poor sam- 200 mL/min and a total purge volume in ple dispersion or reagent deposition on the range 240–880 mL are recommended. equipment and prevent the analyst (B) Use of nitrogen or helium as a from meeting QC requirements, the an- purge gas, provided that the required alyst may attempt to resolve the issue sensitivities for all compounds are by adding a inert surfactant that does met. not affect the chemistry of the method, (C) Sample temperature during the such as Brij-35 or sodium dodecyl sul- purge state. Gentle heating of the sam- fate (SDS), provided that such surfac- ple during purging (e.g., 40 °C) in- tant does not react with or introduce creases purging efficiency of hydro- the target pollutant into the sample philic compounds and may improve (as evidenced by the analysis of method sample-to-sample repeatability because blanks, laboratory control samples, all samples are purged under precisely and spiked samples that also contain the same conditions. such surfactant) and that all require- (D) Trap sorbent. Any trap design is ments of paragraph (b)(1) and (b)(2) of acceptable, provided that the data ac- this section are met. Samples having quired meet all QC criteria. residual chlorine or other halogen (E) Changes to the desorb time. must be dechlorinated prior to the ad- Shortening the desorb time (e.g., from4 dition of such surfactant. minutes to 1 minute) may not affect (xix) The use of gas diffusion (using compound recoveries, and can shorten pH change to convert the analyte to overall cycle time and significantly re- gaseous form and/or heat to separate duce the amount of water introduced an analyte contained in steam from the to the analytical system, thus improv- sample matrix) across a hydrophobic ing the precision of analysis, especially semi-permeable membrane to separate for water-soluble analytes. A desorb the analyte of interest from the sample

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matrix may be used in place of manual Part 136 methods from a consensus or- or automated distillation in methods ganization. For example, Standard for analysis such as ammonia, total cy- Methods contains QA/QC procedures in anide, total Kjeldahl nitrogen, and the Part 1000 section of the Standard total phenols. These procedures do not Methods Compendium. The permittee/ replace the digestion procedures speci- laboratory shall follow these QA/QC fied in the approved methods and must procedures, as described in the method be used in conjunction with those pro- or methods compendium. If the method cedures. lacks QA/QC procedures, the permittee/ (xx) Changes in equipment operating laboratory has the following options to parameters such as the monitoring comply with the QA/QC requirements: wavelength of a colorimeter or the re- (a) Refer to and follow the QA/QC action time and temperature as needed published in the ‘‘equivalent’’ EPA to achieve the chemical reactions de- method for that parameter that has fined in the unmodified CWA method. such QA/QC procedures; For example, molybdenum blue phos- (b) Refer to the appropriate QA/QC phate methods have two absorbance section(s) of an approved part 136 meth- maxima, one at about 660 nm and an- od from a consensus organization com- other at about 880 nm. The former is pendium; about 2.5 times less sensitive than the (c)(1) Incorporate the following latter. Wavelength choice provides a twelve quality control elements, where cost-effective, dilution-free means to applicable, into the laboratory’s docu- increase sensitivity of molybdenum mented standard operating procedure blue phosphate methods. (SOP) for performing compliance anal- (xxi) Interchange of oxidants, such as yses when using an approved part 136 the use of titanium oxide in UV-as- method when the method lacks such sisted automated digestion of TOC and QA/QC procedures. One or more of the total phosphorus, as long as complete twelve QC elements may not apply to a oxidation can be demonstrated. given method and may be omitted if a (xxii) Use of an axially viewed torch written rationale is provided indicating with Method 200.7. why the element(s) is/are inappropriate (c) The permittee must notify their for a specific method. permitting authority of the intent to (i) Demonstration of Capability use a modified method. Such notifica- (DOC); tion should be of the form ‘‘Method xxx (ii) Method Detection Limit (MDL); has been modified within the flexibility (iii) Laboratory reagent blank (LRB), allowed in 40 CFR 136.6.’’ The permittee also referred to as method blank (MB); may indicate the specific paragraph of (iv) Laboratory fortified blank § 136.6 allowing the method modifica- (LFB), also referred to as a spiked tion. Specific details of the modifica- blank, or laboratory control sample tion need not be provided, but must be (LCS); documented in the Standard Operating (v) Matrix spike (MS) and matrix Procedure (SOP) and maintained by spike duplicate (MSD), or laboratory the analytical laboratory that per- fortified matrix (LFM) and LFM dupli- forms the analysis. cate, may be used for suspected matrix [77 FR 29810, May 18, 2012, as amended at 82 interference problems to assess preci- FR 40875, Aug. 28, 2017] sion; (vi) Internal standards (for GC/MS § 136.7 Quality assurance and quality analyses), surrogate standards (for or- control. ganic analysis) or tracers (for The permittee/laboratory shall use radiochemistry); suitable QA/QC procedures when con- (vii) Calibration (initial and con- ducting compliance analyses with any tinuing), also referred to as initial cali- part 136 chemical method or an alter- bration verification (ICV) and con- native method specified by the permit- tinuing calibration verification (CCV); ting authority. These QA/QC proce- (viii) Control charts (or other trend dures are generally included in the an- analyses of quality control results); alytical method or may be part of the (ix) Corrective action (root cause methods compendium for approved analysis);

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(x) QC acceptance criteria; technique. This method describes analytical (xi) Definitions of preparation and conditions for a second gas chromatographic analytical batches that may drive QC column that can be used to confirm measure- frequencies; and ments made with the primary column. Meth- od 624 provides gas chromatograph/mass (xii) Minimum frequency for con- spectrometer (GC/MS) conditions appro- ducting all QC elements. priate for the qualitative and quantitative (2) These twelve quality control ele- confirmation of results for most of the pa- ments must be clearly documented in rameters listed above. the written standard operating proce- 1.3 The method detection limit (MDL, de- dure for each analytical method not fined in Section 12.1) 1 for each parameter is containing QA/QC procedures, where listed in Table 1. The MDL for a specific applicable. wastewater may differ from those listed, de- pending upon the nature of interferences in [77 FR 29813, May 18, 2012] the sample matrix. 1.4 Any modification of this method, be- APPENDIX A TO PART 136—METHODS FOR yond those expressly permitted, shall be con- ORGANIC CHEMICAL ANALYSIS OF sidered as a major modification subject to MUNICIPAL AND INDUSTRIAL WASTE- application and approval of alternate test procedures under 40 CFR 136.4 and 136.5. WATER 1.5 This method is restricted to use by or METHOD 601—PURGEABLE HALOCARBONS under the supervision of analysts experi- enced in the operation of a purge and trap 1. Scope and Application system and a gas chromatograph and in the 1.1 This method covers the determination interpretation of gas chromatograms. Each of 29 purgeable halocarbons. analyst must demonstrate the ability to gen- The following parameters may be deter- erate acceptable results with this method mined by this method: using the procedure described in Section 8.2. 2. Summary of Method Parameter STORET CAS No. No. 2.1 An inert gas is bubbled through a 5-mL Bromodichloromethane ...... 32101 75–27–4 water sample contained in a specially-de- Bromoform ...... 32104 75–25–2 signed purging chamber at ambient tempera- Bromomethane ...... 34413 74–83–9 ture. The halocarbons are efficiently trans- Carbon tetrachloride ...... 32102 56–23–5 ferred from the aqueous phase to the vapor Chlorobenzene ...... 34301 108–90–7 phase. The vapor is swept through a sorbent Chloroethane ...... 34311 75–00–3 2-Chloroethylvinyl ether ...... 34576 100–75–8 trap where the halocarbons are trapped. Chloroform ...... 32106 67–66–3 After purging is completed, the trap is heat- Chloromethane ...... 34418 74–87–3 ed and backflushed with the inert gas to Dibromochloromethane ...... 32105 124–48–1 desorb the halocarbons onto a gas 1,2-Dichlorobenzene ...... 34536 95–50–1 chromatographic column. The gas chro- 1,3-Dichlorobenzene ...... 34566 541–73–1 matograph is temperature programmed to 1,4-Dichlorobenzene ...... 34571 106–46–7 Dichlorodifluoromethane ...... 34668 75–71–8 separate the halocarbons which are then de- 1,1-Dichloroethane ...... 34496 75–34–3 tected with a halide-specific detector. 23 1,2-Dichloroethane ...... 34531 107–06–2 2.2 The method provides an optional gas 1,1-Dichloroethane ...... 34501 75–35–4 chromatographic column that may be help- trans-1,2-Dichloroethene ...... 34546 156–60–5 ful in resolving the compounds of interest 1,2-Dichloropropane ...... 34541 78–87–5 from interferences that may occur. cis-1,3-Dichloropropene ...... 34704 10061–01–5 trans-1,3-Dichloropropene ...... 34699 10061–02–6 3. Interferences Methylene chloride ...... 34423 75–09–2 1,1,2,2-Tetrachloroethane ...... 34516 79–34–5 3.1 Impurities in the purge gas and or- Tetrachloroethene ...... 34475 127–18–4 ganic compounds outgassing from the plumb- 1,1,1-Trichloroethane ...... 34506 71–55–6 1,1,2-Trichloroethane ...... 34511 79–00–5 ing ahead of the trap account for the major- Tetrachloroethene ...... 39180 79–01–6 ity of contamination problems. The analyt- Trichlorofluoromethane ...... 34488 75–69–4 ical system must be demonstrated to be free Vinyl chloride ...... 39715 75–01–4 from contamination under the conditions of the analysis by running laboratory reagent 1.2 This is a purge and trap gas blanks as described in Section 8.1.3. The use chromatographic (GC) method applicable to of non-Teflon plastic tubing, non-Teflon the determination of the compounds listed thread sealants, or flow controllers with rub- above in municipal and industrial discharges ber components in the purge and trap system as provided under 40 CFR 136.1. When this should be avoided. method is used to analyze unfamiliar sam- 3.2 Samples can be contaminated by diffu- ples for any or all of the compounds above, sion of volatile organics (particularly fluoro- compound identifications should be sup- and methylene chloride) through the ported by at least one additional qualitative septum seal ilto the sample during shipment

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and storage. A field reagent blank prepared wash, rinse with tap and distilled water, and from reagent water and carried through the dry at 105 °C for 1 h before use. sampling and handling protocol can serve as 5.2 Purge and trap system—The purge and a check on such contamination. trap system consists of three separate pieces 3.3 Contamination by carry-over can of equipment: a purging device, trap, and occur whenever high level and low level sam- desorber. Several complete systems are now ples are sequentially analyzed. To reduce commercially available. carry-over, the purging device and sample 5.2.1 The purging device must be designed syringe must be rinsed with reagent water to accept 5-mL samples with a water column between sample analyses. Whenever an un- at least 3 cm deep. The gaseous head space usually concentrated sample is encountered, between the water column and the trap must it should be followed by an analysis of rea- have a total volume of less than 15 mL. The gent water to check for cross contamination. purge gas must pass through the water col- For samples containing large amounts of umn as finely divided bubbles with a diame- water-soluble materials, suspended solids, ter of less than 3 mm at the origin. The high boiling compounds or high organohalide purge gas must be introduced no more than levels, it may be necessary to wash out the 5 mm from the base of the water column. purging device with a detergent solution, The purging device illustrated in Figure 1 rinse it with distilled water, and then dry it meets these design criteria. 5.2.2 The trap must be at least 25 cm long in a 105 °C oven between analyses. The trap and have an inside diameter of at least 0.105 and other parts of the system are also sub- in. The trap must be packed to contain the ject to contamination; therefore, frequent following minimum lengths of adsorbents: 1.0 bakeout and purging of the entire system cm of methyl silicone coated packing (Sec- may be required. tion 6.3.3), 7.7 cm of 2,6-diphenylene oxide 4. Safety polymer (Section 6.3.2), 7.7 cm of silica gel (Section 6.3.4), 7.7 cm of coconut 4.1 The toxicity or carcinogenicity of (Section 6.3.1). If it is not necessary to ana- each reagent used in this method has not lyze for dichlorodifluoromethane, the char- been precisely defined; however, each chem- coal can be eliminated, and the polymer sec- ical compound should be treated as a poten- tion lengthened to 15 cm. The minimum tial health hazard. From this viewpoint, ex- specifications for the trap are illustrated in posure to these chemicals must be reduced to Figure 2. the lowest possible level by whatever means 5.2.3 The desorber must be capable of rap- available. The laboratory is responsible for idly heating the trap to 180 °C. The polymer maintaining a current awareness file of section of the trap should not be heated OSHA regulations regarding the safe han- higher than 180 °C and the remaining sec- dling of the chemicals specified in this meth- tions should not exceed 200 °C. The desorber od. A reference file of material data handling illustrated in Figure 2 meets these design sheets should also be made available to all criteria. personnel involved in the chemical analysis. 5.2.4 The purge and trap system may be Additional references to laboratory safety assembled as a separate unit or be coupled to are available and have been identified 46 for a gas chromatograph as illustrated in Fig- the information of the analyst. ures 3 and 4. 4.2 The following parameters covered by 5.3 Gas chromatograph—An analytical this method have been tentatively classified system complete with a temperature pro- as known or suspected, human or mamma- grammable gas chromatograph suitable for lian carcinogens: carbon tetrachloride, chlo- on-column injection and all required acces- roform, 1,4-dichlorobenzene, and vinyl chlo- sories including syringes, analytical col- ride. Primary standards of these toxic com- umns, gases, detector, and strip-chart re- pounds should be prepared in a hood. A corder. A data system is recommended for NIOSH/MESA approved toxic gas respirator measuring peak areas. should be worn when the analyst handles 5.3.1 Column 1—8 ft long × 0.1 in. ID stain- high concentrations of these toxic com- less steel or glass, packed with 1% SP–1000 pounds. on Carbopack B (60/80 mesh) or equivalent. This column was used to develop the method 5. Apparatus and Materials performance statements in Section 12. Guidelines for the use of alternate column 5.1 Sampling equipment, for discrete sam- packings are provided in Section 10.1. pling. 5.3.2 Column 2—6 ft long × 0.1 in. ID stain- 5.1.1 Vial—25-mL capacity or larger, less steel or glass, packed with chemically equipped with a screw cap with a hole in the bonded n-octane on Porasil-C (100/120 mesh) center (Pierce #13075 or equivalent). Deter- or equivalent. gent wash, rinse with tap and distilled water, 5.3.3 Detector—Electrolytic conductivity and dry at 105 °C before use. or microcoulometric detector. These types of 5.1.2 Septum—Teflon-faced silicone detectors have proven effective in the anal- (Pierce #12722 or equivalent). Detergent ysis of wastewaters for the parameters listed

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in the scope (Section 1.1). The electrolytic flask. Allow the flask to stand, unstoppered, conductivity detector was used to develop for about 10 min or until all alcohol wetted the method performance statements in Sec- surfaces have dried. Weigh the flask to the tion 12. Guidelines for the use of alternate learest 0.1 mg. detectors are provided in Section 10.1. 6.5.2 Add the assayed reference material: 5.4 Syringes—5-mL glass hypodermic with 6.5.2.1 Liquid—Using a 100 μL syringe, im- Luerlok tip (two each), if applicable to the mediately add two or more drops of assayed purging device. reference material to the flask, then re- 5.5 Micro syringes—25-μL, 0.006 in. ID nee- weigh. Be sure that the drops fall directly dle. into the alcohol without contacting the neck 5.6 Syringe valve—2-way, with Luer ends of the flask. (three each). 6.5.2.2 Gases—To prepare standards for 5.7 Syringe—5-mL, gas-tight with shut-off any of the six halocarbons that boil below 30 valve. °C (bromomethane, chloroethane, 5.8 Bottle—15-mL, screw-cap, with Teflon chloromethane, dichlorodifluoromethane, cap liner. trichlorofluoromethane, vinyl chloride), fill 5.9 Balance—Analytical, capable of accu- a 5-mL valved gas-tight syringe with the ref- rately weighing 0.0001 g. erence standard to the 5.0-mL mark. Lower the needle to 5 mm above the methanol me- 6. Reagents niscus. Slowly introduce the reference stand- 6.1 Reagent water—Reagent water is de- ard above the surface of the liquid (the heavy fined as a water in which an interferent is gas will rapidly dissolve into the methanol). not observed at the MDL of the parameters 6.5.3 Reweigh, dilute to volume, stopper, of interest. then mix by inverting the flask several 6.1.1 Reagent water can be generated by times. Calculate the concentration in μg/μL passing tap water through a carbon filter bed from the net gain in weight. When compound containing about 1 lb of purity is assayed to be 96% or greater, the (Filtrasorb-300, Calgon Corp., or equivalent). weight can be used without correction to cal- 6.1.2 A water purification system culate the concentration of the stock stand- (Millipore Super-Q or equivalent) may be ard. Commercially prepared stock standards used to generate reagent water. can be used at any concentration if they are 6.1.3 Reagent water may also be prepared certified by the malufacturer or by an inde- by boiling water for 15 min. Subsequently, pendent source. while maintaining the temperature at 90 °C, 6.5.4 Transfer the stock standard solution bubble a contaminant-free inert gas through into a Teflon-sealed screw-cap bottle. Store, the water for 1 h. While still hot, transfer with minimal headspace, at ¥10 to ¥20 °C the water to a narrow mouth screw-cap bot- and protect from light. tle and seal with a Teflon-lined septum and 6.5.5 Prepare fresh standards weekly for cap. the six gases and 2-chloroethylvinyl ether. 6.2 Sodium thiosulfate—(ACS) Granular. All other standards must be replaced after 6.3 Trap Materials: one month, or sooner if comparison with 6.3.1 Coconut charcoal—6/10 mesh sieved check standards indicates a problem. to 26 mesh, Barnabey Cheney, CA–580–26 lot # 6.6 Secondary dilution standards—Using M–2649 or equivalent. stock standard solutions, prepare secondary 6.3.2 2,6-Diphenylene oxide polymer— dilution standards in methanol that contain Tenax, (60/80 mesh), chromatographic grade the compounds of interest, either singly or or equivalent. mixed together. The secondary dilution 6.3.3 Methyl silicone packing—3% OV–1 on standards should be prepared at concentra- Chromosorb-W (60/80 mesh) or equivalent. tions such that the aqueous calibration 6.3.4 Silica gel—35/60 mesh, Davison, standards prepared in Section 7.3.1 or 7.4.1 grade-15 or equivalent. will bracket the working range of the ana- 6.4 Methanol—Pesticide quality or equiv- lytical system. Secondary dilution standards alent. should be stored with minimal headspace and 6.5 Stock standard solutions—Stock should be checked frequently for signs of standard solutions may be prepared from degradation or evaporation, especially just pure standard materials or purchased as cer- prior to preparing calibration standards from tified solutions. Prepare stock standard solu- them. tions in methanol using assayed liquids or 6.7 Quality control check sample con- gases as appropriate. Because of the toxicity centrate—See Section 8.2.1. of some of the organohalides, primary dilu- 7. Calibration tions of these materials should be prepared in a hood. A NIOSH/MESA approved toxic 7.1 Assemble a purge and trap system that gas respirator should be used when the ana- meets the specifications in Section 5.2. Con- lyst handles high concentrations of such ma- dition the trap overnight at 180 °C by terials. backflushing with an inert gas flow of at 6.5.1 Place about 9.8 mL of methanol into least 20 mL/min. Condition the trap for 10 a 10-mL ground glass stoppered volumetric min once daily prior to use.

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7.2 Connect the purge and trap system to standard to 5.0 mL of sample or calibration a gas chromatograph. The gas chro- standard would be equivalent to 30 μg/L. matograph must be operated using tempera- 7.4.3 Analyze each calibration standard ture and flow rate conditions equivalent to according to Section 10, adding 10 μL of in- those given in Table 1. Calibrate the purge ternal standard spiking solution directly to and trap-gas chromatographic system using the syringe (Section 10.4). Tabulate peak either the external standard technique (Sec- height or area responses against concentra- tion 7.3) or the internal standard technique tion for each compound and internal stand- (Section 7.4). ard, and calculate response factors (RF) for 7.3 External standard calibration proce- each compound using Equation 1. dure: 7.3.1 Prepare calibration standards at a ()()ACsis miminum of three concentration levels for RF = μ each parameter by carefully adding 20.0 L of ()()ACis s one or more secondary dilution standards to 100, 500, or 1000 μL of reagent water. A 25-μL Equation 1 syringe with a 0.006 in. ID needle should be where: used for this operation. One of the external As = Response for the parameter to be meas- standards should be at a concentration near, ured. but above, the MDL (Table 1) and the other Ais = Response for the internal standard. concentrations should correspond to the ex- Cis = Concentration of the internal standard. pected range of concentrations found in real Cs = Concentration of the parameter to be samples or should define the working range measured. of the detector. These aqueous standards can If the RF value over the working range is a be stored up to 24 h, if held in sealed vials constant (<10% RSD), the RF can be assumed with zero headspace as described in Section to be invariant and the average RF can be 9.2. If not so stored, they must be discarded used for calculations. Alternatively, the re- after 1 h. sults can be used to plot a calibration curve 7.3.2 Analyze each calibration standard of response ratios, As/Ais, vs. RF. according to Section 10, and tabulate peak 7.5 The working calibration curve, cali- height or area responses versus the con- bration factor, or RF must be verified on centration in the standard. The results can each working day by the measurement of a be used to prepare a calibration curve for QC check sample. each compound. Alternatively, if the ratio of 7.5.1 Prepare the QC check sample as de- response to concentration (calibration fac- scribed in Section 8.2.2. tor) is a constant over the working range 7.5.2 Analyze the QC check sample accord- (<10% relative standard deviation, RSD), lin- ing to Section 10. earity through the origin can be assumed 7.5.3 For each parameter, compare the re- and the average ratio or calibration factor sponse (Q) with the corresponding calibra- can be used in place of a calibration curve. tion acceptance criteria found in Table 2. If 7.4 Internal standard calibration proce- the responses for all parameters of interest dure—To use this approach, the analyst must fall within the designated ranges, analysis of select one or more internal standards that actual samples can begin. If any individual Q are similar in analytical behavior to the falls outside the range, proceed according to compounds of interest. The analyst must fur- Section 7.5.4. ther demonstrate that the measurement of NOTE: The large number of parameters in the internal standard is not affected by Table 2 present a substantial probability method or matrix interferences. Because of that one or more will not meet the calibra- these limitations, no internal standard can tion acceptance criteria when all parameters be suggested that is applicable to all sam- are analyzed. ples. The compounds recommended for use as 7.5.4 Repeat the test only for those pa- surrogate spikes in Section 8.7 have been rameters that failed to meet the calibration used successfully as internal standards, be- acceptance criteria. If the response for a pa- cause of their generally unique retention rameter does not fall within the range in times. this second test, a new calibration curve, 7.4.1 Prepare calibration standards at a calibration factor, or RF must be prepared minimum of three concentration levels for for that parameter according to Section 7.3 each parameter of interest as described in or 7.4. Section 7.3.1. 8. Quality Control 7.4.2 Prepare a spiking solution con- taining each of the internal standards using 8.1 Each laboratory that uses this method the procedures described in Sections 6.5 and is required to operate a formal quality con- 6.6. It is recommended that the secondary di- trol program. The minimum requirements of lution standard be prepared at a concentra- this program consist of an initial demonstra- tion of 15 μg/mL of each internal standard tion of laboratory capability and an ongoing compound. The addition of 10 μL of this analysis of spiked samples to evaluate and

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document data quality. The laboratory must 8.2.2 Prepare a QC check sample to con- maintain records to document the quality of tain 20 μg/L of each parameter by adding 200 data that is generated. Ongoing data quality μL of QC check sample concentrate to 100 mL checks are compared with established per- of reagent water. formance criteria to determine if the results 8.2.3 Analyze four 5-mL aliquots of the of analyses meet the performance character- well-mixed QC check sample according to istics of the method. When results of sample Section 10. spikes indicate atypical method perform- 8.2.4 Calculate the average recovery (X¯ ) in ance, a quality control check standard must μg/L, and the standard deviation of the re- be analyzed to confirm that the measure- covery (s) in μg/L, for each parameter of in- ments were performed in an in-control mode terest using the four results. of operation. 8.2.5 For each parameter compare s and X¯ 8.1.1 The analyst must make an initial, with the corresponding acceptance criteria one-time, demonstration of the ability to for precision and accuracy, respectively, generate acceptable accuracy and precision found in Table 2. If s and X¯ for all param- with this method. This ability is established eters of interest meet the acceptance cri- as described in Section 8.2. teria, the system performance is acceptable 8.1.2 In recognition of advances that are and analysis of actual samples can begin. If occurring in chromatography, the analyst is any individual s exceeds the precision limit permitted certain options (detailed in Sec- or any individual X¯ falls outside the range tion 10.1) to improve the separations or lower for accuracy, then the system performance is the cost of measurements. Each time such a unacceptable for that parameter. modification is made to the method, the ana- NOTE: The large number of parameters in lyst is required to repeat the procedure in Table 2 present a substantial probability Section 8.2. that one or more will fail at least one of the 8.1.3 Each day, the analyst must analyze a acceptance criteria when all parameters are reagent water blank to demonstrate that analyzed. interferences from the analytical system are 8.2.6 When one or more of the parameters under control. tested fail at least one of the acceptance cri- 8.1.4 The laboratory must, on an ongoing teria, the analyst must proceed according to basis, spike and analyze a minimum of 10% Section 8.2.6.1 or 8.2.6.2. of all samples to monitor and evaluate lab- 8.2.6.1 Locate and correct the source of oratory data quality. This procedure is de- the problem and repeat the test for all pa- scribed in Section 8.3. rameters of interest beginning with Section 8.1.5 The laboratory must, on an ongoing 8.2.3. basis, demonstrate through the analyses of 8.2.6.2 Beginning with Section 8.2.3, repeat quality control check standards that the op- the test only for those parameters that eration of the measurement system is in con- failed to meet criteria. Repeated failure, trol. This procedure is described in Section however, will confirm a general problem 8.4. The frequency of the check standard with the measurement system. If this occurs, analyses is equivalent to 10% of all samples locate and correct the source of the problem analyzed but may be reduced if spike recov- and repeat the test for all compounds of in- eries from samples (Section 8.3) meet all terest beginning with Section 8.2.3. specified quality control criteria. 8.3 The laboratory must, on an ongoing 8.1.6 The laboratory must maintain per- basis, spike at least 10% of the samples from formance records to document the quality of each sample site being monitored to assess data that is generated. This procedure is de- accuracy. For laboratories analyzing one to scribed in Section 8.5. ten samples per month, at least one spiked 8.2 To establish the ability to generate sample per month is required. acceptable accuracy and precision, the ana- 8.3.1 The concentration of the spike in the lyst must perform the following operations. sample should be determined as follows: 8.2.1 A quality control (QC) check sample 8.3.1.1 If, as in compliance monitoring, concentrate is required containing each pa- the concentration of a specific parameter in rameter of interest at a concentration of 10 the sample is being checked against a regu- μg/mL in methanol. The QC check sample latory concentration limit, the spike should concentrate must be obtained from the U.S. be at that limit or 1 to 5 times higher than Environmental Protection Agency, Environ- the background concentration determined in mental Monitoring and Support Laboratory Section 8.3.2, whichever concentration would in Cincinnati, Ohio, if available. If not avail- be larger. able from that source, the QC check sample 8.3.1.2 If the concentration of a specific concentrate must be obtained from another parameter in the sample is not being external source. If not available from either checked against a limit specific to that pa- source above, the QC check sample con- rameter, the spike should be at 20 μg/L or 1 centrate must be prepared by the laboratory to 5 times higher than the background con- using stock standards prepared independ- centration determined in Section 8.3.2, ently from those used for calibration. whichever concentration would be larger.

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8.3.2 Analyze one 5-mL sample aliquot to 8.4.3 Compare the percent recovery (Ps) determine the background concentration (B) for each parameter with the corresponding of each parameter. If necessary, prepare a QC acceptance criteria found in Table 2. Only new QC check sample concentrate (Section parameters that failed the test in Section 8.3 8.2.1) appropriate for the background con- need to be compared with these criteria. If centrations in the sample. Spike a second 5- the recovery of any such parameter falls out- mL sample aliquot with 10 μL of the QC side the designated range, the laboratory check sample concentrate and analyze it to performance for that parameter is judged to determine the concentration after spiking be out of control, and the problem must be (A) of each parameter. Calculate each per- immediately identified and corrected. The cent recovery (P) as 100(A¥B)%/T, where T is analytical result for that parameter in the the known true value of the spike. unspiked sample is suspect and may not be 8.3.3 Compare the percent recovery (P) for reported for regulatory compliance purposes. each parameter with the corresponding QC 8.5 As part of the QC program for the lab- acceptance criteria found in Table 2. These oratory, method accuracy for wastewater acceptance criteria were calculated to in- samples must be assessed and records must clude an allowance for error in measurement be maintained. After the analysis of five of both the background and spike concentra- spiked wastewater samples as in Section 8.3, tions, assuming a spike to background ratio calculate the average percent recovery (P¯ ) of 5:1. This error will be accounted for to the and the standard deviation of the percent re- extent that the analyst’s spike to back- covery (s ). Express the accuracy assessment ground ratio approaches 5:1. 7 If spiking was p as a percent recovery interval from P¯ ¥2s to performed at a concentration lower than 20 p P¯ + 2s . If p¯ = 90% and s = 10%, for example, μg/L, the analyst must use either the QC ac- p p the accuracy interval is expressed as 70– ceptance criteria in Table 2, or optional QC 110%. Update the accuracy assessment for acceptance criteria calculated for the spe- each parameter on a regular basis (e.g. after cific spike concentration. To calculate op- each five to ten new accuracy measure- tional acceptance criteria for the recovery of ments). a parameter: (1) Calculate accuracy (X′) using the equation in Table 3, substituting 8.6 It is recommended that the laboratory the spike concentration (T) for C; (2) cal- adopt additional quality assurance practices culate overall precision (S′) using the equa- for use with this method. The specific prac- tion in Table 3, substituting X′ for X¯ ; (3) cal- tices that are most productive depend upon culate the range for recovery at the spike the needs of the laboratory and the nature of concentration as (100 X′/T)±2.44(100 S′/T)%. 7 the samples. Field duplicates may be ana- 8.3.4 If any individual P falls outside the lyzed to assess the precision of the environ- designated range for recovery, that param- mental measurements. When doubt exists eter has failed the acceptance criteria. A over the identification of a peak on the chro- check standard containing each parameter matogram, confirmatory techniques such as that failed the criteria must be analyzed as gas chromatography with a dissimilar col- described in Section 8.4. umn, specific element detector, or mass 8.4 If any parameter fails the acceptance spectrometer must be used. Whenever pos- criteria for recovery in Section 8.3, a QC sible, the laboratory should analyze standard check standard containing each parameter reference materials and participate in rel- that failed must be prepared and analyzed. evant performance evaluation studies. NOTE: The frequency for the required anal- 8.7 The analyst should monitor both the ysis of a QC check standard will depend upon performance of the analytical system and the number of parameters being simulta- the effectiveness of the method in dealing neously tested, the complexity of the sample with each sample matrix by spiking each matrix, and the performance of the labora- sample, standard, and reagent water blank tory. If the entire list of parameters in Table with surrogate halocarbons. A combination 2 must be measured in the sample in Section of bromochloromethane, 2-bromo-1- 8.3, the probability that the analysis of a QC chloropropane, and 1,4-dichlorobutane is rec- check standard will be required is high. In ommended to encompass the range of the this case the QC check standard should be temperature program used in this method. routinely analyzed with the spiked sample. From stock standard solutions prepared as 8.4.1 Prepare the QC check standard by in Section 6.5, add a volume to give 750 μg of adding 10 μL of QC check sample concentrate each surrogate to 45 mL of reagent water (Section 8.2.1 or 8.3.2) to 5 mL of reagent contained in a 50-mL volumetric flask, mix water. The QC check standard needs only to and dilute to volume for a concentration of contain the parameters that failed criteria 15 ng/μL. Add 10 μL of this surrogate spiking in the test in Section 8.3. solution directly into the 5-mL syringe with 8.4.2 Analyze the QC check standard to every sample and reference standard ana- determine the concentration measured (A) of lyzed. Prepare a fresh surrogate spiking solu- each parameter. Calculate each percent re- tion on a weekly basis. If the internal stand- covery (Ps) as 100 (A/T)%, where T is the true ard calibration procedure is being used, the value of the standard concentration. surrogate compounds may be added directly

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to the internal standard spiking solution 10.5 Attach the syringe-syringe valve as- (Section 7.4.2). sembly to the syringe valve on the purging device. Open the syringe valves and inject 9. Sample Collection, Preservation, and the sample into the purging chamber. Handling 10.6 Close both valves and purge the sam- 9.1 All samples must be iced or refrig- ple for 11.0 ±0.1 min at ambient temperature. erated from the time of collection until anal- 10.7 After the 11-min purge time, attach ysis. If the sample contains free or combined the trap to the chromatograph, adjust the chlorine, add sodium thiosulfate preserva- purge and trap system to the desorb mode tive (10 mg/40 mL is sufficient for up to 5 (Figure 4), and begin to temperature pro- ppm Cl2) to the empty sample bottle just gram the gas chromatograph. Introduce the prior to shipping to the sampling site. EPA trapped materials to the GC column by rap- Methods 330.4 and 330.5 may be used for idly heating the trap to 180 °C while measurement of residual chlorine. 8 Field backflushing the trap with an inert gas be- test kits are available for this purpose. tween 20 and 60 mL/min for 4 min. If rapid 9.2 Grab samples must be collected in heating of the trap cannot be achieved, the glass containers having a total volume of at GC column must be used as a secondary trap least 25 mL. Fill the sample bottle just to by cooling it to 30 °C (subambient tempera- overflowing in such a manner that no air ture, if poor peak geometry or random reten- bubbles pass through the sample as the bot- tion time problems persist) instead of the tle is being filled. Seal the bottle so that no initial program temperature of 45 °C air bubbles are entrapped in it. If preserva- 10.8 While the trap is being desorbed into tive has been added, shake vigorously for 1 the gas chromatograph, empty the purging min. Maintain the hermetic seal on the sam- chamber using the sample introduction sy- ple bottle until time of analysis. ringe. Wash the chamber with two 5-mL 9.3 All samples must be analyzed within flushes of reagent water. 14 days of collection. 3 10.9 After desorbing the sample for 4 min, recondition the trap by returning the purge 10. Procedure and trap system to the purge mode. Wait 15 10.1 Table 1 summarizes the recommended s then close the syringe valve on the purging operating conditions for the gas chro- device to begin gas flow through the trap. matograph. Included in this table are esti- The trap temperature should be maintained ° mated retention times and MDL that can be at 180 C After approximately 7 min, turn off achieved under these conditions. An example the trap heater and open the syringe valve to of the separations achieved by Column 1 is stop the gas flow through the trap. When the shown in Figure 5. Other packed columns, trap is cool, the next sample can be ana- chromatographic conditions, or detectors lyzed. may be used if the requirements of Section 10.10 Identify the parameters in the sam- 8.2 are met. ple by comparing the retention times of the 10.2 Calibrate the system daily as de- peaks in the sample chromatogram with scribed in Section 7. those of the peaks in standard 10.3 Adjust the purge gas (nitrogen or he- chromatograms. The width of the retention lium) flow rate to 40 mL/min. Attach the time window used to make identifications trap inlet to the purging device, and set the should be based upon measurements of ac- purge and trap system to purge (Figure 3). tual retention time variations of standards Open the syringe valve located on the purg- over the course of a day. Three times the ing device sample introduction needle. standard deviation of a retention time for a 10.4 Allow the sample to come to ambient compound can be used to calculate a sug- temperature prior to introducing it to the gested window size; however, the experience syringe. Remove the plunger from a 5-mL sy- of the analyst should weigh heavily in the ringe and attach a closed syringe valve. Open interpretation of chromatograms. the sample bottle (or standard) and carefully 10.11 If the response for a peak exceeds pour the sample into the syringe barrel to the working range of the system, prepare a just short of overflowing. Replace the sy- dilution of the sample with reagent water ringe plunger and compress the sample. Open from the aliquot in the second syringe and the syringe valve and vent any residual air reanalyze. while adjusting the sample volume to 5.0 mL. 11. Calculations Since this process of taking an aliquot de- stroys the validity of the sample for future 11.1 Determine the concentration of indi- analysis, the analyst should fill a second sy- vidual compounds in the sample. ringe at this time to protect against possible 11.1.1 If the external standard calibration loss of data. Add 10.0 μL of the surrogate procedure is used, calculate the concentra- spiking solution (Section 8.7) and 10.0 μL of tion of the parameter being measured from the internal standard spiking solution (Sec- the peak response using the calibration tion 7.4.2), if applicable, through the valve curve or calibration factor determined in bore, then close the valve. Section 7.3.2.

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11.1.2 If the internal standard calibration per-Litre-Levels by Gas Chromatography,’’ procedure is used, calculate the concentra- Journal of the American Water Works Associa- tion in the sample using the response factor tion, 66, 739 (1974). (RF) determined in Section 7.4.3 and Equa- 3. Bellar, T.A., and Lichtenberg, J.J. tion 2. ‘‘Semi-Automated Headspace Analysis of Equation 2 Drinking Waters and Industrial Waters for Purgeable Volatile Organic Compounds,’’ Proceedings from Symposium on Measure- ()()AC μ=sis ment of Organic Pollutants in Water and Concentration ( g/L) () Wastewater, American Society for Testing ()ARFis and Materials, STP 686, C.E. Van Hall, edi- where: tor, 1978. 4. ‘‘Carcinogens—Working With Carcino- A = Response for the parameter to be meas- s gens,’’ Department of Health, Education, and ured. Welfare, Public Health Service, Center for Ais = Response for the internal standard. C = Concentration of the internal standard. Disease Control, National Institute for Occu- is pational Safety and Health, Publication No. μ 11.2 Report results in g/L without correc- 77–206, August 1977. tion for recovery data. All QC data obtained 5. ‘‘OSHA Safety and Health Standards, should be reported with the sample results. General Industry’’ (29 CFR part 1910), Occu- 12. Method Performance pational Safety and Health Administration, OSHA 2206 (Revised, January 1976). 12.1 The method detection limit (MDL) is 6. ‘‘Safety in Academic Chemistry Labora- defined as the minimum concentration of a tories,’’ American Chemical Society Publica- substance that can be measured and reported tion, Committee on Chemical Safety, 3rd with 99% confidence that the value is above Edition, 1979. zero. 1 The MDL concentration listed in 7. Provost, L.P., and Elder, R.S. ‘‘Interpre- Table 1 were obtained using reagent water. 11. tation of Percent Recovery Data,’’ American Similar results were achieved using rep- Laboratory, 15, 58–63 (1983). (The value 2.44 resentative wastewaters. The MDL actually used in the equation in Section 8.3.3 is two achieved in a given analysis will vary de- times the value 1.22 derived in this report.) pending on instrument sensitivity and ma- 8. ‘‘Methods 330.4 (Titrimetric, DPD-FAS) trix effects. and 330.5 (Spectrophotometric, DPD) for 12.2 This method is recommended for use Chlorine, Total Residual,’’ Methods for in the concentration range from the MDL to Chemical Analysis of Water and Wastes, EPA 1000 × MDL. Direct aqueous injection tech- 600/4–79–020, U.S. Environmental Protection niques should be used to measure concentra- Agency, Environmental Monitoring and Sup- tion levels above 1000 × MDL. port Laboratory, Cincinnati, Ohio 45268, 12.3 This method was tested by 20 labora- March 1979. tories using reagent water, drinking water, 9. ‘‘EPA Method Study 24, Method 601— surface water, and three industrial Purgeable Halocarbons by the Purge and wastewaters spiked at six concentrations Trap Method,’’ EPA 600/4–84–064, National over the range 8.0 to 500 μg/L. 9 Single oper- Technical Information Service, PB84–212448, ator precision, overall precision, and method Springfield, Virginia 22161, July 1984. accuracy were found to be directly related to 10. ‘‘Method Validation Data for EPA the concentration of the parameter and es- Method 601,’’ Memorandum from B. Potter, sentially independent of the sample matrix. U.S. Environmental Protection Agency, En- Linear equations to describe these relation- vironmental Monitoring and Support Lab- ships are presented in Table 3. oratory, Cincinnati, Ohio 45268, November 10, 1983. References 11. Bellar, T. A., Unpublished data, U.S. 1. 40 CFR part 136, appendix B. Environmental Protection Agency, Environ- 2. Bellar, T.A., and Lichtenberg, J.J. ‘‘De- mental Monitoring and Support Laboratory, termining Volatile Organics at Microgram- Cincinnati, Ohio 45268, 1981.

TABLE 1—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS

Retention time (min) Method detection Parameter μ Column 1 Column 2 limit ( g/L)

Chloromethane ...... 1 .50 5 .28 0 .08 Bromomethane ...... 2 .17 7 .05 1 .18 Dichlorodifluoromethane ...... 2.62 nd 1 .81 Vinyl chloride ...... 2 .67 5 .28 0 .18 Chloroethane ...... 3 .33 8 .68 0 .52 Methylene chloride ...... 5 .25 10 .1 0 .25 Trichlorofluoromethane ...... 7 .18 nd nd

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TABLE 1—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS—Continued

Retention time (min) Method detection Parameter μ Column 1 Column 2 limit ( g/L)

1,1-Dichloroethene ...... 7 .93 7 .72 0.13 1,1-Dichloroethane ...... 9 .30 12.6 0.07 trans-1,2-Dichloroethene ...... 10.1 9.38 0 .10 Chloroform ...... 10 .7 12.1 0.05 1,2-Dichloroethane ...... 11 .4 15.4 0.03 1,1,1-Trichloroethane ...... 12.6 13.1 0 .03 Carbon tetrachloride ...... 13 .0 14 .4 0 .12 Bromodichloromethane ...... 13.7 14 .6 0 .10 1,2-Dichloropropane ...... 14 .9 16 .6 0 .04 cis-1,3-Dichloropropene ...... 15.2 16.6 0 .34 Trichloroethene ...... 15.8 13 .1 0 .12 Dibromochloromethane ...... 16 .5 16 .6 0 .09 1,1,2-Trichloroethane ...... 16.5 18.1 0 .02 trans-1,3-Dichloropropene ...... 16 .5 18.0 0.20 2-Chloroethylvinyl ether ...... 18 .0 nd 0 .13 Bromoform ...... 19 .2 19.2 0.20 1,1,2,2-Tetrachloroethane ...... 21 .6 nd 0 .03 Tetrachloroethene ...... 21 .7 15.0 0.03 Chlorobenzene ...... 24.2 18.8 0 .25 1,3-Dichlorobenzene ...... 34 .0 22 .4 0 .32 1,2-Dichlorobenzene ...... 34 .9 23 .5 0 .15 1,4-Dichlorobenzene ...... 35 .4 22 .3 0 .24 Column 1 conditions: Carbopack B (60/80 mesh) coated with 1% SP–1000 packed in an 8 ft × 0.1 in. ID stainless steel or glass column with helium carrier gas at 40 mL/min flow rate. Column temperature held at 45 °C for 3 min then programmed at 8 °C/min to 220 °C and held for 15 min. Column 2 conditions: Porisil-C (100/120 mesh) coated with n-octane packed in a 6 ft × 0.1 in. ID stainless steel or glass col- umn with helium carrier gas at 40 mL/min flow rate. Column temperature held at 50 °C for 3 min then programmed at 6 °C/min to 170 °C and held for 4 min. nd = not determined.

TABLE 2—CALIBRATION AND QC ACCEPTANCE CRITERIA—METHOD 601 A

¯ Parameter Range for Q Limit for s Range for X Range P, (μg/L) (μg/L) (μg/L) Ps (%)

Bromodichloromethane ...... 15.2–24.8 4.3 10.7–32.0 42–172 Bromoform ...... 14.7–25.3 4.7 5.0–29.3 13–159 Bromomethane ...... 11.7–28.3 7.6 3.4–24.5 D–144 Carbon tetrachloride ...... 13.7–26.3 5.6 11.8–25.3 43–143 Chlorobenzene ...... 14.4–25.6 5.0 10.2–27.4 38–150 Chloroethane ...... 15.4–24.6 4.4 11.3–25.2 46–137 2-Chloroethylvinyl ether ...... 12.0–28.0 8.3 4.5–35.5 14–186 Chloroform ...... 15.0–25.0 4.5 12.4–24.0 49–133 Chloromethane ...... 11.9–28.1 7.4 D–34.9 D–193 Dibromochloromethane ...... 13.1–26.9 6.3 7.9–35.1 24–191 1,2-Dichlorobenzene ...... 14.0–26.0 5.5 1.7–38.9 D–208 1,3-Dichlorobenzene ...... 9.9–30.1 9.1 6.2–32.6 7–187 1,4-Dichlorobenzene ...... 13.9–26.1 5.5 11.5–25.5 42–143 1,1-Dichloroethane ...... 16.8–23.2 3.2 11.2–24.6 47–132 1,2-Dichloroethane ...... 14.3–25.7 5.2 13.0–26.5 51–147 1,1-Dichloroethene ...... 12.6–27.4 6.6 10.2–27.3 28–167 trans-1,2-Dichloroethene ...... 12.8–27.2 6.4 11.4–27.1 38–155 1,2-Dichloropropane ...... 14.8–25.2 5.2 10.1–29.9 44–156 cis-1,3-Dichloropropene ...... 12.8–27.2 7.3 6.2–33.8 22–178 trans-1,3-Dichloropropene ...... 12.8–27.2 7.3 6.2–33.8 22–178 Methylene chloride ...... 15.5–24.5 4.0 7.0–27.6 25–162 1,1,2,2-Tetrachloroethane ...... 9.8–30.2 9.2 6.6–31.8 8–184 Tetrachloroethene ...... 14.0–26.0 5.4 8.1–29.6 26–162 1,1,1-Trichloroethane ...... 14.2–25.8 4.9 10.8–24.8 41–138 1,1,2-Trichloroethane ...... 15.7–24.3 3.9 9.6–25.4 39–136 Trichloroethene ...... 15.4–24.6 4.2 9.2–26.6 35–146 Trichlorofluoromethane ...... 13.3–26.7 6.0 7.4–28.1 21–156 Vinyl chloride ...... 13.7–26.3 5.7 8.2–29.9 28–163 a Criteria were calculated assuming a QC check sample concentration of 20 μg/L. Q = Concentration measured in QC check sample, in μg/L (Section 7.5.3). s = Standard deviation of four recovery measurements, in μg/L (Section 8.2.4). X¯ = Average recovery for four recovery measurements, in μg/L (Section 8.2.4). P, Ps = Percent recovery measured (Section 8.3.2, Section 8.4.2). D = Detected; result must be greater than zero. NOTE: These criteria are based directly upon the method performance data in Table 3. Where necessary, the limits for recov- ery have been broadened to assure applicability of the limits to concentrations below those used to develop Table 3.

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TABLE 3—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 601

Parameter Accuracy, as re- Single analyst pre- Overall precision, covery, X′ (μg/L) cision, sr′ (μg/L) S′ (μg/L)

Bromodichloromethane ...... 1.12C¥1.02 0.11X¯ + 0.04 0.20X¯ + 1.00 Bromoform ...... 0.96C¥2.05 0.12X¯ + 0.58 0.21X¯ + 2.41 Bromomethane ...... 0.76C¥1.27 0.28X¯ + 0.27 0.36X¯ + 0.94 Carbon tetrachloride ...... 0.98C¥1.04 0.15X¯ + 0.38 0.20X¯ + 0.39 Chlorobenzene ...... 1.00C¥1.23 0.15X¯ ¥0.02 0.18X¯ + 1.21 Choroethane ...... 0.99C¥1.53 0.14X¯ ¥0.13 0.17X¯ + 0.63 2-Chloroethylvinyl ether a ...... 1.00C 0.20X¯ 0.35X¯ Chloroform ...... 0.93C¥0.39 0.13X¯ + 0.15 0.19X¯ ¥0.02 Chloromethane ...... 0.77C + 0.18 0.28X¯ ¥0.31 0.52X¯ + 1.31 Dibromochloromethane ...... 0.94C + 2.72 0.11X¯ + 1.10 0.24X¯ + 1.68 1,2-Dichlorobenzene ...... 0.93C + 1.70 0.20X¯ + 0.97 0.13X¯ + 6.13 1,3-Dichlorobenzene ...... 0.95C + 0.43 0.14X¯ + 2.33 0.26X¯ + 2.34 1,4-Dichlorobenzene ...... 0.93C¥0.09 0.15X¯ + 0.29 0.20X¯ + 0.41 1,1-Dichloroethane ...... 0.95C¥1.08 0.09X¯ + 0.17 0.14X¯ + 0.94 1,2-Dichloroethane ...... 1.04C¥1.06 0.11X¯ + 0.70 0.15X¯ + 0.94 1,1-Dichloroethene ...... 0.98C¥0.87 0.21X¯ ¥0.23 0.29X¯ ¥0.40 trans-1,2-Dichloroethene ...... 0.97C¥0.16 0.11X¯ + 1.46 0.17X¯ + 1.46 1,2-Dichloropropane a ...... 1.00C 0.13X¯ 0.23X¯ cis-1,3-Dichloropropene a ...... 1.00C 0.18X¯ 0.32X¯ trans-1,3-Dichloropropene a ...... 1.00C 0.18X¯ 0.32X¯ Methylene chloride ...... 0.91C¥0.93 0.11X¯ + 0.33 0.21X¯ + 1.43 1,1,2,2-Tetrachloroethene ...... 0.95C + 0.19 0.14X¯ + 2.41 0.23X¯ + 2.79 Tetrachloroethene ...... 0.94C + 0.06 0.14X¯ + 0.38 0.18X¯ + 2.21 1,1,1-Trichloroethane ...... 0.90C¥0.16 0.15X¯ + 0.04 0.20X¯ + 0.37 1,1,2-Trichloroethane ...... 0.86C + 0.30 0.13X¯ ¥0.14 0.19X¯ + 0.67 Trichloroethene ...... 0.87C + 0.48 0.13X¯ ¥0.03 0.23X¯ + 0.30 Trichlorofluoromethane ...... 0.89C¥0.07 0.15X¯ + 0.67 0.26X¯ + 0.91 Vinyl chloride ...... 0.97C¥0.36 0.13X¯ + 0.65 0.27X¯ + 0.40 X¯ ′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L. sn′ = Expected single analyst standard deviation of measurements at an average concentration found of X¯ , in μg/L. S1 = Expected interlaboratory standard deviation of measurements at an average concentration found of X¯ , in μg/L. C = True value for the concentration, in μg/L. X¯ = Average recovery found for measurements of samples containing a concentration of C, in μg/L. a Estimates based upon the performance in a single laboratory. 10

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OPTIONAL FOAM TRAP --14MM 0. D. INLET% IN. 0. D.

_...... ,SAMPLE INLET c -2-WAY SYRINGE VALVE --17CM. 20 GAUGE SYRINGE NEEDLE %IN. \; '6MM. 0. D. RUBBER SEPTUM 0. D. EXIT 1/16 IN. O.D. STAINLESS STEEL

13X MOLECULAR SIEVE PURGE GAS FILTER ~ (.) ...0

PURGE GAS FLOW CONTROL

Figure 1 . Purging device.

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PACKING PROCEDURE CONSTRUCTION

GLASS SMM 7 .n. /FOOT ,..--,~JOMPRESSION WOOL T RESISTANCE FITTING NUT ACTIVATED WIRE WRAPPED AND FERRULES CHARCOAL 7.7CI SOLID (DOUBLE LAYER) THERMOCOUPLE/ CONTROLLER SENSOR GRADE 15. 1 ELECTRONIC SILICA GEL7'7Cr TEMPERATURE CONTROL 7.n./f00T AND PYROMETER ;•,,.;._, ... RESISTANCE T WIRE WRAPPED TENAX 7.7 CMl ~~:: SOLID (SINGLE LAYER) JOI ov 1 . ~it SCM G~ASS-WOOL1 CMT .. SMM TRAP INLET

- Figure 2. Trap packings and construction to include desorb capability

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CARRIER GAS FLOW CONTROL LIQUID____ INJECTION PORTS_.. COLUMN OVEN PRESSURE REGULATOR 1 - CONFIRMATORY COLUMN ~ 1 TO DETECTOR 1.!:::=~[;~~~11_._ __..... 1 --ANALYTICAL COLUMN

PURGE GAS FLOW CONTROL

13X MOLECULAR SIEVE FILTER .

... Note:ALL LINES BETWEEN ~· TRAP AND GC > L..:J't-- PURGING l::::====;jj' DEVICE SHOULD BE HEATED TO so-c Figure 3. Purge and trap system-purge mode.

CARRIER GAS FLOW CONTROL PRESSURE REGULATOR~ = ...~~~

PURGE GAS FLOW CONTROL'

Note: .... All LINES BETWEEN ifl . PURGING TRAP AND GC >·-DEVICE l.!::::====:di SHOULD BE HEATED TO 80°C. Figure 4. Purge and trap system - desorb mode.

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Pt. 136, App. A, Meth. 601 40 CFR Ch. I (7–1–19 Edition)

< <

::::; ::::; < <

_, _,

("') ("')

c:n c:n

0 0 0 0

4-DICHLOROBENZENE 4-DICHLOROBENZENE 1, 1, 2~ 2~

o"' o"'

CD CD ("') ("')

na:::o< na:::o< ~ ~

.., ..,

;:!Z("') ;:!Z("')

-> ->

< <

r- ...... :!!:: :!!::

~~~ ~~~ 2-CHLOROTOLUENE 2-CHLOROTOLUENE

...., ....,

.., ..,

o:O o:O _, _,

""l> ""l> ("') ("')

::;;~n ::;;~n

0 0 m~Z m~Z

w w >""o >""o

0 0 0 0

O::t>O O::t>O

co co ......

"T''.!.a "T''.!.a ,... ,...

BROMOBENZENE BROMOBENZENE n-.. n-.. r-

l> l> 0(1) 0(1)

:I:~~ :I:~~

~~:.:. ~~:.:.

EXANE EXANE 1-CHLOROH 1-CHLOROH

>z >z _, _,

3!: 3!: ::t> ::t> ("') ("')

CHLOROBENZENE CHLOROBENZENE ~ ~ c:->c: c:->c: m m

_,Or­

m::co m::co

0 0 -on -on 2-TETRACHLOROETHANE 2-TETRACHLOROETHANE 2. 2. 1, 1, 1, 1,

3-TRJCHLOROPROPANE 3-TRJCHLOROPROPANE :::=~§::,_:1~, :::=~§::,_:1~, 2. 2.

Ul Ul '":::::=====----

:I :I

0 0

TETRACHLOROETHANE TETRACHLOROETHANE 1,1,1.2-

0' 0' .. ..

Ill Ill

2-DIBROMOETHANE 2-DIBROMOETHANE 1, 1,

n n

0 0

1,3-DICHLOROPROPENE 1,3-DICHLOROPROPENE - trans trans

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~ ~ · · cis cis 1.3-DICHLDROPROPENE 1.3-DICHLDROPROPENE

CD' CD'

0' 0' ~===~~;:--.....:.1· ~===~~;:--.....:.1· 2-DICHLOROPROPANE 2-DICHLOROPROPANE

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TRICHLOROETHANE TRICHLOROETHANE - c: c: ":::=====--1.1,1-

, ,

0 0

3 3

trans-1,2-DICHLOROETHENE trans-1,2-DICHLOROETHENE

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D) D) CHLOROETHENE CHLOROETHENE 1,1-DJ 1,1-DJ 3 3

a a

~ ~

n n

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CHLOROETHANE CHLOROETHANE

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~ ~ I» I» CHLOROMETHANE CHLOROMETHANE BROMOMETHANE BROMOMETHANE .. ..

=· =· c: c: "TI "TI

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METHOD 602—PURGEABLE AROMATICS umn. The gas chromatograph is temperature programmed to separate the aromatics 1. Scope and Application which are then detected with a 1.1 This method covers the determination photoionization detector. 23 of various purgeable aromatics. The fol- 2.2 The method provides an optional gas lowing parameters may be determined by chromatographic column that may be help- this method: ful in resolving the compounds of interest from interferences that may occur. STORET Parameter No. CAS No. 3. Interferences

Benzene ...... 34030 71–43–2 3.1 Impurities in the purge gas and or- Chlorobenzene ...... 34301 108–90–7 ganic compounds outgassing from the plumb- 1,2-Dichlorobenzene ...... 34536 95–50–1 ing ahead of the trap account for the major- 1,3-Dichlorobenzene ...... 34566 541–73–1 ity of contamination problems. The analyt- 1,4-Dichlorobenzene ...... 34571 106–46–7 ical system must be demonstrated to be free Ethylbenzene ...... 34371 100–41–4 Toluene ...... 34010 108–88–3 from contamination under the conditions of the analysis by running laboratory reagent 1.2 This is a purge and trap gas blanks as described in Section 8.1.3. The use chromatographic (GC) method applicable to of non-Teflon plastic tubing, non-Teflon the determination of the compounds listed thread sealants, or flow controllers with rub- above in municipal and industrial discharges ber components in the purge and trap system as provided under 40 CFR 136.1. When this should be avoided. method is used to analyze unfamiliar sam- 3.2 Samples can be contaminated by diffu- ples for any or all of the compounds above, sion of volatile organics through the septum compound identifications should be sup- seal into the sample during shipment and ported by at least one additional qualitative storage. A field reagent blank prepared from technique. This method describes analytical reagent water and carried through the sam- conditions for a second gas chromatographic pling and handling protocol can serve as a column that can be used to confirm measure- check on such contamination. ments made with the primary column. Meth- 3.3 Contamination by carry-over can od 624 provides gas chromatograph/mass occur whenever high level and low level sam- spectrometer (GC/MS) conditions appro- ples are sequentially analyzed. To reduce priate for the qualitative and quantitative carry-over, the purging device and sample confirmation of results for all of the param- syringe must be rinsed with reagent water eters listed above. between sample analyses. Whenever an un- 1.3 The method detection limit (MDL, de- usually concentrated sample is encountered, fined in Section 12.1) 1 for each parameter is it should be followed by an analysis of rea- listed in Table 1. The MDL for a specific gent water to check for cross contamination. wastewater may differ from those listed, de- For samples containing large amounts of pending upon the nature of interferences in water-soluble materials, suspended solids, the sample matrix. high boiling compounds or high aromatic 1.4 Any modification of this method, be- levels, it may be necessary to wash the purg- yond those expressly permitted, shall be con- ing device with a detergent solution, rinse it sidered as a major modification subject to with distilled water, and then dry it in an application and approval of alternate test oven at 105 °C between analyses. The trap procedures under 40 CFR 136.4 and 136.5. and other parts of the system are also sub- 1.5 This method is restricted to use by or ject to contamination; therefore, frequent under the supervision of analysts experi- bakeout and purging of the entire system enced in the operation of a purge and trap may be required. system and a gas chromatograph and in the interpretation of gas chromatograms. Each 4. Safety analyst must demonstrate the ability to gen- 4.1 The toxicity or carcinogenicity of erate acceptable results with this method each reagent used in this method has not using the procedure described in Section 8.2. been precisely defined; however, each chem- ical compound should be treated as a poten- 2. Summary of Method tial health hazard. From this viewpoint, ex- 2.1 An inert gas is bubbled through a 5-mL posure to these chemicals must be reduced to water sample contained in a specially-de- the lowest possible level by whatever means signed purging chamber at ambient tempera- available. The laboratory is responsible for ture. The aromatics are efficiently trans- maintaining a current awareness file of ferred from the aqueous phase to the vapor OSHA regulations regarding the safe han- phase. The vapor is swept through a sorbent dling of the chemicals specified in this meth- trap where the aromatics are trapped. After od. A reference file of material data handling purging is completed, the trap is heated and sheets should also be made available to all backflushed with the inert gas to desorb the personnel involved in the chemical analysis. aromatics onto a gas chromatographic col- Additional references to laboratory safety

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are available and have been identified 46 for 5.3 Gas chromatograph—An analytical the information of the analyst. system complete with a temperature pro- 4.2 The following parameters covered by grammable gas chromatograph suitable for this method have been tentatively classified on-column injection and all required acces- as known or suspected, human or mamma- sories including syringes, analytical col- lian carcinogens: benzene and 1,4- umns, gases, detector, and strip-chart re- dichlorobenzene. Primary standards of these corder. A data system is recommended for toxic compounds should be prepared in a measuring peak areas. hood. A NIOSH/MESA approved toxic gas 5.3.1 Column 1—6 ft long × 0.082 in. ID respirator should be worn when the analyst stainless steel or glass, packed with 5% SP– handles high concentrations of these toxic 1200 and 1.75% Bentone-34 on Supelcoport compounds. (100/120 mesh) or equivalent. This column was used to develop the method performance 5. Apparatus and Materials statements in Section 12. Guidelines for the 5.1 Sampling equipment, for discrete sam- use of alternate column packings are pro- pling. vided in Section 10.1. 5.1.1 Vial]25-mL capacity or larger, 5.3.2 Column 2—8 ft long × 0.1 in ID stain- equipped with a screw cap with a hole in the less steel or glass, packed with 5% 1,2,3- center (Pierce #13075 or equivalent). Deter- Tris(2-cyanoethoxy)propane on Chromosorb gent wash, rinse with tap and distilled water, W-AW (60/80 mesh) or equivalent. and dry at 105 °C before use. 5.3.3 Detector—Photoionization detector 5.1.2 Septum—Teflon-faced silicone (h-Nu Systems, Inc. Model PI–51–02 or equiv- (Pierce #12722 or equivalent). Detergent alent). This type of detector has been proven wash, rinse with tap and distilled water, and effective in the analysis of wastewaters for dry at 105 °C for 1 h before use. the parameters listed in the scope (Section 5.2 Purge and trap system—The purge and 1.1), and was used to develop the method per- trap system consists of three separate pieces formance statements in Section 12. Guide- of equipment: A purging device, trap, and lines for the use of alternate detectors are desorber. Several complete systems are now provided in Section 10.1. commercially available. 5.4 Syringes—5-mL glass hypodermic with 5.2.1 The purging device must be designed Luerlok tip (two each), if applicable to the to accept 5-mL samples with a water column purging device. at least 3 cm deep. The gaseous head space 5.5 Micro syringes—25-μL, 0.006 in. ID nee- between the water column and the trap must dle. have a total volume of less than 15 mL. The 5.6 Syringe valve—2-way, with Luer ends purge gas must pass through the water col- (three each). umn as finely divided bubbles with a diame- 5.7 Bottle—15-mL, screw-cap, with Teflon ter of less than 3 mm at the origin. The cap liner. purge gas must be introduced no more than 5.8 Balance—Analytical, capable of accu- 5 mm from the base of the water column. rately weighing 0.0001 g. The purging device illustrated in Figure 1 6. Reagents meets these design criteria. 5.2.2 The trap must be at least 25 cm long 6.1 Reagent water—Reagent water is de- and have an inside diameter of at least 0.105 fined as a water in which an interferent is in. not observed at the MDL of the parameters 5.2.2.1 The trap is packed with 1 cm of of interest. methyl silicone coated packing (Section 6.1.1 Reagent water can be generated by 6.4.2) and 23 cm of 2,6-diphenylene oxide poly- passing tap water through a carbon filter bed mer (Section 6.4.1) as shown in Figure 2. This containing about 1 lb of activated carbon trap was used to develop the method per- (Filtrasorb-300, Calgon Corp., or equivalent). formance statements in Section 12. 6.1.2 A water purification system 5.2.2.2 Alternatively, either of the two (Millipore Super-Q or equivalent) may be traps described in Method 601 may be used, used to generate reagent water. although water vapor will preclude the meas- 6.1.3 Reagent water may also be prepared urement of low concentrations of benzene. by boiling water for 15 min. Subsequently, 5.2.3 The desorber must be capable of rap- while maintaining the temperature at 90 °C, idly heating the trap to 180 °C. The polymer bubble a contaminant-free inert gas through section of the trap should not be heated the water for 1 h. While still hot, transfer higher than 180 °C and the remaining sec- the water to a narrow mouth screw-cap bot- tions should not exceed 200 °C. The desorber tle and seal with a Teflon-lined septum and illustrated in Figure 2 meets these design cap. criteria. 6.2 Sodium thiosulfate—(ACS) Granular. 5.2.4 The purge and trap system may be 6.3 Hydrochloric acid (1 + 1)—Add 50 mL assembled as a separate unit or be coupled to of concentrated HCl (ACS) to 50 mL of rea- a gas chromatograph as illustrated in Fig- gent water. ures 3, 4, and 5. 6.4 Trap Materials:

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6.4.1 2,6-Diphenylene oxide polymer— dition the trap overnight at 180 °C by Tenax, (60/80 mesh), chromatographic grade backflushing with an inert gas flow of at or equivalent. least 20 mL/min. Condition the trap for 10 6.4.2 Methyl silicone packing—3% OV–1 on min once daily prior to use. Chromosorb-W (60/80 mesh) or equivalent. 7.2 Connect the purge and trap system to 6.5 Methanol—Pesticide quality or equiv- a gas chromatograph. The gas chro- alent. matograph must be operated using tempera- 6.6 Stock standard solutions—Stock ture and flow rate conditions equivalent to standard solutions may be prepared from those given in Table 1. Calibrate the purge pure standard materials or purchased as cer- and trap-gas chromatographic system using tified solutions. Prepare stock standard solu- either the external standard technique (Sec- tions in methanol using assayed liquids. Be- tion 7.3) or the internal standard technique cause of the toxicity of benzene and 1,4- (Section 7.4). dichlorobenzene, primary dilutions of these 7.3 External standard calibration proce- materials should be prepared in a hood. A dure: NIOSH/MESA approved toxic gas respirator 7.3.1 Prepare calibration standards at a should be used when the analyst handles minimum of three concentration levels for high concentrations of such materials. each parameter by carefully adding 20.0 μL of 6.6.1 Place about 9.8 mL of methanol into one or more secondary dilution standards to a 10–mL ground glass stoppered volumetric 100, 500, or 1000 mL of reagent water. A 25-μL flask. Allow the flask to stand, unstoppered, syringe with a 0.006 in. ID needle should be for about 10 min or until all alcohol wetted used for this operation. One of the external surfaces have dried. Weigh the flask to the standards should be at a concentration near, nearest 0.1 mg. but above, the MDL (Table 1) and the other 6.6.2 Using a 100–μL syringe, immediately concentrations should correspond to the ex- add two or more drops of assayed reference material to the flask, then reweigh. Be sure pected range of concentrations found in real that the drops fall directly into the alcohol samples or should define the working range without contacting the neck of the flask. of the detector. These aqueous standards 6.6.3 Reweigh, dilute to volume, stopper, must be prepared fresh daily. then mix by inverting the flask several 7.3.2 Analyze each calibration standard times. Calculate the concentration in μg/μL according to Section 10, and tabulate peak from the net gain in weight. When compound height or area responses versus the con- purity is assayed to be 96% or greater, the centration in the standard. The results can weight can be used without correction to cal- be used to prepare a calibration curve for culate the concentration of the stock stand- each compound. Alternatively, if the ratio of ard. Commercially prepared stock standards response to concentration (calibration fac- can be used at any concentration if they are tor) is a constant over the working range certified by the manufacturer or by an inde- (<10% relative standard deviation, RSD), lin- pendent source. earity through the origin can be assumed 6.6.4 Transfer the stock standard solution and the average ratio or calibration factor into a Teflon-sealed screw-cap bottle. Store can be used in place of a calibration curve. at 4 °C and protect from light. 7.4 Internal standard calibration proce- 6.6.5 All standards must be replaced after dure—To use this approach, the analyst must one month, or sooner if comparison with select one or more internal standards that check standards indicates a problem. are similar in analytical behavior to the 6.7 Secondary dilution standards—Using compounds of interest. The analyst must fur- stock standard solutions, prepare secondary ther demonstrate that the measurement of dilution standards in methanol that contain the internal standard is not affected by the compounds of interest, either singly or method or matrix interferences. Because of mixed together. The secondary dilution these limitations, no internal standard can standards should be prepared at concentra- be suggested that is applicable to all sam- tions such that the aqueous calibration ples. The compound, a,a,a,-trifluorotoluene, standards prepared in Section 7.3.1 or 7.4.1 recommended as a surrogate spiking com- will bracket the working range of the ana- pound in Section 8.7 has been used success- lytical system. Secondary solution standards fully as an internal standard. must be stored with zero headspace and 7.4.1 Prepare calibration standards at a should be checked frequently for signs of minimum of three concentration levels for degradation or evaporation, especially just each parameter of interest as described in prior to preparing calibration standards from Section 7.3.1. them. 7.4.2 Prepare a spiking solution con- 6.8 Quality control check sample con- taining each of the internal standards using centrate—See Section 8.2.1. the procedures described in Sections 6.6 and 6.7. It is recommended that the secondary di- 7. Calibration lution standard be prepared at a concentra- 7.1 Assemble a purge and trap system that tion of 15 μg/mL of each internal standard meets the specifications in Section 5.2. Con- compound. The addition of 10 μl of this

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standard to 5.0 mL of sample or calibration with this method. This ability is established standard would be equivalent to 30 μg/L. as described in Section 8.2. 7.4.3 Analyze each calibration standard 8.1.2 In recognition of advances that are according to Section 10, adding 10 μL of in- occurring in chromatography, the analyst is ternal standard spiking solution directly to permitted certain options (detailed in Sec- the syringe (Section 10.4). Tabulate peak tion 10.1) to improve the separations or lower height or area responses against concentra- the cost of measurements. Each time such a tion for each compound and internal stand- modification is made to the method, the ana- ard, and calculate response factors (RF) for lyst is required to repeat the procedure in each compound using Equation 1. Section 8.2. RF = (As)(Cis (Ais)(Cs) 8.1.3 Each day, the analyst must analyze a Equation 1 reagent water blank to demonstrate that where: interferences from the analytical system are As = Response for the parameter to be meas- under control. ured. 8.1.4 The laboratory must, on an ongoing Ais = Response for the internal standard. basis, spike and analyze a minimum of 10% Cis = Concentration of the internal standard of all samples to monitor and evaluate lab- Cs = Concentration of the parameter to be oratory data quality. This procedure is de- measured. scribed in Section 8.3. If the RF value over the working range is a 8.1.5 The laboratory must, on an ongoing constant (<10% RSD), the RF can be assumed basis, demonstrate through the analyses of to be invariant and the average RF can be quality control check standards that the op- used for calculations. Alternatively, the re- eration of the measurement system is in con- sults can be used to plot a calibration curve trol. This procedure is described in Section of response ratios, As/Ais, vs. RF. 8.4. The frequency of the check standard 7.5 The working calibration curve, cali- analyses is equivalent to 10% of all samples bration factor, or RF must be verified on analyzed but may be reduced if spike recov- each working day by the measurement of a eries from samples (Section 8.3) meet all QC check sample. specified quality control criteria. 7.5.1 Prepare the QC check sample as de- 8.1.6 The laboratory must maintain per- scribed in Section 8.2.2. formance records to document the quality of 7.5.2 Analyze the QC check sample accord- data that is generated. This procedure is de- ing to Section 10. scribed in Section 8.5. 7.5.3 For each parameter, compare the re- 8.2 To establish the ability to generate sponse (Q) with the corresponding calibra- acceptable accuracy and precision, the ana- tion acceptance criteria found in Table 2. If lyst must perform the following operations. the responses for all parameters of interest fall within the designated ranges, analysis of 8.2.1 A quality control (QC) check sample actual samples can begin. If any individual Q concentrate is required containing each pa- falls outside the range, a new calibration rameter of interest at a concentration of 10 μ curve, calibration factor, or RF must be pre- g/mL in methanol. The QC check sample pared for that parameter according to Sec- concentrate must be obtained from the U.S. tion 7.3 or 7.4. Environmental Protection Agency, Environ- mental Monitoring and Support Laboratory 8. Quality Control in Cincinnati, Ohio, if available. If not avail- able from that source, the QC check sample 8.1 Each laboratory that uses this method concentrate must be obtained from another is required to operate a formal quality con- external source. If not available from either trol program. The minimum requirements of source above, the QC check sample con- this program consist of an initial demonstra- centrate must be prepared by the laboratory tion of laboratory capability and an ongoing using stock standards prepared independ- analysis of spiked samples to evaluate and ently from those used for calibration. document data quality. The laboratory must maintain records to document the quality of 8.2.2 Prepare a QC check sample to con- μ data that is generated. Ongoing data quality tain 20 g/L of each parameter by adding 200 μ checks are compared with established per- L of QC check sample concentrate to 100 mL formance criteria to determine if the results of reagant water. of analyses meet the performance character- 8.2.3 Analyze four 5-mL aliquots of the istics of the method. When results of sample well-mixed QC check sample according to spikes indicate atypical method perform- Section 10. ance, a quality control check standard must 8.2.4 Calculate the average recovery (X¯ ) in be analyzed to confirm that the measure- μg/L, and the standard deviation of the re- ments were performed in an in-control mode covery (s) in μg/L, for each parameter of in- of operation. terest using the four results. 8.1.1 The analyst must make an initial, 8.2.5 For each parameter compare s and X¯ one-time, demonstration of the ability to with the corresponding acceptance criteria generate acceptable accuracy and precision for precision and accuracy, respectively,

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found in Table 2. If s and X¯ for all param- acceptance criteria were calculated to in- eters of interest meet the acceptance cri- clude an allowance for error in measurement teria, the system performance is acceptable of both the background and spike concentra- and analysis of actual samples can begin. If tions, assuming a spike to background ratio any individual s exceeds the precision limit of 5:1. This error will be accounted for to the or any individual X¯ falls outside the range extent that the analyst’s spike to back- for accuracy, the system performance is un- ground ratio approaches 5:1. 7 If spiking was acceptable for that parameter. performed at a concentration lower than 20 NOTE: The large number of parameters in μg/L, the analyst must use either the QC ac- Table 2 present a substantial probability ceptance criteria in Table 2, or optional QC that one or more will fail at least one of the acceptance criteria calculated for the spe- acceptance criteria when all parameters are cific spike concentration. To calculate op- analyzed. tional acceptance criteria for the recovery of 8.2.6 When one or more of the parameters a parameter: (1) Calculate accuracy (X′) tested fail at least one of the acceptance cri- using the equation in Table 3, substituting teria, the analyst must proceed according to the spike concentration (T) for C; (2) cal- Section 8.2.6.1 or 8.2.6.2. culate overall precision (S′) using the equa- 8.2.6.1 Locate and correct the source of tion in Table 3, substituting X′ for X¯ ; (3) cal- the problem and repeat the test for all pa- culate the range for recovery at the spike rameters of interest beginning with Section concentration as (100 X′/T) ±2.44(100 S′/T)%. 7 8.2.3. 8.3.4 If any individual P falls outside the 8.2.6.2 Beginning with Section 8.2.3, repeat designated range for recovery, that param- the test only for those parameters that eter has failed the acceptance criteria. A failed to meet criteria. Repeated failure, check standard containing each parameter however, will confirm a general problem that failed the criteria must be analyzed as with the measurement system. If this occurs, described in Section 8.4. locate and correct the source of the problem 8.4 If any parameter fails the acceptance and repeat the test for all compounds of in- criteria for recovery in Section 8.3, a QC terest beginning with Section 8.2.3. check standard containing each parameter 8.3 The laboratory must, on an ongoing that failed must be prepared and analyzed. basis, spike at least 10% of the samples from each sample site being monitored to assess NOTE: The frequency for the required anal- accuracy. For laboratories analyzing one to ysis of a QC check standard will depend upon ten samples per month, at least one spiked the number of parameters being simulta- sample per month is required. neously tested, the complexity of the sample 8.3.1 The concentration of the spike in the matrix, and the performance of the labora- sample should be determined as follows: tory. 8.3.1.1 If, as in compliance monitoring, 8.4.1 Prepare the QC check standard by the concentration of a specific parameter in adding 10 μL of QC check sample concentrate the sample is being checked against a regu- (Section 8.2.1 or 8.3.2) to 5 mL of reagent latory concentration limit, the spike should water. The QC check standard needs only to be at that limit or 1 to 5 times higher than contain the parameters that failed criteria the background concentration determined in in the test in Section 8.3. Section 8.3.2, whichever concentration would 8.4.2 Analyze the QC check standard to be larger. determine the concentration measured (A) of 8.3.1.2 If the concentration of a specific each parameter. Calculate each percent re- parameter in the sample is not being covery (Ps) as 100 (A/T)%, where T is the true checked against a limit specific to that pa- value of the standard concentration. rameter, the spike should be at 20 μg/L or 1 8.4.3 Compare the percent recovery (Ps) to 5 times higher than the background con- for each parameter with the corresponding centration determined in Section 8.3.2, QC acceptance criteria found in Table 2. Only whichever concentration would be larger. parameters that failed the test in Section 8.3 8.3.2 Analyze one 5-mL sample aliquot to need to be compared with these criteria. If determine the background concentration (B) the recovery of any such parameter falls out- of each parameter. If necessary, prepare a side the designated range, the laboratory new QC check sample concentrate (Section performance for that parameter is judged to 8.2.1) appropriate for the background con- be out of control, and the problem must be centrations in the sample. Spike a second 5- immediately identified and corrected. The mL sample aliquot with 10 μL of the QC analytical result for that parameter in the check sample concentrate and analyze it to unspiked sample is suspect and may not be determine the concentration after spiking reported for regulatory compliance purposes. (A) of each parameter. Calculate each per- 8.5 As part of the QC program for the lab- cent recovery (P) as 100(A¥B)%/T, where T is oratory, method accuracy for wastewater the known true value of the spike. samples must be assessed and records must 8.3.3 Compare the percent recovery (P) for be maintained. After the analysis of five each parameter with the corresponding QC spiked wastewater samples as in Section 8.3, acceptance criteria found in Table 2. These calculate the average percent recovery (P¯ )

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and the standard deviation of the percent re- tain the hermetic seal on the sample bottle covery (sp). Express the accuracy assessment until time of analysis. ¯ as a percent recovery interval from P¥2sp to 9.3 All samples must be analyzed within ¯ ¯ 3 P + 2sp. If P = 90% and sp = 10%, for example, 14 days of collection. the accuracy interval is expressed as 70– 110%. Update the accuracy assessment for 10. Procedure each parameter on a regular basis (e.g. after 10.1 Table 1 summarizes the recommended each five to ten new accuracy measure- operating conditions for the gas chro- ments). matograph. Included in this table are esti- 8.6 It is recommended that the laboratory mated retention times and MDL that can be adopt additional quality assurance practices achieved under these conditions. An example for use with this method. The specific prac- of the separations achieved by Column 1 is tices that are most productive depend upon shown in Figure 6. Other packed columns, the needs of the laboratory and the nature of chromatographic conditions, or detectors the samples. Field duplicates may be ana- may be used if the requirements of Section lyzed to assess the precision of the environ- 8.2 are met. mental measurements. When doubt exists 10.2 Calibrate the system daily as de- over the identification of a peak on the chro- scribed in Section 7. matogram, confirmatory techniques such as gas chromatography with a dissimilar col- 10.3 Adjust the purge gas (nitrogen or he- umn, specific element detector, or mass lium) flow rate to 40 mL/min. Attach the spectrometer must be used. Whenever pos- trap inlet to the purging device, and set the sible, the laboratory should analyze standard purge and trap system to purge (Figure 3). reference materials and participate in rel- Open the syringe valve located on the purg- evant performance evaluation studies. ing device sample introduction needle. 8.7 The analyst should monitor both the 10.4 Allow the sample to come to ambient performance of the analytical system and temperature prior to introducing it to the the effectiveness of the method in dealing syringe. Remove the plunger from a 5-mL sy- with each sample matrix by spiking each ringe and attach a closed syringe valve. Open sample, standard, and reagent water blank the sample bottle (or standard) and carefully with surrogate compounds (e.g. a, a, a,- pour the sample into the syringe barrel to trifluorotoluene) that encompass the range just short of overflowing. Replace the sy- of the temperature program used in this ringe plunger and compress the sample. Open method. From stock standard solutions pre- the syringe valve and vent any residual air pared as in Section 6.6, add a volume to give while adjusting the sample volume to 5.0 mL. 750 μg of each surrogate to 45 mL of reagent Since this process of taking an aliquot de- water contained in a 50-mL volumetric flask, stroys the validity of the sample for future mix and dilute to volume for a concentration analysis, the analyst should fill a second sy- of 15 mg/μL. Add 10 μL of this surrogate spik- ringe at this time to protect against possible ing solution directly into the 5-mL syringe loss of data. Add 10.0 μL of the surrogate with every sample and reference standard spiking solution (Section 8.7) and 10.0 μL of analyzed. Prepare a fresh surrogate spiking the internal standard spiking solution (Sec- solution on a weekly basis. If the internal tion 7.4.2), if applicable, through the valve standard calibration procedure is being used, bore, then close the valve. the surrogate compounds may be added di- 10.5 Attach the syringe-syringe valve as- rectly to the internal standard spiking solu- sembly to the syringe valve on the purging tion (Section 7.4.2). device. Open the syringe valves and inject the sample into the purging chamber. 9. Sample Collection, Preservation, and 10.6 Close both valves and purge the sam- Handling ple for 12.0 ±0.1 min at ambient temperature. 9.1 The samples must be iced or refrig- 10.7 After the 12-min purge time, dis- erated from the time of collection until anal- connect the purging device from the trap. ysis. If the sample contains free or combined Dry the trap by maintaining a flow of 40 mL/ chlorine, add sodium thiosulfate preserva- min of dry purge gas through it for 6 min tive (10 mg/40 mL is sufficient for up to 5 (Figure 4). If the purging device has no provi- ppm Cl2) to the empty sample bottle just sion for bypassing the purger for this step, a prior to shipping to the sampling site. EPA dry purger should be inserted into the device Method 330.4 or 330.5 may be used for meas- to minimize moisture in the gas. Attach the urement of residual chlorine. 8 Field test kits trap to the chromatograph, adjust the purge are available for this purpose. and trap system to the desorb mode (Figure 9.2 Collect about 500 mL of sample in a 5), and begin to temperature program the gas clean container. Adjust the pH of the sample chromatograph. Introduce the trapped mate- to about 2 by adding 1 + 1 HCl while stirring. rials to the GC column by rapidly heating Fill the sample bottle in such a manner that the trap to 180 °C while backflushing the trap no air bubbles pass through the sample as with an inert gas between 20 and 60 mL/min the bottle is being filled. Seal the bottle so for 4 min. If rapid heating of the trap cannot that no air bubbles are entrapped in it. Main- be achieved, the GC column must be used as

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a secondary trap by cooling it to 30 °C (sub- 11.2 Report results in μg/L without correc- ambient temperature, if poor peak geometry tion for recovery data. All QC data obtained and random retention time problems persist) should be reported with the sample results. instead of the initial program temperature of 50 °C. 12. Method Performance 10.8 While the trap is being desorbed into 12.1 The method detection limit (MDL) is the gas chromatograph column, empty the defined as the minimum concentration of a purging chamber using the sample introduc- substance that can be measured and reported tion syringe. Wash the chamber with two 5- with 99% confidence that the value is above mL flushes of reagent water. zero. 1 The MDL concentrations listed in 10.9 After desorbing the sample for 4 min, Table 1 were obtained using reagent water. 9 recondition the trap by returning the purge Similar results were achieved using rep- and trap system to the purge mode. Wait 15 resentative wastewaters. The MDL actually s, then close the syringe valve on the purg- achieved in a given analysis will vary de- ing device to begin gas flow through the pending on instrument sensitivity and ma- trap. The trap temperature should be main- trix effects. tained at 180 °C. After approximately 7 min, 12.2 This method has been demonstrated turn off the trap heater and open the syringe to be applicable for the concentration range valve to stop the gas flow through the trap. from the MDL to 100 × MDL. 9 Direct aqueous When the trap is cool, the next sample can injection techniques should be used to meas- be analyzed. ure concentration levels above 1000 × MDL. 10.10 Identify the parameters in the sam- 12.3 This method was tested by 20 labora- ple by comparing the retention times of the tories using reagent water, drinking water, peaks in the sample chromatogram with surface water, and three industrial those of the peaks in standard wastewaters spiked at six concentrations chromatograms. The width of the retention over the range 2.1 to 550 μg/L. 9 Single oper- time window used to make identifications ator precision, overall precision, and method should be based upon measurements of ac- accuracy were found to be directly related to tual retention time variations of standards the concentration of the parameter and es- over the course of a day. Three times the sentially independent of the sample matrix. standard deviation of a retention time for a Linear equations to describe these relation- compound can be used to calculate a sug- ships are presented in Table 3. gested window size; however, the experience of the analyst should weigh heavily in the References interpretation of chromatograms. 10.11 If the response for a peak exceeds 1. 40 CFR part 136, appendix B. the working range of the system, prepare a 2. Lichtenberg, J.J. ‘‘Determining Volatile dilution of the sample with reagent water Organics at Microgram-per-Litre-Levels by from the aliquot in the second syringe and Gas Chromatography,’’ Journal American reanalyze. Water Works Association, 66, 739 (1974). 3. Bellar, T.A., and Lichtenberg, J.J. 11. Calculations ‘‘Semi-Automated Headspace Analysis of 11.1 Determine the concentration of indi- Drinking Waters and Industrial Waters for vidual compounds in the sample. Purgeable Volatile Organic Compounds,’’ 11.1.1 If the external standard calibration Proceedings of Symposium on Measurement procedure is used, calculate the concentra- of Organic Pollutants in Water and Waste- tion of the parameter being measured from water. American Society for Testing and Ma- the peak response using the calibration terials, STP 686, C.E. Van Hall, editor, 1978. curve or calibration factor determined in 4. ‘‘Carcinogens—Working with Carcino- Section 7.3.2. gens,’’ Department of Health, Education, and 11.1.2 If the internal standard calibration Welfare, Public Health Service, Center for procedure is used, calculate the concentra- Disease Control, National Institute for Occu- tion in the sample using the response factor pational Safety and Health. Publication No. (RF) determined in Section 7.4.3 and Equa- 77–206, August 1977. tion 2. 5. ‘‘OSHA Safety and Health Standards, General Industry,’’ (29 CFR part 1910), Occu- ()()AC pational Safety and Health Administration, Concentration (μ= g/L) sis OSHA 2206 (Revised, January 1976). () 6. ‘‘Safety in Academic Chemistry Labora- ()ARFis tories,’’ American Chemical Society Publica- Equation 2 tion, Committee on Safety, 3rd Edition, 1979. where: 7. Provost, L.P., and Elder, R.S. ‘‘Interpre- As = Response for the parameter to be meas- tation of Percent Recovery Data,’’ American ured. Laboratory, 15, 58-63 (1983). (The value 2.44 Ais = Response for the internal standard. used in the equation in Section 8.3.3. is two Cis = Concentration of the internal standard. times the value 1.22 derived in this report.) 95

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8.‘‘Methods 330.4 (Titrimetric, DPD-FAS) TABLE 1—CHROMATOGRAPHIC CONDITIONS AND and 330.5 (Spectrophotometric, DPD) for METHOD DETECTION LIMITS—Continued Chlorine, Total Residual,’’ Methods for Chemical Analysis of Water and Wastes, Retention time (min) Method EPA–600/4–79–020, U.S. Environmental Pro- detection Parameter limit (μg/ tection Agency, Office of Research and De- Column 1 Column 2 L) velopment, Environmental Monitoring and Support Laboratory, Cincinnati, Ohio 45268. Toluene ...... 5 .75 4 .25 0 .2 March 1979. Ethylbenzene ...... 8 .25 6 .25 0 .2 9. ‘‘EPA Method Study 25, Method 602, Chlorobenzene ...... 9 .17 8.02 0 .2 Purgeable Aromatics,’’ EPA 600/4–84–042, Na- 1,4-Dichlorobenzene ..... 16 .8 16 .2 0 .3 tional Technical Information Service, PB84– 1,3-Dichlorobenzene ..... 18 .2 15 .0 0 .4 196682, Springfield, Virginia 22161, May 1984. 1,2-Dichlorobenzene ..... 25 .9 19 .4 0 .4 Column 1 conditions: Supelcoport (100/120 mesh) coated TABLE 1—CHROMATOGRAPHIC CONDITIONS AND with 5% SP–1200/1.75% Bentone-34 packed in a 6 ft × 0.085 in. ID stainless steel column with helium carrier gas at 36 mL/ METHOD DETECTION LIMITS min flow rate. Column temperature held at 50 °C for 2 min then programmed at 6 °C/min to 90 °C for a final hold. Retention time (min) Method Column 2 conditions: Chromosorb W-AW (60/80 mesh) detection coated with 5% 1,2,3-Tris(2-cyanoethyoxy)propane packed in Parameter μ × Column 1 Column 2 limit ( g/ a 6 ft 0.085 in. ID stainless steel column with helium carrier L) gas at 30 mL/min flow rate. Column temperature held at 40 °C for 2 min then programmed at 2 °C/min to 100 °C for a Benzene ...... 3.33 2 .75 0 .2 final hold.

TABLE 2—CALIBRATION AND QC ACCEPTANCE CRITERIA—METHOD 602 A

¯ Parameter Range for Q Limit for Range for X Range for (μg/L) s (μg/L) (μg/L) P, Ps(%)

Benzene ...... 15.4–24.6 4.1 10.0–27.9 39–150 Chlorobenzene ...... 16.1–23.9 3.5 12.7–25.4 55–135 1,2-Dichlorobenzene ...... 13.6–26.4 5.8 10.6–27.6 37–154 1,3-Dichlorobenzene ...... 14.5–25.5 5.0 12.8–25.5 50–141 1,4-Dichlorobenzene ...... 13.9–26.1 5.5 11.6–25.5 42–143 Ethylbenzene ...... 12.6–27.4 6.7 10.0–28.2 32–160 Toluene ...... 15.5–24.5 4.0 11.2–27.7 46–148 Q = Concentration measured in QC check sample, in μg/L (Section 7.5.3). s = Standard deviation of four recovery measurements, in μg/L (Section 8.2.4). X¯ = Average recovery for four recovery measurements, in μg/L (Section 8.2.4). Ps, P = Percent recovery measured (Section 8.3.2, Section 8.4.2). a Criteria were calculated assuming a QC check sample concentration of 20 μg/L. Note: These criteria are based directly upon the method performance data in Table 3. Where necessary, the limits for recovery have been broadened to assure applicability of the limits to concentrations below those used to develop Table 3.

TABLE 3—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 602

Accuracy, as Single analyst Overall preci- Parameter recovery, X′ precision, s′ ′ μ (μg/L) (μg/L) sion, S ( g/L)

Benzene ...... 0.92C + 0.57 0.09X¯ + 0.59 0.21X¯ + 0.56 Chlorobenzene ...... 0.95C + 0.02 0.09X¯ + 0.23 0.17X¯ + 0.10 1,2-Dichlorobenzene ...... 0.93C + 0.52 0.17X¯ ¥0.04 0.22X¯ + 0.53 1,3-Dichlorobenzene ...... 0.96C¥0.05 0.15X¯ ¥0.10 0.19X¯ + 0.09 1,4-Dichlorobenzene ...... 0.93C¥0.09 0.15X¯ + 0.28 0.20X¯ + 0.41 Ethylbenzene ...... 0.94C + 0.31 0.17X¯ + 0.46 0.26X¯ + 0.23 Toluene ...... 0.94C + 0.65 0.09X¯ + 0.48 0.18X¯ + 0.71 X′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L. S′ = Expected single analyst standard deviation of measurements at an average concentration found of X¯ , in X μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X¯ , in μg/L. C = True value for the Concentration, in μg/L. X¯ = Average recovery found for measurements of samples containing a concentration of C, in μg/L.

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OPTIONAL FOAM TRAP --14MM 0. D. INLET Y4 IN. 0. D.

1 I I I 1 I .....--sAMPLE INLET I I I I c -2-WAY SYRINGE VALVE --17CM. 20 GAUGE SYRINGE NEEDLE Y4 IN. · "-sMM. 0. D. RUBBER SEPTUM O, D. EXIT 1/16 IN. O.D. I STAINLESS STEEL

13X MOLECULAR SIEVE PURGE GAS FILTER

PURGE GAS FLOW CONTROL

Figure 1. Purging device.

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CONSTRUCTION PACKING PROCEDURE COMPRESSION FITTING GLASS 5MM WOOL -NUT AND FERRULES 14fT.7A/FOOT RESISTANCE ;r.,. WIRE WRAPPED SOLID ,·•.. !-:. THERMOCOUPLE/ ·.:/· ,!).- CONTROLLER -~t'r. SENSOR :l?J,· ELECTRONIC .TEN AX 23CM ft. TEMPERATURE :, CONTROL AND PYROMETER ~ TUBING 25CM. -~}J 0.105 IN. I.D. 3% OV-1 ~~>- 0.125 IN. O.D. STAINLESS STEa GLASS WOOL 1 5MMCMI TRAP INLET Figure 2. Trap packings and construction to include desorb capability.

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_ Confirmatory Column To Detector ---Analytical Column

Purge Gas Flow Control "-

,. Heater Control

Note: All Lines Between Trap and GC Should be Heated to -sooc

Figure 3. Purge and trap system - purge mode.

Liquid Injection Ports Carrier Gas Flow Control ,... ____'I_.. Column Oven f _Confirmatory Column I To Detector IL.::::~~Bir""""!''!""!" __... l --Analytical Column l!:::::::!:::llllf4A1 Valve-3 Optional 4-Port Column Selection Valve Purge Gas Trap Inlet (Tenax End) Flow Control Resistance Wire /Heater Control

13X Molecular Sieve Filter

Note: All Lines Between Trap and GC Should be Heated to 80°C

Figure 4. Purge and trap system-dry mode.

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Carrier Gas Flow Control Liquid Injection Ports r------.- Column Oven I _Confirmatory Column I To Detector lb~~t:Tlr______,l ----Analytical Column 1.!::::!:::1'+;,

Purge Gas Flow Control '\...... -Heater Control

13X Molecular Sieve Filter

Note: All Lines Between Trap and GC Should be Heated to 80°C

Figure 5. Purge and trap system-desorb mode.

Cllc Cll Column: 5% SP 1200/1.75% Bentone- 34 cN on Supelcoport Cll lXI Cll Program: 50°C for 2 min, &OC/min to 900C ~ Detector: Photoionization, 10.2 V Cll :I ~0 :::~1- 0 -e 0 Cllc :I Cll !E Nc 1- 1l e 0 :2 ".; 1.) i5 r't...

40 6 8 2 10 12 14 16 18 20 22 24 26 28.

Retention Time, Min.

Figure 6. Gas chromatogram of purgeable aromatics.

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METHOD 603—ACROLEIN AND ACRYLONITRILE separate the analytes which are then de- tected with a flame ionization detector. 23 1. Scope and Application 2.2 The method provides an optional gas 1.1 This method covers the determination chromatographic column that may be help- of acrolein and acrylonitrile. The following ful in resolving the compounds of interest parameters may be determined by this meth- from the interferences that may occur. od: 3. Interferences STORET Parameter No. CAS No. 3.1 Impurities in the purge gas and or- ganic compound outgassing from the plumb- Acrolein ...... 34210 107–02–8 ing of the trap account for the majority of Acrylonitrile ...... 34215 107–13–1 contamination problems. The analytical sys- tem must be demonstrated to be free from contamination under the conditions of the 1.2 This is a purge and trap gas chromatographic (GC) method applicable to analysis by running laboratory reagent the determination of the compounds listed blanks as described in Section 8.1.3. The use above in municipal and industrial discharges of non-Teflon plastic tubing, non-Teflon as provided under 40 CFR 136.1. When this thread sealants, or flow controllers with rub- method is used to analyze unfamiliar sam- ber components in the purge and trap system ples for either or both of the compounds should be avoided. above, compound identifications should be 3.2 Samples can be contaminated by diffu- supported by at least one additional quali- sion of volatile organics through the septum tative technique. This method describes ana- seal into the sample during shipment and lytical conditions for a second gas storage. A field reagent blank prepared from chromatographic column that can be used to reagent water and carried through the sam- confirm measurements made with the pri- pling and handling protocol can serve as a mary column. Method 624 provides gas chro- check on such contamination. matograph/mass spectrometer (GC/MS) con- 3.3 Contamination by carry-over can ditions appropriate for the qualitative and occur whenever high level and low level sam- quantitative confirmation of results for the ples are sequentially analyzed. To reduce parameters listed above, if used with the carry-over, the purging device and sample purge and trap conditions described in this syringe must be rinsed between samples with method. reagent water. Whenever an unusually con- 1.3 The method detection limit (MDL, de- centrated sample is encountered, it should be fined in Section 12.1) 1 for each parameter is followed by an analysis of reagent water to listed in Table 1. The MDL for a specific check for cross contamination. For samples wastewater may differ from those listed, de- containing large amounts of water-soluble pending upon the nature of interferences in materials, suspended solids, high boiling the sample matrix. compounds or high analyte levels, it may be 1.4 Any modification of this method, be- necessary to wash the purging device with a yond those expressly permitted, shall be con- detergent solution, rinse it with distilled sidered as a major modification subject to water, and then dry it in an oven at 105 °C application and approval of alternate test between analyses. The trap and other parts procedures under 40 CFR 136.4 and 136.5. of the system are also subject to contamina- 1.5 This method is restricted to use by or tion, therefore, frequent bakeout and purg- under the supervision of analysts experi- ing of the entire system may be required. enced in the operation of a purge and trap system and a gas chromatograph and in the 4. Safety interpretation of gas chromatograms. Each 4.1 The toxicity or carcinogenicity of analyst must demonstrate the ability to gen- each reagent used in this method has not erate acceptable results with this method been precisely defined; however, each chem- using the procedure described in Section 8.2. ical compound should be treated as a poten- tial health hazard. From this view point, ex- 2. Summary of Method posure to these chemicals must be reduced to 2.1 An inert gas is bubbled through a 5-mL the lowest possible level by whatever means water sample contained in a heated purging available. The laboratory is responsible for chamber. Acrolein and acrylonitrile are maintaining a current awareness file of transferred from the aqueous phase to the OSHA regulations regarding the safe han- vapor phase. The vapor is swept through a dling of the chemicals specified in this meth- sorbent trap where the analytes are trapped. od. A reference file of material data handling After the purge is completed, the trap is sheets should also be made available to all heated and backflushed with the inert gas to personnel involved in the chemical analysis. desorb the compound onto a gas Additional references to laboratory safety chromatographic column. The gas chro- are available and have been identified 46 for matograph is temperature programmed to the information of the analyst.

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5. Apparatus and Materials sories including syringes, analytical col- umns, gases, detector, and strip-chart re- 5.1 Sampling equipment, for discrete sam- corder. A data system is recommended for pling. measuring peak areas. 5.1.1 Vial—25-mL capacity or larger, 5.4.1 Column 1—10 ft long × 2 mm ID glass equipped with a screw cap with a hole in the or stainless steel, packed with Porapak-QS center (Pierce #13075 or equivalent). Deter- (80/100 mesh) or equivalent. This column was gent wash, rinse with tap and distilled water, used to develop the method performance and dry at 105 °C before use. statements in Section 12. Guidelines for the 5.1.2 Septum—Teflon-faced silicone use of alternate column packings are pro- (Pierce #12722 or equivalent). Detergent vided in Section 10.1. wash, rinse with tap and distilled water and × dry at 105 °C for 1 h before use. 5.4.2 Column 2—6 ft long 0.1 in. ID glass 5.2 Purge and trap system—The purge and or stainless steel, packed with Chromosorb trap system consists of three separate pieces 101 (60/80 mesh) or equivalent. of equipment: a purging device, trap, and 5.4.3 Detector—Flame ionization detector. desorber. Several complete systems are now This type of detector has proven effective in commercially available. the analysis of wastewaters for the param- 5.2.1 The purging device must be designed eters listed in the scope (Section 1.1), and to accept 5-mL, samples with a water column was used to develop the method performance at least 3 cm deep. The gaseous head space statements in Section 12. Guidelines for the between the water column and the trap must use of alternate detectors are provided in have a total volume of less than 15 mL. The Section 10.1. purge gas must pass through the water col- 5.5 Syringes—5-mL, glass hypodermic umn as finely divided bubbles with a diame- with Luerlok tip (two each). μ ter of less than 3 mm at the origin. The 5.6 Micro syringes—25- L, 0.006 in. ID nee- purge gas must be introduced no more than dle. 5 mm from the base of the water column. 5.7 Syringe valve—2-way, with Luer ends The purging device must be capable of being (three each). heated to 85 °C within 3.0 min after transfer 5.8 Bottle—15-mL, screw-cap, with Teflon of the sample to the purging device and cap liner. being held at 85 ±2 °C during the purge cycle. 5.9 Balance—Analytical, capable of accu- The entire water column in the purging de- rately weighing 0.0001 g. vice must be heated. Design of this modifica- 6. Reagents tion to the standard purging device is op- tional, however, use of a water bath is sug- 6.1 Reagent water—Reagent water is de- gested. fined as a water in which an interferent is 5.2.1.1 Heating mantle—To be used to heat not observed at the MDL of the parameters water bath. of interest. 5.2.1.2 Temperature controller—Equipped 6.1.1 Reagent water can be generated by with thermocouple/sensor to accurately con- passing tap water through a carbon filter bed trol water bath temperature to ±2 °C. The containing about 1 lb of activated carbon purging device illustrated in Figure 1 meets (Filtrasorb-300, Calgon Corp., or equivalent). these design criteria. 6.1.2 A water purification system 5.2.2 The trap must be at least 25 cm long (Millipore Super-Q or equivalent) may be and have an inside diameter of at least 0.105 used to generate reagent water. in. The trap must be packed to contain 1.0 6.1.3 Regent water may also be prepared cm of methyl silicone coated packing (Sec- by boiling water for 15 min. Subsequently, tion 6.5.2) and 23 cm of 2,6-diphenylene oxide while maintaining the temperature at 90 °C, polymer (Section 6.5.1). The minimum speci- bubble a contaminant-free inert gas through fications for the trap are illustrated in Fig- the water for 1 h. While still hot, transfer ure 2. the water to a narrow mouth screw-cap bot- 5.2.3 The desorber must be capable of rap- tle and seal with a Teflon-lined septum and idly heating the trap to 180 °C, The desorber cap. illustrated in Figure 2 meets these design 6.2 Sodium thiosulfate—(ACS) Granular. criteria. 6.3 Sodium hydroxide solution (10 N)— 5.2.4 The purge and trap system may be Dissolve 40 g of NaOH (ACS) in reagent water assembled as a separate unit as illustrated in and dilute to 100 mL. Figure 3 or be coupled to a gas chro- 6.4 Hydrochloric acid (1 + 1)—Slowly, add matograph. 50 mL of concentrated HCl (ACS) to 50 mL of 5.3 pH paper—Narrow pH range, about 3.5 reagent water. to 5.5 (Fisher Scientific Short Range Alkacid 6.5 Trap Materials: No. 2, #14–837–2 or equivalent). 6.5.1 2,6-Diphenylene oxide polymer— 5.4 Gas chromatograph—An analytical Tenax (60/80 mesh), chromatographic grade system complete with a temperature pro- or equivalent. grammable gas chromatograph suitable for 6.5.2 Methyl silicone packing—3% OV–1 on on-column injection and all required acces- Chromosorb-W (60/80 mesh) or equivalent.

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6.6 Stock standard solutions—Stock and trap-gas chromatographic system using standard solutions may be prepared from either the external standard technique (Sec- pure standard materials or purchased as cer- tion 7.3) or the internal standard technique tified solutions. Prepare stock standard solu- (Section 7.4). tions in reagent water using assayed liquids. 7.3 External standard calibration proce- Since acrolein and acrylonitrile are dure: lachrymators, primary dilutions of these 7.3.1 Prepare calibration standards at a compounds should be prepared in a hood. A minimum of three concentration levels for NIOSH/MESA approved toxic gas respirator each parameter by carefully adding 20.0 μL of should be used when the analyst handles one or more secondary dilution standards to high concentrations of such materials. 100, 500, or 1000 mL of reagent water. A 25-μL 6.6.1 Place about 9.8 mL of reagent water syringe with a 0.006 in. ID needle should be into a 10-mL ground glass stoppered volu- used for this operation. One of the external metric flask. For acrolein standards the rea- standards should be at a concentration near, gent water must be adjusted to pH 4 to 5. but above, the MDL and the other concentra- Weight the flask to the nearest 0.1 mg. tions should correspond to the expected 6.6.2 Using a 100-μL syringe, immediately range of concentrations found in real sam- add two or more drops of assayed reference ples or should define the working range of material to the flask, then reweigh. Be sure the detector. These standards must be pre- that the drops fall directly into the water pared fresh daily. without contacting the neck of the flask. 7.3.2 Analyze each calibration standard 6.6.3 Reweigh, dilute to volume, stopper, according to Section 10, and tabulate peak then mix by inverting the flask several height or area responses versus the con- times. Calculate the concentration in μg/μL centration of the standard. The results can from the net gain in weight. When compound be used to prepare a calibration curve for purity is assayed to be 96% or greater, the each compound. Alternatively, if the ratio of weight can be used without correction to cal- response to concentration (calibration fac- culate the concentration of the stock tor) is a constant over the working range staldard. Optionally, stock standard solu- (<10% relative standard deviation, RSD), lin- tions may be prepared using the pure stand- earity through the origin can be assumed ard material by volumetrically measuring and the average ratio or calibration factor the appropriate amounts and determining can be used in place of a calibration curve. the weight of the material using the density 7.4 Internal standard calibration proce- of the material. Commercially prepared dure—To use this approach, the analyst must stock standards may be used at any con- select one or more internal standards that centration if they are certified by the are similar in analytical behavior to the manufactaurer or by an independent source. compounds of interest. The analyst must fur- 6.6.4 Transfer the stock standard solution ther demonstrate that the measurement of into a Teflon-sealed screw-cap bottle. Store the internal standard is not affected by at 4 °C and protect from light. method or matrix interferences. Because of 6.6.5 Prepare fresh standards daily. these limitations, no internal standard can 6.7 Secondary dilution standards—Using be suggested that is applicable to all sam- stock standard solutions, prepare secondary ples. dilution standards in reagent water that con- 7.4.1 Prepare calibration standards at a tain the compounds of interest, either singly minimum of three concentration levels for or mixed together. The secondary dilution each parameter of interest as described in standards should be prepared at concentra- Section 7.3.1. tions such that the aqueous calibration 7.4.2 Prepare a spiking solution con- standards prepared in Section 7.3.1 or 7.4.1 taining each of the internal standards using will bracket the working range of the ana- the procedures described in Sections 6.6 and lytical system. Secondary dilution standards 6.7. It is recommended that the secondary di- should be prepared daily and stored at 4 °C. lution standard be prepared at a concentra- 6.8 Quality control check sample con- tion of 15 μg/mL of each internal standard centrate—See Section 8.2.1. compound. The addition of 10 μL of this standard to 5.0 mL of sample or calibration 7. Calibration standard would be equivalent to 30 μg/L. 7.1 Assemble a purge and trap system that 7.4.3 Analyze each calibration standard meets the specifications in Section 5.2. Con- according to Section 10, adding 10 μL of in- dition the trap overnight at 180 °C by ternal standard spiking solution directly to backflushing with an inert gas flow of at the syringe (Section 10.4). Tabulate peak least 20 mL/min. Condition the trap for 10 height or area responses against concentra- min once daily prior to use. tion for each compound and internal stand- 7.2 Connect the purge and trap system to ard, and calculate response factors (RF) for a gas chromatograph. The gas chro- each compound using Equation 1. matograph must be operated using tempera- RF = (A )(C (A )(C ) ture and flow rate conditions equivalent to s is is s those given in Table 1. Calibrate the purge Equation 1

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where: interferences from the analytical system are under control. As = Response for the parameter to be meas- ured. 8.1.4 The laboratory must, on an ongoing basis, spike and analyze a minimum of 10% Ais = Response for the internal standard. of all samples to monitor and evaluate lab- Cis = Concentration of the internal standard. oratory data quality. This procedure is de- Cs = Concentration of the parameter to be measured. scribed in Section 8.3. 8.1.5 The laboratory must, on an ongoing If the RF value over the working range is a basis, demonstrate through the analyses of constant (<10% RSD), the RF can be assumed quality control check standards that the op- to be invariant and the average RF can be eration of the measurement system is in con- used for calculations. Alternatively, the re- trol. This procedure is described in Section sults can be used to plot a calibration curve 8.4. The frequency of the check standard of response ratios, A /A , vs. RF. s is analyses is equivalent to 10% of all samples 7.5 The working calibration curve, cali- analyzed but may be reduced if spike recov- bration factor, or RF must be verified on eries from samples (Section 8.3) meet all each working day by the measurement of a specified quality control criteria. QC check sample. 8.1.6 The laboratory must maintain per- 7.5.1 Prepare the QC check sample as de- formance records to document the quality of scribed in Section 8.2.2. data that is generated. This procedure is de- 7.5.2 Analyze the QC check sample accord- scribed in Section 8.5. ing to Section 10. 8.2 To establish the ability to generate 7.5.3 For each parameter, compare the re- acceptable accuracy and precision, the ana- sponse (Q) with the corresponding calibra- lyst must perform the following operations. tion acceptance criteria found in Table 2. If 8.2.1 A quality control (QC) check sample the responses for all parameters of interest concentrate is required containing each pa- fall within the designated ranges, analysis of rameter of interest at a concentration of 25 actual samples can begin. If any individual Q μg/mL in reagent water. The QC check sam- falls outside the range, a new calibration ple concentrate must be obtained from the curve, calibration factor, or RF must be pre- U.S. Environmental Protection Agency, En- pared for that parameter according to Sec- vironmental Monitoring and Support Lab- tion 7.3 or 7.4. oratory in Cincinnati, Ohio, if available. If 8. Quality Control not available from that source, the QC check sample concentrate must be obtained from 8.1 Each laboratory that uses this method another external source. If not available is required to operate a formal quality con- from either source above, the QC check sam- trol program. The minimum requirements of ple concentrate must be prepared by the lab- this program consist of an initial demonstra- oratory using stock standards prepared inde- tion of laboratory capability and an ongoing pendently from those used for calibration. analysis of spiked samples to evaluate and 8.2.2 Prepare a QC check sample to con- document data quality. The laboratory must tain 50 μg/L of each parameter by adding 200 maintain records to document the quality of μL of QC check sample concentrate to 100 mL data that is generated. Ongoing data quality of reagent water. checks are compared with established per- 8.2.3 Analyze four 5-mL aliquots of the formance criteria to determine if the results well-mixed QC check sample according to of analyses meet the performance character- Section 10. istics of the method. When results of sample 8.2.4 Calculate the average recovery (X¯ ) in spikes indicate atypical method perform- μg/L, and the standard deviation of the re- ance, a quality control check standard must covery (s) in μg/L, for each parameter using be analyzed to confirm that the measure- the four results. ments were performed in an in-control mode 8.2.5 For each parameter compare s and X¯ of operation. with the corresponding acceptance criteria 8.1.1 The analyst must make an initial, for precision and accuracy, respectively, one-time, demonstration of the ability to found in Table 3. If s and X¯ for all param- generate acceptable accuracy and precision eters of interest meet the acceptance cri- with this method. This ability is established teria, the system performance is acceptable as described in Section 8.2. and analysis of actual samples can begin. If 8.1.2 In recognition of advances that are either s exceeds the precision limit or X¯ falls occurring in chromatography, the analyst is outside the range for accuracy, the system permitted certain options (detailed in Sec- performance is unacceptable for that param- tion 10.1) to improve the separations or lower eter. Locate and correct the source of the the cost of measurements. Each time such a problem and repeat the test for each com- modification is made to the method, the ana- pound of interest. lyst is required to repeat the procedure in 8.3 The laboratory must, on an ongoing Section 8.2. basis, spike at least 10% of the samples from 8.1.3 Each day, the analyst must analyze a each sample site being monitored to assess reagent water blank to demonstrate that accuracy. For laboratories analyzing one to

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ten samples per month, at least one spiked covery (Ps) as 100 (A/T)%, where T is the true sample per month is required. value of the standard concentration. 8.3.1 The concentration of the spike in the 8.4.3 Compare the percent recovery (Ps) sample should be determined as follows: for each parameter with the corresponding 8.3.1.1 If, as in compliance monitoring, QC acceptance criteria found in Table 3. Only the concentration of a specific parameter in parameters that failed the test in Section 8.3 the sample is being checked against a regu- need to be compared with these criteria. If latory concentration limit, the spike should the recovery of any such parameter falls out- be at that limit or 1 to 5 times higher than side the designated range, the laboratory the background concentration determined in performance for that parameter is judged to Section 8.3.2, whichever concentration would be out of control, and the problem must be be larger. immediately identified and corrected. The 8.3.1.2 If the concentration of a specific analytical result for that parameter in the parameter in the sample is not being unspiked sample is suspect and may not be checked against a limit specific to that pa- reported for regulatory compliance purposes. rameter, the spike should be at 50 μg/L or 1 8.5 As part of the QC program for the lab- to 5 times higher than the background con- oratory, method accuracy for wastewater centration determined in Section 8.3.2, samples must be assessed and records must whichever concentration would be larger. be maintained. After the analysis of five 8.3.2 Analyze one 5-mL sample aliquot to spiked wastewater samples as in Section 8.3, determine the background concentration (B) calculate the average percent recovery (P¯ ) of each parameter. If necessary, prepare a and the standard deviation of the percent re- new QC check sample concentrate (Section covery (sp). Express the accuracy assessment ¯ 8.2.1) appropriate for the background con- as a percent recovery interval from P¥2sp to ¯ ¯ centrations in the sample. Spike a second 5- P + 2sp. If P = 90% and sp = 10%, for example, mL sample aliquot with 10 μL of the QC the accuracy interval is expressed as 70– check sample concentrate and analyze it to 110%. Update the accuracy assessment for determine the concentration after spiking each parameter on a regular basis (e.g. after (A) of each parameter. Calculate each per- each five to ten new accuracy measure- cent recovery (P) as 100(A¥B)%/T, where T is ments). the known true value of the spike. 8.6 It is recommended that the laboratory 8.3.3 Compare the percent recovery (P) for adopt additional quality assurance practices each parameter with the corresponding QC for use with this method. The specific prac- acceptance criteria found in Table 3. These tices that are most productive depend upon acceptance criteria were calculated to in- the needs of the laboratory and the nature of clude an allowance for error in measurement the samples. Field duplicates may be ana- of both the background and spike concentra- lyzed to assess the precision of the environ- tions, assuming a spike to background ratio mental measurements. When doubt exists of 5:1. This error will be accounted for to the over the identification of a peak on the chro- extent that the analyst’s spike to back- matogram, confirmatory techniques such as ground ratio approaches 5:1. 7 gas chromatography with a dissimilar col- 8.3.4 If any individual P falls outside the umn or mass spectrometer must be used. designated range for recovery, that param- Whenever possible, the laboratory should eter has failed the acceptance criteria. A analyze standard reference materials and check standard containing each parameter participate in relevant performance evalua- that failed the criteria must be analyzed as tion studies. described in Section 8.4. 8.4 If any parameter fails the acceptance 9. Sample Collection, Preservation, and criteria for recovery in Section 8.3, a QC Handling check standard containing each parameter 9.1 All samples must be iced or refrig- that failed must be prepared and analyzed. erated from the time of collection until anal- NOTE: The frequency for the required anal- ysis. If the sample contains free or combined ysis of a QC check standard will depend upon chlorine, add sodium thiosulfate preserva- the number of parameters being simulta- tive (10 mg/40 mL is sufficient for up to 5 neously tested, the complexity of the sample ppm Cl2) to the empty sample bottle just matrix, and the performance of the labora- prior to shipping to the sampling site. EPA tory. Methods 330.4 and 330.5 may be used for 8.4.1 Prepare the QC check standard by measurement of residual chlorine. 8 Field adding 10 μL of QC check sample concentrate test kits are available for this purpose. (Section 8.2.1 or 8.3.2) to 5 mL of reagent 9.2 If acrolein is to be analyzed, collect water. The QC check standard needs only to about 500 mL of sample in a clean glass con- contain the parameters that failed criteria tainer. Adjust the pH of the sample to 4 to 5 in the test in Section 8.3. using acid or base, measuring with narrow 8.4.2 Analyze the QC check standard to range pH paper. Samples for acrolein anal- determine the concentration measured (A) of ysis receiving no pH adjustment must be each parameter. Calculate each percent re- analyzed within 3 days of sampling.

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9.3 Grab samples must be collected in ringe. Wash the chamber with two 5-mL glass containers having a total volume of at flushes of reagent water. least 25 mL. Fill the sample bottle just to 10.9 After desorbing the sample for 1.5 overflowing in such a manner that no air min, recondition the trap by returning the bubbles pass through the sample as the bot- purge and trap system to the purge mode. tle is being filled. Seal the bottle so that no Wait 15 s then close the syringe valve on the air bubbles are entrapped in it. If preserva- purging device to begin gas flow through the tive has been added, shake vigorously for 1 trap. The trap temperature should be main- min. Maintain the hermetic seal on the sam- tained at 210 °C. After approximately 7 min, ple bottle until time of analysis. turn off the trap heater and open the syringe 9.4 All samples must be analyzed within valve to stop the gas flow through the trap. 14 days of collection. 3 When the trap is cool, the next sample can be analyzed. 10. Procedure 10.10 Identify the parameters in the sam- 10.1 Table 1 summarizes the recommended ple by comparing the retention times of the operating conditions for the gas chro- peaks in the sample chromatogram with matograph. Included in this table are esti- those of the peaks in standard mated retention times and MDL that can be chromatograms. The width of the retention achieved under these conditions. An example time window used to make identifications of the separations achieved by Column 1 is should be based upon measurements of ac- shown in Figure 5. Other packed columns, tual retention time variations of standards chromatographic conditions, or detectors over the course of a day. Three times the may be used if the requirements of Section standard deviation of a retention time for a 8.2 are met. compound can be used to calculate a sug- 10.2 Calibrate the system daily as de- gested window size; however, the experience scribed in Section 7. of the analyst should weigh heavily in the 10.3 Adjust the purge gas (nitrogen or he- interpretation of chromatograms. lium) flow rate to 20 mL-min. Attach the trap inlet to the purging device, and set the 11. Calculations purge and trap system to purge (Figure 3). 11.1 Determine the concentration of indi- Open the syringe valve located on the purg- vidual compounds in the sample. ing device sample introduction needle. 11.1.1 If the external standard calibration 10.4 Remove the plunger from a 5-mL sy- procedure is used, calculate the concentra- ringe and attach a closed syringe valve. Open tion of the parameter being measured from the sample bottle (or standard) and carefully the peak response using the calibration pour the sample into the syringe barrel to curve or calibration factor determined in just short of overflowing. Replace the sy- Section 7.3.2. ringe plunger and compress the sample. Open 11.1.2 If the internal standard calibration the syringe valve and vent any residual air procedure is used, calculate the concentra- while adjusting the sample volume to 5.0 mL. tion in the sample using the response factor Since this process of taking an aliquot de- (RF) determined in Section 7.4.3 and Equa- stroys the validity of the sample for future tion 2. analysis, the analyst should fill a second sy- ringe at this time to protect against possible ()()AC loss of data. Add 10.0 μL of the internal Concentration (μ= g/L) sis standard spiking solution (Section 7.4.2), if ()() applicable, through the valve bore then close ARFis the valve. Equation 2 10.5 Attach the syringe-syringe valve as- sembly to the syringe valve on the purging where: device. Open the syringe valves and inject As = Response for the parameter to be meas- the sample into the purging chamber. ured. 10.6 Close both valves and purge the sam- A = Response for the internal standard. ± ± ° is ple for 15.0 0.1 min while heating at 85 2 C. C = Concentration of the internal standard. 10.7 After the 15-min purge time, attach is μ the trap to the chromatograph, adjust the 11.2 Report results in g/L without correc- purge and trap system to the desorb mode tion for recovery data. All QC data obtained (Figure 4), and begin to temperature pro- should be reported with the sample results. gram the gas chromatograph. Introduce the 12. Method Performance trapped materials to the GC column by rap- idly heating the trap to 180 °C while 12.1 The method detection limit (MDL) is backflushing the trap with an inert gas be- defined as the minimum concentration of a tween 20 and 60 mL/min for 1.5 min. substance that can be measured and reported 10.8 While the trap is being desorbed into with 99% confidence that the value is above the gas chromatograph, empty the purging zero. 1 The MDL concentrations listed in chamber using the sample introduction sy- Table 1 were obtained using reagent water. 9

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The MDL actually achieved in a given anal- tion, Committee on Chemical Safety, 3rd ysis will vary depending on instrument sen- Edition, 1979. sitivity and matrix effects. 7. Provost, L.P., and Elder, R.S. ‘‘Interpre- 12.2 This method is recommended for the tation of Percent Recovery Data,’’ American concentration range from the MDL to 1,000 × Laboratory, 15, 58–63 (1983). MDL. Direct aqueous injection techniques 8. ‘‘Methods 330.4 (Titrimetric, DPD-FAS) should be used to measure concentration lev- and 330.5 (Spectrophotometric, DPD) for els above 1,000 × MDL. Chlorine, Total Residual,’’ Methods for 12.3 In a single laboratory (Battelle-Co- Chemical Analysis of Water and Wastes, lumbus), the average recoveries and standard EPA–600/4–79–020, U.S. Environmental Pro- deviations presented in Table 2 were ob- tained. 9 Seven replicate samples were ana- tection Agency, Environmental Monitoring lyzed at each spike level. and Support Laboratory, Cincinnati, Ohio 45268, March 1979. References 9. ‘‘Evaluation of Method 603 (Modified),’’ 1. 40 CFR part 136, appendix B. EPA–600/4–84–ABC, National Technical Infor- 2. Bellar, T.A., and Lichtenberg, J.J. ‘‘De- mation Service, PB84–, Springfield, Virginia termining Volatile Organics at Microgram- 22161, Nov. 1984. per-Litre-Levels by Gas Chromatography,’’ Journal American Water Works Association, 66, TABLE 1—CHROMATOGRAPHIC CONDITIONS AND 739 (1974). METHOD DETECTION LIMITS 3. ‘‘Evaluate Test Procedures for Acrolein and Acrylonitrile,’’ Special letter report for Retention time (min) Method Parameter detection EPA Project 4719–A, U.S. Environmental Column 1 Column 2 limit (μg/L) Protection Agency, Environmental Moni- toring and Support Laboratory, Cincinnati, Acrolein ...... 10 .6 8 .2 0 .7 Ohio 45268, 27 June 1979. Acrylonitrile ...... 12.7 9 .8 0 .5 4. ‘‘Carcinogens—Working With Carcino- Column 1 conditions: Porapak-QS (80/100 mesh) packed in gens,’’ Department of Health, Education, and a 10 ft × 2 mm ID glass or stainless steel column with helium Welfare, Public Health Service, Center for carrier gas at 30 mL/min flow rate. Column temperature held Disease Control, National Institute for Occu- isothermal at 110 °C for 1.5 min (during desorption), then heated as rapidly as possible to 150 °C and held for 20 min; pational Safety and Health, Publication No. column bakeout at 190 °C for 10 min. 9 77–206, August 1977. Column 2 conditions: Chromosorb 101 (60/80 mesh) 5. ‘‘OSHA Safety and Health Standards, packed in a 6 ft. × 0.1 in. ID glass or stainless steel column General Industry,’’ (29 CFR part 1910), Occu- with helium carrier gas at 40 mL/min flow rate. Column tem- perature held isothermal at 80 °C for 4 min, then programmed pational Safety and Health Administration, at 50 °C/min to 120 °C and held for 12 min. OSHA 2206 (Revised, January 1976). 6. ‘‘Safety in Academic Chemistry Labora- tories,’’ American Chemical Society Publica-

TABLE 2—SINGLE LABORATORY ACCURACY AND PRECISION—METHOD 603

Spike Average Standard Average Parameter Sample conc. recovery deviation percent matrix (μg/L) (μg/L) (μg/L) recovery

Acrolein ...... RW 5.0 5.2 0.2 104 RW 50.0 51.4 0.7 103 POTW 5.0 4.0 0.2 80 POTW 50.0 44.4 0.8 89 IW 5.0 0.1 0.1 2 IW 100.0 9.3 1.1 9 Acrylonitrile ...... RW 5.0 4.2 0.2 84 RW 50.0 51.4 1.5 103 POTW 20.0 20.1 0.8 100 POTW 100.0 101.3 1.5 101 IW 10.0 9.1 0.8 91 IW 100.0 104.0 3.2 104 RW = Reagent water. POTW = Prechlorination secondary effluent from a municipal sewage treatment plant. IW = Industrial wastewater containing an unidentified acrolein reactant.

TABLE 3—CALIBRATION AND QC ACCEPTANCE CRITERIA—METHOD 603 A

Parameter Range for Q Limit for Range for X Range for (μg/L) S (μg/L) (μg/L) P, Ps (%)

Acrolein ...... 45.9–54.1 4.6 42.9–60.1 88–118

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TABLE 3—CALIBRATION AND QC ACCEPTANCE CRITERIA—METHOD 603 A—Continued

Parameter Range for Q Limit for Range for X Range for (μg/L) S (μg/L) (μg/L) P, Ps (%)

Acrylonitrile ...... 41.2–58.8 9.9 33.1–69.9 71–135

a = Criteria were calculated assuming a QC check sample concentration of 50 μg/L. 9 Q = Concentration measured in QC check sample, in μg/L (Section 7.5.3). s = Standard deviation of four recovery measurements, in μg/L (Section 8.2.4). X = Average recovery for four recovery measurements, in μg/L (Section 8.2.4). P, Ps = Percent recovery measured (Section 8.3.2, Section 8.4.2).

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THERMOCOUPLE/ CONTROLLER ;:? SENSOR ELECTRONIC TENAX 23CM ·t: TEMPERATURE CONTROL %. AND :ti. PYROMETER ~{ TUBING 25CM. ·~~ 0.105 IN. 1.0. 0.125 IN. 0.0. 3% OV-11CMI ~{: STAINLESS STEEL GLASS WOOL 5MM ' TRAP INLET - ·- Figure 2. Trap packings and construction to include desorb capability.

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13X MOLECULAR GC INJECTION SIEVE FILTER PORT

HEATED WATER BATH

Figure 3. Purge and trap system-purge mode.

13X MOLECULAR GC INJECTION SIEVE FILTER PORT

Figure 4. Purge and trap system-desorb mode.

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METHOD 604—PHENOLS ples for any or all of the compounds above, compound identifications should be sup- 1. Scope and Application ported by at least one additional qualitative 1.1 This method covers the determination technique. This method describes analytical of phenol and certain substituted phenols. conditions for derivatization, cleanup, and The following parameters may be deter- electron capture detector gas chroma- mined by this method: tography (ECDGC) that can be used to con- firm measurements made by FIDGC. Method STORET 625 provides gas chromatograph/mass spec- Parameter No. CAS No. trometer (GC/MS) conditions appropriate for 4-Chloro-3-methylphenol ...... 34452 59–50–7 the qualitative and quantitative confirma- 2–-Chlorophenol ...... 34586 95–57–8 tion of results for all of the parameters list- 2,4-Dichlorophenol ...... 34601 120–83–2 ed above, using the extract produced by this 2,4-Dimethylphenol ...... 34606 105–67–9 2,4-Dinitrophenol ...... 34616 51–28–5 method. 2-Methyl-4,6-dinitrophenol ...... 34657 534–52–1 1.3 The method detection limit (MDL, de- 2-Nitrophenol ...... 34591 88–75–5 fined in Section 14.1) 1 for each parameter is 4-Nitrophenol ...... 34646 100–02–7 listed in Table 1. The MDL for a specific Pentachlorophenol ...... 39032 87–86–5 wastewater may differ from those listed, de- Phenol ...... 34694 108–95–2 2,4,6-Trichlorophenol ...... 34621 88–06–2 pending upon the nature of interferences in the sample matrix. The MDL listed in Table 1.2 This is a flame ionization detector gas 1 for each parameter was achieved with a chromatographic (FIDGC) method applicable flame ionization detector (FID). The MDLs to the determination of the compounds listed that were achieved when the derivatization above in municipal and industrial discharges cleanup and electron capture detector (ECD) as provided under 40 CFR 136.1. When this were employed are presented in Table 2. method is used to analyze unfamiliar sam-

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1.4 Any modification of this method, be- 3.1.2 The use of high purity reagents and yond those expressly permitted, shall be con- solvents helps to minimize interference prob- sidered as a major modification subject to lems. Purification of solvents by distillation application and approval of alternate test in all-glass systems may be required. procedures under 40 CFR 136.4 and 136.5. 3.2 Matrix interferences may be caused by 1.5 This method is restricted to use by or contaminants that are coextracted from the under the supervision of analysts experi- sample. The extent of matrix interferences enced in the use of a gas chromatograph and will vary considerably from source to source, in the interpretation of gas chromatograms. depending upon the nature and diversity of Each analyst must demonstrate the ability the industrial complex or municipality being to generate acceptable results with this sampled. The derivatization cleanup proce- method using the procedure described in Sec- dure in Section 12 can be used to overcome tion 8.2. many of these interferences, but unique sam- ples may require additional cleanup ap- 2. Summary of Method proaches to achieve the MDL listed in Tables 2.1 A measured volume of sample, ap- 1 and 2. proximately 1-L, is acidified and extracted 3.3 The basic sample wash (Section 10.2) with methylene chloride using a separatory may cause significantly reduced recovery of funnel. The methylene chloride extract is phenol and 2,4-dimethylphenol. The analyst dried and exchanged to 2-propanol during must recognize that results obtained under concentration to a volume of 10 mL or less. these conditions are minimum concentra- The extract is separated by gas chroma- tions. tography and the phenols are then measured 4. Safety with an FID. 2 2.2 A preliminary sample wash under 4.1 The toxicity or carcinogenicity of basic conditions can be employed for samples each reagent used in this mothod has not having high general organic and organic base been precisely defined; however, each chem- interferences. ical compound should be treated as a poten- 2.3 The method also provides for a tial health hazard. From this viewpoint, ex- derivatization and column chromatography posure to these chemicals must be reduced to cleanup procedure to aid in the elimination the lowest possible level by whatever means of interferences. 23 The derivatives are ana- available. The laboratory is responsible for lyzed by ECDGC. maintaining a current awareness file of OSHA regulations regarding the safe han- 3. Interferences dling of the chemicals specified in this meth- 3.1 Method interferences may be caused od. A reference file of material data handling by contaminants in solvents, reagents, glass- sheets should also be made available to all ware, and other sample processing hardware personnel involved in the chemical analysis. that lead to discrete artifacts and/or ele- Additional references to laboratory safety vated baselines in gas chromatograms. All of are available and have been identified 57 for these materials must be routinely dem- the information of analyst. onstrated to be free from interferences under 4.2 Special care should be taken in han- the conditions of the analysis by running dling pentafluorobenzyl bromide, which is a laboratory reagent blanks as described in lachrymator, and 18-crown-6-ether, which is Section 8.1.3. highly toxic. 3.1.1 Glassware must be scrupulously 5. Apparatus and Materials cleaned. 4 Clean all glassware as soon as pos- sible after use by rinsing with the last sol- 5.1 Sampling equipment, for discrete or vent used in it. Solvent rinsing should be fol- composite sampling. lowed by detergent washing with hot water, 5.1.1 Grab sample bottle—1–L or 1-qt, and rinses with tap water and distilled amber glass, fitted with a screw cap lined water. The glassware should then be drained with Teflon. Foil may be substituted for Tef- dry, and heated in a muffle furnace at 400 °C lon if the sample is not corrosive. If amber for 15 to 30 min. Some thermally stable ma- bottles are not available, protect samples terials, such as PCBs, may not be eliminated from light. The bottle and cap liner must be by this treatment. Solvent rinses with ace- washed, rinsed with acetone or methylene tone and pesticide quality hexane may be chloride, and dried before use to minimize substituted for the muffle furnace heating. contamination. Thorough rinsing with such solvents usually 5.1.2 Automatic sampler (optional)—The eliminates PCB interference. Volumetric sampler must incorporate glass sample con- ware should not be heated in a muffle fur- tainers for the collection of a minimum of nace. After drying and cooling, glassware 250 mL of sample. Sample containers must be should be sealed and stored in a clean envi- kept refrigerated at 4 °C and protected from ronment to prevent any accumulation of light during compositing. If the sampler uses dust or other contaminants. Store inverted a peristaltic pump, a minimum length of or capped with aluminum foil. compressible silicone rubber tubing may be

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used. Before use, however, the compressible tive in the analysis of wastewaters for tubing should be thoroughly rinsed with derivatization products of the parameters methanol, followed by repeated rinsings with listed in the scope (Section 1.1), and was used distilled water to minimize the potential for to develop the method performance state- contamination of the sample. An integrating ments in Section 14. Guidelines for the use of flow meter is required to collect flow propor- alternate column packings are provided in tional composites. Section 11.1. 5.2 Glassware (All specifications are sug- 5.6.3 Detectors—Flame ionization and gested. Catalog numbers are included for il- electron capture detectors. The FID is used lustration only.): when determining the parent phenols. The 5.2.1 Separatory funnel—2–L, with Teflon ECD is used when determining the stopcock. derivatized phenols. Guidelines for the use of 5.2.2 Drying column—Chromatographic alternatve detectors are provided in Section column, 400 mm long × 19 mm ID, with coarse 11.1. frit filter disc. 5.2.3 Chromatographic column—100 mm 6. Reagents long × 10 mm ID, with Teflon stopcock. 6.1 Reagent water—Reagent water is de- 5.2.4 Concentrator tube, Kuderna-Dan- fined as a water in which an interferent is ish—10-mL, graduated (Kontes K–570050–1025 not observed at the MDL of the parameters or equivalent). Calibration must be checked of interest. at the volumes employed in the test. Ground 6.2 Sodium hydroxide solution (10 N)— glass stopper is used to prevent evaporation Dissolve 40 g of NaOH (ACS) in reagent water of extracts. and dilute to 100 mL. 5.2.5 Evaporative flask, Kuderna-Danish— 6.3 Sodium hydroxide solution (1 N)—Dis- 500-mL (Kontes K–570001–0500 or equivalent). solve 4 g of NaOH (ACS) in reagent water and Attach to concentrator tube with springs. dilute to 100 mL. 5.2.6 Snyder column, Kuderna-Danish— 6.4 Sodium sulfate—(ACS) Granular, an- Three-ball macro (Kontes K–503000–0121 or hydrous. Purify by heating at 400 °C for 4 h equivalent). in a shallow tray. 5.2.7 Snyder column, Kuderna-Danish— 6.5 Sodium thiosulfate—(ACS) Granular. Two-ball micro (Kontes K–569001–0219 or 6.6 Sulfuric acid (1 + 1)—Slowly, add 50 equivalent). mL of H2SO4 (ACS, sp. gr. 1.84) to 50 mL of 5.2.8 Vials—10 to 15-mL, amber glass, with reagent water. Teflon-lined screw cap. 6.7 Sulfuric acid (1 N)—Slowly, add 58 mL 5.2.9 Reaction flask—15 to 25-mL round of H2SO4 (ACS, sp. gr. 1.84) to reagent water bottom flask, with standard tapered joint, and dilute to 1 L. fitted with a water-cooled condenser and U- 6.8 Potassium —(ACS) Pow- shaped drying tube containing granular cal- dered. cium chloride. 6.9 Pentafluorobenzyl bromide (a- 5.3 Boiling chips—Approximately 10/40 Bromopentafluorotoluene)—97% minimum mesh. Heat to 400 °C for 30 min or Soxhlet ex- purity. tract with methylene chloride. NOTE: This chemical is a lachrymator. (See 5.4 Water bath—Heated, with concentric Section 4.2.) ring cover, capable of temperature control 6.10 18-crown-6-ether (1,4,7,10,13,16- (±2 °C). The bath should be used in a hood. Hexaoxacyclooctadecane)—98% minimum 5.5 Balance—Analytical, capable of accu- purity. rately weighting 0.0001 g. NOTE: This chemical is highly toxic. 5.6 Gas chromatograph—An analytical 6.11 Derivatization reagent—Add 1 mL of system complete with a temperature pro- pentafluorobenzyl bromide and 1 g of 18- grammable gas chromatograph suitable for crown-6-ether to a 50-mL volumetric flask on-column injection and all required acces- and dilute to volume with 2-propanol. Pre- sories including syringes, analytical col- pare fresh weekly. This operation should be umns, gases, detector, and strip-chart re- carried out in a hood. Store at 4 °C and pro- corder. A data system is recommended for tect from light. measuring peak areas. 6.12 Acetone, hexane, methanol, meth- 5.6.1 Column for underivatized phenols— ylene chloride, 2-propanol, toluene—Pes- 1.8 m long × 2 mm ID glass, packed with 1% ticide quality or equivalent. SP–1240DA on Supelcoport (80/100 mesh) or 6.13 Silica gel—100/200 mesh, Davison, equivalent. This column was used to develop grade-923 or equivalent. Activate at 130 °C the method performance statements in Sec- overnight and store in a desiccator. tion 14. Guidelines for the use of alternate 6.14 Stock standard solutions (1.00 μg/ column packings are provided in Section μL)—Stock standard solutions may be pre- 11.1. pared from pure standard materials or pur- 5.6.2 Column for derivatized phenols—1.8 chased as certified solutions. m long × 2 mm ID glass, packed with 5% OV– 6.14.1 Prepare stock standard solutions by 17 on Chromosorb W-AW-DMCS (80/100 mesh) accurately weighing about 0.0100 g of pure or equivalent. This column has proven effec- material. Dissolve the material in 2-propanol

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and dilute to volume in a 10-mL volumetric affected by method or matrix interferences. flask. Larger volumes can be used at the con- Because of these limitations, no internal venience of the analyst. When compound pu- standard can be suggested that is applicable rity is assayed to be 96% or greater, the to all samples. weight can be used without correction to cal- 7.3.1 Prepare calibration standards at a culate the concentration of the stock stand- minimum of three concentration levels for ard. Commercially prepared stock standards each parameter of interest by adding vol- can be used at any concentration if they are umes of one or more stock standards to a certified by the manufacturer or by an inde- volumetric flask. To each calibration stand- pendent source. ard, add a known constant amount of one or 6.14.2 Transfer the stock standard solu- more internal standards, and dilute to vol- tions into Teflon-sealed screw-cap bottles. ume with 2-propanol. One of the standards Store at 4 °C and protect from light. Stock should be at a concentration near, but above, standard solutions should be checked fre- the MDL and the other concentrations quently for signs of degradation or evapo- should correspond to the expected range of ration, especially just prior to preparing concentrations found in real samples or calibration standards from them. should define the working range of the detec- 6.14.3 Stock standard solutions must be tor. replaced after six months, or sooner if com- 7.3.2 Using injections of 2 to 5 μL, analyze parison with check standards indicates a each calibration standard according to Sec- problem. tion 11 and tabulate peak height or area re- 6.15 Quality control check sample con- sponses against concentration for each com- centrate—See Section 8.2.1. pound and internal standard. Calculate re- sponse factors (RF) for each compound using 7. Calibration Equation 1. 7.1 To calibrate the FIDGC for the RF = (A )(C (A )(C ) anaylsis of underivatized phenols, establish s is is s gas chromatographic operating conditions Equation 1 equivalent to those given in Table 1. The gas where: chromatographic system can be calibrated As = Response for the parameter to be meas- using the external standard technique (Sec- ured. tion 7.2) or the internal standard technique Ais = Response for the internal standard. (Section 7.3). Cis = Concentration of the internal standard 7.2 External standard calibration proce- (μg/L). dure for FIDGC: C = Concentration of the parameter to be 7.2.1 Prepare calibration standards at a s measured (μg/L). minimum of three concentration levels for each parameter of interest by adding vol- If the RF value over the working range is umes of one or more stock standards to a a constant (<10% RSD), the RF can be as- volumetric flask and diluting to volume with sumed to be invariant and the average RF 2-propanol. One of the external standards can be used for calculations. Alternatively, should be at a concentration near, but above, the results can be used to plot a calibration the MDL (Table 1) and the other concentra- curve of response ratios, As/Ais, vs. RF. tions should correspond to the expected 7.4 The working calibration curve, cali- range of concentrations found in real sam- bration factor, or RF must be verified on ples or should define the working range of each working day by the measurement of one the detector. or more calibration standards. If the re- 7.2.2 Using injections of 2 to 5 μl, analyze sponse for any parameter varies from the each calibration standard according to Sec- predicted response by more than ±15%, a new tion 11 and tabulate peak height or area re- calibration curve must be prepared for that sponses against the mass injected. The re- compound. sults can be used to prepare a calibration 7.5 To calibrate the ECDGC for the anal- curve for each compound. Alternatively, if ysis of phenol derivatives, establish gas the ratio of response to amount injected chromatographic operating conditions equiv- (calibration factor) is a constant over the alent to those given in Table 2. working range (<10% relative standard devi- 7.5.1 Prepare calibration standards at a ation, RSD), linearity through the origin can minimum of three concentration levels for be assumed and the average ratio or calibra- each parameter of interest by adding vol- tion factor can be used in place of a calibra- umes of one or more stock standards to a tion curve. volumetric flask and diluting to volume with 7.3 Internal standard calibration proce- 2-propanol. One of the external standards dure for FIDGC—To use this approach, the should be at a concentration near, but above, analyst must select one or more internal the MDL (Table 2) and the other concentra- standards that are similar in analytical be- tions should correspond to the expected havior to the compounds of interest. The an- range of concentrations found in real sam- alyst must further demonstrate that the ples or should define the working range of measurement of the internal standard is not the detector.

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7.5.2 Each time samples are to be trol. This procedure is described in Section derivatized, simultaneously treat a 1-mL ali- 8.4. The frequency of the check standard quot of each calibration standard as de- analyses is equivalent to 10% of all samples scribed in Section 12. analyzed but may be reduced if spike recov- 7.5.3 After derivatization, analyze 2 to 5 eries from samples (Section 8.3) meet all μL of each column eluate collected according specified quality control criteria. to the method beginning in Section 12.8 and 8.1.6 The laboratory must maintain per- tabulate peak height or area responses formance records to document the quality of against the calculated equivalent mass of data that is generated. This procedure is de- underivatized phenol injected. The results scribed in Section 8.5. can be used to prepare a calibration curve for 8.2 To establish the ability to generate each compound. acceptable accuracy and precision, the ana- 7.6 Before using any cleanup procedure, lyst must perform the following operations. the analyst must process a series of calibra- 8.2.1 A quality control (QC) check sample tion standards through the procedure to vali- concentrate is required containing each pa- date elution patterns and the absence of rameter of interest at a concentration of 100 interferences from the reagents. μg/mL in 2-propanol. The QC check sample concentrate must be obtained from the U.S. 8. Quality Control Environmental Protection Agency, Environ- 8.1 Each laboratory that uses this method mental Monitoring and Support Laboratory is required to operate a formal quality con- in Cincinnati, Ohio, if available. If not avail- trol program. The minimum requirements of able from that source, the QC check sample this program consist of an initial demonstra- concentrate must be obtained from another tion of laboratory capability and an ongoing external source. If not available from either analysis of spiked samples to evaluate and source above, the QC check sample con- document data quality. The laboratory must centrate must be prepared by the laboratory maintain records to document the quality of using stock standards prepared independ- data that is generated. Ongoing data quality ently from those used for calibration. checks are compared with established per- 8.2.2 Using a pipet, prepare QC check sam- formance criteria to determine if the results ples at a concentration of 100 μg/L by adding of analyses meet the performance character- 1.00 mL of QC check sample concentrate to istics of the method. When results of sample each of four 1-L aliquots of reagent water. spikes indicate atypical method perform- 8.2.3 Analyze the well-mixed QC check ance, a quality control check standard must samples according to the method beginning be analyzed to confirm that the measure- in Section 10. ¯ ments were performed in an in-control mode 8.2.4 Calculate the average recovery (X) in of operation. μg/L, and the standard deviation of the re- 8.1.1 The analyst must make an initial, covery (s) in μg/L, for each parameter using one-time, demonstration of the ability to the four results. ¯ generate acceptable accuracy and precision 8.2.5 For each parameter compare s and X with this method. This ability is established with the corresponding acceptance criteria as described in Section 8.2. for precision and accuracy, respectively, 8.1.2 In recognition of advances that are found in Table 3. If s and X¯ for all param- occurring in chromatography, the analyst is eters of interest meet the acceptance cri- permitted certain options (detailed in Sec- teria, the system performance is acceptable tions 10.6 and 11.1) to improve the separa- and analysis of actual samples can begin. If tions or lower the cost of measurements. any individual s exceeds the precision limit Each time such a modification is made to or any individual X¯ falls outside the range the method, the analyst is required to repeat for accuracy, the system performance is un- the procedure in Section 8.2. acceptable for that parameter. 8.1.3 Before processing any samples the NOTE: The large number of parameters in analyst must analyze a reagent water blank Talbe 3 present a substantial probability to demonstrate that interferences from the that one or more will fail at least one of the analytical system and glassware are under acceptance criteria when all parameters are control. Each time a set of samples is ex- analyzed. tracted or reagents are changed a reagent 8.2.6 When one or more of the parameters water blank must be processed as a safe- tested fail at least one of the acceptance cri- guard against laboratory contamination. teria, the analyst must proceed according to 8.1.4 The laboratory must, on an ongoing Section 8.2.6.1 or 8.2.6.2. basis, spike and analyze a minimum of 10% 8.2.6.1 Locate and correct the source of of all samples to monitor and evaluate lab- the problem and repeat the test for all pa- oratory data quality. This procedure is de- rameters of interest beginning with Section scribed in Section 8.3. 8.2.2. 8.1.5 The laboratory must, on an ongoing 8.2.6.2 Beginning with Section 8.2.2, repeat basis, demonstrate through the analyses of the test only for those parameters that quality control check standards that the op- failed to meet criteria. Repeated failure, eration of the measurement system is in con- however, will confirm a general problem

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with the measurement system. If this occurs, tion in Table 4, substituting X′ for X¯ ; (3) cal- locate and correct the source of the problem culate the range for recovery at the spike and repeat the test for all compounds of in- concentration as (100 X′/T)±2.44(100 S′/T)%. 8 terest beginning with Section 8.2.2. 8.3.4 If any individual P falls outside the 8.3 The laboratory must, on an ongoing designated range for recovery, that param- basis, spike at least 10% of the samples from eter has failed the acceptance criteria. A each sample site being monitored to assess check standard containing each parameter accuracy. For laboratories analyzing one to that failed the criteria must be analyzed as ten samples per month, at least one spiked described in Section 8.4. sample per month is required. 8.4 If any parameter fails the acceptance 8.3.1 The concentration of the spike in the criteria for recovery in Section 8.3, a QC sample should be determined as follows: check standard containing each parameter 8.3.1.1 If, as in compliance monitoring, that failed must be prepared and analyzed. the concentration of a specific parameter in NOTE: The frequency for the required anal- the sample is being checked against a regu- ysis of a QC check standard will depend upon latory concentration limit, the spike should the number of parameters being simulta- be at that limit or 1 to 5 times higher than neously tested, the complexity of the sample the background concentration determined in matrix, and the performance of the labora- Section 8.3.2, whichever concentration would tory. be larger. 8.3.1.2 If the concentration of a specific 8.4.1 Prepare the QC check standard by parameter in the sample is not being adding 1.0 mL of QC check sample con- checked against a limit specific to that pa- centrate (Section 8.2.1 or 8.3.2) to 1 L of rea- rameter, the spike should be at 100 μg/L or 1 gent water. The QC check standard needs to 5 times higher than the background con- only to contain the parameters that failed centration determined in Section 8.3.2, criteria in the test in Section 8.3. whichever concentration would be larger. 8.4.2 Analyze the QC check standard to 8.3.1.3 If it is impractical to determine determine the concentration measured (A) of background levels before spiking (e.g., max- each parameter. Calculate each percent re- imum holding times will be exceeded), the covery (Ps) as 100 (A/T)%, where T is the true spike concentration should be (1) the regu- value of the standard concentration. latory concentration limit, if any, or, if 8.4.3 Compare the percent recovery (Ps) none, (2) the larger of either 5 times higher for each parameter with the corresponding than the expected background concentration QC acceptance criteria found in Table 3. Only or 100 μg/L. parameters that failed the test in Section 8.3 8.3.2 Analyze one sample aliquot to deter- need to be compared with these criteria. If mine the background concentration (B) of the recovery of any such parameter falls out- each parameter. If necessary, prepare a new side the designated range, the laboratory QC check sample concentrate (Section 8.2.1) performance for that parameter is judged to appropriate for the background concentra- be out of control, and the problem must be tions in the sample. Spike a second sample immediately identified and corrected. The aliquot with 1.0 mL of the QC check sample analytical result for that parameter in the concentrate and analyze it to determine the unspiked sample is suspect and may not be concentration after spiking (A) of each pa- reported for regulatory compliance purposes. rameter. Calculate each percent recovery (P) 8.5 As part of the QC program for the lab- as 100(A¥B)%/T, where T is the known true oratory, method accuracy for wastewater value of the spike. samples must be assessed and records must 8.3.3 Compare the percent recovery (P) for be maintained. After the analysis of five each parameter with the corresponding QC spiked wastewater samples as in Section 8.3, acceptance criteria found in Table 3. These calculate the average percent recovery (P¯ ) acceptance criteria were calculated to in- and the standard deviation of the percent re- clude an allowance for error in measurement covery (sp). Express the accuracy assessment ¯ of both the background and spike concentra- as a percent recovery interval from P¥2sp to ¯ ¯ tions, assuming a spike to background ratio P + 2sp. If P = 90% and sp = 10%, for example, of 5:1. This error will be accounted for to the the accuracy interval is expressed as 70– extent that the analyst’s spike to back- 110%. Update the accuracy assessment for ground ratio approaches 5:1. 8 If spiking was each parameter on a regular basis (e.g. after performed at a concentration lower than 100 each five to ten new accuracy measure- μg/L, the analyst must use either the QC ac- ments). ceptance criteria in Table 3, or optional QC 8.6. It is recommended that the labora- acceptance criteria calculated for the spe- tory adopt additional quality assurance cific spike concentration. To calculate op- practices for use with this method. The spe- tional acceptance criteria for the recovery of cific practices that are most productive de- a parameter: (1) Calculate accuracy (X′) pend upon the needs of the laboratory and using the equation in Table 4, substituting the nature of the samples. Field duplicates the spike concentration (T) for C; (2) cal- may be analyzed to assess the precision of culate overall precision (S′) using the equa- the environmental measurements. When

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doubt exists over the identification of a peak periodic venting to release excess pressure. on the chromatogram, confirmatory tech- Allow the organic layer to separate from the niques such as gas chromatography with a water phase for a minimum of 10 min. If the dissimilar column, specific element detector, emulsion interface between layers is more or mass spectrometer must be used. When- than one-third the volume of the solvent ever possible, the laboratory should analyze layer, the analyst must employ mechanical standard reference materials and participate techniques to complete the phase separation. in relevant performance evaluation studies. The optimum technique depends upon the sample, but may include stirring, filtration 9. Sample Collection, Preservation, and of the emulsion through glass wool, cen- Handling trifugation, or other physical methods. Col- 9.1 Grab samples must be collected in lect the methylene chloride extract in a 250- glass containers. Conventional sampling mL Erlenmeyer flask. practices 9 should be followed, except that 10.5 Add a second 60-mL volume of meth- the bottle must not be prerinsed with sample ylene chloride to the sample bottle and re- before collection. Composite samples should peat the extraction procedure a second time, be collected in refrigerated glass containers combining the extracts in the Erlenmeyer flask. Perform a third extraction in the same in accordance with the requirements of the manner. program. Automatic sampling equipment 10.6 Assemble a Kuderna-Danish (K-D) must be as free as possible of Tygon tubing concentrator by attaching a 10-mL concen- and other potential sources of contamina- trator tube to a 500-mL evaporative flask. tion. Other concentration devices or techniques 9.2 All samples must be iced or refrig- may be used in place of the K-D concentrator erated at 4 °C from the time of collection if the requirements of Section 8.2 are met. until extraction. Fill the sample bottles and, 10.7 Pour the combined extract through a if residual chlorine is present, add 80 mg of solvent-rinsed drying column containing sodium thiosulfate per liter of sample and about 10 cm of anhydrous sodium sulfate, mix well. EPA Methods 330.4 and 330.5 may and collect the extract in the K-D concen- be used for measurement of residual chlo- trator. Rinse the Erlenmeyer flask and col- 10 rine. Field test kits are available for this umn with 20 to 30 mL of methylene chloride purpose. to complete the quantitative transfer. 9.3 All samples must be extracted within 7 10.8 Add one or two clean boiling chips to days of collection and completely analyzed the evaporative flask and attach a three-ball 2 within 40 days of extraction. Snyder column. Prewet the Snyder column 10. Sample Extraction by adding about 1 mL of methylene chloride to the top. Place the K-D apparatus on a hot 10.1 Mark the water meniscus on the side water bath (60 to 65 °C) so that the concen- of sample bottle for later determination of trator tube is partially immersed in the hot sample volume. Pour the entire sample into water, and the entire lower rounded surface a 2-L separatory funnel. of the flask is bathed with hot vapor. Adjust 10.2 For samples high in organic content, the vertical position of the apparatus and the analyst may solvent wash the sample at the water temperature as required to com- basic pH as prescribed in Sections 10.2.1 and plete the concentration in 15 to 20 min. At 10.2.2 to remove potential method inter- the proper rate of distillation the balls of the ferences. Prolonged or exhaustive contact column will actively chatter but the cham- with solvent during the wash may result in bers will not flood with condensed solvent. low recovery of some of the phenols, notably When the apparent volume of liquid reaches phenol and 2,4-dimethylphenol. For rel- 1 mL, remove the K-D apparatus and allow it atively clean samples, the wash should be to drain and cool for at least 10 min. omitted and the extraction, beginning with 10.9 Increase the temperature of the hot Section 10.3, should be followed. water bath to 95 to 100 °C. Remove the 10.2.1 Adjust the pH of the sample to 12.0 Synder column and rinse the flask and its or greater with sodium hydroxide solution. lower joint into the concentrator tube with 1 10.2.2 Add 60 mL of methylene chloride to to 2 mL of 2-propanol. A 5-mL syringe is rec- the sample by shaking the funnel for 1 min ommended for this operation. Attach a two- with periodic venting to release excess pres- ball micro-Snyder column to the concen- sure. Discard the solvent layer. The wash trator tube and prewet the column by adding can be repeated up to two additional times if about 0.5 mL of 2-propanol to the top. Place significant color is being removed. the micro-K-D apparatus on the water bath 10.3 Adjust the sample to a pH of 1 to 2 so that the concentrator tube is partially with sulfuric acid. immersed in the hot water. Adjust the 10.4 Add 60 mL of methylene chloride to vertical position of the apparatus and the the sample bottle, seal, and shake 30 s to water temperature as required to complete rinse the inner surface. Transfer the solvent concentration in 5 to 10 min. At the proper to the separatory funnel and extract the rate of distillation the balls of the column sample by shaking the funnel for 2 min. with will actively chatter but the chambers will

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not flood. When the apparent volume of liq- gested window size; however, the experience uid reaches 2.5 mL, remove the K-D appa- of the analyst should weigh heavily in the ratus and allow it to drain and cool for at interpretation of chromatograms. least 10 min. Add an additional 2 mL of 2- 11.6 If the response for a peak exceeds the propanol through the top of the micro-Sny- working range of the system, dilute the ex- der column and resume concentrating as be- tract and reanalyze. fore. When the apparent volume of liquid 11.7 If the measurement of the peak re- reaches 0.5 mL, remove the K-D apparatus sponse is prevented by the presence of inter- and allow it to drain and cool for at least 10 ferences, an alternative gas chromatographic min. procedure is required. Section 12 describes a 10.10 Remove the micro-Snyder column derivatization and column chromatographic and rinse its lower joint into the concen- procedure which has been tested and found trator tube with a minimum amount of 2- to be a practical means of analyzing phenols propanol. Adjust the extract volume to 1.0 in complex extracts. mL. Stopper the concentrator tube and store refrigerated at 4 °C if further processing will 12. Derivatization and Electron Capture not be performed immediately. If the extract Detector Gas Chromatography will be stored longer than two days, it should 12.1 Pipet a 1.0-mL aliquot of the 2-pro- be transferred to a Teflon-sealed screw-cap panol solution of standard or sample extract vial. If the sample extract requires no fur- ther cleanup, proceed with FIDGC analysis into a glass reaction vial. Add 1.0 mL of (Section 11). If the sample requires further derivatizing reagent (Section 6.11). This cleanup, proceed to Section 12. amount of reagent is sufficient to derivatize 10.11 Determine the original sample vol- a solution whose total phenolic content does ume by refilling the sample bottle to the not exceed 0.3 mg/mL. mark and transferring the liquid to a 1000- 12.2 Add about 3 mg of potassium car- mL graduated cylinder. Record the sample bonate to the solution and shake gently. volume to the nearest 5 mL. 12.3 Cap the mixture and heat it for 4 h at 80 °C in a hot water bath. 11. Flame Ionization Detector Gas 12.4 Remove the solution from the hot Chromatography water bath and allow it to cool. 12.5 Add 10 mL of hexane to the reaction 11.1 Table 1 summarizes the recommended flask and shake vigorously for 1 min. Add 3.0 operating conditions for the gas chro- mL of distilled, deionized water to the reac- matograph. Included in this table are reten- tion times and MDL that can be achieved tion flask and shake for 2 min. Decant a por- under these conditions. An example of the tion of the organic layer into a concentrator separations achieved by this column is tube and cap with a glass stopper. shown in Figure 1. Other packed or capillary 12.6 Place 4.0 g of silica gel into a (open-tubular) columns, chromatographic chromatographic column. Tap the column to conditions, or detectors may be used if the settle the silica gel and add about 2 g of an- requirements of Section 8.2 are met. hydrous sodium sulfate to the top. 11.2 Calibrate the system daily as de- 12.7 Preelute the column with 6 mL of scribed in Section 7. hexane. Discard the eluate and just prior to 11.3 If the internal standard calibration exposure of the sodium sulfate layer to the procedure is used, the internal standard air, pipet onto the column 2.0 mL of the must be added to the sample extract and hexane solution (Section 12.5) that contains mixed thoroughly immediately before injec- the derivatized sample or standard. Elute the tion into the gas chromatograph. column with 10.0 mL of hexane and discard 11.4 Inject 2 to 5 μL of the sample extract the eluate. Elute the column, in order, with: or standard into the gas chromatograph 10.0 mL of 15% toluene in hexane (Fraction using the solvent-flush technique. 11 Smaller 1); 10.0 mL of 40% toluene in hexane (Frac- (1.0 μL) volumes may be injected if auto- tion 2); 10.0 mL of 75% toluene in hexane matic devices are employed. Record the vol- (Fraction 3); and 10.0 mL of 15% 2-propanol ume injected to the nearest 0.05 μL, and the in toluene (Fraction 4). All elution mixtures resulting peak size in area or peak height are prepared on a volume: volume basis. units. Elution patterns for the phenolic derivatives 11.5 Identify the parameters in the sample are shown in Table 2. Fractions may be com- by comparing the retention times of the bined as desired, depending upon the specific peaks in the sample chromatogram with phenols of interest or level of interferences. those of the peaks in standard 12.8 Analyze the fractions by ECDGC. chromatograms. The width of the retention Table 2 summarizes the recommended oper- time window used to make identifications ating conditions for the gas chromatograph. should be based upon measurements of ac- Included in this table are retention times tual retention time variations of standards and MDL that can be achieved under these over the course of a day. Three times the conditions. An example of the separations standard deviation of a retention time for a achieved by this column is shown in Figure compound may be used to calculate a sug- 2.

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12.9 Calibrate the system daily with a Equation 4 minimum of three aliquots of calibration where: standards, containing each of the phenols of A = Mass of underivatized phenol represented interest that are derivatized according to by area of peak in sample chromatogram, Section 7.5. determined from calibration curve in 12.10 Inject 2 to 5 μL of the column frac- Section 7.5.3 (ng). tions into the gas chromatograph using the μ μ Vi = Volume of eluate injected ( L). solvent-flush technique. Smaller (1.0 L) vol- V = Total volume of column eluate or com- umes can be injected if automatic devices t bined fractions from which Vi was taken are employed. Record the volume injected to (μL). the nearest 0.05 μL, and the resulting peak Vs = Volume of water extracted in Section size in area or peak height units. If the peak 10.10 (mL). response exceeds the linear range of the sys- B = Total volume of hexane added in Section tem, dilute the extract and reanalyze. 12.5 (mL). 13. Calculations C = Volume of hexane sample solution added to cleanup column in Section 12.7 (mL). 13.1 Determine the concentration of indi- D = Total volume of 2-propanol extract prior vidual compounds in the sample analyzed by to derivatization (mL). FIDGC (without derivatization) as indicated E = Volume of 2-propanol extract carried below. through derivatization in Section 12.1 13.1.1 If the external standard calibration (mL). procedure is used, calculate the amount of 13.3 Report results in μg/L without correc- material injected from the peak response tion for recovery data. All QC data obtained using the calibration curve or calibration should be reported with the sample results. factor determined in Section 7.2.2. The con- centration in the sample can be calculated 14. Method Performance from Equation 2. 14.1 The method detection limit (MDL) is () defined as the minimum concentration of a AV()t substance that can be measured and reported Concentration (μ= g/L) with 99% confidence that the value is above ()VVis() zero. 1 The MDL concentrations listed in Ta- bles 1 and 2 were obtained using reagent Equation 2 water. 12 Similar results were achieved using where: representative wastewaters. The MDL actu- A = Amount of material injected (ng). ally achieved in a given analysis will vary depending on instrument sensitivity and ma- Vi = Volume of extract injected (μL). Vt = Volume of total extract (μL). trix effects. Vs = Volume of water extracted (mL). 14.2 This method was tested by 20 labora- 13.1.2 If the internal standard calibration tories using reagent water, drinking water, procedure is used, calculate the concentra- surface water, and three industrial wastewaters spiked as six concentrations tion in the sample using the response factor over the range 12 to 450 μg/L. 13 Single oper- (RF) determined in Section 7.3.2 and Equa- ator precision, overall precision, and method tion 3. accuracy were found to be directly related to the concentration of the parameter and es- ()()AI μ= ss sentially independent of the sample matrix. Concentration ( g/L) () Linear equations to describe these relation- ()ARFVis() o ships for a flame ionization detector are pre- sented in Table 4. Equation 3 where: References As = Response for the parameter to be meas- 1. 40 CFR part 136, appendix B. ured. 2. ‘‘Determination of Phenols in Industrial Ais = Response for the internal standard. and Municipal Wastewaters,’’ EPA 600/4–84– Is = Amount of internal standard added to ABC, National Technical Information Serv- each extract (μg). ice, PBXYZ, Springfield, Virginia 22161, No- Vo = Volume of water extracted (L). vember 1984. 13.2 Determine the concentration of indi- 3. Kawahara, F. K. ‘‘Microdetermination of vidual compounds in the sample analyzed by Derivatives of Phenols and Mercaptans by derivatization and ECDGC according to Means of Electron Capture Gas Chroma- Equation 4. tography,’’ Analytical Chemistry, 40, 1009 (1968). ()AV()()() BD 4. ASTM Annual Book of Standards, Part Concentration (μ= g/L) t 31, D3694–78. ‘‘Standard Practices for Prepa- ()VVCE()()() ration of Sample Containers and for Preser- is vation of Organic Constituents,’’ American

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Society for Testing and Materials, Philadel- 12. ‘‘Development of Detection Limits, phia. EPA Method 604, Phenols,’’ Special letter re- 5. ‘‘Carcinogens—Working With Carcino- port for EPA Contract 68–03–2625, U.S. Envi- gens,’’ Department of Health, Education, and ronmental Protection Agency, Environ- Welfare, Public Health Service, Center for mental Monitoring and Support Laboratory, Disease Control, National Institute for Occu- Cincinnati, Ohio 45268. pational Safety and Health, Publication No. 13. ‘‘EPA Method Study 14 Method 604-Phe- 77–206, August 1977. nols,’’ EPA 600/4–84–044, National Technical 6. ‘‘OSHA Safety and Health Standards, Information Service, PB84–196211, Spring- General Industry,’’ (29 CFR part 1910), Occu- field, Virginia 22161, May 1984. pational Safety and Health Administration, OSHA 2206 (Revised, January 1976). 7. ‘‘Safety in Academic Chemistry Labora- TABLE 1—CHROMATOGRAPHIC CONDITIONS AND tories,’’ American Chemical Society Publica- METHOD DETECTION LIMITS tion, Committee on Chemical Safety, 3rd Edition, 1979. Method de- Parameter Retention tection limit 8. Provost, L. P., and Elder, R. S. ‘‘Inter- time (min) (μg/L) pretation of Percent Recovery Data,’’ Amer- ican Laboratory, 15, 58–63 (1983). (The value 2-Chlorophenol ...... 1.70 0 .31 2.44 used in the equation in Section 8.3.3 is 2-Nitrophenol ...... 2.00 0 .45 two times the value 1.22 derived in this re- Phenol ...... 3.01 0 .14 port.) 2,4-Dimethylphenol ...... 4.03 0.32 9. ASTM Annual Book of Standards, Part 2,4-Dichlorophenol ...... 4.30 0 .39 31, D3370–76. ‘‘Standard Practices for Sam- 2,4,6-Trichlorophenol ...... 6.05 0 .64 pling Water,’’ American Society for Testing 4-Chloro-3-methylphenol ...... 7.50 0 .36 and Materials, Philadelphia. 2,4-Dinitrophenol ...... 10.00 13 .0 10. ‘‘Methods 330.4 (Titrimetric, DPD-FAS) 2-Methyl-4,6-dinitrophenol ...... 10.24 16.0 and 330.5 (Spectrophotometric, DPD) for Pentachlorophenol ...... 12.42 7.4 Chlorine, Total Residual,’’ Methmds for 4-Nitrophenol ...... 24.25 2 .8 Chemical Analysis of Water and Wastes, Column conditions: Supelcoport (80/100 mesh) coated with EPA–600/4–79–020, U.S. Environmental Pro- 1% SP–1240DA packed in a 1.8 m long × 2 mm ID glass col- tection Agency, Environmental Monitoring umn with nitrogen carrier gas at 30 mL/min flow rate. Column temperature was 80 °C at injection, programmed immediately and Support Laboratory, Cincinnati, Ohio at 8 °C/min to 150 °C final temperature. MDL were deter- 45268, March 1979. mined with an FID. 11. Burke, J. A. ‘‘Gas Chromatography for Pesticide Residue Analysis; Some Practical Aspects,’’ Journal of the Association of Official Analytical Chemists, 48, 1037 (1965).

TABLE 2—SILICA GEL FRACTIONATION AND ELECTRON CAPTURE GAS CHROMATOGRAPHY OF PFBB DERIVATIVES

Percent recovery by frac- a Method Parent compound tion Retention detection time (min) μ 1 2 3 4 limit ( g/L)

2-Chlorophenol ...... 90 1 ...... 3.3 0 .58 2-Nitrophenol ...... 9 90 9.1 0.77 Phenol ...... 90 10 ...... 1.8 2.2 2,4-Dimethylphenol ...... 95 7 ...... 2.9 0 .63 2,4-Dichlorophenol ...... 95 1 ...... 5.8 0.68 2,4,6-Trichlorophenol ...... 50 50 ...... 7.0 0 .58 4-Chloro-3-methylphenol ...... 84 14 ...... 4.8 1.8 Pentachlorophenol ...... 75 20 ...... 28.8 0 .59 4-Nitrophenol ...... 1 90 14.0 0 .70 Column conditions: Chromosorb W-AW-DMCS (80/100 mesh) coated with 5% OV–17 packed in a 1.8 m long × 2.0 mm ID glass column with 5% methane/95% argon carrier gas at 30 mL/min flow rate. Column temperature held isothermal at 200 °C. MDL were determined with an ECD. a Eluant composition: Fraction 1—15% toluene in hexane. Fraction 2—40% toluene in hexane. Fraction 3—75% toluene in hexane. Fraction 4—15% 2-propanol in toluene.

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TABLE 3—QC ACCEPTANCE CRITERIA—METHOD 604

Test Limit for s Range for X¯ Range for Parameter conc. μ μ P, Ps (per- (μg/L) ( g/L) ( g/L) cent)

4-Chloro-3-methylphenol ...... 100 16.6 56.7–113.4 49–122 2-Chlorophenol ...... 100 27.0 54.1–110.2 38–126 2,4-Dichlorophenol ...... 100 25.1 59.7–103.3 44–119 2,4-Dimethylphenol ...... 100 33.3 50.4–100.0 24–118 4,6-Dinitro-2-methylphenol ...... 100 25.0 42.4–123.6 30–136 2,4-Dinitrophenol ...... 100 36.0 31.7–125.1 12–145 2-Nitrophenol ...... 100 22.5 56.6–103.8 43–117 4-Nitrophenol ...... 100 19.0 22.7–100.0 13–110 Pentachlorophenol ...... 100 32.4 56.7–113.5 36–134 Phenol ...... 100 14.1 32.4–100.0 23–108 2,4,6-Trichlorophenol ...... 100 16.6 60.8–110.4 53–119 s—Standard deviation of four recovery measurements, in μg/L (Section 8.2.4). X¯ —Average recovery for four recovery measurements, in μg/L (Section 8.2.4). P, Ps—Percent recovery measured (Section 8.3.2, Section 8.4.2). NOTE: These criteria are based directly upon the method performance data in Table 4. Where necessary, the limits for recov- ery have been broadened to assure applicability of the limits to concentrations below those used to develop Table 4.

TABLE 4—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 604

Single Analyst Accuracy, as re- ′ μ Overall precision, Parameter ′ μ precision, sr ( g/ ′ μ covery, X ( g/L) L) S ( g/L)

4-Chloro-3-methylphenol ...... 0.87C–1.97 0.11X¯ –0.21 0.16X¯ + 1.41 2-Chlorophenol ...... 0.83C–0.84 0.18X¯ + 0.20 0.21X¯ + 0.75 2,4-Dichlorophenol ...... 0.81C + 0.48 0.17X¯ –0.02 0.18X¯ + 0.62 2,4-Dimethylphenol ...... 0.62C–1.64 0.30X¯ –0.89 0.25X¯ + 0.48 4,6-Dinitro-2-methylphenol ...... 0.84C–1.01 0.15X¯ + 1.25 0.19X¯ + 5.85 2,4-Dinitrophenol ...... 0.80C–1.58 0.27X¯ –1.15 0.29X¯ + 4.51 2-Nitrophenol ...... 0.81C–0.76 0.15X¯ + 0.44 0.14X¯ + 3.84 4-Nitrophenol ...... 0.46C + 0.18 0.17X¯ + 2.43 0.19X¯ + 4.79 Pentachlorophenol ...... 0.83C + 2.07 0.22X¯ –0.58 0.23X¯ + 0.57 Phenol ...... 0.43C + 0.11 0.20X¯ –0.88 0.17X¯ + 0.77 2,4,6-Trichlorophenol ...... 0.86C–0.40 0.10X¯ + 0.53 0.13X¯ + 2.40 X′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L. sr′ = Expected single analyst standard deviation of measurements at an average concentration found of X¯ , in μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X¯ , in μg/L. C = True value for the concentration, in μg/L. X¯ = Average recovery found for measurements of samples containing a concentration of C, in μg/L.

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COLUMN: 1% SP-1240DA ON SUPELCOPORT PROGRAM: 80°C AT INJECTION. IMMEDIATE 8°C/MIN TO 150°C DETECTOR: FLAME IONIZATION

0 16 20 28 RETENTION TIME, MIN.

Figure 1. Gas chromatogram of phenols.

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METHOD 605—BENZIDINES as provided under 40 CFR 136.1. When this method is used to analyze unfamiliar sam- 1. Scope and Application ples for the compounds above, identifications 1.1 This method covers the determination should be supported by at least one addi- of certain benzidines. The following param- tional qualitative technique. This method eters can be determined by this method: describes electrochemical conditions at a second potential which can be used to con- Parameter Storet No CAS No. firm measurements made with this method. Method 625 provides gas chromatograph/mass Benzidine ...... 39120 92–87–5 spectrometer (GC/MS) conditions appro- 3,3′-Dichlorobenzidine ...... 34631 91–94–1 priate for the qualitative and quantitative confirmation of results for the parameters 1.2 This is a high performance liquid chro- listed above, using the extract produced by matography (HPLC) method applicable to this method. the determination of the compounds listed 1.3 The method detection limit (MDL, de- above in municipal and industrial discharges fined in Section 14.1) 1 for each parameter is

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listed in Table 1. The MDL for a specific 3.1.2 The use of high purity reagents and wastewater may differ from those listed, de- solvents helps to minimize interference prob- pending upon the nature of the interferences lems. Purification of solvents by distillation in the sample matrix. in all-glass systems may be required. 1.4 Any modification of this method, be- 3.2 Matrix interferences may be caused by yond those expressly permitted, shall be con- contaminants that are co-extracted from the sidered as a major modification subject to sample. The extent of matrix interferences application and approval of alternate test will vary considerably from source to source, procedures under 40 CFR 136.4 and 136.5. depending upon the nature and diversity of 1.5 This method is restricted to use by or the industrial complex or municipality being under the supervision of analysts experi- sampled. The cleanup procedures that are in- enced in the use of HPLC instrumentation herent in the extraction step are used to and in the interpretation of liquid overcome many of these interferences, but chromatograms. Each analyst must dem- unique samples may require additional onstrate the ability to generate acceptable cleanup approaches to achieve the MDL list- results with this method using the procedure ed in Table 1. described in Section 8.2. 3.3 Some dye plant effluents contain large amounts of components with retention times 2. Summary of Method closed to benzidine. In these cases, it has been found useful to reduce the electrode po- 2.1 A measured volume of sample, ap- tential in order to eliminate interferences proximately 1–L, is extracted with chloro- and still detect benzidine. (See Section 12.7.) form using liquid-liquid extractions in a separatory funnel. The chloroform extract is 4. Safety extracted with acid. The acid extract is then 4.1 The toxicity or carcinogenicity of neutralized and extracted with chloroform. each reagent used in this method has not The final chloroform extract is exchanged to been precisely defined; however, each chem- methanol while being concentrated using a ical compound should be treated as a poten- rotary evaporator. The extract is mixed with tial health harzard. From this viewpoint, ex- buffer and separated by HPLC. The benzidine posure to these chemicals must be reduced to compounds are measured with an electro- the lowest possible level by whatever means chemical detector. 2 available. The laboratory is responsible for 2.2 The acid back-extraction acts as a maintaining a current awareness file of general purpose cleanup to aid in the elimi- OSHA regulations regarding the safe han- nation of interferences. dling of the chemicals specified in this meth- od. A reference file of material data handling 3. Interferences sheets should also be made available to all 3.1 Method interferences may be caused personnel involved in the chemical analysis. by contaminants in solvents, reagents, glass- Additional references to laboratory safety ware, and other sample processing hardware are available and have been identified 46 for that lead to discrete artifacts and/or ele- the information of the analyst. vated baselines in chromatograms. All of 4.2 The following parameters covered by these materials must be routinely dem- this method have been tentatively classified onstrated to be free from interferences under as known or suspected, human or mamma- the conditions of the analysis by running lian carcinogens: benzidine and 3,3′- laboratory reagent blanks as described in dichlorobenzidine. Primary standards of Section 8.1.3. these toxic compounds should be prepared in 3.1.1 Glassware must be scrupulously a hood. A NIOSH/MESA approved toxic gas cleaned. 3 Clean all glassware as soon as pos- respirator should be worn when the analyst sible after use by rinsing with the last sol- handles high concentrations of these toxic vent used in it. Solvent rinsing should be fol- compounds. lowed by detergent washing with hot water, 4.3 Exposure to chloroform should be and rinses with tap water and distilled minimized by performing all extractions and water. The glassware should then be drained extract concentrations in a hood or other dry, and heated in a muffle furnace at 400 °C well-ventiliated area. for 15 to 30 min. Some thermally stable ma- 5. Apparatus and Materials terials may not be eliminated by this treat- ment. Solvent rinses with acetone and pes- 5.1 Sampling equipment, for discrete or ticide quality hexane may be substituted for composite sampling. the muffle furnace heating. Volumetric ware 5.1.1 Grab sample bottle—1–L or 1-qt, should not be heated in a muffle furnace. amber glass, fitted with a screw cap lined After drying and cooling, glassware should with Teflon. Foil may be substituted for Tef- be sealed and stored in a clean environment lon if the sample is not corrosive. If amber to prevent any accumulation of dust or other bottles are not available, protect samples contaminants. Store inverted or capped with from light. The bottle and cap liner must be aluminum foil. washed, rinsed with acetone or methylene

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chloride, and dried before use to minimize 6.2 Sodium hydroxide solution (5 N)—Dis- contamination. solve 20 g of NaOH (ACS) in reagent water 5.1.2 Automatic sampler (optional)—The and dilute to 100 mL. sampler must incorporate glass sample con- 6.3 Sodium hydroxide solution (1 M)—Dis- tainers for the collection of a minimum of solve 40 g of NaOH (ACS) in reagent water 250 mL of sample. Sample containers must be and dilute to 1 L. kept refrigerated at 4 °C and protected from 6.4 Sodium thiosulfate—(ACS) Granular. light during compositing. If the sampler uses 6.5 Sodium tribasic phosphate (0.4 M)— a peristaltic pump, a minimum length of Dissolve 160 g of trisodium phosphate deca- compressible silicone rubber tubing may be hydrate (ACS) in reagent water and dilute to used. Before use, however, the compressible 1 L. tubing should be thoroughly rinsed with 6.6 Sulfuric acid (1 + 1)—Slowly, add 50 methanol, followed by repeated rinsings with mL of H SO (ACS, sp. gr. 1.84) to 50 mL of distilled water to minimize the potential for 2 4 reagent water. contamination of the sample. An integrating flow meter is required to collect flow propor- 6.7 Sulfuric acid (1 M)—Slowly, add 58 mL tional composites. of H2SO4 (ACS, sp. gr. 1.84) to reagent water 5.2 Glassware (All specifications are sug- and dilute to 1 L. gested): 6.8 Acetate buffer (0.1 M, pH 4.7)—Dissolve 5.2.1 Separatory funnels—2000, 1000, and 5.8 mL of glacial acetic acid (ACS) and 13.6 g 250-mL, with Teflon stopcock. of sodium acetate trihydrate (ACS) in rea- 5.2.2 Vials—10 to 15-mL, amber glass, with gent water which has been purified by filtra- Teflon-lined screw cap. tion through a RO–4 Millipore System or 5.2.3 Rotary evaporator. equivalent and dilute to 1 L. 5.2.4 Flasks—Round bottom, 100–mL, with 6.9 , chloroform (preserved 24/40 joints. with 1% ethanol), methanol—Pesticide qual- 5.2.5 Centrifuge tubes—Conical, grad- ity or equivalent. uated, with Teflon-lined screw caps. 6.10 Mobile phase—Place equal volumes of 5.2.6 Pipettes—Pasteur, with bulbs. filtered acetonitrile (Millipore type FH filter 5.3 Balance—Analytical, capable of accu- or equivalent) and filtered acetate buffer rately weighing 0.0001 g. (Millipore type GS filter or equivalent) in a 5.4 High performance liquid chro- narrow-mouth, glass container and mix thor- matograph (HPLC)—An analytical system oughly. Prepare fresh weekly. Degas daily by complete with column supplies, high pres- sonicating under vacuum, by heating and sure syringes, detector, and compatible re- stirring, or by purging with helium. corder. A data system is recommended for 6.11 Stock standard solutions (1.00 μg/ measuring peak areas and retention times. μL)—Stock standard solutions may be pre- 5.4.1 Solvent delivery system—With pulse pared from pure standard materials or pur- damper, Altex 110A or equivalent. chased as certified solutions. 5.4.2 Injection valve (optional)—Waters 6.11.1 Prepare stock standard solutions by U6K or equivalent. accurately weighing about 0.0100 g of pure 5.4.3 Electrochemical detector—Bio- material. Dissolve the material in methanol analytical Systems LC–2A with glassy car- and dilute to volume in a 10–mL volumetric bon electrode, or equivalent. This detector flask. Larger volumes can be used at the con- has proven effective in the analysis of venience of the analyst. When compound pu- wastewaters for the parameters listed in the rity is assayed to be 96% or greater, the scope (Section 1.1), and was used to develop weight can be used without correction to cal- the method performance statements in Sec- culate the concentration of the stock stand- tion 14. Guidelines for the use of alternate ard. Commercially prepared stock standards detectors are provided in Section 12.1. can be used at any concentration if they are 5.4.4 Electrode polishing kit—Princeton certified by the manufacturer or by an inde- Applied Research Model 9320 or equivalent. pendent source. 5.4.5 Column—Lichrosorb RP–2, 5 micron 6.11.2 Transfer the stock standard solu- particle diameter, in a 25 cm × 4.6 mm ID tions into Teflon-sealed screw-cap bottles. stainless steel column. This column was used Store at 4 °C and protect from light. Stock to develop the method performance state- standard solutions should be checked fre- ments in Section 14. Guidelines for the use of quently for signs of degradation or evapo- alternate column packings are provided in ration, especially just prior to preparing Section 12.1. calibration standards from them. 6.11.3 Stock standard solutions must be 6. Reagents replaced after six months, or sooner if com- 6.1 Reagent water—Reagent water is de- parison with check standards indicates a fined as a water in which an interferent is problem. not observed at the MDL of the parameters 6.12 Quality control check sample con- of interest. centrate—See Section 8.2.1.

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7. Calibration sponse factors (RF) for each compound using Equation 1. 7.1 Establish chromatographic operating conditions equivalent to those given in Table RF = (As)(Cis (Ais)(Cs) 1. The HPLC system can be calibrated using the external standard technique (Section 7.2) Equation 1 or the internal standard technique (Section where: 7.3). As = Response for the parameter to be meas- 7.2 External standard calibration proce- ured. dure: Ais = Response for the internal standard. 7.2.1 Prepare calibration standards at a Cis = Concentration of the internal standard minimum of three concentration levels for (μg/L). each parameter of interest by adding vol- Cs = Concentration of the parameter to be umes of one or more stock standards to a measured (μg/L). volumetric flask and diluting to volume with If the RF value over the working range is mobile phase. One of the external standards a constant (<10% RSD), the RF can be as- should be at a concentration near, but above, sumed to be invariant and the average RF the MDL (Table 1) and the other concentra- can be used for calculations. Alternatively, tions should correspond to the expected the results can be used to plot a calibration range of concentrations found in real sam- curve of response ratios, As/Ais, vs. RF. ples or should define the working range of 7.4 The working calibration curve, cali- the detector. bration factor, or RF must be verified on 7.2.2 Using syringe injections of 5 to 25 μL each working day by the measurement of one or a constant volume injection loop, analyze or more calibration standards. If the re- each calibration standard according to Sec- sponse for any parameter varies from the tion 12 and tabulate peak height or area re- predicted response by more than ±15%, a new sponses against the mass injected. The re- calibration curve must be prepared for that sults can be used to prepare a calibration compound. If serious loss of response occurs, curve for each compound. Alternatively, if polish the electrode and recalibrate. the ratio of response to amount injected 7.5 Before using any cleanup procedure, (calibration factor) is a constant over the the analyst must process a series of calibra- working range (<10% relative standard devi- tion standards through the procedure to vali- ation, RSD), linearity through the origin can date elution patterns and the absence of be assumed and the average ratio or calibra- interferences from the reagents. tion factor can be used in place of a calibra- 8. Quality Control tion curve. 7.3 Internal standard calibration proce- 8.1 Each laboratory that uses this method dure—To use this approach, the analyst must is required to operate a formal quality con- select one or more internal standards that trol program. The minimum requirements of are similar in analytical behavior to the this program consist of an initial demonstra- compounds of interest. The analyst must fur- tion of laboratory capability and an ongoing ther demonstrate that the measurement of analysis of spiked samples to evaluate and the internal standard is not affected by document data quality. The laboratory must method or matrix interferences. Because of maintain records to document the quality of these limitations, no internal standard can data that is generated. Ongoing data quality be suggested that is applicable to all sam- checks are compared with established per- ples. formance criteria to determine if the results 7.3.1 Prepare calibration standards at a of analyses meet the performance character- minimum of three concentration levels for istics of the method. When results of sample each parameter of interest by adding vol- spikes indicate atypical method perform- umes of one or more stock standards to a ance, a quality control check standard must volumetric flask. To each calibration stand- be analyzed to confirm that the measure- ard, add a known constant amount of one or ments were performed in an in-control mode more internal standards, and dilute to vol- of operation. ume with mobile phase. One of the standards 8.1.1 The analyst must make an initial, should be at a concentration near, but above, one-time, demonstration of the ability to the MDL and the other concentrations generate acceptable accuracy and precision should correspond to the expected range of with this method. This ability is established concentrations found in real samples or as described in Section 8.2. should define the working range of the detec- 8.1.2 In recognition of advances that are tor. occurring in chromatography, the analyst is 7.3.2 Using syringe injections of 5 to 25 μL permitted certain options (detailed in Sec- or a constant volume injection loop, analyze tions 10.9, 11.1, and 12.1) to improve the sepa- each calibration standard according to Sec- rations or lower the cost of measurements. tion 12 and tabulate peak height or area re- Each time such a modification is made to sponses against concentration for each com- the method, the analyst is required to repeat pound and internal standard. Calculate re- the procedure in Section 8.2.

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8.1.3 Before processing any samples, the acceptable for that parameter. Locate and analyst must analyze a reagent water blank correct the source of the problem and repeat to demonstrate that interferences from the the test for all parameters of interest begin- analytical system and glassware are under ning with Section 8.2.2. control. Each time a set of samples is ex- 8.3 The laboratory must, on an ongoing tracted or reagents are changed, a reagent basis, spike at least 10% of the samples from water blank must be processed as a safe- each sample site being monitored to assess guard against laboratory contamination. accuracy. For laboratories analyzing one to 8.1.4 The laboratory must, on an ongoing ten samples per month, at least one spiked basis, spike and analyze a minimum of 10% sample per month is required. of all samples to monitor and evaluate lab- 8.3.1 The concentration of the spike in the oratory data quality. This procedure is de- sample should be determined as follows: scribed in Section 8.3. 8.1.5 The laboratory must, on an ongoing 8.3.1.1 If, as in compliance monitoring, basis, demonstrate through the analyses of the concentration of a specific parameter in quality control check standards that the op- the sample is being checked against a regu- eration of the measurement system is in con- latory concentration limit, the spike should trol. This procedure is described in Section be at that limit or 1 to 5 times higher than 8.4. The frequency of the check standard the background concentration determined in analyses is equivalent to 10% of all samples Section 8.3.2, whichever concentration would analyzed but may be reduced if spike recov- be larger. eries from samples (Section 8.3) meet all 8.3.1.2 If the concentration of a specific specified quality control criteria. parameter in the sample is not being 8.1.6 The laboratory must maintain per- checked against a limit specific to that pa- formance records to document the quality of rameter, the spike should be at 50 μg/L or 1 data that is generated. This procedure is de- to 5 times higher than the background con- scribed in Section 8.5. centration determined in Section 8.3.2, 8.2 To establish the ability to generate whichever concentration would be larger. acceptable accuracy and precision, the ana- 8.3.1.3 If it is impractical to determine lyst must perform the following operations. background levels before spiking (e.g., max- 8.2.1 A quality control (QC) check sample imum holding times will be exceeded), the concentrate is required containing benzidine spike concentration should be (1) the regu- and/or 3,3′-dichlorobenzidine at a concentra- latory concentration limit, if any; or, if none μ tion of 50 g/mL each in methanol. The QC (2) the larger of either 5 times higher than check sample concentrate must be obtained the expected background concentration or 50 from the U.S. Environmental Protection μg/L. Agency, Environmental Monitoring and Sup- 8.3.2 Analyze one sample aliquot to deter- port Laboratory in Cincinnati, Ohio, if avail- mine the background concentration (B) of able. If not available from that source, the each parameter. If necessary, prepare a new QC check sample concentrate must be ob- QC check sample concentrate (Section 8.2.1) tained from another external source. If not appropriate for the background concentra- available from either source above, the QC tions in the sample. Spike a second sample check sample concentrate must be prepared aliquot with 1.0 mL of the QC check sample by the laboratory using stock standards pre- pared independently from those used for cali- concentrate and analyze it to determine the bration. concentration after spiking (A) of each pa- 8.2.2 Using a pipet, prepare QC check sam- rameter. Calculate each percent recovery (P) ¥ ples at a concentration of 50 μg/L by adding as 100(A B)%/T, where T is the known true 1.00 mL of QC check sample concentrate to value of the spike. each of four 1–L-L aliquots of reagent water. 8.3.3 Compare the percent recovery (P) for 8.2.3 Analyze the well-mixed QC check each parameter with the corresponding QC samples according to the method beginning acceptance criteria found in Table 2. These in Section 10. acceptance criteria were calculated to in- 8.2.4 Calculate the average recovery (X¯ ) in clude an allowance for error in measurement μg/L, and the standard deviation of the re- of both the background and spike concentra- covery (s) in μg/L, for each parameter using tions, assuming a spike to background ratio the four results. of 5:1. This error will be accounted for to the 8.2.5 For each parameter compare s and X¯ extent that the analyst’s spike to back- with the corresponding acceptance criteria ground ratio approaches 5:1. 7 If spiking was for precision and accuracy, respectively, performed at a concentration lower than 50 found in Table 2. If s and X¯ for all param- μg/L, the analyst must use either the QC ac- eters of interest meet the acceptance cri- ceptance criteria in Table 2, or optional QC teria, the system performance is acceptable acceptance criteria calculated for the spe- and analysis of actual samples can begin. If cific spike concentration. To calculate op- any individual s exceeds the precision limit tional acceptance criteria for the recovery of or any individual X¯ falls outside the range a parameter: (1) Calculate accuracy (X′) for accuracy, the system performance is un- using the equation in Table 3, substituting

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the spike concentration (T) for C; (2) cal- lyzed to assess the precision of the environ- culate overall precision (S′) using the equa- mental measurements. When doubt exists tion in Table 3, substituting X′ for X¯ ; (3) cal- over the identification of a peak on the chro- culate the range for recovery at the spike matogram, confirmatory techniques such as concentration as (100 X′/T)±2.44(100 S′/T)%. 7 HPLC with a dissimilar column, gas chroma- 8.3.4 If any individual P falls outside the tography, or mass spectrometer must be designated range for recovery, that param- used. Whenever possible, the laboratory eter has failed the acceptance criteria. A should analyze standard reference materials check standard containing each parameter and participate in relevant performance that failed the criteria must be analyzed as evaluation studies. described in Section 8.4. 8.4 If any parameter fails the acceptance 9. Sample Collection, Preservation, and criteria for recovery in Section 8.3, a QC Handling check standard containing each parameter 9.1 Grab samples must be collected in that failed must be prepared and analyzed. glass containers. Conventional sampling NOTE: The frequency for the required anal- practices 8 should be followed, except that ysis of a QC check standard will depend upon the bottle must not be prerinsed with sample the number of parameters being simulta- before collection. Composite samples should neously tested, the complexity of the sample be collected in refrigerated glass containers matrix, and the performance of the labora- in accordance with the requirements of the tory. program. Automatic sampling equipment 8.4.1 Prepare the QC check standard by must be as free as possible of Tygon tubing adding 1.0 mL of QC check sample con- and other potential sources of contamina- centrate (Sections 8.2.1 or 8.3.2) to 1 L of rea- tion. gent water. The QC check standard needs 9.2 All samples must be iced or refrig- only to contain the parameters that failed erated at 4 °C and stored in the dark from criteria in the test in Section 8.3. the time of collection until extraction. Both 8.4.2 Analyze the QC check standard to benzidine and 3,3′-dichlorobenzidine are eas- determine the concentration measured (A) of ily oxidized. Fill the sample bottles and, if each parameter. Calculate each percent re- residual chlorine is present, add 80 mg of so- dium thiosulfate per liter of sample and mix covery (Ps) as 100 (A/T)%, where T is the true value of the standard concentration. well. EPA Methods 330.4 and 330.5 may be used for measurement of residual chlorine. 9 8.4.3 Compare the percent recovery (Ps) for each parameter with the corresponding Field test kits are available for this purpose. QC acceptance criteria found in Table 2. Only After mixing, adjust the pH of the sample to parameters that failed the test in Section 8.3 a range of 2 to 7 with sulfuric acid. need to be compared with these criteria. If 9.3 If 1,2-diphenylhydrazine is likely to be the recovery of any such parameter falls out- present, adjust the pH of the sample to 4.0 side the designated range, the laboratory ±0.2 to prevent rearrangement to benzidine. performance for that parameter is judged to 9.4 All samples must be extracted within 7 be out of control, and the problem must be days of collection. Extracts may be held up immediately identified and corrected. The to 7 days before analysis, if stored under an analytical result for that parameter in the inert (oxidant free) atmosphere. 2 The ex- unspiked sample is suspect and may not be tract should be protected from light. reported for regulatory compliance purposes. 10. Sample Extraction 8.5 As part of the QC program for the lab- oratory, method accuracy for wastewater 10.1 Mark the water meniscus on the side samples must be assessed and records must of the sample bottle for later determination be maintained. After the analysis of five of sample volume. Pour the entire sample spiked wastewater samples as in Section 8.3, into a 2–L separatory funnel. Check the pH calculate the average percent recovery (P¯ ) of the sample with wide-range pH paper and and the standard deviation of the percent re- adjust to within the range of 6.5 to 7.5 with covery (sp). Express the accuracy assessment sodium hydroxide solution or sulfuric acid. ¯ as a percent recovery interval from P¥2sp to 10.2 Add 100 mL of chloroform to the sam- ¯ ¯ P + 2sp. If P = 90% and sp = 10%, for example, ple bottle, seal, and shake 30 s to rinse the the accuracy interval is expressed as 70– inner surface. (Caution: Handle chloroform 110%. Update the accuracy assessment for in a well ventilated area.) Transfer the sol- each parameter on a regular basis (e.g. after vent to the separatory funnel and extract each five to ten new accuracy measure- the sample by shaking the funnel for 2 min ments). with periodic venting to release excess pres- 8.6 It is recommended that the laboratory sure. Allow the organic layer to separate adopt additional quality assurance practices from the water phase for a minimum of 10 for use with this method. The specific prac- min. If the emulsion interface between lay- tices that are most productive depend upon ers is more than one-third the volume of the the needs of the laboratory and the nature of solvent layer, the analyst must employ me- the samples. Field duplicates may be ana- chanical techniques to complete the phase

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separation. The optimum technique depends performed immediately. If the extract will upon the sample, but may include stirring, be stored longer than two days, it should be filtration of the emulsion through glass transferred to a Teflon-sealed screw-cap vial. wool, centrifugation, or other physical meth- If the sample extract requires no further ods. Collect the chloroform extract in a 250- cleanup, proceed with HPLC analysis (Sec- mL separatory funnel. tion 12). If the sample requires further clean- 10.3 Add a 50-mL volume of chloroform to up, proceed to Section 11. the sample bottle and repeat the extraction 10.12 Determine the original sample vol- procedure a second time, combining the ex- ume by refilling the sample bottle to the tracts in the separatory funnel. Perform a mark and transferring the liquid to a 1,000- third extraction in the same manner. mL graduated cylinder. Record the sample 10.4 Separate and discard any aqueous volume to the nearest 5 mL. layer remaining in the 250-mL separatory funnel after combining the organic extracts. 11. Cleanup and Separation Add 25 mL of 1 M sulfuric acid and extract 11.1 Cleanup procedures may not be nec- the sample by shaking the funnel for 2 min. essary for a relatively clean sample matrix. Transfer the aqueous layer to a 250-mL beak- If particular circumstances demand the use er. Extract with two additional 25-mL por- of a cleanup procedure, the analyst first tions of 1 M sulfuric acid and combine the must demonstrate that the requirements of acid extracts in the beaker. Section 8.2 can be met using the method as 10.5 Place a stirbar in the 250-mL beaker revised to incorporate the cleanup proce- and stir the acid extract while carefully add- dure. ing 5 mL of 0.4 M sodium tribasic phosphate. While monitoring with a pH meter, neu- 12. High Performance Liquid Chromatography tralize the extract to a pH between 6 and 7 by dropwise addition of 5 N sodium hydroxide 12.1 Table 1 summarizes the recommended solution while stirring the solution vigor- operating conditions for the HPLC. Included ously. Approximately 25 to 30 mL of 5 N so- in this table are retention times, capacity dium hydroxide solution will be required and factors, and MDL that can be achieved under it should be added over at least a 2-min pe- these conditions. An example of the separa- riod. Do not allow the sample pH to exceed 8. tions achieved by this HPLC column is 10.6 Transfer the neutralized extract into shown in Figure 1. Other HPLC columns, a 250-mL separatory funnel. Add 30 mL of chromatographic conditions, or detectors chloroform and shake the funnel for 2 min. may be used if the requirements of Section Allow the phases to separate, and transfer 8.2 are met. When the HPLC is idle, it is ad- the organic layer to a second 250-mL sepa- visable to maintain a 0.1 mL/min flow ratory funnel. through the column to prolong column life. 10.7 Extract the aqueous layer with two 12.2 Calibrate the system daily as de- additional 20-mL aliquots of chloroform as scribed in Section 7. before. Combine the extracts in the 250-mL 12.3 If the internal standard calibration separatory funnel. procedure is being used, the internal stand- 10.8 Add 20 mL of reagent water to the ard must be added to the sample extract and combined organic layers and shake for 30 s. mixed thoroughly immediately before injec- 10.9 Transfer the organic extract into a tion into the instrument. 100-mL round bottom flask. Add 20 mL of 12.4 Inject 5 to 25 μL of the sample extract methanol and concentrate to 5 mL with a ro- or standard into the HPLC. If constant vol- tary evaporator at reduced pressure and 35 ume injection loops are not used, record the °C. An aspirator is recommended for use as volume injected to the nearest 0.05 μL, and the source of vacuum. Chill the receiver with the resulting peak size in area or peak ice. This operation requires approximately 10 height units. min. Other concentration techniques may be 12.5 Identify the parameters in the sample used if the requirements of Section 8.2 are by comparing the retention times of the met. peaks in the sample chromatogram with 10.10 Using a 9-in. Pasteur pipette, trans- those of the peaks in standard fer the extract to a 15-mL, conical, screw-cap chromatograms. The width of the retention centrifuge tube. Rinse the flask, including time window used to make identifications the entire side wall, with 2-mL portions of should be based upon measurements of ac- methanol and combine with the original ex- tual retention time variations of standards tract. over the course of a day. Three times the 10.11 Carefully concentrate the extract to standard deviation of a retention time for a 0.5 mL using a gentle stream of nitrogen compound can be used to calculate a sug- while heating in a 30 °C water bath. Dilute to gested window size; however, the experience 2 mL with methanol, reconcentrate to 1 mL, of the analyst should weigh heavily in the and dilute to 5 mL with acetate buffer. Mix interpretation of chromatograms. the extract thoroughly. Cap the centrifuge 12.6 If the response for a peak exceeds the tube and store refrigerated and protected working range of the system, dilute the ex- from light if further processing will not be tract with mobile phase and reanalyze.

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12.7 If the measurement of the peak re- over the concentration range from 7 × MDL sponse for benzidine is prevented by the pres- to 3000 × MDL. 10 ence of interferences, reduce the electrode 14.3 This method was tested by 17 labora- potential to + 0.6 V and reanalyze. If the ben- tories using reagent water, drinking water, zidine peak is still obscured by interferences, surface water, and three industrial further cleanup is required. wastewaters spiked at six concentrations over the range 1.0 to 70 μg/L. 11 Single oper- 13. Calculations ator precision, overall precision, and method 13.1 Determine the concentration of indi- accuracy were found to be directly related to vidual compounds in the sample. the concentration of the parameter and es- 13.1.1 If the external standard calibration sentially independent of the sample matrix. procedure is used, calculate the amount of Linear equations to describe these relation- material injected from the peak response ships are presented in Table 3. using the calibration curve or calibration factor determined in Section 7.2.2. The con- References centration in the sample can be calculated 1. 40 CFR part 136, appendix B. from Equation 2. 2. ‘‘Determination of Benzidines in Indus- trial and Muncipal Wastewaters,’’ EPA 600/4– ()AV() μ= t 82–022, National Technical Information Serv- Concentration ( g/L) ice, PB82–196320, Springfield, Virginia 22161, ()VVis() April 1982. Equation 2 3. ASTM Annual Book of Standards, Part 31, D3694–78. ‘‘Standard Practices for Prepa- where: ration of Sample Containers and for Preser- A = Amount of material injected (ng). vation of Organic Constituents,’’ American Vi = Volume of extract injected (μL). Society for Testing and Materials, Philadel- Vt = Volume of total extract (μL). phia. Vs = Volume of water extracted (mL). 4. ‘‘Carcinogens—Working With Carcino- 13.1.2 If the internal standard calibration gens,’’ Department of Health, Education, and procedure is used, calculate the concentra- Welfare, Public Health Service, Center for tion in the sample using the response factor Disease Control, National Institute for Occu- (RF) determined in Section 7.3.2 and Equa- pational Safety and Health, Publication No. tion 3. 77–206, August 1977. 5. ‘‘OSHA Safety and Health Standards, ()()AI General Industry,’’ (29 CFR part 1910), Occu- μ= ss Concentration ( g/L) () pational Safety and Health Administration, ()ARFVis() o OSHA 2206 (Revised, January 1976). 6. ‘‘Safety in Academic Chemistry Labora- Equation 3 tories,’’ American Chemical Society Publica- where: tion, Committee on Chemical Safety, 3rd Edition, 1979. As = Response for the parameter to be meas- ured. 7. Provost, L.P., and Elder, R.S. ‘‘Interpre- Ais = Response for the internal standard. tation of Percent Recovery Data,’’ American Is = Amount of internal standard added to Laboratory, 15, 58–63 (1983). (The value 2.44 each extract (μg). used in the equation in Section 8.3.3 is two Vo = Volume of water extracted (L). times the value 1.22 derived in this report.) 13.2 Report results in μg/L without correc- 8. ASTM Annual Book of Standards, Part tion for recovery data. All QC data obtained 31, D3370–76. ‘‘Standard Practices for Sam- should be reported with the sample results. pling Water,’’ American Society for Testing and Materials, Philadelphia. 14. Method Performance 9. ‘‘Methods 330.4 (Titrimetric, DPD-FAS) 14.1 The method detection limit (MDL) is and 330.5 (Spectrophotometric, DPD) for defined as the minimum concentration of a Chlorine Total Residual,’’ Methods for substance that can be measured and reported Chemical Analysis of Water and Wastes, with 99% confidence that the value is above EPA–600/4–79–020, U.S. Environmental Pro- zero. 1 The MDL concentrations listed in tection Agency, Environmental Monitoring Table 1 were obtained using reagent water. 10 and Support Laboratory, Cincinnati, Ohio Similar results were achieved using rep- 45268, March 1979. resentative wastewaters. The MDL actually 10. ‘‘EPA Method Study 15, Method 605 achieved in a given analysis will vary de- (Benzidines),’’ EPA 600/4–84–062, National pending on instrument sensitivity and ma- Technical Information Service, PB84–211176, trix effects. Springfield, Virginia 22161, June 1984. 14.2 This method has been tested for lin- 11. ‘‘EPA Method Validation Study 15, earity of spike recovery from reagent water Method 605 (Benzidines),’’ Report for EPA and has been demonstrated to be applicable Contract 68–03–2624 (In preparation).

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TABLE 1—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS

Column ca- Method de- Parameter Retention pacity factor tection limit time (min) (k′) (μg/L)

Benzidine ...... 6.1 1.44 0.08 3,3′-Dichlorobenzidine ...... 12.1 3.84 0.13 HPLC Column conditions: Lichrosorb RP–2, 5 micron particle size, in a 25 cm × 4.6 mm ID stainless steel column. Mobile Phase: 0.8 mL/min of 50% acetonitrile/50% 0.1M pH 4.7 acetate buffer. The MDL were determined using an electrochemical de- tector operated at + 0.8 V.

TABLE 2—QC ACCEPTANCE CRITERIA—METHOD 605

Test Range for μ Limit for s Range for Parameter conc. ( g/ μ ¯ μ P, Ps L) ( g/L) X ( g/L) (percent)

Benzidine ...... 50 18.7 9.1–61.0 D–140 3.3′-Dichlorobenzidine ...... 50 23.6 18.7–50.0 5–128 s = Standard deviation of four recovery measurements, in μg/L (Section 8.2.4). X¯ = Average recovery for four recovery measurements, in μg/L (Section 8.2.4). P, Ps = Percent recovery measured (Section 8.3.2, Section 8.4.2). D = Detected; result must be greater than zero. Note: These criteria are based directly upon the method performance data in Table 3. Where necessary, the limits for recovery have been broadened to assure applicability of the limits to concentrations below those used to develop Table 3.

TABLE 3—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 605

Accuracy, as Single analyst ′ Overall preci- Parameter recovery, precision, sr ′ μ X′(μg/L) (μg/L) sion, S ( g/L)

Benzidine ...... 0.70C + 0.06 0.28X¯ + 0.19 0.40X¯ + 0.18 3,3′-Dichlorobenzidine ...... 0.66C + 0.23 0.39X¯ ¥0.05 0.38X¯ + 0.02 X′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L. sr′ = Expected single analyst standard deviation of measurements at an average concentration found of X¯ , in μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X¯ , in μg/L. C = True value for the concentration, in μg/L. X¯ = Average recovery found for measurements of samples containing a concentration of C, in μg/L.

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COLUMN: LICHROSORB RP-2 MOBILE PHASE: 50% ACETONITRILE IN ACETATE BUFFER DETECTOR: ELECTROCHEMICAL AT+ 0.8 V

UJ UJz z 0 c ;:::; ;:::; z z UJ UJ = = 0 a: 0 ..J J: uc c.,1 cw)

6 12 RETENTION TIME, MIN. Figure 1. Liquid chromatogram of benzidines.

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METHOD 606—PHTHALATE ESTER 2. Summary of Method 1. Scope and Application 2.1 A measured volume of sample, ap- proximately 1–L, is extracted with meth- 1.1 This method covers the determination ylene chloride using a separatory funnel. The of certain phthalate esters. The following pa- methylene chloride extract is dried and ex- rameters can be determined by this method: changed to hexane during concentration to a volume of 10 mL or less. The extract is sepa- STORET Parameter No. CAS No. rated by gas chromatography and the phthalate esters are then measured with an Bis(2-ethylhexyl) phthalate ...... 39100 117–81–7 electron capture detector. 2 Butyl benzyl phthalate ...... 34292 85–68–7 2.2 Analysis for phthalates is especially Di-n-butyl phthalate ...... 39110 84–74–2 complicated by their ubiquitous occurrence Diethyl phthalate ...... 34336 84–66–2 in the environment. The method provides Dimethyl phthalate ...... 34341 131–11–3 Di-n-octyl phthalate ...... 34596 117–84–0 Florisil and alumina column cleanup proce- dures to aid in the elimination of inter- 1.2 This is a gas chromatographic (GC) ferences that may be encountered. method applicable to the determination of 3. Interferences the compounds listed above in municipal and industrial discharges as provided under 40 3.1 Method interferences may be caused CFR 136.1. When this method is used to ana- by contaminants in solvents, reagents, glass- lyze unfamiliar samples for any or all of the ware, and other sample processing hardware compounds above, compound identifications that lead to discrete artifacts and/or ele- should be supported by at least one addi- vated baselines in gas chromatograms. All of tional qualitative technique. This method these materials must be routinely dem- describes analytical conditions for a second onstrated to be free from interferences under gas chromatographic column that can be the conditions of the analysis by running used to confirm measurements made with laboratory reagent blanks as described in the primary column. Method 625 provides gas Section 8.1.3. chromatograph/mass spectrometer (GC/MS) 3.1.1 Glassware must be scrupulously conditions appropriate for the qualitative cleaned. 3 Clean all glassware as soon as pos- and quantitative confirmation of results for sible after use by rinsing with the last sol- all of the parameters listed above, using the vent used in it. Solvent rinsing should be fol- extract produced by this method. lowed by detergent washing with hot water, 1.3 The method detection limit (MDL, de- and rinses with tap water and distilled fined in Section 14.1) 1 for each parameter is water. The glassware should then be drained listed in Table 1. The MDL for a specific dry, and heated in a muffle furnace at 400 °C wastewater may differ from those listed, de- for 15 to 30 min. Some thermally stable ma- pending upon the nature of interferences in terials, such as PCBs, may not be eliminated the sample matrix. by this treatment. Solvent rinses with ace- 1.4 The sample extraction and concentra- tone and pesticide quality hexane may be tion steps in this method are essentially the substituted for the muffle furnace heating. same as in Methods 608, 609, 611, and 612. Thorough rinsing with such solvents usually Thus, a single sample may be extracted to eliminates PCB interference. Volumetric measure the parameters included in the ware should not be heated in a muffle fur- scope of each of these methods. When clean- nace. After drying and cooling, glassware up is required, the concentration levels must should be sealed and stored in a clean envi- be high enough to permit selecting aliquots, ronment to prevent any accumulation of as necessary, to apply appropriate cleanup dust or other contaminants. Store inverted procedures. The analyst is allowed the lati- or capped with aluminum foil. tude, under Section 12, to select 3.1.2 The use of high purity reagents and chromatographic conditions appropriate for solvents helps to minimize interference prob- the simultaneous measurement of combina- lems. Purification of solvents by distillation tions of these parameters. in all-glass systems may be required. 1.5 Any modification of this method, be- 3.2 Phthalate esters are contaminants in yond those expressly permitted, shall be con- many products commonly found in the lab- sidered as a major modification subject to oratory. It is particularly important to avoid application and approval of alternate test the use of because phthalates are procedures under 40 CFR 136.4 and 136.5. commonly used as plasticizers and are easily 1.6 This method is restricted to use by or extracted from plastic materials. Serious under the supervision of analysts experi- phthalate contamination can result at any enced in the use of a gas chromatograph and time, if consistent quality control is not in the interpretation of gas chromatograms. practiced. Great care must be experienced to Each analyst must demonstrate the ability prevent such contamination. Exhaustive to generate acceptable results with this cleanup of reagents and glassware may be re- method using the procedure described in Sec- quired to eliminate background phthalate tion 8.2. contamination. 45

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3.3 Matrix interferences may be caused by 5.2.3 Chromatographic column—300 mm contaminants that are co-extracted from the long × 10 mm ID, with Teflon stopcock and sample. The extent of matrix interferences coarse frit filter disc at bottom (Kontes K– will vary considerably from source to source, 420540–0213 or equivalent). depending upon the nature and diversity of 5.2.4 Concentrator tube, Kuderna-Dan- the industrial complex or municipality being ish—10–mL, graduated (Kontes K–570050–1025 sampled. The cleanup procedures in Section or equivalent). Calibration must be checked 11 can be used to overcome many of these at the volumes employed in the test. Ground interferences, but unique samples may re- glass stopper is used to prevent evaporation quire additional cleanup approaches to of extracts. achieve the MDL listed in Table 1. 5.2.5 Evaporative flask, Kuderna-Danish— 500–mL (Kontes K–570001–0500 or equivalent). 4. Safety Attach to concentrator tube with springs. 4.1 The toxicity or carcinogenicity of 5.2.6 Snyder column, Kuderna-Danish— each reagent used in this method has not Three-ball macro (Kontes K–503000–0121 or been precisely defined; however, each chem- equivalent). 5.2.7 Snyder column, Kuderna-Danish— ical compound should be treated as a poten- Two-ball micro (Kontes K–569001–0219 or tial health hazard. From this viewpoint, ex- equivalent). posure to these chemicals must be reduced to 5.2.8 Vials—10 to 15–mL, amber glass, with the lowest possible level by whatever means Teflon-lined screw cap. available. The laboratory is responsible for 5.3 Boiling chips—Approximately 10/40 maintaining a current awareness file of mesh. Heat to 400 °C for 30 min or Soxhlet ex- OSHA regulations regarding the safe han- tract with methylene chloride. dling of the chemicals specified in this meth- 5.4 Water bath—Heated, with concentric od. A reference file of material data handling ring cover, capable of temperature control sheets should also be made available to all (±2 °C). The bath should be used in a hood. personnel involved in the chemical analysis. 5.5 Balance—Analytical, capable of accu- Additional references to laboratory safety rately weighing 0.0001 g. are available and have been identified 68 for 5.6 Gas chromatograph—An analytical the information of the analyst. system complete with gas chromatograph suitable for on-column injection and all re- 5. Apparatus and Materials quired accessories including syringes, ana- 5.1 Sampling equipment, for discrete or lytical columns, gases, detector, and strip- composite sampling. chart recorder. A data system is rec- 5.1.1 Grab sample bottle—1–L or 1–qt, ommended for measuring peak areas. amber glass, fitted with a screw cap lined 5.6.1 Column 1—1.8 m long × 4 mm ID with Teflon. Foil may be substituted for Tef- glass, packed with 1.5% SP–2250/1.95% SP– lon if the sample is not corrosive. If amber 2401 Supelcoport (100/120 mesh) or equivalent. bottles are not available, protect samples This column was used to develop the method from light. The bottle and cap liner must be performance statemelts in Section 14. Guide- washed, rinsed with acetone or methylene lines for the use of alternate column chloride, and dried before use to minimize packings are provided in Section 12.1. contamination. 5.6.2 Column 2—1.8 m long × 4 mm ID 5.1.2 Automatic sampler (optional)—The glass, packed with 3% OV–1 on Supelcoport sampler must incorporate glass sample con- (100/120 mesh) or equivalent. tainers for the collection of a minimum of 5.6.3 Detector—Electron capture detector. 250 mL of sample. Sample containers must be This detector has proven effective in the kept refrigerated at 4 °C and protected from analysis of wastewaters for the parameters light during compositing. If the sampler uses listed in the scope (Section 1.1), and was used a peristaltic pump, a minimum length of to develop the method performance state- compressible silicone rubber tubing may be ments in Section 14. Guidelines for the use of used. Before use, however, the compressible alternate detectors are provided in Section tubing should be thoroughly rinsed with 12.1. methanol, followed by repeated rinsings with distilled water to minimize the potential for 6. Reagents contamination of the sample. An integrating 6.1 Reagent water—Reagent water is de- flow meter is required to collect flow propor- fined as a water in which an interferent is tional composites. not observed at the MDL of the parameters 5.2 Glassware (All specifications are sug- of interest. gested. Catalog numbers are included for il- 6.2 Acetone, hexane, isooctane, methylene lustration only). chloride, methanol—Pesticide quality or 5.2.1 Separatory funnel—2–L, with Teflon equivalent. stopcock. 6.3 Ethyl ether—nanograde, redistilled in 5.2.2 Drying column—Chromatographic glass if necessary. column, approximately 400 mm long × 19 mm 6.3.1 Ethyl ether must be shown to be free ID, with coarse frit filter disc. of peroxides before it is used as indicated by

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EM Laboratories Quant test strips. (Avail- 6.8 Quality control check sample con- able from Scientific Products Co., Cat. No. centrate—See Section 8.2.1. P1126–8, and other suppliers.) 6.3.2 Procedures recommended for re- 7. Calibration moval of peroxides are provided with the test strips. After cleanup, 20 mL of ethyl alcohol 7.1 Establish gas chromatograph oper- preservative must be added to each liter of ating conditions equivalent to those given in ether. Table 1. The gas chromatographic system 6.4 Sodium sulfate—(ACS) Granular, an- can be calibrated using the external standard hydrous. Several levels of purification may technique (Section 7.2) or the internal stand- be required in order to reduce background ard technique (Section 7.3). phthalate levels to an acceptable level: 1) 7.2 External standard calibration proce- Heat 4 h at 400 °C in a shallow tray, 2) Heat dure: 16 h at 450 to 500 °C in a shallow tray, 3) 7.2.1 Prepared calibration standards at a Soxhlet extract with methylene chloride for minimum of three concentration levels for 48 h. each parameter of interest by adding vol- 6.5 Florisil—PR grade (60/100 mesh). Pur- umes of one or more stock standards to a chase activated at 1250 °F and store in the volumetric flask and diluting to volume with dark in glass containers with ground glass isooctane. One of the external standards stoppers or foil-lined screw caps. To prepare for use, place 100 g of Florisil into a 500-mL should be at a concentration near, but above, beaker and heat for approximately 16 h at 40 the MDL (Table 1) and the other concentra- °C. After heating transfer to a 500-mL rea- tions should correspond to the expected gent bottle. Tightly seal and cool to room range of concentrations found in real sam- temperature. When cool add 3 mL of reagent ples or should define the working range of water. Mix thoroughly by shaking or rolling the detector. for 10 min and let it stand for at least 2 h. 7.2.2 Using injections of 2 to 5 μL, analyze Keep the bottle sealed tightly. each calibration standard according to Sec- 6.6 Alumina—Neutral activity Super I, tion 12 and tabulate peak height or area re- W200 series (ICN Life Sciences Group, No. sponses against the mass injected. The re- 404583). To prepare for use, place 100 g of alu- sults can be used to prepare a calibration mina into a 500-mL beaker and heat for ap- curve for each compound. Alternatively, if ° proximately 16 h at 400 C. After heating the ratio of response to amount injected transfer to a 500-mL reagent bottle. Tightly (calibration factor) is a constant over the seal and cool to room temperature. When working range (<10% relative standard devi- cool add 3 mL of reagent water. Mix thor- oughly by shaking or rolling for 10 min and ation, RSD), linearity through the origin can let it stand for at least 2 h. Keep the bottle be assumed and the average ratio or calibra- sealed tightly. tion factor can be used in place of a calibra- 6.7 Stock standard solutions (1.00 μg/μL)— tion curve. Stock standard solutions can be prepared 7.3 Internal standard calibration proce- from pure standard materials or purchased dure—To use this approach, the analyst must as certified solutions. select one or more internal standards that 6.7.1 Prepare stock standard solutions by are similar in analytical behavior to the accurately weighing about 0.0100 g of pure compounds of interest. The analyst must fur- material. Dissolve the material in isooctane ther demonstrate that the measurement of and dilute to volume in a 10-mL volumetric the internal standard is not affected by flask. Larger volumes can be used at the con- method or matrix interferences. Because of venience of the analyst. When compound pu- these limitations, no internal standard can rity is assayed to be 96% or greater, the be suggested that is applicable to all sam- weight can be used without correction to cal- ples. culate the concentration of the stock stand- 7.3.1 Prepare calibration standards at a ard. Commercially prepared stock standards can be used at any concentration if they are minimum of three concentration levels for certified by the manufacturer or by an inde- each parameter of interest by adding vol- pendent source. umes of one or more stock standards to a 6.7.2 Transfer the stock standard solu- volumetric flash. To each calibration stand- tions into Teflon-sealed screw-cap bottles. ard, add a known constant amount of one or Store at 4 °C and protect from light. Stock more internal standards, and dilute to vol- standard solutions should be checked fre- ume with isooctane. One of the standards quently for signs of degradation or evapo- should be at a concentration near, but above, ration, especially just prior to preparing the MDL and the other concentrations calibration standards from them. should correspond to the expected range of 6.7.3 Stock standard solutions must be re- concentrations found in real samples or placed after six months, or sooner if com- should define the working range of the detec- parison with check standards indicates a tor. problem.

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7.3.2 Using injections of 2 to 5 μL, analyze rations or lower the cost of measurements. each calibration standard according to Sec- Each time such a modification is made to tion 12 and tabulate peak height or area re- the method, the analyst is required to repeat sponses against concentration for each com- the procedure in Section 8.2. pound and internal standard. Calculate re- 8.1.3 Before processing any samples, the sponse factors (RF) for each compound using analyst must analyze a reagent water blank Equation 1. to demonstrate that interferences from the analytical system and glassware are under RF = (As)(Cis (Ais)(Cs) control. Each time a set of samples is ex- Equation 1 tracted or reagents are changed, a reagent water blank must be processed as a safe- where: guard against laboratory contamination. A = Response for the parameter to be meas- s 8.1.4 The laboratory must, on an ongoing ured. basis, spike and analyze a minimum of 10% A = Response for the internal standard. is of all samples to monitor and evaluate lab- C = Concentration of the internal standard is oratory data quality. This procedure is de- (μg/L). scribed in Section 8.3. C = Concentration of the parameter to be s 8.1.5 The laboratory must, on an ongoing measured (μg/L). basis, demonstrate through the analyses of If the RF value over the working range is quality control check standards that the op- a constant (<10% RSD), the RF can be as- eration of the measurement system is in con- sumed to be invariant and the average RF trol. This procedure is described in Section can be used for calculations. Alternatively, 8.4. The frequency of the check standard the results can be used to plot a calibration analyses is equivalent to 10% of all samples curve of response ratios, As/Ais, vs. RF. analyzed but may be reduced if spike recov- 7.4 The working calibration curve, cali- eries from samples (Section 8.3) meet all bration factor, or RF must be verified on specified quality control criteria. each working day by the measurement of one 8.1.6 The laboratory must maintain per- or more calibration standards. If the re- formance records to document the quality of sponse for any parameter varies from the data that is generated. This procedure is de- predicted response by more than ±15%, a new scribed in Section 8.5. calibration curve must be prepared for that 8.2 To establish the ability to generate compound. acceptable accuracy and precision, the ana- 7.5 Before using any cleanup procedure, lyst must perform the following operations. the analyst must process a series of calibra- 8.2.1 A quality contrml (QC) check sample tion standards through the procedure to vali- concentrate is required containing each pa- date elution patterns and the absence of rameter of interest at the following con- interferences from the reagents. centrations in acetone: butyl benzyl phthal- ate, 10 μg/mL; bis(2-ethylhexyl) phthalate, 50 8. Quality Control μg/mL; di-n-octyl phthalate, 50 μg/mL; any 8.1 Each laboratory that uses this method other phthlate, 25 μg/mL. The QC check sam- is required to operate a formal quality con- ple concentrate must be obtained from the trol program. The minimum requirements of U.S. Environmental Protection Agancy, En- this program consist of an initial demonstra- vironmental Monitoring and Support Lab- tion of laboratory capability and an ongoing oratory in Cincinnati, Ohio, if available. If analysis of spiked samples to evaluate and not available from that source, the QC check document data quality. The laboratory must sample concentrate must be obtained from maintain records to document the quality of another external source. If not available data that is generated. Ongoing data quality from either source above, the QC check sam- checks are compared with established per- ple concentrate must be prepared by the lab- formance criteria to determine if the results oratory using stock standards prepared inde- of analyses meet the performance character- pendently from those used for calibration. istics of the method. When results of sample 8.2.2 Using a pipet, prepare QC check sam- spikes indicate atypical method perform- ples at the test concentrations shown in ance, a quality control check standard must Table 2 by adding 1.00 mL of QC check sam- be analyzed to confirm that the measure- ple concentrate to each of four 1–L aliquots ments were performed in an in-control mode of reagent water. of operation. 8.2.3 Analyze the well-mixed QC check 8.1.1 The analyst must make an initial, samples according to the method beginning one-time, demonstration of the ability to in Section 10. generate acceptable accuracy and precision 8.2.4 Calculate the average recovery (X¯ ) in with this method. This ability is established μg/L, and the standard deviation of the re- as described in Section 8.2. covery (s) in μg/L, for each parameter using 8.1.2 In recognition of advances that are the four results. occurring in chromatography, the analyst is 8.2.5 For each parameter compare s and X¯ permitted certain options (detailed in Sec- with the corresponding acceptance criteria tions 10.4, 11.1, and 12.1) to improve the sepa- for precision and accuracy, respectively,

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found in Table 2. If s and X¯ for all param- lyst must use either the QC acceptance cri- eters of interest meet the acceptance cri- teria in Table 2, or optional QC acceptance teria, the system performance is acceptable criteria calculated for the specific spike con- and analysis of actual samples can begin. If centration. To calculate optional acceptance any individual s exceeds the precision limit criteria for the recovery of a parameter: (1) or any individual X¯ falls outside the range Calculate accuracy (X′) using the equation in for accuracy, the system performance is un- Table 3, substituting the spike concentration acceptable for that parameter. Locate and (T) for C; (2) calculate overall precision (S′) correct the source of the problem and repeat using the equation in Table 3, substituting X′ the test for all parameters of interest begin- for X¯ ; (3) calculate the range for recovery at ning with Section 8.2.2. the spike concentration as (100 X′/T)±2.44(100 8.3 The laboratory must, on an ongoing S′/T)%. 9 basis, spike at least 10% of the samples from 8.3.4 If any individual P falls outside the each sample site being monitored to assess designated range for recovery, that param- accuracy. For laboratories analyzing one to eter has failed the acceptance criteria. A ten samples per month, at least one spiked check standard containing each parameter sample per month is required. that failed the criteria must be analyzed as 8.3.1 The concentration of the spike in the described in Section 8.4. sample should be determined as follows: 8.4 If any parameter fails the acceptance 8.3.1.1 If, as in compliance monitoring, criteria for recovery in Section 8.3, a QC the concentration of a specific parameter in check standard containing each parameter the sample is being checked against a regu- that failed must be prepared and analyzed. latory concentration limit, the spike should NOTE: The frequency for the required anal- be at that limit or 1 to 5 times higher than ysis of a QC check standard will depend upon the background concentration determined in the number of parameters being simulta- Section 8.3.2, whichever concentration would neously tested, the complexity of the sample be larger. matrix, and the performance of the labora- 8.3.1.2 If the concentration of a specific tory. parameter in the sample is not being 8.4.1 Prepare the QC check standard by checked against a limit specific to that pa- adding 1.0 mL of QC check sample con- rameter, the spike should be at the test con- centrate (Section 8.2.1 or 8.3.2) to 1 L of rea- centration in Section 8.2.2 or 1 to 5 times gent water. The QC check standard needs higher than the background concentration only to contain the parameters that failed determined in Section 8.3.2, whichever con- criteria in the test in Section 8.3. centration would be larger. 8.4.2 Analyze the QC check standard to 8.3.1.3 If it is impractical to determine determine the concentration measured (A) of background levels before spiking (e.g., max- each parameter. Calculate each percent re- imum holding times will be exceeded), the covery (Ps) as 100 (A/T)%, where T is the true spike concentration should be (1) the regu- value of the standard concentration. latory concentration limit, if any; or, if none 8.4.3 Compare the percent recovery (Ps) (2) the larger of either 5 times higher than for each parameter with the corresponding the expected background concentration or QC acceptance criteria found in Table 2. Only the test concentration in Section 8.2.2. parameters that failed the test in Section 8.3 8.3.2 Analyze one sample aliquot to deter- need to be compared with these criteria. If mine the background concentration (B) of the recovery of any such parameter falls out- each parameter. If necessary, prepare a new side the designated range, the laboratory QC check sample concentrate (Section 8.2.1) performance for that parameter is judged to appropriate for the background concentra- be out of control, and the problem must be tions in the sample. Spike a second sample immediately identified and corrected. The aliquot with 1.0 mL of the QC check sample analytical result for that parameter in the concentrate and analyze it to determine the unspiked sample is suspect and may not be concentration after spiking (A) of each pa- reported for regulatory compliance purposes. rameter. Calculate each percent recovery (P) 8.5 As part of the QC program for the lab- as 100(A-B)%/T, where T is the known true oratory, method accuracy for wastewater value of the spike. samples must be assessed and records must 8.3.3 Compare the percent recovery (P) for be maintained. After the analysis of five each parameter with the corresponding QC spiked wastewater samples as in Section 8.3, acceptance criteria found in Table 2. These calculate the average percent recovery (P¯ ) acceptance criteria were calculated to in- and the standard deviation of the percent re- clude an allowance for error in measurement covery (sp). Express the accuracy assessment ¯ of both the background and spike concentra- as a percent recovery interval from P¥2sp to ¯ ¯ tions, assuming a spike to background ratio P + 2sp. If P = 90% and sp = 10%, for example, of 5:1. This error will be accounted for to the the accuracy interval is expressed as 70– extent that the analyst’s spike to back- 110%. Update the accuracy assessment for ground ratio approaches 5:1. 9 If spiking was each parameter on a regular basis (e.g. after performed at a concentration lower than the each five to ten new accuracy measure- test concentration in Section 8.2.2, the ana- ments).

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8.6 It is recommended that the laboratory 10.4 Assemble a Kuderna-Danish (K-D) adopt additional quality assurance practices concentrator by attaching a 10-mL concen- for use with this method. The specific prac- trator tube to a 500-mL evaporative flask. tices that are most productive depend upon Other concentrator devices or techniques the needs of the laboratory and the nature of may be used in place of the K-D concentrator the samples. Field duplicates may be ana- if the requirements of Section 8.2 are met. lyzed to assess the precision of the environ- 10.5 Pour the combined extract through a mental measurements. When doubt exists solvent-rinsed drying column containing over the identification of a peak on the chro- about 10 cm of anhydrous sodium sulfate, matogram, confirmatory techniques such as and collect the extract in the K-D concen- gas chromatography with a dissimilar col- trator. Rinse the Erlenmeyer flask and col- umn, specific element detector, or mass umn with 20 to 30 mL of methylene chloride spectrometer must be used. Whenever pos- to complete the quantitative transfer. sible, the laboratory should analyze standard 10.6 Add one or two clean boiling chips to reference materials and participate in rel- the evaporative flask and attach a three-ball evant performance evaluation studies. Snyder column. Prewet the Snyder column by adding about 1 mL of methylene chloride 9. Sample Collection, Preservation, and to the top. Place the K-D apparatus on a hot Handling water bath (60 to 65 °C) so that the concen- trator tube is partially immersed in the hot 9.1 Grab samples must be collected in water, and the entire lower rounded surface glass containers. Conventional sampling of the flask is bathed with hot vapor. Adjust practices 10 should be followed, except that the vertical position of the apparatus and the bottle must not be prerinsed with sample the water temperature as required to com- before collection. Composite samples should plete the concentration in 15 to 20 min. At be collected in refrigerated glass containers the proper rate of distillation the balls of the in accordance with the requirements of the column will actively chatter but the cham- program. Automatic sampling equipment bers will not flood with condensed solvent. must be as free as possible of Tygon tubing When the apparent volume of liquid reaches and other potential sources of contamina- 1 mL, remove the K-D apparatus and allow it tion. to drain and cool for at least 10 min. 9.2 All samples must be iced or refrig- 10.7 Increase the temperature of the hot ° erated at 4 C from the time of collection water bath to about 80 °C. Momentarily re- until extraction. move the Snyder column, add 50 mL of 9.3 All samples must be extracted within 7 hexane and a new boiling chip, and reattach days of collection and completely analyzed the Snyder column. Concentrate the extract 2 within 40 days of extraction. as in Section 10.6, except use hexane to 10. Sample Extraction prewet the column. The elapsed time of con- centration should be 5 to 10 min. 10.1 Mark the water meniscus on the side 10.8 Remove the Snyder column and rinse of the sample bottle for later determination the flask and its lower joint into the concen- of sample volume. Pour the entire sample trator tube with 1 to 2 mL of hexane. A 5-mL into a 2–L separatory funnel. syringe is recommended for this operation. 10.2 Add 60 mL of methylene chloride to Adjust the extract volume to 10 mL. Stopper the sample bottle, seal, and shake 30 s to the concentrator tube and store refrigerated rinse the inner surface. Transfer the solvent if further processing will not be performed to the separatory funnel and extract the immediately. If the extract will be stored sample by shaking the funnel for 2 min. with longer than two days, it should be trans- periodic venting to release excess pressure. ferred to a Teflon-sealed screw-cap vial. If Allow the organic layer to separate from the the sample extract requires no further clean- water phase for a minimum of 10 min. If the up, proceed with gas chromatographic anal- emulsion interface between layers is more ysis (Section 12). If the sample requires fur- than one-third the volume of the solvent ther cleanup, proceed to Section 11. layer, the analyst must employ mechanical 10.9 Determine the original sample vol- techniques to complete the phrase separa- ume by refilling the sample bottle to the tion. The optimum technique depends upon mark and transferring the liquid to a 1000- the sample, but may include stirring, filtra- mL graduated cylinder. Record the sample tion of the emulsion through glass wool, cen- volume to the nearest 5 mL. trifugation, or other physical methods. Col- lect the methylene chloride extract in a 250- 11. Cleanup and Separation mL Erlenmeyer flask. 11. Cleanup procedures may not be nec- 10.3 Add a second 60-mL volume of meth- essary for a relatively clean sample matrix. ylene chloride to the sample bottle and re- If particular circumstances demand the use peat the extraction procedure a second time, of a cleanup procedure, the analyst may use combining the extracts in the Erlenmeyer either procedure below or any other appro- flask. Perform a third extraction in the same priate procedure. However, the analyst first manner. must demonstrate that the requirements of

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Section 8.2 can be met using the method as hexane and continue the elution of the col- revised to incorporate the cleanup proce- umn. Discard this hexane eluate. dure. 11.4.3 Next, elute the column with 140 mL 11.2 If the entire extract is to be cleaned of 20% ethyl ether in hexane (V/V) into a 500- up by one of the following procedures, it mL K-D flask equipped with a 10–mL concen- must be concentrated to 2.0 mL. To the con- trator type. Concentrate the collected frac- centrator tube in Section 10.8, add a clean tion as in Section 10.6. No solvent exchange boiling chip and attach a two-ball micro- is necessary. Adjust the volume of the Snyder column. Prewet the column by add- cleaned up extract to 10 mL in the concen- ing about 0.5 mL of hexane to the top. Place trator tube and analyze by gas chroma- the micro-K-D apparatus on a hot water bath tography (Section 12). (80 °C) so that the concentrator tube is par- tially immersed in the hot water. Adjust the 12. Gas Chromatography vertical position of the apparatus and the 12.1 Table 1 summarizes the recommended water temperature as required to complete operating conditions for the gas chro- the concentration in 5 to 10 min. At the prop- matograph. Included in this table are reten- er rate of distillation the balls of the column tion times and MDL that can be achieved will actively chatter but the chambers will under these conditions. Examples of the sep- not flood. When the apparent volume of liq- arations achieved by Column 1 are shown in uid reaches about 0.5 mL, remove the K-D Figures 1 and 2. Other packed or capillary apparatus and allow it to drain and cool for (open-tubular) columns, chromatographic at least 10 min. Remove the micro-Snyder conditions, or detectors may be used if the column and rinse its lower joint into the requirements of Section 8.2 are met. concentrator tube with 0.2 mL of hexane. Ad- 12.2 Calibrate the system daily as de- just the final volume to 2.0 mL and proceed scribed in Section 7. with one of the following cleanup procedures. 12.3 If the internal standard calibration 11.3 Florisil column cleanup for phthalate procedure is being used, the internal staldard esters: 11.3.1 Place 10 g of Florisil into a must be added to the sample extract and chromatographic column. Tap the column to mixed thoroughly immediately before injec- settle the Florisil and add 1 cm of anhydrous tion into the gas chromatograph. μ sodium sulfate to the top. 12.4 Inject 2 to 5 L of the sample extract 11.3.2 Preelute the column with 40 mL of or standard into the gas-chromatograph 11 hexane. The rate for all elutions should be using the solvent-flush technique. Smaller μ about 2 mL/min. Discard the eluate and just (1.0 L) volumes may be injected if auto- prior to exposure of the sodium sulfate layer matic devices are employed. Record the vol- μ to the air, quantitatively transfer the 2-mL ume injected to the nearest 0.05 L, and the sample extract onto the column using an ad- resulting peak size in area or peak height ditional 2 mL of hexane to complete the units. transfer. Just prior to exposure of the so- 12.5 Identify the parameters in the sample dium sulfate layer to the air, add 40 mL of by comparing the retention times of the hexane and continue the elution of the col- peaks in the sample chromatogram with umn. Discard this hexane eluate. those of the peaks in standard 11.3.3 Next, elute the column with 100 mL chromatograms. The width of the retention of 20% ethyl ether in hexane (V/V) into a 500- time window used to make identifications mL K-D flask equipped with a 10-mL concen- should be based upon measurements of ac- trator tube. Concentrate the collected frac- tual retention time variations of standards tion as in Section 10.6. No solvent exchange over the course of a day. Three times the is necessary. Adjust the volume of the standard deviation of a retention time for a cleaned up extract to 10 mL in the concen- compound can be used to calculate a sug- trator tube and analyze by gas chroma- gested window size; however, the experience tography (Section 12). of the analyst should weigh heavily in the 11.4 Alumina column cleanup for phthal- interpretation of chromatograms. ate esters: 12.6 If the response for a peak exceeds the 11.4.1 Place 10 g of alumina into a working range of the system, dilute the ex- chromatographic column. Tap the column to tract and reanalyze. settle the alumina and add 1 cm of anhy- 12.7 If the measurement of the peak re- drous sodium sulfate to the top. sponse is prevented by the presence of inter- 11.4.2 Preelute the column with 40 mL of ferences, further cleanup is required. hexane. The rate for all elutions should be 13. Calculations about 2 mL/min. Discard the eluate and just prior to exposure of the sodium sulfate layer 13.1 Determine the concentration of indi- to the air, quantitatively transfer the 2-mL vidual compounds in the sample. sample extract onto the column using an ad- 13.1.1 If the external standard calibration ditional 2 mL of hexane to complete the procedure is used, calculate the amount of transfer. Just prior to exposure of the so- material injected from the peak response dium sulfate layer to the air, add 35 mL of using the calibration curve or calibration

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factor determined in Section 7.2.2. The con- sentially independent of the sample matrix. centration in the sample can be calculated Linear equations to describe these relation- from Equation 2. ships are presented in Table 3. ()AV() References μ= t Concentration ( g/L) () 1. 40 CFR part 136, appendix B. ()VVsi 2. ‘‘Determination of Phthalates in Indus- trial and Muncipal Wastewaters,’’ EPA 600/4– Equation 2 81–063, National Technical Information Serv- where: ice, PB81–232167, Springfield, Virginia 22161, A = Amount of material injected (ng). July 1981. Vi = Volume of extract injected (μL). 3. ASTM Annual Book of Standards, Part Vt = Volume of total extract (μL). 31, D3694–78. ‘‘Standard Practices for Prepa- Vs = Volume of water extracted (mL). ration of Sample Containers and for Preser- 13.1.2 If the internal standard calibration vation of Organic Constituents,’’ American procedure is used, calculate the concentra- Society for Testing and Materials, Philadel- tion in the sample using the response factor phia. (RF) determined in Section 7.3.2 and Equa- 4. Giam, C.S., Chan, H.S., and Nef, G.S. tion 3. ‘‘Sensitive Method for Determination of Phthalate Ester Plasticizers in Open-Ocean ()()AI Biota Samples,’’ Analytical Chemistry, 47, 2225 Concentration (μ= g/L) ss (1975). ()ARFV()() 5. Giam, C.S., and Chan, H.S. ‘‘Control of is o Blanks in the Analysis of Phthalates in Air Equation 3 and Ocean Biota Samples,’’ U.S. National Bureau of Standards, Special Publication where: 442, pp. 701–708, 1976. A = Response for the parameter to be meas- s 6. ‘‘Carcinogens—Working with Carcino- ured. gens,’’ Department of Health, Education, and A = Response for the internal standard. is Welfare, Public Health Service, Center for Is = Amount of internal standard added to each extract (μg). Disease Control, National Institute for Occu- V = Volume of water extracted (L). pational Safety and Health, Publication No. o 77–206, August 1977. 13.2 Report results in μg/L without correc- 7. ‘‘OSHA Safety and Health Standards, tion for recovery data. All QC data obtained General Industry,’’ (29 CFR part 1910), Occu- should be reported with the sample results. pational Safety and Health Administration, 14. Method Performance OSHA 2206 (Revised, January 1976). 8. ‘‘Safety in Academic Chemistry Labora- 14.1 The method detection limit (MDL) is tories,’’ American Chemical Society Publica- defined as the minimum concentration of a tion, Committee on Chemical Safety, 3rd substance that can be measured and reported Edition, 1979. with 99% confidence that the value is above 9. Provost L.P., and Elder, R.S. ‘‘Interpre- zero. 1 The MDL concentrations listed in tation of Percent Recovery Data,’’ American Table 1 were obtained using reagent water. 12 Laboratory, 15, 58–63 (1983). (The value 2.44 Similar results were achieved using rep- used in the equation in Section 8.3.3 is two resentative wastewaters. The MDL actually times the value 1.22 derived in this report.) achieved in a given analysis will vary de- pending on instrument sensitivity and ma- 10. ASTM Annual Book of Standards, Part trix effects. 31, D3370–76. ‘‘Standard Practices for Sam- 14.2 This method has been tested for lin- pling Water,’’ American Society for Testing earity of spike recovery from reagent water and Materials, Philadelphia. and has been demonstrated to be applicable 11. Burke, J.A. ‘‘Gas Chromatography for over the concentration range from 5 × MDL Pesticide Residue Analysis; Some Practical to 1000 × MDL with the following exceptions: Aspects,’’ Journal of the Association of Official dimethyl and diethyl phthalate recoveries at Analytical Chemists, 48, 1037 (1965). 1000 × MDL were low (70%); bis-2-ethylhexyl 12. ‘‘Method Detection Limit and Analyt- and di-n-octyl phthalate recoveries at 5 × ical Curve Studies, EPA Methods 606, 607, MDL were low (60%). 12 and 608,’’ Special letter report for EPA Con- 14.3 This method was tested by 16 labora- tract 68–03–2606, U.S. Environmental Protec- tories using reagent water, drinking water, tion Agency, Environmental Monitoring and surface water, and three industrial Support Laboratory, Cincinnati, Ohio 45268, wastewaters spiked at six concentrations June 1980. over the range 0.7 to 106 μg/L. 13 Single oper- 13. ‘‘EPA Method Study 16 Method 606 ator precision, overall precision, and method (Phthalate Esters),’’ EPA 600/4–84–056, Na- accuracy were found to be directly related to tional Technical Information Service, PB84– the concentration of the parameter and es- 211275, Springfield, Virginia 22161, June 1984.

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TABLE 1—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS

Retention time (min) Method de- Parameter tection limit Column 1 Column 2 (μg/L)

Dimethyl phthalate ...... 2 .03 0.95 0 .29 Diethyl phthalate ...... 2 .82 1 .27 0.49 Di-n-butyl phthalate ...... 8.65 3 .50 0 .36 Butyl benzyl phthalate ...... a 6 .94 a 5 .11 0 .34 Bis(2-ethylhexyl) phthalate ...... a 8 .92 a 10 .5 2 .0 Di-n-octyl phthalate ...... a 16 .2 a 18 .0 3 .0 Column 1 conditions: Supelcoport (100/120 mesh) coated with 1.5% SP–2250/1.95% SP–2401 packed in a 1.8 m long × 4 mm ID glass column with 5% methane/95% argon carrier gas at 60 mL/min flow rate. Column temperature held isothermal at 180 °C, except where otherwise indicated. Column 2 conditions: Supelcoport (100/120 mesh) coated with 3% OV–1 packed in a 1.8 m long × 4 mm ID glass column with 5% methane/95% argon carrier gas at 60 mL/min flow rate. Column temperature held isothermal at 200 °C, except where other- wise indicated. a 220 °C column temperature.

TABLE 2—QC ACCEPTANCE CRITERIA—METHOD 606

Test Range for μ Limit for s Range for Parameter conc. ( g/ μ ¯ μ P, Ps L) ( g/L) X ( g/L) (percent)

Bis(2-ethylhexyl) phthalate ...... 50 38.4 1.2–55.9 D–158 Butyl benzyl phthalate ...... 10 4.2 5.7–11.0 30–136 Di-n-butyl phthalate ...... 25 8.9 10.3–29.6 23–136 Diethyl phthalate ...... 25 9.0 1.9–33.4 D–149 Dimethyl phathalate ...... 25 9.5 1.3–35.5 D–156 Di-n-octyl phthalate ...... 50 13.4 D–50.0 D–114 s = Standard deviation of four recovery measurements, in μg/L (Section 8.2.4). X¯ = Average recovery for four recovery measurements, in μg/L (Section 8.2.4). P, Ps = Percent recovery measured (Section 8.3.2, Section 8.4.2). D = Detected; result must be greater than zero. Note: These criteria are based directly upon the method performance data in Table 3. Where necessary, the limits for recovery have been broadened to assure applicability of the limits to concentrations below those used to develop Table 3.

TABLE 3—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 606

Accuracy, as Single analyst ′ ′ Overall preci- Parameter recovery, X precision, sr ′ μ (μg/L) (μg/L) sion, S ( g/L)

Bis(2-ethylhexyl) phthalate ...... 0.53C + 2.02 0.80X¯ ¥2.54 0.73X¯ ¥0.17 Butyl benzyl phthalate ...... 0.82C + 0.13 0.26X¯ + 0.04 0.25X¯ + 0.07 Di-n-butyl phthalate ...... 0.79C + 0.17 0.23X¯ + 0.20 0.29X¯ + 0.06 Diethyl phthalate ...... 0.70C + 0.13 0.27X¯ + 0.05 0.45X¯ + 0.11 Dimethyl phthalate ...... 0.73C + 0.17 0.26X¯ + 0.14 0.44X¯ + 0.31 Di-n-octyl phthalate ...... 0.35C¥0.71 0.38X¯ + 0.71 0.62X¯ + 0.34 X¯ ′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L. sr′ = Expected single analyst standard deviation of measurements at an average concentration found of X¯ , in μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X¯ , in μg/L. C = True value for the concentration, in μg/L. X¯ = Average recovery found for measurements of samples containing a concentration of C, in μg/L.

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COLUMN: 1.5% SP-2250/1.95% SP-2401 ON SUPELCOPORT TEMPERATURE: 180°C DETECTOR; ELECTRON CAPTURE

w.... C( -I C( i: :X: w~ D. 1-C( C-1 -I -IC( ~i: 1-:t: i= • :X:: D. IT D._. -I> c >:t: w-i:~ c:~:c

0 2 4 6 8 10 12 RETENTION TIME, MIN.

Figure 1 . Gas chromatogram of phthalates.

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COLUMN: 1.5% SP-2250/1.95% SP-2401 ON SUPELCOPORT TEMPERATURE: 220"C DETECTOR: ELECTRON CAPTURE .....w .....< < :r::..... :r:: w..... w A. < ..... :::i ..... > < <..... :oc :r:: < w ..... :r:: :r:: :r:: ...... A. :r:: > ..... A. :r::..... > ..... w ..... > u N N. 0 2 en c. w iii ...... = Q ~ =::;)

0 4 8 12 16 18 RETENTION TIME, MIN.

Figure 2. Gas chromatogram of phthalates.

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METHOD 607—NITROSAMINES of 10 mL or less. After the extract has been exchanged to methanol, it is separated by 1. Scope and Application gas chromatography and the parameters are 1.1 This method covers the determination then measured with a nitrogen-phosphorus of certain nitrosamines. The following pa- detector. 4 rameters can be determined by this method: 2.2 The method provides Florisil and alu- mina column cleanup procedures to separate Parameter Storet No. CAS No. diphenylamine from the nitrosamines and to aid in the elimination of interferences that N-Nitrosodimethylamine ...... 34438 62–75–9 may be encountered. N-Nitrosodiphenylamine ...... 34433 86–30–6 N-Nitrosodi-n-propylamine ...... 34428 621–64–7 3. Interferences 1.2 This is a gas chromatographic (GC) 3.1 Method interferences may be caused method applicable to the determination of by contaminants in solvents, reagents, glass- the parameters listed above in municipal and ware, and other sample processing hardware industrial discharges as provided under 40 that lead to discrete artifacts and/or ele- CFR 136.1. When this method is used to ana- vated baselines in gas chromatograms. All of lyze unfamiliar samples for any or all of the these materials must be routinely dem- compmunds above, compound identifications onstrated to be free from interferences under should be supported by at least one addi- the conditions of the analysis by running tional qualitative technique. This method laboratory reagent blanks as described in describes analytical conditimns for a second Section 8.1.3. gas chromatographic column that can be 3.1.1 Glassware must be scrupulously used to confirm measurements made with cleaned. 5 Clean all glassware as soon as pos- the primary column. Method 625 provides gas sible after use by rinsing with the last sol- chromatograph/mass spectrometer (GC/MS) vent used in it. Solvent rinsing should be fol- conditions appropriate for the qualitative lowed by detergent washing with hot water, and quantitative confirmation of results for and rinses with tap water and distilled N-nitrosodi-n-propylamine. In order to con- water. The glassware should then be drained ° firm the presence of N- dry, and heated in a muffle furnace at 400 C nitrosodiphenylamine, the cleanup procedure for 15 to 30 min. Solvent rinses with acetone specified in Section 11.3 or 11.4 must be used. and pesticide quality hexane may be sub- In order to confirm the presence of N- stituted for the muffle furnace heating. Vol- nitrosodimethylamine by GC/MS, Column 1 umetric ware should not be heated in a muf- of this method must be substituted for the fle furnace. After drying and cooling, glass- ware should be sealed and stored in a clean column recommended in Method 625. Con- environment to prevent any accumulation of firmation of these parameters using GC-high dust or other contaminants. Store inverted resolution mass spectrometry or a Thermal or capped with aluminum foil. Energy Analyzer is also recommended. 12 3.1.2 The use of high purity reagents and 1.3 The method detection limit (MDL, de- solvents helps to minimize interference prob- fined in Section 14.1) 3 for each parameter is lems. Purification of solvents by distillation listed in Table 1. The MDL for a specific in all-glass systems may be required. wastewater may differ from those listed, de- 3.2 Matrix interferences may be caused by pending upon the nature of interferences in contaminants that are co-extracted from the the sample matrix. sample. The extent of matrix interferences 1.4 Any modification of this method, be- will vary considerably from source to source, yond those expressly permitted, shall be con- depending upon the nature and diversity of sidered as a major modification subject to the industrial complex or municipality being application and approval of alternate test sampled. The cleanup procedures in Section procedures under 40 CFR 136.4 and 136.5. 11 can be used to overcome many of these 1.5 This method is restricted to use by or interferences, but unique samples may re- under the supervision of analysts experi- quire additional cleanup approaches to enced in the use of a gas chromatograph and achieve the MDL listed in Table 1. in the interpretation of gas chromatograms. 3.3 N-Nitrosodiphenylamine is re- Each analyst must demonstrate the ability ported 6M9 to undergo transnitrosation reac- to generate acceptable results with this tions. Care must be exercised in the heating method using the procedure described in Sec- or concentrating of solutions containing this tion 8.2. compound in the presence of reactive 2. Summary of Method amines. 3.4 The sensitive and selective Thermal 2.1 A measured volume of sample, ap- Energy Analyzer and the reductive Hall de- proximately 1–L, is extracted with meth- tector may be used in place of the nitrogen- ylene chloride using a separatory funnel. The phosphorus detector when interferences are methylene chloride extract is washed with encountered. The Thermal Energy Analyzer dilute hydrochloric acid to remove free offers the highest selectivity of the non-MS amines, dried, and concentrated to a volume detectors.

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4. Safety glass stopper is used to prevent evaporation of extracts. 4.1 The toxicity or carcinogenicity of 5.2.4 Evaporative flask, Kuderna-Danish— each reagent used in this method has not 500-mL (Kontes K–570001–0500 or equivalent). been precisely defined; however, each chem- Attach to concentrator tube with springs. ical compound should be treated as a poten- 5.2.5 Snyder column, Kuderna-Danish— tial health hazard. From this viewpoint, ex- Three-ball macro (Kontes K–503000–0121 or posure to these chemicals must be reduced to equivalent). the lowest possible level by whatever means 5.2.6 Snyder column, Kuderna-Danish— available. The laboratory is responsible for Two-ball micro (Kontes K–569001–0219 or maintaining a current awareness file of equivalent). OSHA regulations regarding the safe han- 5.2.7 Vials—10 to 15-mL, amber glass, with dling of the chemicals specified in this meth- Teflon-lined screw cap. od. A reference file of material data handling 5.2.8 Chromatographic column—Approxi- sheets should also be made available to all mately 400 mm long × 22 mm ID, with Teflon personnel involved in the chemical analysis. stopcock and coarse frit filter disc at bottom Additional references to laboratory safety (Kontes K–420540–0234 or equivalent), for use are available and have been identified 10M12 in Florisil column cleanup procedure. for the information of the analyst. 5.2.9 Chromatographic column—Approxi- 4.2 These nitrosamines are known car- mately 300 mm long × 10 mm ID, with Teflon cinogens, 13M17 therefore, utmost care must stopcock and coarse frit filter disc at bottom be exercised in the handling of these mate- (Kontes K–420540–0213 or equivalent), for use rials. Nitrosamine reference standards and in alumina column cleanup procedure. standard solutions should be handled and 5.3 Boiling chips—Approximately 10/40 prepared in a ventilated glove box within a mesh. Heat to 400 °C for 30 min or Soxhlet ex- properly ventilated room. tract with methylene chloride. 5.4 Water bath—Heated, with concentric 5. Apparatus and Materials ring cover, capable of temperature control (±2 °C). The bath should be used in a hood. 5.1 Sampling equipment, for discrete or 5.5 Balance—Analytical, capable of accu- composite sampling. rately weighing 0.0001 g. 5.1.1 Grab sample bottle—1–L or 1-qt, 5.6 Gas chromatograph—An analytical amber glass, fitted with a screw cap lined system complete with gas chromatograph with Teflon. Foil may be substituted for Tef- suitable for on-column injection and all re- lon if the sample is not corrosive. If amber quired accessories including syringes, ana- bottles are not available, protect samples lytical columns, gases, detector, and strip- from light. The bottle and cap liner must be chart recorder. A data system is rec- washed, rinsed with acetone or methylene ommended for measuring peak areas. chloride, and dried before use to minimize 5.6.1 Column 1—1.8 m long × 4 mm ID contamination. glass, packed with 10% Carbowax 20 M/2% 5.1.2 Automatic sampler (optional)—The KOH on Chromosorb W-AW (80/100 mesh) or sampler must incorporate glass sample con- equivalent. This column was used to develop tainers for the collection of a minimum of the method performance statements in Sec- 250 mL of sample. Sample containers must be tion 14. Guidelines for the use of alternate kept refrigerated at 4 °C and protected from column packings are provided in Section light during compositing. If the sampler uses 12.2. a peristaltic pump, a minimum length of 5.6.2 Column 2—1.8 m long × 4 mm ID compressible silicone rubber tubing may be glass, packed with 10% SP–2250 on Supel- used. Before use, however, the compressible coport (100/120 mesh) or equivalent. tubing should be thoroughly rinsed with 5.6.3 Detector—Nitrogen-phosphorus, re- methanol, followed by repeated rinsings with ductive Hall, or Thermal Energy Analyzer distilled water to minimize the potential for detector. 12 These detectors have proven ef- contamination of the sample. An integrating fective in the analysis of wastewaters for the flowmeter is required to collect flow propor- parameters listed in the scope (Section 1.1). tional composites. A nitrogen-phosphorus detector was used to 5.2 Glassware (All specifications are sug- develop the method performance statements gested. Catalog numbers are included for il- in Section 14. Guidelines for the use of alter- lustration only.): nate detectors are provided in Section 12.2. 5.2.1 Separatory funnels—2–L and 250–mL, with Teflon stopcock. 6. Reagents 5.2.2 Drying column—Chromatographic 6.1 Reagent water—Reagent water is de- column, approximately 400 mm long × 19 mm fined as a water in which an interferent is ID, with coarse frit filter disc. not observed at the MDL of the parameters 5.2.3 Concentrator tube, Kuderna-Dan- of interest. ish—10-mL, graduated (Kontes K–570050–1025 6.2 Sodium hydroxide solution (10 N)— or equivalent). Calibration must be checked Dissolve 40 g of NaOH (ACS) in reagent water at the volumes employed in the test. Ground and dilute to 100 ml.

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6.3 Sodium thiosulfate—(ACS) Granular. 6.11.3 Stock standard solutions must be 6.4 Sulfuric acid (1 + 1)—Slowly, add 50 replaced after six months, or sooner if com- mL of H2SO4 (ACS, sp. gr. 1.84) to 50 mL of parison with check standards indicates a reagent water. problem. 6.5 Sodium sulfate—(ACS) Granular, an- 6.12 Quality control check sample con- hydrous. Purify by heating at 400 °C for 4 h centrate—See Section 8.2.1. in a shallow tray. 6.6 Hydrochloric acid (1 + 9)—Add one vol- 7. Calibration ume of concentrated HCl (ACS) to nine vol- 7.1 Establish gas chromatographic oper- umes of reagent water. ating conditions equivalent to those given in 6.7 Acetone, methanol, methylene chlo- Table 1. The gas chromatographic system ride, pentane—Pesticide quality or equiva- can be calibrated using the external standard lent. technique (Section 7.2) or the internal stand- 6.8 Ethyl ether—Nanograde, redistilled in ard technique (Section 7.3). glass if necessary. 7.2 External standard calibration proce- 6.8.1 Ethyl ether must be shown to be free dure: of peroxides before it is used as indicated by 7.2.1 Prepare calibration standards at a EM Laboratories Quant test strips. (Avail- minimum of three concentration levels for able from Scientific Products Co., Cat No. each parameter of interest by adding vol- P1126–8, and other suppliers.) umes of one or more stock standards to a 6.8.2 Procedures recommended for re- volumetric flask and diluting to volume with moval of peroxides are provided with the test methanol. One of the external standards strips. After cleanup, 20 mL of ethyl alcohol should be at a concentration near, but above, preservative must be added to each liter of the MDL (Table 1) and the other concentra- ether. tions should correspond to the expected 6.9 Florisil—PR grade (60/100 mesh). Pur- range of concentrations found in real sam- chase activated at 1250 °F and store in the ples or should define the working range of dark in glass containers with ground glass the detector. stoppers or foil-lined screw caps. Before use, 7.2.2 Using injections of 2 to 5 μL, analyze activate each batch at least 16 h at 130 °C in each calibration standard according to Sec- a foil-covered glass container and allow to tion 12 and tabulate peak height or area re- cool. sponses against the mass injected. The re- 6.10 Alumina—Basic activity Super I, sults can be used to prepare a calibration W200 series (ICN Life Sciences Group, No. curve for each compound. Alternatively, if 404571, or equivalent). To prepare for use, the ratio of response to amount injected place 100 g of alumina into a 500-mL reagent (calibration factor) is a constant over the bottle and add 2 mL of reagent water. Mix working range (<10% relative standard devi- the alumina preparation thoroughly by ation, RSD), linearity through the origin can shaking or rolling for 10 min and let it stand be assumed and the average ratio or calibra- for at least 2 h. The preparation should be tion factor can be used in place of a calibra- homogeneous before use. Keep the bottle tion curve. sealed tightly to ensure proper activity. 7.3 Internal standard calibration proce- 6.11 Stock standard solutions (1.00 μg/ dure—To use this approach, the analyst must μL)—Stock standard solutions can be pre- select one or more internal standards that pared from pure standard materials or pur- are similar in analytical behavior to the chased as certified solutions. compounds of interest. The analyst must fur- 6.11.1 Prepare stock standard solutions by ther demonstrate that the measurement of accurately weighing about 0.0100 g of pure the internal standard is not affected by material. Dissolve the material in methanol method or matrix interferences. Because of and dilute to volume in a 10-mL volumetric these limitations, no internal standard can flask. Larger volumes can be used at the con- be suggested that is applicable to all sam- venience of the analyst. When compound pu- ples. rity is assayed to be 96% or greater, the 7.3.1 Prepare calibration standards at a weight can be used without correction to cal- minimum of three concentration levels for culate the concentration of the stock stand- each parameter of interest by adding vol- ard. Commercially prepared stock standards umes of one or more stock standards to a can be used at any concentration if they are volumetric flask. To each calibration stand- certified by the manufacturer or by an inde- ard, add a known constant amount of one or pendent source. more internal standards, and dilute to vol- 6.11.2 Transfer the stock standard solu- ume with methanol. One of the standards tions into Teflon-sealed screw-cap bottles. should be at a concentration near, but above, Store at 4 °C and protect from light. Stock the MDL and the other concentrations standard solutions should be checked fre- should correspond to the expected range of quently for signs of degradation or evapo- concentrations found in real samples or ration, especially just prior to preparing should define the working range of the detec- calibration standards from them. tor.

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7.3.2 Using injections of 2 to 5 μL, analyze rations or lower the cost of measurements. each calibration standard according to Sec- Each time such a modification is made to tion 12 and tabulate peak height or area re- the method, the analyst is required to repeat sponses against concentration for each com- the procedure in Section 8.2. pound and internal standard. Calculate re- 8.1.3 Before processing any samples, the sponse factors (RF) for each compound using analyst must analyze a reagent water blank Equation 1. to demonstrate that interferences from the analytical system and glassware are under RF = (As)(Cis (Ais)(Cs) control. Each time a set of samples is ex- Equation 1 tracted or reagents are changed, a reagent water blank must be processed as a safe- where: guard against laboratory contamination. A = Response for the parameter to be meas- s 8.1.4 The laboratory must, on an ongoing ured. basis, spike and analyze a minimum of 10% A = Response for the internal standard. is of all samples to monitor and evaluate lab- C = Concentration of the internal standard is oratory data quality. This procedure is de- (μg/L). scribed in Section 8.3. C = Concentration of the parameter to be s 8.1.5 The laboratory must, on an ongoing measured (μg/L). basis, demonstrate through the analyses of If the RF value over the working range is quality control check standards that the op- a constant (<10% RSD), the RF can be as- eration of the measurement system is in con- sumed to be invariant and the average RF trol. This procedure is described in Section can be used for calculations. Alternatively, 8.4. The frequency of the check standard the results can be used to plot a calibration analyses is equivalent to 10% of all samples curve of response ratios, As/Ais, vs. RF. analyzed but may be reduced if spike recov- 7.4 The working calibration curve, cali- eries from samples (Section 8.3) meet all bration factor, or RF must be verified on specified quality control criteria. each working day by the measurement of one 8.1.6 The laboratory must maintain per- or more calibration standards. If the re- formance records to document the quality of sponse for any parameter varies from the data that is generated. This procedure is de- predicted response by more than ±15%, a new scribed in Section 8.5. calibration curve must be prepared for that 8.2 To establish the ability to generate compound. acceptable accuracy and precision, the ana- 7.5 Before using any cleanup procedure, lyst must perform the following operations. the analyst must process a series of calibra- 8.2.1 A quality control (QC) check sample tion standards through the procedure to vali- concentrate is required containing each pa- date elution patterns and the absence of rameter of interest at a concentration of 20 interferences from the reagents. μg/mL in methanol. The QC check sample concentrate must be obtained from the U.S. 8. Quality Control Environmental Protection Agency, Environ- 8.1 Each laboratory that uses this method mental Monitoring and Support Laboratory is required to operate a formal quality con- in Cincinnati, Ohio, if available. If not avail- trol program. The minimum requirements of able from that source, the QC check sample this program consist of an initial demonstra- concentrate must be obtained from another tion of laboratory capability and an ongoing external source. If not available from either analysis of spiked samples to evaluate and source above, the QC check sample con- document data quality. The laboratory must centrate must be prepared by the laboratory maintain records to document the quality of using stock standards prepared independ- data that is generated. Ongoing data quality ently from those used for calibration. checks are compared with established per- 8.2.2 Using a pipet, prepare QC check sam- formance criteria to determine if the results ples at a concentration of 20 μg/L by adding of analyses meet the performance character- 1.00 mL of QC check sample concentrate to istics of the method. When results of sample each of four 1–L aliquots of reagent water. spikes indicate atypical method perform- 8.2.3 Analyze the well-mixed QC check ance, a quality control check standard must samples according to the method beginning be analyzed to confirm that the measure- in Section 10. ments were performed in an in-control mode 8.2.4 Calculate the average recovery (X¯ ) in of operation. μg/L, and the standard deviation of the re- 8.1.1 The analyst must make an initial, covery (s) in μg/L, for each parameter using one-time, demonstration of the ability to the four results. generate acceptable accuracy and precision 8.2.5 For each parameter compare s and X¯ with this method. This ability is established with the corresponding acceptance criteria as described in Section 8.2. for precision and accuracy, respectively, 8.1.2 In recognition of advances that are found in Table 2. If s and X¯ for all param- occurring in chromatography, the analyst is eters of interest meet the acceptance cri- permitted certain options (detailed in Sec- teria, the system performance is acceptable tions 10.4, 11.1, and 12.2) to improve the sepa- and analysis of actual samples can begin. If

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any individual s exceeds the precision limit using the equation in Table 3, substituting or any individual X¯ falls outside the range the spike concentration (T) for C; (2) cal- for accuracy, the system performance is un- culate overall precision (S′) using the equa- acceptable for that parameter. Locate and tion in Table 3, substituting X′ for X¯ ; (3) cal- correct the source of the problem and repeat culate the range for recovery at the spike the test for all parameters of interest begin- concentration as (100 X′/T) ±2.44(100 S′/T)%. 18 ning with Section 8.2.2. 8.3.4 If any individual P falls outside the 8.3 The laboratory must, on an ongoing designated range for recovery, that param- basis, spike at least 10% of the samples from eter has failed the acceptance criteria. A each sample site being monitored to assess check standard containing each parameter accuracy. For laboratories analyzing one to that failed the criteria must be analyzed as ten samples per month, at least one spiked described in Section 8.4. sample per month is required. 8.4 If any parameter fails the acceptance 8.3.1 The concentration of the spike in the criteria for recovery in Section 8.3, a QC sample should be determined as follows: check standard containing each parameter 8.3.1.1 If, as in compliance monitoring, that failed must be prepared and analyzed. the concentration of a specific parameter in NOTE: The frequency for the required anal- the sample is being checked against a regu- ysis of a QC check standard will depend upon latory concentration limit, the spike should the number of parameters being simulta- be at that limit or 1 to 5 times higher than neously tested, the complexity of the sample the background concentration determined in matrix, and the performance of the labora- Section 8.3.2, whichever concentration would tory. be larger. 8.4.1 Prepare the QC check standard by 8.3.1.2 If the concentration of a specific adding 1.0 mL of QC check sample con- parameter in the sample is not being centrate (Section 8.2.1 or 8.3.2) to 1 L of rea- checked against a limit specific to that pa- rameter, the spike should be at 20 μg/L or 1 gent water. The QC check standard needs to 5 times higher than the background con- only to contain the parameters that failed centration determined in Section 8.3.2, criteria in the test in Section 8.3. whichever concentration would be larger. 8.4.2 Analyze the QC check standard to 8.3.1.3 If it is impractical to determine determine the concentration measured (A) of background levels before spiking (e.g., max- each parameter. Calculate each percent re- imum holding times will be exceeded), the covery (Ps) as 100 (A/T)%, where T is the true spike concentration should be (1) the regu- value of the standard concentration. latory concentration limit, if any; or, if none 8.4.3 Compare the percent recovery (Ps) (2) the larger of either 5 times higher than for each parameter with the corresponding the expected background concentration or 20 QC acceptance criteria found in Table 2. Only μg/L. parameters that failed the test in Section 8.3 8.3.2 Analyze one sample aliquot to deter- need to be compared with these criteria. If mine the background concentration (B) of the recovery of any such parameter falls out- each parameter. If necessary, prepare a new side the designated range, the laboratory QC check sample concentrate (Section 8.2.1) performance for that parameter is judged to appropriate for the background concentra- be out of control, and the problem must be tions in the sample. Spike a second sample immediately identified and corrected. The aliquot with 1.0 mL of the QC check sample analytical result for that parameter in the concentrate and analyze it to determine the unspiked sample is suspect and may not be concentration after spiking (A) of each pa- reported for regulatory compliance purposes. rameter. Calculate each percent recovery (P) 8.5 As part of the QC program for the lab- as 100(A¥B)%/T, where T is the known true oratory, method accuracy for wastewater value of the spike. samples must be assessed and records must 8.3.3 Compare the percent recovery (P) for be maintained. After the analysis of five each parameter with the corresponding QC spiked wastewater samples as in Section 8.3, acceptance criteria found in Table 2. These calculate the average percent recovery (P¯ ) acceptance criteria were caluclated to in- and the standard deviation of the percent re- clude an allowance for error in measurement covery (sp). Express the accuracy assessment ¯ of both the background and spike concentra- as a percent recovery interval from P¥2sp to ¯ ¯ tions, assuming a spike to background ratio P + 2sp. If P = 90% and sp = 10%, for example, of 5:1. This error will be accounted for to the the accuracy interval is expressed as 70– extent that the analyst’s spike to back- 110%. Update the accuracy assessment for ground ratio approaches 5:1. 18 If spiking was each parameter on a regular basis (e.g. after performed at a concentration lower than 20 each five to ten new accuracy measure- μg/L, the analyst must use either the QC ac- ments). ceptance criteria in Table 2, or optional QC 8.6 It is recommended that the laboratory acceptance criteria caluclated for the spe- adopt additional quality assurance practices cific spike concentration. To calculate op- for use with this method. The specific prac- tional acceptance criteria for the recovery of tices that are most productive depend upon a parameter: (1) Calculate accuracy (X′) the needs of the laboratory and the nature of

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the samples. Field duplicates may be ana- sample, but may include stirring, filtration lyzed to assess the precision of the environ- of the emulsion through glass wool, cen- mental measurements. When doubt exists trifugation, or other physical methods. Col- over the identification of a peak on the chro- lect the methylene chloride extract in a 250- matogram, confirmatory techniques such as mL Erlenmeyer flask. gas chromatography with a dissimilar col- 10.3 Add a second 60-mL volume of meth- umn, specific element detector, or mass ylene chloride to the sample bottle and re- spectrometer must be used. Whenever pos- peat the extraction procedure a second time, sible, the laboratory should analyze standard combining the extracts in the Erlenmeyer reference materials and participate in rel- flask. Perform a third extraction in the same evant performance evaluation studies. manner. 10.4 Assemble a Kuderna-Danish (K-D) 9. Sample Collection, Preservation, and concentrator by attaching a 10-mL concen- Handling trator tube to a 500-mL evaporative flask. 9.1 Grab samples must be collected in Other concentration devices or techniques glass containers. Conventional sampling may be used in place of the K-D concentrator if the requirements of Section 8.2 are met. practices 19 should be followed, except that 10.5 Add 10 mL of hydrochloric acid to the the bottle must not be prerinsed with sample combined extracts and shake for 2 min. before collection. Composite samples should Allow the layers to separate. Pour the com- be collected in refrigerated glass containers bined extract through a solvent-rinsed dry- in accordance with the requirements of the ing column containing about 10 cm of anhy- program. Automatic sampling equipment drous sodium sulfate, and collect the extract must be as free as possible of Tygon tubing in the K-D concentrator. Rinse the Erlen- and other potential sources of contamina- meyer flask and column with 20 to 30 mL of tion. methylene chloride to complete the quan- 9.2 All samples must be iced or refrig- ° titative transfer. erated at 4 C from the time of collection 10.6 Add one or two clean boiling chips to until extraction. Fill the sample bottles and, the evaporative flask and attach a three-ball if residual chlorine is present, add 80 mg of Snyder column. Prewet the Snyder column sodium thiosulfate per liter of sample and by adding about 1 mL of methylene chloride mix well. EPA Methods 330.4 and 330.5 may to the top. Place the K-D apparatus on a hot be used for measurement of residual chlo- water bath (60 to 65 °C) so that the concen- 20 rine. Field test kits are available for this trator tube is partially immersed in the hot purpose. If N-nitrosodiphenylamine is to be water, and the entire lower rounded surface determined, adjust the sample pH to 7 to 10 of the flask is bathed with hot vapor. Adjust with sodium hydroxide solution or sulfuric the vertical position of the apparatus and acid. the water temperature as required to com- 9.3 All samples must be extracted within 7 plete the concentration in 15 to 20 min. At days of collection and completely analyzed the proper rate of distillation the balls of the within 40 days of extraction. 4 column will actively chatter but the cham- 9.4 Nitrosamines are known to be light bers will not flood with condensed solvent. sensitive. 7 Samples should be stored in When the apparent volume of liquid reaches amber or foil-wrapped bottles in order to 1 mL, remove the K-D apparatus and allow it minimize photolytic decomposition. to drain and cool for at least 10 min. 10.7 Remove the Snyder column and rinse 10. Sample Extraction the flask and its lower joint into the concen- 10.1 Mark the water meniscus on the side trator tube with 1 to 2 mL of methylene of the sample bottle for later determination chloride. A 5-mL syringe is recommended for of sample volume. Pour the entire sample this operation. Stopper the concentrator into a 2–L separatory funnel. Check the pH tube and store refrigerated if further proc- of the sample with wide-range pH paper and essing will not be performed immediately. If adjust to within the range of 5 to 9 with so- the extract will be stored longer than two dium hydroxide solution or sulfuric acid. days, it should be transferred to a Teflon- 10.2 Add 60 mL of methylene chloride to sealed screw-cap vial. If N- the sample bottle, seal, and shake 30 s to nitrosodiphenylamine is to be measured by rinse the inner surface. Transfer the solvent gas chromatography, the analyst must first to the separatory funnel and extract the use a cleanup column to eliminate sample by shaking the funnel for 2 min with diphenylamine interference (Section 11). If periodic venting to release excess pressure. N-nitrosodiphenylamine is of no interest, the Allow the organic layer to separate from the analyst may proceed directly with gas water phase for a minimum of 10 min. If the chromatographic analysis (Section 12). emulsion interface between layers is more 10.8 Determine the original sample vol- than one-third the volume of the solvent ume by refilling the sample bottle to the layer, the analyst must employ mechanical mark and transferring the liquid to a 1000- techniques to complete the phase separation. mL graduated cylinder. Record the sample The optimum technique depends upon the volume to the nearest 5 mL.

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11. Cleanup and Separation When the apparatus is cool, remove the Sny- der column and rinse the flask and its lower 11.1 Cleanup procedures may not be nec- joint into the concentrator tube with 1 to 2 essary for a relatively clean sample matrix. mL of pentane. Analyze by gas chroma- If particular circumstances demand the use tography (Section 12). of a cleanup procedure, the analyst may use 11.4 Alumina column cleanup for either procedure below or any other appro- nitrosamines: priate procedure. However, the analyst first 11.4.1 Place 12 g of the alumina prepara- must demonstrate that the requirements of tion (Section 6.10) into a 10-mm ID Section 8.2 can be met using the method as chromatographic column. Tap the column to revised to incorporate the cleanup proce- settle the alumina and add 1 to 2 cm of anhy- dure. Diphenylamine, if present in the origi- drous sodium sulfate to the top. nal sample extract, must be separated from 11.4.2 Preelute the column with 10 mL of the nitrosamines if N-nitrosodiphenylamine ethyl ether/pentane (3 + 7)(V/V). Discard the is to be determined by this method. eluate (about 2 mL) and just prior to expo- 11.2 If the entire extract is to be cleaned sure of the sodium sulfate layer to the air, up by one of the following procedures, it quantitatively transfer the 2 mL sample ex- must be concentrated to 2.0 mL. To the con- tract onto the column using an additional 2 centrator tube in Section 10.7, add a clean mL of pentane to complete the transfer. boiling chip and attach a two-ball micro- 11.4.3 Just prior to exposure of the sodium Snyder column. Prewet the column by add- sulfate layer to the air, add 70 mL of ethyl ing about 0.5 mL of methylene chloride to ether/pentane (3 + 7)(V/V). Discard the first the top. Place the micr-K-D apparatus on a 10 mL of eluate. Collect the remainder of the hot water bath (60 to 65 °C) so that the con- eluate in a 500–mL K-D flask equipped with a centrator tube is partially immersed in the 10 mL concentrator tube. This fraction con- hot water. Adjust the vertical position of the tains N-nitrosodiphenylamine and probably a apparatus and the water temperature as re- small amount of N-nitrosodi-n-propylamine. quired to complete the concentration in 5 to 11.4.4 Next, elute the column with 60 mL 10 min. At the proper rate of distillation the of ethyl ether/pentane (1 + 1)(V/V), collecting balls of the column will actively chatter but the eluate in a second K-D flask equipped the chambers will not flood. When the appar- with a 10–mL concentrator tube. Add 15 mL ent volume of liquid reaches about 0.5 mL, of methanol to the K-D flask. This fraction remove the K-D apparatus and allow it to will contain N-nitrosodimethylamine, most drain and cool for at least 10 min. Remove of the N-nitrosodi-n-propylamine and any the micro-Snyder column and rinse its lower diphenylamine that is present. joint into the concentrator tube with 0.2 mL 11.4.5 Concentrate both fractions as in of methylene chloride. Adjust the final vol- Section 10.6, except use pentane to prewet ume to 2.0 mL and proceed with one of the the column. When the apparatus is cool, re- following cleanup procedures. move the Snyder column and rinse the flask 11.3 Florisil column cleanup for and its lower joint into the concentrator nitrosamines: tube with 1 to 2 mL of pentane. Analyze the 11.3.1 Place 22 g of activated Florisil into fractions by gas chromatography (Section a 22-mm ID chromatographic column. Tap 12). the column to settle the Florisil and add about 5 mm of anhydrous sodium sulfate to 12. Gas Chromatography the top. 12.1 N-nitrosodiphenylamine completely 11.3.2 Preelute the column with 40 mL of reacts to form diphenylamine at the normal ethyl ether/pentane (15 + 85)(V/V). Discard operating temperatures of a GC injection the eluate and just prior to exposure of the port (200 to 250 °C). Thus, N- sodium sulfate layer to the air, quan- nitrosodiphenylamine is chromatographed titatively transfer the 2-mL sample extract and detected as diphenylamine. Accurate de- onto the column using an additional 2 mL of termination depends on removal of pentane to complete the transfer. diphenylamine that may be present in the 11.3.3 Elute the column with 90 mL of original extract prior to GC analysis (See ethyl ether/pentane (15 + 85)(V/V) and discard Section 11). the eluate. This fraction will contain the 12.2 Table 1 summarizes the recommended diphenylamine, if it is present in the extract. operating conditions for the gas chro- 11.3.4 Next, elute the column with 100 mL matograph. Included in this table are reten- of acetone/ethyl ether (5 + 95)(V/V) into a 500- tion times and MDL that can be achieved mL K-D flask equipped with a 10-mL concen- under these conditions. Examples of the sep- trator tube. This fraction will contain all of arations achieved by Column 1 are shown in the nitrosamines listed in the scope of the Figures 1 and 2. Other packed or capillary method. (open-tubular) columns, chromatographic 11.3.5 Add 15 mL of methanol to the col- conditions, or detectors may be used if the lected fraction and concentrate as in Section requirements of Section 8.2 are met. 10.6, except use pentane to prewet the col- 12.3 Calibrate the system daily as de- umn and set the water bath at 70 to 75 °C. scribed in Section 7.

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12.4 If the extract has not been subjected using the calibration curve or calibration to one of the cleanup procedures in Section factor determined in Section 7.2.2. The con- 11, it is necessary to exchange the solvent centration in the sample can be calculated from methylene chloride to methanol before from Equation 2. the thermionic detector can be used. To a 1 to 10-mL volume of methylene chloride ex- ()AV() tract in a concentrator tube, add 2 mL of Concentration (μ= g/L) t methanol and a clean boiling chip. Attach a ()VV() two-ball micro-Snyder column to the con- is centrator tube. Prewet the column by adding Equation 2 about 0.5 mL of methylene chloride to the top. Place the micro-K-D apparatus on a where: boiling (100 °C) water bath so that the con- A = Amount of material injected (ng). μ centrator tube is partially immersed in the Vi = Volume of extract injected ( L). μ hot water. Adjust the vertical position of the Vt = Volume of total extract ( L). apparatus and the water temperature as re- Vs = Volume of water extracted (mL). quired to complete the concentration in 5 to 13.1.2 If the internal standard calibration 10 min. At the proper rate of distillation the procedure is used, calculate the concentra- balls of the column will actively chatter but tion in the sample using the response factor the chambers will not flood. When the appar- (RF) determined in Section 7.3.2 and Equa- ent volume of liquid reaches about 0.5 mL, tion 3. remove the K-D apparatus and allow it to drain and cool for at least 10 min. Remove ()()ACsis the micro-Snyder column and rinse its lower RF = joint into the concentrator tube with 0.2 mL ()()ACis s of methanol. Adjust the final volume to 2.0 mL. Equation 3 12.5 If the internal standard calibration where: procedure is being used, the internal stand- A = Response for the parameter to be meas- ard must be added to the sample extract and s ured. mixed thoroughly immediately before injec- A = Response for the internal standard. tion into the gas chromatograph. is I = Amount of internal standard added to 12.6 Inject 2 to 5 μL of the sample extract s each extract (μg). or standard into the gas chromatograph V = Volume of water extracted (L). using the solvent-flush technique. 21 Smaller o (1.0 μL) volumes may be injected if auto- 13.2 Report results in μg/L without correc- matic devices are employed. Record the vol- tion for recovery data. All QC data obtained ume injected to the nearest 0.05 μL, and the should be reported with the sample results. resulting peak size in area or peak height 14. Method Performance units. 12.7 Identify the parameters in the sample 14.1 The method detection limit (MDL) is by comparing the retention times of the defined as the minimum concentration of a peaks in the sample chromatogram with substance that can be measured and reported those of the peaks in standard with 99% confidence that the value is above chromatograms. The width of the retention zero. 3 The MDL concentrations listed in time window used to make identifications Table 1 were obtained using reagent water. 22 should be based upon measurements of ac- Similar results were achieved using rep- tual retention time variations of standards resentative wastewaters. The MDL actually over the course of a day. Three times the achieved in a given analysis will vary de- standard deviation of a retention time for a pending on instrument sensitivity and ma- compound can be used to calculate a sug- trix effects. gested window size; however, the experience 14.2 This method has been tested for lin- of the analyst should weigh heavily in the earity of spike recovery from reagent water interpretation of chromatograms. and has been demonstrated to be applicable 12.8 If the response for a peak exceeds the over the concentration range from 4 × MDL working range of the system, dilute the ex- to 1000 × MDL. 22 tract and reanalyze. 14.3 This method was tested by 17 labora- 12.9 If the measurement of the peak re- tories using reagent water, drinking water, sponse is prevented by the presence of inter- surface water, and three industrial ferences, further cleanup is required. wastewaters spiked at six concentrations over the range 0.8 to 55 μg/L. 23 Single oper- 13. Calculations ator precision, overall precision, and method 13.1 Determine the concentration of indi- accuracy were found to be directly related to vidual compounds in the sample. the concentration of the parameter and es- 13.1.1 If the external standard calibration sentially independent of the sample matrix. procedure is used, calculate the amount of Linear equations to describe these relation- material injected from the peak response ships are presented in Table 3.

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References 12. ‘‘Safety in Academic Chemistry Labora- tories,’’ American Chemical Society Publica- 1. Fine, D.H., Lieb, D., and Rufeh, R. tion, Committee on Chemical Safety, 3rd ‘‘Principle of Operation of the Thermal En- Edition, 1979. ergy Analyzer for the Trace Analysis of 13. Lijinsky, W. ‘‘How Nitrosamines Cause Volatile and Non-volatile N-nitroso Com- Cancer,’’ New Scientist, 73, 216 (1977). pounds,’’ Journal of Chromatography, 107, 351 14. Mirvish, S.S. ‘‘N-Nitroso compounds: (1975). Their Chemical and in vivo Formation and 2. Fine, D.H., Hoffman, F., Rounbehler, Possible Importance as Environmental Car- D.P., and Belcher, N.M. ‘‘Analysis of N-nitro- cinogens,’’ J. Toxicol. Environ. Health, 3, 1267 so Compounds by Combined High Perform- (1977). ance Liquid Chromatography and Thermal 15. ‘‘Reconnaissance of Environmental Lev- Energy Analysis,’’ Walker, E.A., Bogovski, els of Nitrosamines in the Central United P. and Griciute, L., Editors, N-nitroso Com- States,’’ EPA–330/1–77–001, National Enforce- pounds—Analysis and Formation, Lyon, ment Investigations Center, U.S. Environ- International Agency for Research on Cancer mental Protection Agency (1977). (IARC Scientific Publications No. 14), pp. 43– 16. ‘‘Atmospheric Nitrosamine Assessment 50 (1976). Report,’’ Office of Air Quality Planning and 3. 40 CFR part 136, appendix B. Standards, U.S. Environmental Protection 4. ‘‘Determination of Nitrosamines in In- Agency, Research Triangle Park, North dustrial and Municipal Wastewaters,’’ EPA Carolina (1976). 600/4–82–016, National Technical Information 17. ‘‘Scientific and Technical Assessment Service, PB82–199621, Springfield, Virginia Report on Nitrosamines,’’ EPA–660/6–7–001, 22161, April 1982. Office of Research and Development, U.S. 5. ASTM Annual Book of Standards, Part Environmental Protection Agency (1976). 31, D3694–78. ‘‘Standard Practices for Prepa- 18. Provost, L.P., and Elder, R.S. ‘‘Inter- ration of Sample Containers and for Preser- pretation of Percent Recovery Data,’’ Amer- vation of Organic Constituents,’’ American ican Laboratory, 15, 58–63 (1983). (The value Society for Testing and Materials, Philadel- 2.44 used in the equation in Section 8.3.3 is phia. two times the value of 1.22 derived in this re- 6. Buglass, A.J., Challis, B.C., and Osborn, port.) M.R. ‘‘Transnitrosation and Decomposition 19. ASTM Annual Book of Standards, Part of Nitrosamines,’’ Bogovski, P. and Walker, 31, D3370–76. ‘‘Standard Practices for Sam- E.A., Editors, N-nitroso Compounds in the pling Water,’’ American Society for Testing Environment, Lyon, International Agency and Materials, Philadelphia. for Research on Cancer (IARC Scientific 20. ‘‘Methods 330.4 (Titrimetric, DPD-FAS) Publication No. 9), pp. 94–100 (1974). and 330.5 (Spectrophotometric, DPD) for 7. Burgess, E.M., and Lavanish, J.M. ‘‘Pho- Chlorine, Total Residual,’’ Methods for tochemical Decomposition of N- Chemical Analysis of Water and Wastes, nitrosamines,’’ Tetrahedon Letters, 1221 (1964) EPA–600/4–79–020, U.S. Environmental Pro- 8. Druckrey, H., Preussmann, R., tection Agency, Environmental Monitoring Ivankovic, S., and Schmahl, D. ‘‘Organotrope and Support Laboratory, Cincinnati, Ohio Carcinogene Wirkungen bei 65 Verschiedenen 45268, March 1979. N-NitrosoVerbindungen an BD-Ratten,’’ Z. 21. Burke, J. A. ‘‘Gas Chromatography for Krebsforsch., 69, 103 (1967). Pesticide Residue Analysis; Some Practical 9. Fiddler, W. ‘‘The Occurrence and Deter- Aspects,’’ Journal of the Association of Official mination of N-nitroso Compounds,’’ Toxicol. Analytical Chemists, 48, 1037 (1965). Appl. Pharmacol., 31, 352 (1975). 22. ‘‘Method Detection Limit and Analyt- 10. ‘‘Carcinogens—Working With Carcino- ical Curve Studies EPA Methods 606, 607, and gens,’’ Department of Health, Education, and 608,’’ Special letter report for EPA Contract Welfare, Public Health Service, Center for 68–03–2606, U.S. Environmental Protection Disease Control, National Institute for Occu- Agency, Environmental Monitoring and Sup- pational Safety and Health, Publication No. port Laboratory, Cincinnati, Ohio 45268, 77–206, August 1977. June 1980. 11. ‘‘OSHA Safety and Health Standards, 23. ‘‘EPA Method Study 17 Method 607— General Industry,’’ (29 CFR Part 1910), Occu- Nitrosamines,’’ EPA 600/4–84–051, National pational Safety and Health Administration, Technical Information Service, PB84–207646, OSHA 2206 (Revised, January 1976). Springfield, Virginia 22161, June 1984.

TABLE 1—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS

Retention time (min) Method de- Parameter tection limit Column 1 Column 2 (μg/L)

N-Nitrosodimethylamine ...... 4.1 0.88 0.15 N-Nitrosodi-n-propylamine ...... 12.1 4.2 .46

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TABLE 1—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS—Continued

Retention time (min) Method de- Parameter tection limit Column 1 Column 2 (μg/L)

N-Nitrosodiphenylamine a ...... b 12.8 c 6.4 .81 Column 1 conditions: Chromosorb W-AW (80/100 mesh) coated with 10% Carbowax 20 M/2% KOH packed in a 1.8 m long × 4mm ID glass column with helium carrier gas at 40 mL/min flow rate. Column temperature held isothermal at 110 °C, except where otherwise indicated. Column 2 conditions: Supelcoport (100/120 mesh) coated with 10% SP–2250 packed in a 1.8 m long × 4 mm ID glass column with helium carrier gas at 40 mL/min flow rate. Column temperature held isothermal at 120 °C, except where otherwise indicated. a Measured as diphenylamine. b 220 °C column temperature. c 210 °C column temperature.

TABLE 2—QC ACCEPTANCE CRITERIA—METHOD 607

Test conc. Limit for s Range for X¯ Range for Parameter μ μ μ P, Ps (per- ( g/L) ( g/L) ( g/L) cent)

N-Nitrosodimethylamine ...... 20 3.4 4.6–20.0 13–109 N-Nitrosodiphenyl ...... 20 6.1 2.1–24.5 D–139 N-Nitrosodi-n-propylamine ...... 20 5.7 11.5–26.8 45–146 s = Standard deviation for four recovery measurements, in μg/L (Section 8.2.4). X¯ = Average recovery for four recovery measurements, in μg/L (Section 8.2.4). P, Ps = Percent recovery measured (Section 8.3.2, Section 8.4.2). D = Detected; result must be greater than zero. NOTE: These criteria are based directly upon the method performance data in Table 3. Where necessary, the limits for recov- ery have been broadened to assure applicability of the limits to concentrations below those used to develop Table 3.

TABLE 3—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 607

Accuracy, as Single analyst ′ ′ Overall preci- Parameter recovery, X precision, sr ′ μ (μg/L) (μg/L) sion, S ( g/L)

N-Nitrosodimethylamine ...... 0.37C + 0.06 0.25X¯ ¥0.04 0.25X¯ + 0.11 N-Nitrosodiphenylamine ...... 0.64C + 0.52 0.36X¯ ¥1.53 0.46X¯ ¥0.47 N-Nitrosodi-n-propylamine ...... 0.96C¥0.07 0.15X¯ + 0.13 0.21X¯ + 0.15 X′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L. sr′ = Expected single analyst standard deviation of measurements at an average concentration found of X¯ , in μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X¯ , in μg/L. C = True value for the concentration, in μg/L. X¯ = Average recovery found for measurements of samples containing a concentration of C, in μg/L.

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COLUMN: 10% CARBOWAX 20M I 2% KOH ON CHROMOSORB W-AW TEMPERATURE: 110°C DETECTOR: PHOSPHORUS/NITROGEN w 2 :::!: <(.... > ::J: 1- w :::!: w c 2 0 U) :::!: 0 a: s !::: >a. 0 2 a: 2' a. c' c' 0 U) 0 a: !::: 2 2'

2 4 6 8 10 12 14 RETENTION TIME, MIN.

Figure 1. Gas chromatogram of nitrosamines.

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COLUMN: 10% CARBOWAX 20M /2% KOH ON CHROMOSORB W-AW TEMPERATURE: 220°C DETECTOR: PHOSPHORUS/NITROGEN

0 2 4 6 8 10 12 14 16 18 RETENTION TIME, MIN.

Figure 2. Gas chromatogram of N-nitrosodiphenylamine as diphenylamine.

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METHOD 608.3—ORGANOCHLORINE PESTICIDES be by the individual isomers, or as the sum AND PCBS BY GC/HSD of the concentrations of these isomers, as re- quested or required by a regulatory/control 1. Scope and Application authority or in a permit. Technical 1.1 This method is for determination of Chlordane is listed in Table 2 and may be organochlorine pesticides and poly- used in cases where historical reporting has chlorinated biphenyls (PCBs) in industrial only been the Technical Chlordane. discharges and other environmental samples Toxaphene and the PCBs have been moved by gas chromatography (GC) combined with from Table 1 to Table 2 (Additional a halogen-specific detector (HSD; e.g., elec- Analytes) to distinguish these analytes from tron capture, electrolytic conductivity), as the analytes required in quality control tests provided under 40 CFR 136.1. This revision is (Table 1). QC acceptance criteria for based on a previous protocol (Reference 1), Toxaphene and the PCBs have been retained on the revision promulgated October 26, 1984, in Table 4 and may continue to be applied if on an inter-laboratory method validation desired, or if these analytes are requested or study (Reference 2), and on EPA Method 1656 required by a regulatory/control authority or (Reference 16). The analytes that may be in a permit. Method 1668C (Reference 17) may qualitatively and quantitatively determined be useful for determination of PCBs as indi- using this method and their CAS Registry vidual chlorinated biphenyl congeners, and numbers are listed in Table 1. Method 1699 (Reference 18) may be useful for 1.2 This method may be extended to de- determination of the pesticides listed in this termine the analytes listed in Table 2. How- method. However, at the time of writing of ever, extraction or gas chromatography chal- this revision, Methods 1668C and 1699 had not lenges for some of these analytes may make been approved for use at 40 CFR part 136. quantitative determination difficult. 1.6 Method detection limits (MDLs; Ref- 1.3 When this method is used to analyze erence 3) for the analytes in Tables 1 and unfamiliar samples for an analyte listed in some of the analytes in Table 2 are listed in Table 1 or Table 2, analyte identification those tables. These MDLs were determined must be supported by at least one additional in reagent water (Reference 3). Advances in qualitative technique. This method gives an- analytical technology, particularly the use alytical conditions for a second GC column of capillary (open-tubular) columns, allowed that can be used to confirm and quantify laboratories to routinely achieve MDLs for measurements. Additionally, Method 625.1 the analytes in this method that are 2–10 provides gas chromatograph/mass spectrom- times lower than those in the version pro- eter (GC/MS) conditions appropriate for the mulgated in 1984. The MDL for an analyte in qualitative confirmation of results for the a specific wastewater may differ from those analytes listed in Tables 1 and 2 using the listed, depending upon the nature of inter- extract produced by this method, and Meth- ferences in the sample matrix. od 1699 (Reference 18) provides high resolu- 1.6.1 EPA has promulgated this method at tion GC/MS conditions for qualitative con- 40 CFR part 136 for use in wastewater com- firmation of results using the original sam- pliance monitoring under the National Pol- ple. When such methods are used to confirm lutant Discharge Elimination System the identifications of the target analytes, (NPDES). The data reporting practices de- the quantitative results should be derived scribed in section 15.6 are focused on such from the procedure with the calibration monitoring needs and may not be relevant to range and sensitivity that are most appro- other uses of the method. priate for the intended application. 1.6.2 This method includes ‘‘reporting lim- 1.4 The large number of analytes in Ta- its’’ based on EPA’s ‘‘minimum level’’ (ML) bles 1 and 2 makes testing difficult if all concept (see the glossary in section 23). Ta- analytes are determined simultaneously. bles 1 and 2 contain MDL values and ML val- Therefore, it is necessary to determine and ues for many of the analytes. perform quality control (QC) tests for the 1.7 The separatory funnel and continuous ‘‘analytes of interest’’ only. The analytes of liquid-liquid sample extraction and con- interest are those required to be determined centration steps in this method are essen- by a regulatory/control authority or in a per- tially the same as those steps in Methods mit, or by a client. If a list of analytes is not 606, 609, 611, and 612. Thus, a single sample specified, the analytes in Table 1 must be de- may be extracted to measure the analytes termined, at a minimum, and QC testing included in the scope of each of these meth- must be performed for these analytes. The ods. Samples may also be extracted using a analytes in Table 1 and some of the analytes disk-based solid-phase extraction (SPE) pro- in Table 2 have been identified as Toxic Pol- cedure developed by the 3M Corporation and lutants (40 CFR 401.15), expanded to a list of approved by EPA as an Alternate Test Pro- Priority Pollutants (40 CFR part 423, appen- cedure (ATP) for wastewater analyses in 1995 dix A). (Reference 20). 1.5 In this revision to Method 608, 1.8 This method is performance-based. It Chlordane has been listed as the alpha- and may be modified to improve performance gamma- isomers in Table 1. Reporting may (e.g., to overcome interferences or improve

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the accuracy of results) provided all per- tion process in a given 24-hour period, to a formance requirements are met. maximum of 20 samples—see Glossary for de- 1.8.1 Examples of allowed method modi- tailed definition), as described in section 8.5. fications are described at 40 CFR 136.6. Other Specific selection of reagents and purifi- examples of allowed modifications specific to cation of solvents by distillation in all-glass this method are described in section 8.1.2. systems may be required. Where possible, 1.8.2 Any modification beyond those ex- labware is cleaned by extraction or solvent pressly permitted at 40 CFR 136.6 or in sec- rinse, or baking in a kiln or oven. tion 8.1.2 of this method shall be considered 3.2 Glassware must be scrupulously a major modification subject to application cleaned (Reference 4). Clean all glassware as and approval of an alternate test procedure soon as possible after use by rinsing with the under 40 CFR 136.4 and 136.5. last solvent used in it. Solvent rinsing 1.8.3 For regulatory compliance, any should be followed by detergent washing modification must be demonstrated to with hot water, and rinses with tap water produce results equivalent or superior to re- and reagent water. The glassware should sults produced by this method when applied then be drained dry, and heated at 400 °C for to relevant wastewaters (section 8.1.2). 15–30 minutes. Some thermally stable mate- 1.9 This method is restricted to use by or rials, such as PCBs, may require higher tem- under the supervision of analysts experi- peratures and longer baking times for re- enced in the use of GC/HSD. The laboratory moval. Solvent rinses with pesticide quality must demonstrate the ability to generate ac- acetone, hexane, or other solvents may be ceptable results with this method using the substituted for heating. Do not heat volu- procedure in section 8.2. metric labware above 90 °C. After drying and 1.10 Terms and units of measure used in cooling, store inverted or capped with sol- this method are given in the glossary at the vent-rinsed or baked aluminum foil in a end of the method. clean environment to prevent accumulation of dust or other contaminants. 2. Summary of Method 3.3 Interferences by phthalate esters can pose a major problem in pesticide analysis 2.1 A measured volume of sample, the when using the electron capture detector. amount required to meet an MDL or report- The phthalate esters generally appear in the ing limit (nominally 1–L), is extracted with chromatogram as large late eluting peaks, methylene chloride using a separatory fun- especially in the 15 and 50% fractions from nel, a continuous liquid/liquid extractor, or Florisil®. Common flexible plastics contain disk-based solid-phase extraction equipment. varying amounts of phthalates that may be The extract is dried and concentrated for extracted or leached from such materials cleanup, if required. After cleanup, or if during laboratory operations. Cross contami- cleanup is not required, the extract is ex- nation of clean glassware routinely occurs changed into an appropriate solvent and con- when plastics are handled during extraction centrated to the volume necessary to meet steps, especially when solvent-wetted sur- the required compliance or detection limit, faces are handled. Interferences from and analyzed by GC/HSD. phthalates can best be minimized by avoid- 2.2 Qualitative identification of an ing use of non-fluoropolymer plastics in the analyte in the extract is performed using the laboratory. Exhaustive cleanup of reagents retention times on dissimilar GC columns. and glassware may be required to eliminate Quantitative analysis is performed using the background phthalate contamination (Ref- peak areas or peak heights for the analyte erences 5 and 6). Interferences from phthal- on the dissimilar columns with either the ex- ate esters can be avoided by using a ternal or internal standard technique. microcoulometric or electrolytic conduc- ® 2.3 Florisil , alumina, a C18 solid-phase tivity detector. cleanup, and an elemental cleanup 3.4 Matrix interferences may be caused by procedure are provided to aid in elimination contaminants co-extracted from the sample. of interferences that may be encountered. The extent of matrix interferences will vary Other cleanup procedures may be used if considerably from source to source, depend- demonstrated to be effective for the analytes ing upon the nature and diversity of the in- in a wastewater matrix. dustrial complex or municipality being sam- pled. Interferences extracted from samples 3. Contamination and Interferences high in total organic carbon (TOC) may re- 3.1 Solvents, reagents, glassware, and sult in elevated baselines, or by enhancing or other sample processing lab ware may yield suppressing a signal at or near the retention artifacts, elevated baselines, or matrix inter- time of an analyte of interest. Analyses of ferences causing misinterpretation of the matrix spike and matrix spike duplicate chromatograms. All materials used in the (Section 8.3) may be useful in identifying analysis must be demonstrated free from matrix interferences, and the cleanup proce- contamination and interferences by running dures in Section 11 may aid in eliminating blanks initially and with each extraction these interferences. EPA has provided guid- batch (samples started through the extrac- ance that may aid in overcoming matrix

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interferences (Reference 7); however, unique chloride, and dried before use to minimize samples may require additional cleanup ap- contamination. proaches to achieve the MDLs listed in Ta- 5.1.2 Automatic sampler (optional)—The bles 1 and 2. sampler must use a glass or fluoropolymer container and tubing for sample collection. 4. Safety If the sampler uses a peristaltic pump, a 4.1 Hazards associated with each reagent minimum length of compressible silicone used in this method have not been precisely rubber tubing may be used. Before use, rinse defined; however, each the compressible tubing thoroughly with should be treated as a potential health haz- methanol, followed by repeated rinsing with ard. From this viewpoint, exposure to these reagent water to minimize the potential for chemicals must be reduced to the lowest pos- sample contamination. An integrating flow sible level by whatever means available. The meter is required to collect flow propor- laboratory is responsible for maintaining a tional composites. The sample container ≤ ° current awareness file of OSHA regulations must be kept refrigerated at 6 C and pro- regarding the safe handling of the chemicals tected from light during compositing. specified in this method. A reference file of 5.2. Lab ware. safety data sheets (SDSs, OSHA, 29 CFR 5.2.1 Extraction. 1910.12009(g)) should also be made available 5.2.1.1 pH measurement. to all personnel involved in sample handling 5.2.1.1.1 pH meter, with combination glass and chemical analysis. Additional references electrode. to laboratory safety are available and have 5.2.1.1.2 pH paper, wide range (Hydrion been identified (References 8 and 9) for the Papers, or equivalent). information of the analyst. 5.2.1.2 Separatory funnel—Size appro- 4.2 The following analytes covered by this priate to hold the sample and extraction sol- method have been tentatively classified as vent volumes, equipped with fluoropolymer known or suspected human or mammalian stopcock. carcinogens: 4,4′-DDT, 4,4′-DDD, the BHCs, 5.2.1.3 Continuous liquid-liquid extrac- and the PCBs. Primary standards of these tor—Equipped with fluoropolymer or glass toxic analytes should be prepared in a chem- connecting joints and stopcocks requiring no ical fume hood, and a NIOSH/MESA approved lubrication. (Hershberg-Wolf Extractor, Ace toxic gas respirator should be worn when Glass Company, Vineland, NJ, or equiva- high concentrations are handled. lent.) 4.3 This method allows the use of hydro- 5.2.1.3.1 Round-bottom flask, 500-mL, with gen as a carrier gas in place of helium (sec- heating mantle. tion 5.8.2). The laboratory should take the 5.2.1.3.2 Condenser, Graham, to fit extrac- necessary precautions in dealing with hydro- tor. gen, and should limit hydrogen flow at the 5.2.1.4 Solid-phase extractor—90-mm filter source to prevent buildup of an explosive apparatus (Figure 2) or multi-position mani- mixture of hydrogen in air. fold. NOTE: The approved ATP for solid-phase 5. Apparatus and Materials extraction is limited to disk-based extrac- NOTE: Brand names and suppliers are for il- tion media and associated peripheral equip- lustration purposes only. No endorsement is ment. implied. Equivalent performance may be 5.2.1.4.1 Vacuum system—Capable of achieved using equipment and materials achieving 0.1 bar (25 inch) Hg (house vacuum, other than those specified here. Dem- vacuum pump, or water aspirator), equipped onstrating that the equipment and supplies with shutoff valve and vacuum gauge. used in the laboratory achieve the required 5.2.1.4.2 Vacuum trap—Made from 500-mL performance is the responsibility of the lab- sidearm flask fitted with single-hole rubber oratory. Suppliers for equipment and mate- stopper and glass tubing. rials in this method may be found through 5.2.2 Filtration. an on-line search. Please do not contact EPA 5.2.2.1 Glass powder funnel, 125- to 250-mL. for supplier information. 5.2.2.2 Filter paper for above, Whatman 41, 5.1 Sampling equipment, for discrete or or equivalent. composite sampling. 5.2.2.3 Prefiltering aids—90-mm 1-μm glass 5.1.1 Grab sample bottle—Amber glass fiber filter or Empore® Filter Aid 400. bottle large enough to contain the necessary 5.2.3 Drying column. sample volume (nominally 1 L), fitted with a 5.2.3.1 Chromatographic column—Ap- fluoropolymer-lined screw cap. Foil may be proximately 400 mm long x 15 mm ID, with substituted for fluoropolymer if the sample fluoropolymer stopcock and coarse frit filter is not corrosive. If amber bottles are not disc (Kontes or equivalent). available, protect samples from light. Unless 5.2.3.2 Glass wool—Pyrex, extracted with pre-cleaned, the bottle and cap liner must be methylene chloride or baked at 450 °C for 1 washed, rinsed with acetone or methylene hour minimum.

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5.2.4 Column for Florisil® or alumina decyl (C18) bonded silica uniformly enmeshed cleanup—Approximately 300 mm long x 10 in a matrix of inert PTFE fibrils (3M mm ID, with fluoropolymer stopcock. (This Empore® or equivalent). The disks should column is not required if cartridges con- not contain any organic compounds, either taining Florisil® are used.) from the PTFE or the bonded silica, which 5.2.5 Concentration/evaporation. will leach into the methylene chloride NOTE: Use of a solvent recovery system eluant. One liter of reagent water should with the K–D or other solvent evaporation pass through the disks in 2–5 minutes, using apparatus is strongly recommended. a vacuum of at least 25 inches of mercury. 5.2.5.1 Kuderna-Danish concentrator. NOTE: Extraction disks from other manu- 5.2.5.1.1 Concentrator tube, Kuderna-Dan- facturers may be used in this procedure, pro- ish—10-mL, graduated (Kontes or equiva- vided that they use the same solid-phase ma- lent). Calibration must be checked at the terials (i.e., octadecyl bonded silica). Disks volumes employed for extract volume meas- of other diameters also may be used, but urement. A ground-glass stopper is used to may adversely affect the flow rate of the prevent evaporation of extracts. sample through the disk. 5.2.5.1.2 Evaporative flask, Kuderna-Dan- 5.3 Vials. ish—500-mL (Kontes or equivalent). Attach 5.3.1 Extract storage—10- to 15-mL, amber to concentrator tube with connectors. glass, with fluoropolymer-lined screw cap. 5.2.5.1.3 Snyder column, Kuderna/Danish— 5.3.2 GC autosampler—1- to 5-mL, amber Three-ball macro (Kontes or equivalent). glass, with fluoropolymer-lined screw- or 5.2.5.1.4 Snyder column—Two-ball micro crimp-cap, to fit GC autosampler. (Kontes or equivalent). 5.4 Balances. 5.2.5.1.5 Water bath—Heated, with concen- 5.4.1 Analytical—Capable of accurately tric ring cover, capable of temperature con- weighing 0.1 mg. ± ° trol ( 2 C), installed in a hood using appro- 5.4.2 Top loading—Capable of weighing 10 priate engineering controls to limit exposure mg. to solvent vapors. 5.5 Sample cleanup. 5.2.5.2 Nitrogen evaporation device— 5.5.1 Oven—For baking and storage of ad- Equipped with heated bath that can be main- sorbents, capable of maintaining a constant tained at an appropriate temperature for the temperature (±5 °C) in the range of 105–250 °C. solvent and analytes. (N-Evap, 5.5.2 Muffle furnace—Capable of cleaning Organomation Associates, Inc., or equiva- glassware or baking sodium sulfate in the lent). range of 400–450 °C. 5.2.5.3 Rotary evaporator—Buchi/ 5.5.3 Vacuum system and cartridges for Brinkman-American Scientific or equiva- solid-phase cleanup (see Section 11.2). lent, equipped with a variable temperature 5.5.3.1 Vacuum system—Capable of water bath, vacuum source with shutoff achieving 0.1 bar (25 in.) Hg (house vacuum, valve at the evaporator, and vacuum gauge. vacuum pump, or water aspirator), equipped 5.2.5.3.1 A recirculating water pump and with shutoff valve and vacuum gauge. chiller are recommended, as use of tap water 5.5.3.2 VacElute Manifold (Analytichem for cooling the evaporator wastes large vol- International, or equivalent). umes of water and can lead to inconsistent 5.5.3.3 Vacuum trap—Made from 500-mL performance as water temperatures and pres- sidearm flask fitted with single-hole rubber sures vary. stopper and glass tubing. 5.2.5.3.2 Round-bottom flask—100-mL and 5.5.3.4 Rack for holding 50-mL volumetric 500-mL or larger, with ground-glass fitting flasks in the manifold. compatible with the rotary evaporator 5.5.3.5 Cartridge—Mega Bond Elute, Non- NOTE: This equipment is used to prepare polar, C18 Octadecyl, 10 g/60 mL copper foil or copper powder for removing (Analytichem International or equivalent), sulfur from sample extracts (see Section used for solid-phase cleanup of sample ex- 6.7.4). tracts (see Section 11.2). 5.2.5.4 Automated concentrator—Equipped 5.5.4 Sulfur removal tube—40- to 50-mL with glassware sufficient to concentrate 3– bottle, test tube, or Erlenmeyer flask with 400 mL extract to a final volume of 1–10 mL fluoropolymer-lined screw cap. under controlled conditions of temperature 5.6 Centrifuge apparatus. and nitrogen flow (Turbovap, or equivalent). 5.6.1 Centrifuge—Capable of rotating 500- Follow manufacturer’s directions and re- mL centrifuge bottles or 15-mL centrifuge quirements. tubes at 5,000 rpm minimum. 5.2.5.5 Boiling chips—Glass, silicon car- 5.6.2 Centrifuge bottle—500-mL, with bide, or equivalent, approximately 10/40 screw cap, to fit centrifuge. mesh. Heat at 400 °C for 30 minutes, or sol- 5.6.3 Centrifuge tube—15-mL, with screw vent rinse or Soxhlet extract with methylene cap, to fit centrifuge. chloride. 5.7 Miscellaneous lab ware—Graduated 5.2.6 Solid-phase extraction disks—90-mm cylinders, pipettes, beakers, volumetric extraction disks containing 2 g of 8-μm octa- flasks, vials, syringes, and other lab ware

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necessary to support the operations in this must have software that allows searching GC method. data for specific analytes, and for plotting 5.8 Gas chromatograph—Dual-column responses versus time. Software must also be with simultaneous split/splitless, tempera- available that allows integrating peak areas ture programmable split/splitless (PTV), or or peak heights in selected retention time on-column injection; temperature program windows and calculating concentrations of with isothermal holds, and all required ac- the analytes. cessories including syringes, analytical col- umns, gases, and detectors. An autosampler 6. Reagents and Standards is highly recommended because it injects 6.1 pH adjustment. volumes more reproducibly than manual in- 6.1.1 Sodium hydroxide solutions. jection techniques. Alternatively, two sepa- 6.1.1.1 Concentrated (10 M)—Dissolve 40 g rate single-column gas chromatographic sys- of NaOH (ACS) in reagent water and dilute tems may be employed. to 100 mL. 5.8.1 Example columns and operating con- 6.1.1.2 Dilute (1 M)—Dissolve 40 g NaOH in ditions. 1 L of reagent water. 5.8.1.1 DB–608 (or equivalent), 30-m long x 6.1.2 Sulfuric acid (1+1)—Slowly add 50 μ 0.53-mm ID fused-silica capillary, 0.83- m mL of H2SO4 (ACS, sp. gr. 1.84) to 50 mL of film thickness. reagent water. 5.8.1.2 DB–1701 (or equivalent), 30-m long x 6.1.3 Hydrochloric acid—Reagent grade, 6 0.53-mm ID fused-silica capillary, 1.0-μm film N. thickness. 6.2 Sodium thiosulfate—(ACS) granular. 5.8.1.3 Suggested operating conditions 6.3 Sodium sulfate—Sodium sulfate, rea- used to meet the retention times shown in gent grade, granular anhydrous (Baker or Table 3 are: equivalent), rinsed with methylene chloride, (a) Carrier gas flow rate: Approximately 7 baked in a shallow tray at 450 °C for 1 hour mL/min, minimum, cooled in a desiccator, and stored (b) Initial temperature: 150 °C for 0.5 in a pre-cleaned glass bottle with screw cap minute, which prevents moisture from entering. If, (c) Temperature program: 150–270 °C at 5 °C/ after heating, the sodium sulfate develops a min, and noticeable grayish cast (due to the presence (d) Final temperature: 270 °C, until trans- of carbon in the crystal matrix), that batch Permethrin elutes. of reagent is not suitable for use and should NOTE: Other columns, internal diameters, be discarded. Extraction with methylene film thicknesses, and operating conditions chloride (as opposed to simple rinsing) and may be used, provided that the performance baking at a lower temperature may produce requirements in this method are met. How- sodium sulfate suitable for use. ever, the column pair chosen must have dis- 6.4 Reagent water—Reagent water is de- similar phases/chemical properties in order fined as water in which the analytes of inter- to separate the compounds of interest in dif- est and interfering compounds are not ob- ferent retention time order. Columns that served at the MDLs of the analytes in this only differ in the length, ID, or film thick- method. ness, but use the same stationary phase do 6.5 Solvents—Methylene chloride, ace- not qualify as ‘‘dissimilar.’’ tone, methanol, hexane, acetonitrile, and 5.8.2 Carrier gas—Helium or hydrogen. isooctane, high purity pesticide quality, or Data in the tables in this method were ob- equivalent, demonstrated to be free of the tained using helium carrier gas. If hydrogen analytes and interferences (section 3). Purifi- is used, analytical conditions may need to be cation of solvents by distillation in all-glass adjusted for optimum performance, and cali- systems may be required. bration and all QC tests must be performed NOTE: The standards and final sample ex- with hydrogen carrier gas. See Section 4.3 tracts must be prepared in the same final for precautions regarding the use of hydro- solvent. gen as a carrier gas. 6.6 Ethyl ether—Nanograde, redistilled in 5.8.3 Detector—Halogen-specific detector glass if necessary. Ethyl ether must be (electron capture detector [ECD], electro- shown to be free of peroxides before use, as lytic conductivity detector [ELCD], or equiv- indicated by EM Laboratories Quant test alent). The ECD has proven effective in the strips (available from Scientific Products Co. analysis of wastewaters for the analytes list- and other suppliers). Procedures rec- ed in Tables 1 and 2, and was used to develop ommended for removal of peroxides are pro- the method performance data in Section 17 vided with the test strips. After removal of and Tables 4 and 5. peroxides, add 20 mL of ethyl alcohol pre- 5.8.4 Data system—A computer system servative to each liter of ether. must be interfaced to the GC that allows 6.7 Materials for sample cleanup. continuous acquisition and storage of data 6.7.1 Florisil®—PR grade (60/100 mesh), ac- from the detectors throughout the tivated at 650–700 °C, stored in the dark in a chromatographic program. The computer glass container with fluoropolymer-lined

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screw cap. Activate each batch immediately pearance to be effective. Copper foil or pow- prior to use for 16 hours minimum at 130 °C der that has oxidized may be reactivated in a foil-covered glass container and allow to using the procedure described above. cool. Alternatively, 500 mg cartridges (J.T. 6.7.4.2 Tetrabutylammonium sulfite (TBA Baker, or equivalent) may be used. sulfite)—Prepare as described below. 6.7.1.1 Cartridge certification—Each car- 6.7.4.2.1 Tetrabutylammonium hydrogen tridge lot must be certified to ensure recov- sulfate, [CH3(CH2)3]4NHSO4. ery of the analytes of interest and removal 6.7.4.2.2 Sodium sulfite, Na2SO3. of 2,4,6-trichlorophenol. To make the test 6.7.4.2.3 Dissolve approximately 3 g mixture, add the trichlorophenol solution tetrabutylammonium hydrogen sulfate in 100 (section 6.7.1.3) to the same standard used to mL of reagent water in an amber bottle with prepare the Quality Control Check Sample fluoropolymer-lined screw cap. Extract with (section 6.8.3). Transfer the mixture to the three 20-mL portions of hexane and discard column and dry the column. Pre-elute with the hexane extracts. three 10-mL portions of elution solvent, dry- 6.7.4.2.4 Add 25 g sodium sulfite to produce ing the column between elutions. Elute the a saturated solution. Store at room tempera- cartridge with 10 mL each of methanol and ture. Replace after 1 month. water, as in section 11.2.3.3. 6.7.5 —Reagent grade, 6.7.1.2 Concentrate the eluant to per sec- prepare at 5% (w/v) solution in reagent tion 10.3.3, exchange to isooctane or hexane water. per section 10.3.3, and inject 1.0 μL of the 6.8 Stock standard solutions—Stock concentrated eluant into the GC using the standard solutions may be prepared from procedure in section 12. The recovery of all pure materials, or purchased as certified so- analytes (including the unresolved GC peaks) lutions. Traceability must be to the National shall be within the ranges for calibration Institute of Standards and Technology verification (section 13.6 and Table 4), the re- (NIST) or other national or international covery of trichlorophenol shall be less than standard, when available. Stock solution 5%, and no peaks interfering with the target concentrations alternative to those below analytes shall be detected. Otherwise the may be used. Because of the toxicity of some Florisil cartridge is not performing properly of the compounds, primary dilutions should and the cartridge lot shall be rejected. be prepared in a hood, and a NIOSH/MESA 6.7.1.3 Florisil cartridge calibration solu- approved toxic gas respirator should be worn tion—2,4,6-Trichlorophenol, 0.1 μg/mL in ace- when high concentrations of neat materials tone. are handled. The following procedure may be 6.7.2 SPE elution solvent—Methylene used to prepare standards from neat mate- chloride:acetonitrile:hexane (50:3:47). rials. 6.7.3 Alumina, neutral, Brockman Activ- 6.8.1 Accurately weigh about 0.0100 g of ity I, 80–200 mesh (Fisher Scientific certified, pure material in a 10-mL volumetric flask. or equivalent). Heat in a glass bottle for 16 Dilute to volume in pesticide quality hexane, hours at 400 to 450 °C. Seal and cool to room isooctane, or other suitable solvent. Larger temperature. Add 7% (w/w) reagent water volumes may be used at the convenience of and mix for 10 to 12 hours. Keep bottle tight- the laboratory. When compound purity is as- ly sealed. sayed to be 96% or greater, the weight may 6.7.4 Sulfur removal. be used without correction to calculate the 6.7.4.1 Copper foil or powder—Fisher, Alfa concentration of the stock standard. Com- Aesar, or equivalent. Cut copper foil into ap- mercially prepared stock standards may be proximately 1-cm squares. Copper must be used at any concentration if they are cer- activated before it may be used, as described tified by the manufacturer or by an inde- below. pendent source. 6.7.4.1.1 Place the quantity of copper 6.8.1.1 Unless stated otherwise in this needed for sulfur removal (section 11.5.1.3) in method, store non-aqueous standards in a ground-glass-stoppered Erlenmeyer flask fluoropolymer-lined screw-cap, or heat- or bottle. Cover the foil or powder with sealed, glass containers, in the dark at ¥20 methanol. to ¥10 °C. Store aqueous standards; e.g., the 6.7.4.1.2 Add HCl dropwise (0.5–1.0 mL) aqueous LCS (section 8.4), in the dark at ≤6 while swirling, until the copper brightens. °C, but do not freeze. 6.7.4.1.3 Pour off the methanol/HCl and 6.8.1.2 Standards prepared by the labora- rinse 3 times with reagent water to remove tory may be stored for up to one year, except all traces of acid, then 3 times with acetone, when comparison with QC check standards then 3 times with hexane. indicates that a standard has degraded or be- 6.7.4.1.4 For copper foil, cover with hexane come more concentrated due to evaporation, after the final rinse. Store in a stoppered or unless the laboratory has data on file to flask under nitrogen until used. For the pow- prove stability for a longer period. Commer- der, dry on a rotary evaporator. Store in a cially prepared standards may be stored stoppered flask under nitrogen until used. until the expiration date provided by the Inspect the copper foil or powder before each vendor, except when comparison with QC use. It must have a bright, non-oxidized ap- check standards indicates that a standard

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has degraded or become more concentrated tions to cover two groups of analytes). Table due to evaporation, or unless the laboratory 7 provides information on dividing the target has data from the vendor on file to prove sta- analytes into separate calibration mixtures bility for a longer period. that should minimize or eliminate co- 6.8.2 Calibration solutions—It is nec- elutions. This table is provided solely as essary to prepare calibration solutions for guidance, based on the GC columns sug- the analytes of interest (section 1.4) only gested in this method. If an analyte listed in using an appropriate solvent (isooctane or Table 7 is not an analyte of interest in a hexane may be used). Whatever solvent is given laboratory setting, then it need not be used, both the calibration standards and the included in a calibration mixture. final sample extracts must use the same sol- vent. Other analytes may be included as de- NOTE: Many commercially available stand- sired. ards are divided into separate mixtures to 6.8.2.1 Prepare calibration standards for address this issue. the single-component analytes of interest (c) If co-elutions occur in analysis of a and surrogates at a minimum of three con- sample, a co-elution on one column is ac- centration levels (five are suggested) by add- ceptable so long as effective separation of ing appropriate volumes of one or more the co-eluting compounds can be achieved on stock standards to volumetric flasks. One of the second column. the calibration standards should be at a con- 6.8.2.2 Multi-component analytes (e.g., centration at or below the ML specified in PCBs as Aroclors, and Toxaphene). Table 1, or 2, or as specified by a regulatory/ 6.8.2.2.1 A standard containing a mixture control authority or in a permit. The ML of Aroclor 1016 and Aroclor 1260 will include value may be rounded to a whole number many of the peaks represented in the other that is more convenient for preparing the Aroclor mixtures. As a result, a multi-point standard, but must not exceed the ML value initial calibration employing a mixture of listed in Tables 1 or 2 for those analytes Aroclors 1016 and 1260 at three to five con- which list ML values. Alternatively, the lab- centrations should be sufficient to dem- oratory may establish an ML for each onstrate the linearity of the detector re- analyte based on the concentration of the sponse without the necessity of performing lowest calibration standard in a series of standards produced by the laboratory or ob- multi-point initial calibrations for each of tained from a commercial vendor, again, pro- the seven Aroclors. In addition, such a mix- vided that the ML does not exceed the ML in ture can be used as a standard to dem- Table 1 and 2, and provided that the result- onstrate that a sample does not contain ing calibration meets the acceptance criteria peaks that represent any one of the Aroclors. in section 7.5.2 based on the RSD, RSE, or R2. This standard can also be used to determine (a) The other concentrations should cor- the concentrations of either Aroclor 1016 or respond to the expected range of concentra- Aroclor 1260, should they be present in a tions found in real samples or should define sample. Therefore, prepare a minimum of the working range of the GC system. A min- three calibration standards containing equal imum of six concentration levels is required concentrations of both Aroclor 1016 and for a second order, non-linear (e.g., quad- Aroclor 1260 by dilution of the stock stand- ratic; ax2 + bx + c = 0) calibration (section ard with isooctane or hexane. The concentra- 7.5.2 or 7.6.2). Calibrations higher than sec- tions should correspond to the expected ond order are not allowed. A separate stand- range of concentrations found in real sam- ard near the MDL may be analyzed as a ples and should bracket the linear range of check on sensitivity, but should not be in- the detector. cluded in the linearity assessment. The sol- 6.8.2.2.2 Single standards of each of the vent for the standards must match the final other five Aroclors are required to aid the solvent for the sample extracts (e.g., iso- analyst in pattern recognition. Assuming octane or hexane). that the Aroclor 1016/1260 standards described NOTE: The option for non-linear calibration in Section 6.8.2.2.1 have been used to dem- may be necessary to address specific instru- onstrate the linearity of the detector, these mental techniques. However, it is not EPA’s single standards of the remaining five intent to allow non-linear calibration to be Aroclors also may be used to determine the used to compensate for detector saturation calibration factor for each Aroclor. Prepare or to avoid proper instrument maintenance. a standard for each of the other Aroclors. (b) Given the number of analytes included The concentrations should generally cor- in this method, it is highly likely that some respond to the mid-point of the linear range will coelute on one or both of the GC col- of the detector, but lower concentrations umns used for the analysis. Divide the may be employed at the discretion of the an- analytes into two or more groups and pre- alyst based on project requirements. pare separate calibration standards for each 6.8.2.2.3 For Toxaphene, prepare a min- group, at multiple concentrations (e.g., a imum of three calibration standards con- five-point calibration will require ten solu- taining Toxaphene by dilution of the stock

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standard with isooctane or hexane. The con- and meets all QC acceptance criteria with centrations should correspond to the ex- the alternative or additional internal stand- pected range of concentrations found in real ard(s) as an integral part of this method. samples and should bracket the linear range 6.8.6 Surrogate solution—Prepare a solu- of the detector. tion containing one or more surrogates at a 6.8.3 Quality Control (QC) Check Sample concentration of 2 μg/mL in acetone. Poten- Concentrate—Prepare one or more mid-level tial surrogates include: dibutyl chlorendate standard mixtures (concentrates) in acetone (DBC), tetrachloro-m-xylene (TCMX), 4,4′- (or other water miscible solvent). The con- dibromobiphenyl, or decachlorobiphenyl. Al- centrate is used as the spiking solution with ternative surrogates and concentrations may which to prepare the Demonstration of Capa- be used, provided the laboratory performs all bilities (DOC) samples, the Laboratory Con- QC tests and meets all QC acceptance cri- trol Sample (LCS), and Matrix Spike (MS) teria with the alternative surrogate(s) as an and Matrix Spike Duplicate (MSD) samples integral part of this method. If the internal described in section 8. If prepared by the lab- standard calibration technique is used, do oratory (as opposed the purchasing it from a not use the internal standard as a surrogate. commercial supplier), the concentrate must 6.8.7 DDT and endrin decomposition be prepared independently from the stand- (breakdown) solution—Prepare a solution ards used for calibration, but may be pre- containing endrin at a concentration of 50 pared from the same source as the second- ng/mL and 4,4’-DDT at a concentration of 100 source standard used for calibration ng/mL, in isooctane or hexane. A 1-μL injec- verification (section 7.7). Regardless of the tion of this standard will contain 50 source, the concentrate must be in a water- picograms (pg) of endrin and 100 pg of DDT. miscible solvent, as noted above. The con- The concentration of the solution may be ad- centrate is used to prepare the DOC and LCS justed by the laboratory to accommodate (sections 8.2.1 and 8.4) and MS/MSD samples other injection volumes such that the same (section 8.3). Depending on the analytes of masses of the two analytes are introduced interest for a given sample (see Section 1.4), into the instrument. multiple solutions and multiple LCS or MS/ MSD samples may be required to account for 7. Calibration co-eluting analytes. However, a co-elution on 7.1 Establish gas chromatographic oper- one column is acceptable so long as effective ating conditions equivalent to those in Sec- separation of the co-eluting compounds can tion 5.8.1 and Footnote 2 to Table 3. Alter- be achieved on the second column. In addi- native temperature program and flow rate tion, the concentrations of the MS/MSD sam- conditions may be used. The system may be ples should reflect any relevant compliance calibrated using the external standard tech- limits for the analytes of interest, as de- nique (section 7.5) or the internal standard scribed in section 8.3.1. If a custom spiking technique (section 7.6). It is necessary to solution is required for a specific discharge calibrate the system for the analytes of in- (section 8.3.1), prepare it separately from the terest (section 1.4) only. DOC and LCS solution. 7.2 Separately inject the mid-level cali- NOTE: Some commercially available stand- bration standard for each calibration mix- ards are divided into separate mixtures to ture. Store the retention time on each GC address the co-elution issue. column. 6.8.4 Calibration Verification Standards— 7.3 Injection of calibration solutions—In- In order to verify the results of the initial ject a constant volume in the range of 0.5 to calibration standards, prepare one or more 2.0 μL of each calibration solution into the mid-level standard mixtures in isooctane or GC column/detector pairs. An alternative hexane, using standards obtained from a sec- volume (see Section 12.3) may be used pro- ond source (different manufacturer or dif- vided all requirements in this method are ferent certified lot from the calibration met. Beginning with the lowest level mix- standards). These standards will be analyzed ture and proceeding to the highest level mix- to verify the accuracy of the calibration ture may limit the risk of carryover from (sections 7.7 and 13.6.2). As with the QC sam- one standard to the next, but other se- ple concentrate in section 6.8.3, multiple so- quences may be used. An instrument blank lutions may be required to address co- should be analyzed after the highest stand- elutions among all of the analytes. ard to demonstrate that there is no carry- 6.8.5 Internal standard solution—If the in- over within the system for this calibration ternal standard calibration technique is to range. be used, prepare pentachloronitrobenzene 7.4 For each analyte, compute, record, (PCNB) at a concentration of 10 μg/mL in and store, as a function of the concentration ethyl acetate. Alternative and multiple in- injected, the retention time and peak area ternal standards; e.g., tetrachloro-m-xylene, on each column/detector system. If multi- 4,4′-dibromobiphenyl, and/or component analytes are to be analyzed, store decachlorobiphenyl may be used provided the retention time and peak area for the that the laboratory performs all QC tests three to five exclusive (unique large) peaks

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for each PCB or technical chlordane. Use 7.5.1 From the calibration data (Section four to six peaks for toxaphene. 7.4), calculate the calibration factor (CF) for 7.5 External standard calibration. each analyte at each concentration accord- ing to the following equation:

Where: the regression must be weighted inversely proportional to concentration. The coeffi- Cs = Concentration of the analyte in the standard (ng/mL) cient of determination (R2) of the weighted As = Peak height or area regression must be greater than 0.920. Alter- For multi-component analytes, choose a natively, the relative standard error (Ref- series of characteristic peaks for each erence 10) may be used as an acceptance cri- analyte (3 to 5 for each Aroclor, 4 to 6 for terion. As with the RSD, the RSE must be toxaphene) and calculate individual calibra- less than 20%. If an RSE less than 20% can- tion factors for each peak. Alternatively, for not be achieved for a quadratic regression, toxaphene, sum the areas of all of the peaks system performance is unacceptable and the in the standard chromatogram and use the system must be adjusted and re-calibrated. summed area to determine the calibration NOTE: Regression calculations are not in- factor. (If this alternative is used, the same cluded in this method because the calcula- approach must be used to quantitate the tions are cumbersome and because many GC/ analyte in the samples.) ECD data systems allow selection of weight- 7.5.2 Calculate the mean (average) and ed regression for calibration and calculation relative standard deviation (RSD) of the of analyte concentrations. calibration factors. If the RSD is less than 20%, linearity through the origin can be as- 7.6 Internal standard calibration. sumed and the average CF can be used for 7.6.1 From the calibration data (Section calculations. Alternatively, the results can 7.4), calculate the response factor (RF) for be used to fit a linear or quadratic regression each analyte at each concentration accord- of response, As, vs. concentration Cs. If used, ing to the following equation:

Where: of determination of the weighted regression must be greater than 0.920. Alternatively, As = Response for the analyte to be meas- ured. the relative standard error (Reference 10) A = Response for the internal standard. may be used as an acceptance criterion. As is with the RSD, the RSE must be less than C = Concentration of the internal standard is 15%. If an RSE less than 15% cannot be (ng/mL) achieved for a quadratic regression, system Cs = Concentration of the analyte to be performance is unacceptable and the system measured (ng/mL). must be adjusted and re-calibrated. 7.6.2 Calculate the mean (average) and 7.7 The working calibration curve, CF, or relative standard deviation (RSD) of the re- RF must be verified immediately after cali- sponse factors. If the RSD is less than 15%, bration and at the beginning and end of each linearity through the origin can be assumed 24-hour shift by the analysis of a mid-level and the average RF can be used for calcula- calibration standard. The calibration tions. Alternatively, the results can be used verification standard(s) must be obtained to prepare a calibration curve of response ra- from a second manufacturer or a manufac- tios, As/Ais, vs. concentration ratios, Cs/Cis, turer’s batch prepared independently from for the analyte. A minimum of six con- the batch used for calibration (Section 6.8.4). centration levels is required for a non-linear Requirements for calibration verification are (e.g., quadratic) regression. If used, the re- given in Section 13.6 and Table 4. Alter- gression must be weighted inversely propor- natively, calibration verification may be tional to concentration, and the coefficient performed after a set number of injections

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(e.g., every 20 injections), to include injec- overcome matrix interferences, the labora- tion of extracts of field samples, QC samples, tory is permitted certain options (section 1.8 instrument blanks, etc. (i.e., it is based on and 40 CFR 136.6(b) [Reference 12]) to im- the number of injections performed, not sam- prove separations or lower the costs of meas- ple extracts). The time for the injections urements. These options may include alter- may not exceed 24 hours. native extraction (e.g., other solid-phase ex- NOTE: The 24-hour shift begins after anal- traction materials and formats), concentra- ysis of the combined QC standard (calibra- tion, and cleanup procedures, and changes in tion verification) and ends 24 hours later. GC columns (Reference 12). Alternative de- The ending calibration verification standard terminative techniques, such as the substi- is run immediately after the last sample run tution of spectroscopic or immunoassay during the 24-hour shift, so the beginning techniques, and changes that degrade meth- and ending calibration verifications are out- od performance, are not allowed. If an ana- side of the 24-hour shift. If calibration lytical technique other than the techniques verification is based on the number of injec- specified in this method is used, that tech- tions instead of time, then the ending nique must have a specificity equal to or verification standard for one group of injec- greater than the specificity of the techniques tions may be used as the beginning in this method for the analytes of interest. verification for the next group of injections. The laboratory is also encouraged to partici- 7.8 Florisil® calibration—The column pate in performance evaluation studies (see cleanup procedure in Section 11.3 utilizes section 8.8). Florisil column chromatography. Florisil® 8.1.2.1 Each time a modification listed from different batches or sources may vary above is made to this method, the laboratory in adsorptive capacity. To standardize the is required to repeat the procedure in section amount of Florisil® which is used, use of the 8.2. If the detection limit of the method will lauric acid value (Reference 11) is suggested. be affected by the change, the laboratory is The referenced procedure determines the ad- required to demonstrate that the MDLs (40 sorption from a hexane solution of lauric CFR part 136, appendix B) are lower than acid (mg) per g of Florisil®. The amount of one-third the regulatory compliance limit or Florisil® to be used for each column is cal- as low as the MDLs in this method, which- culated by dividing 110 by this ratio and mul- ever are greater. If calibration will be af- tiplying by 20 g. If cartridges containing fected by the change, the instrument must Florisil® are used, then this step is not nec- be recalibrated per section 7. Once the modi- essary. fication is demonstrated to produce results equivalent or superior to results produced by 8. Quality Control this method as written, that modification 8.1 Each laboratory that uses this method may be used routinely thereafter, so long as is required to operate a formal quality assur- the other requirements in this method are ance program. The minimum requirements met (e.g., matrix spike/matrix spike dupli- of this program consist of an initial dem- cate recovery and relative percent dif- onstration of laboratory capability and on- ference). going analysis of spiked samples and blanks 8.1.2.1.1 If an allowed method modifica- to evaluate and document data quality. The tion, is to be applied to a specific discharge, laboratory must maintain records to docu- the laboratory must prepare and analyze ma- ment the quality of data generated. Ongoing trix spike/matrix spike duplicate (MS/MSD) data quality checks are compared with es- samples (section 8.3) and LCS samples (sec- tablished performance criteria to determine tion 8.4). The laboratory must include surro- if the results of analyses meet performance gates (Section 8.7) in each of the samples. requirements of this method. A quality con- The MS/MSD and LCS samples must be for- trol check standard (LCS, section 8.4) must tified with the analytes of interest (section be prepared and analyzed with each batch of 1.4). If the modification is for nationwide samples to confirm that the measurements use, MS/MSD samples must be prepared from were performed in an in-control mode of op- a minimum of nine different discharges (See eration. A laboratory may develop its own section 8.1.2.1.2), and all QC acceptance cri- performance criteria (as QC acceptance cri- teria in this method must be met. This eval- teria), provided such criteria are as or more uation only needs to be performed once other restrictive than the criteria in this method. than for the routine QC required by this 8.1.1 The laboratory must make an initial method (for example it could be performed demonstration of the capability (IDC) to by the vendor of an alternative material) but generate acceptable precision and recovery any laboratory using that specific material with this method. This demonstration is de- must have the results of the study available. tailed in Section 8.2. On a continuing basis, This includes a full data package with the the laboratory must repeat demonstration of raw data that will allow an independent re- capability (DOC) at least annually. viewer to verify each determination and cal- 8.1.2 In recognition of advances that are culation performed by the laboratory (see occurring in analytical technology, and to section 8.1.2.2.5, items (a)–(q)).

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8.1.2.1.2 Sample matrices on which MS/ (f) Laboratory control sample analysis MSD tests must be performed for nationwide (section 8.4). use of an allowed modification: 8.1.2.2.5 Data that will allow an inde- (a) Effluent from a publicly owned treat- pendent reviewer to validate each deter- ment works (POTW). mination by tracing the instrument output (b) ASTM D5905 Standard Specification for (peak height, area, or other signal) to the Substitute Wastewater. final result. These data are to include: (c) Sewage sludge, if sewage sludge will be (a) Sample numbers and other identifiers. in the permit. (b) Extraction dates. (d) ASTM D1141 Standard Specification for (c) Analysis dates and times. Substitute Ocean Water, if ocean water will (d) Analysis sequence/run chronology. be in the permit. (e) Sample weight or volume (section 10). (e) Untreated and treated wastewaters up to a total of nine matrix types (see https:// (f) Extract volume prior to each cleanup www.epa.gov/eg/industrial-effluent-guidelines step (sections 10 and 11). for a list of industrial categories with exist- (g) Extract volume after each cleanup step ing effluent guidelines). (section 11). (i) At least one of the above wastewater (h) Final extract volume prior to injection matrix types must have at least one of the (sections 10 and 12). following characteristics: (i) Injection volume (sections 12.3 and 13.2). (A) Total suspended solids greater than 40 (j) Sample or extract dilution (section mg/L. 15.4). (B) Total dissolved solids greater than 100 (k) Instrument and operating conditions. mg/L. (l) Column (dimensions, material, etc.). (C) Oil and grease greater than 20 mg/L. (m) Operating conditions (temperatures, (D) NaCl greater than 120 mg/L. flow rates, etc.). (E) CaCO3 greater than 140 mg/L. (n) Detector (type, operating conditions, (ii) The interim acceptance criteria for etc.). MS, MSD recoveries that do not have recov- (o) Chromatograms and other recordings of ery limits in Table 4 or developed in section raw data. 8.3.3, and for surrogates that do not have re- (p) Quantitation reports, data system out- covery limits developed in section 8.6, must puts, and other data to link the raw data to be no wider than 60–140%, and the relative the results reported. percent difference (RPD) of the concentra- (q) A written Standard Operating Proce- tions in the MS and MSD that do not have dure (SOP). RPD limits in Table 4 or developed in section 8.1.2.2.6 Each individual laboratory wish- 8.3.3, must be less than 30%. Alternatively, ing to use a given modification must perform the laboratory may use the laboratory’s in- the start-up tests in section 8.1.2 (e.g., DOC, house limits if they are tighter. (f) A proficiency testing (PT) sample from MDL), with the modification as an integral a recognized provider, in addition to tests of part of this method prior to applying the the nine matrices (section 8.1.2.1.1). modification to specific discharges. Results 8.1.2.2 The laboratory must maintain of the DOC must meet the QC acceptance cri- records of modifications made to this meth- teria in Table 5 for the analytes of interest od. These records include the following, at a (section 1.4), and the MDLs must be equal to minimum: or lower than the MDLs in Tables 1 and 2 for 8.1.2.2.1 The names, titles, and business the analytes of interest. street addresses, telephone numbers, and 8.1.3 Before analyzing samples, the lab- email addresses, of the analyst(s) that per- oratory must analyze a blank to dem- formed the analyses and modification, and of onstrate that interferences from the analyt- the quality control officer that witnessed ical system, lab ware, and reagents, are and will verify the analyses and modifica- under control. Each time a batch of samples tions. is extracted or reagents are changed, a blank 8.1.2.2.2 A list of analytes, by name and must be extracted and analyzed as a safe- CAS Registry number. guard against laboratory contamination. Re- 8.1.2.2.3 A narrative stating reason(s) for quirements for the blank are given in section the modifications. 8.5. 8.1.2.2.4 Results from all quality control 8.1.4 The laboratory must, on an ongoing (QC) tests comparing the modified method to basis, spike and analyze samples to monitor this method, including: and evaluate method and laboratory per- (a) Calibration (section 7). formance on the sample matrix. The proce- (b) Calibration verification (section 13.6). dure for spiking and analysis is given in sec- (c) Initial demonstration of capability (sec- tion 8.3. tion 8.2). 8.1.5 The laboratory must, on an ongoing (d) Analysis of blanks (section 8.5). basis, demonstrate through analysis of a (e) Matrix spike/matrix spike duplicate quality control check sample (laboratory analysis (section 8.3). control sample, LCS; on-going precision and

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recovery sample, OPR) that the measure- percent recovery for each analyte using the ment system is in control. This procedure is four results. described in Section 8.4. 8.2.5 For each analyte, compare s and X 8.1.6 The laboratory should maintain per- with the corresponding acceptance criteria formance records to document the quality of for precision and recovery in Table 4. For data that is generated. This procedure is analytes in Table 2 that are not listed in given in section 8.7. Table 4, QC acceptance criteria must be de- 8.1.7 The large number of analytes tested veloped by the laboratory. EPA has provided in performance tests in this method present guidance for development of QC acceptance a substantial probability that one or more criteria (References 12 and 13). If s and X for will fail acceptance criteria when all all analytes of interest meet the acceptance analytes are tested simultaneously, and a re- criteria, system performance is acceptable test (reanalysis) is allowed if this situation and analysis of blanks and samples can should occur. If, however, continued re-test- begin. If any individual s exceeds the preci- ing results in further repeated failures, the sion limit or any individual X falls outside laboratory should document the failures and the range for recovery, system performance either avoid reporting results for the is unacceptable for that analyte. analytes that failed or report the problem NOTE: The large number of analytes in Ta- and failures with the data. A QC failure does bles 1 and 2 present a substantial probability not relieve a discharger or permittee of re- that one or more will fail at least one of the porting timely results. acceptance criteria when many or all 8.2 Demonstration of capability (DOC)— analytes are determined simultaneously. To establish the ability to generate accept- 8.2.6 When one or more of the analytes able recovery and precision, the laboratory tested fail at least one of the acceptance cri- must perform the DOC in sections 8.2.1 teria, repeat the test for only the analytes through 8.2.6 for the analytes of interest ini- that failed. If results for these analytes pass, tially and in an on-going manner at least an- system performance is acceptable and anal- nually. The laboratory must also establish ysis of samples and blanks may proceed. If MDLs for the analytes of interest using the one or more of the analytes again fail, sys- MDL procedure at 40 CFR part 136, appendix tem performance is unacceptable for the B. The laboratory’s MDLs must be equal to analytes that failed the acceptance criteria. or lower than those listed in Tables 1 or 2, or Correct the problem and repeat the test (sec- lower than one-third the regulatory compli- tion 8.2). See section 8.1.7 for disposition of ance limit, whichever is greater. For MDLs repeated failures. not listed in Tables 1 or 2, the laboratory NOTE: To maintain the validity of the test must determine the MDLs using the MDL and re-test, system maintenance and/or ad- procedure at 40 CFR part 136, appendix B justment is not permitted between this pair under the same conditions used to determine of tests. the MDLs for the analytes listed in Tables 1 8.3 Matrix spike and matrix spike dupli- and 2. When analyzing the PCBs as Aroclors, cate (MS/MSD)—The purpose of the MS/MSD it is only necessary to establish an MDL for requirement is to provide data that dem- one of the multi-component analytes (e.g., onstrate the effectiveness of the method as PCB 1254), or the mixture of Aroclors 1016 applied to the samples in question by a given and 1260 may be used to establish MDLs for laboratory, and both the data user (dis- all of the Aroclors. Similarly, MDLs for charger, permittee, regulated entity, regu- other multi-component analytes (e.g., latory/control authority, customer, other) Chlordanes) may be determined using only and the laboratory share responsibility for one of the major components. All procedures provision of such data. The data user should used in the analysis, including cleanup pro- identify the sample and the analytes of in- cedures, must be included in the DOC. terest (section 1.4) to be spiked and provide 8.2.1 For the DOC, a QC check sample con- sufficient sample volume to perform MS/ centrate containing each analyte of interest MSD analyses. The laboratory must, on an (section 1.4) is prepared in a water-miscible ongoing basis, spike at least 5% of the sam- solvent using the solution in section 6.8.3. ples in duplicate from each discharge being NOTE: QC check sample concentrates are monitored to assess accuracy (recovery and no longer available from EPA. precision). If direction cannot be obtained 8.2.2 Using a pipet or syringe, prepare four from the data user, the laboratory must QC check samples by adding an appropriate spike at least one sample in duplicate per ex- volume of the concentrate and of the surro- traction batch of up to 20 samples with the gate(s) to each of four 1–L aliquots of rea- analytes in Table 1. Spiked sample results gent water. Swirl or stir to mix. should be reported only to the data user 8.2.3 Extract and analyze the well-mixed whose sample was spiked, or as requested or QC check samples according to the method required by a regulatory/control authority, beginning in section 10. or in a permit. 8.2.4 Calculate the average percent recov- 8.3.1. If, as in compliance monitoring, the ery (X) and the standard deviation (s) of the concentration of a specific analyte will be

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checked against a regulatory concentration 8.3.2 Analyze one sample aliquot to deter- limit, the concentration of the spike should mine the background concentration (B) of be at that limit; otherwise, the concentra- the each analyte of interest. If necessary to tion of the spike should be one to five times meet the requirement in section 8.3.1, pre- higher than the background concentration pare a new check sample concentrate (sec- determined in section 8.3.2, at or near the tion 8.2.1) appropriate for the background midpoint of the calibration range, or at the concentration. Spike and analyze two addi- concentration in the LCS (section 8.4) which- tional sample aliquots of the same volume as ever concentration would be larger. When no the original sample, and determine the con- centrations after spiking (A and A ) of each information is available, the mid-point of 1 2 analyte. Calculate the percent recoveries (P the calibration may be used. 1 and P2) as:

where T is the known true value of the spike. Also calculate the relative percent dif- ference (RPD) between the concentrations (A1 and A2):

8.3.3 Compare the percent recoveries (P1 80% of the analytes tested in the MS/MSD and P2) and the RPD for each analyte in the must have in-house QC acceptance criteria MS/MSD aliquots with the corresponding QC that are tighter than those in Table 4 and acceptance criteria for recovery (P) and RPD the remaining analytes (those not included in Table 4. in the 80%) must meet the acceptance cri- (a) If any individual P falls outside the des- teria in Table 4. If an in-house QC limit for ignated range for recovery in either aliquot, the RPD is greater than the limit in Table 4, or the RPD limit is exceeded, the result for then the limit in Table 4 must be used. Simi- the analyte in the unspiked sample is sus- larly, if an in-house lower limit for recovery pect and may not be reported or used for per- is below the lower limit in Table 4, then the mitting or regulatory compliance. See sec- lower limit in Table 4 must be used, and if an tion 8.1.7 for disposition of failures. in-house upper limit for recovery is above (b) For analytes in Table 2 not listed in the upper limit in Table 4, then the upper Table 4, QC acceptance criteria must be de- limit in Table 4 must be used. The labora- veloped by the laboratory. EPA has provided tory must evaluate surrogate recovery data guidance for development of QC acceptance in each sample against its in-house surrogate criteria (References 12 and 13). recovery limits. The laboratory may use 60 8.3.4 After analysis of a minimum of 20 -140% as interim acceptance criteria for sur- MS/MSD samples for each target analyte and rogate recoveries until in-house limits are surrogate, and if the laboratory chooses to developed. Alternatively, surrogate recovery develop and apply optional in-house QC lim- limits may be developed from laboratory its, the laboratory should calculate and control charts. In-house QC acceptance cri- apply the optional in-house QC limits for re- teria must be updated at least every two covery and RPD of future MS/MSD samples years. (Section 8.3). The optional in-house QC lim- 8.4 Laboratory control sample (LCS)—A its for recovery are calculated as the mean QC check sample (laboratory control sample, observed recovery ±3 standard deviations, LCS; on-going precision and recovery sam- and the upper QC limit for RPD is calculated ple, OPR) containing each single-component as the mean RPD plus 3 standard deviations analyte of interest (section 1.4) must be ex- of the RPDs. The in-house QC limits must be tracted, concentrated, and analyzed with updated at least every two years and re-es- each extraction batch of up to 20 samples tablished after any major change in the ana- (section 3.1) to demonstrate acceptable re- lytical instrumentation or process. At least covery of the analytes of interest from a

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clean sample matrix. If multi-peak analytes reagent water must be the same as the nomi- are required, extract and prepare at least one nal volume used for the sample, the DOC as an LCS for each batch. Alternatively, the (Section 8.2), the blank (section 8.5), and the laboratory may set up a program where MS/MSD (section 8.3). Also add a volume of multi-peak LCS is rotated with a single-peak the surrogate solution (section 6.8.6). LCS. 8.4.2 Analyze the LCS prior to analysis of 8.4.1 Prepare the LCS by adding QC check samples in the extraction batch (Section 3.1). sample concentrate (sections 6.8.3 and 8.2.1) Determine the concentration (A) of each to reagent water. Include all analytes of in- analyte. Calculate the percent recovery as: terest (section 1.4) in the LCS. The volume of

where T is the true value of the concentra- ture LCS samples (section 8.4). Limits for re- tion in the LCS. covery in the LCS should be calculated as 8.4.3 For each analyte, compare the per- the mean recovery ±3 standard deviations. A cent recovery (P) with its corresponding QC minimum of 80% of the analytes tested for in acceptance criterion in Table 4. For analytes the LCS must have QC acceptance criteria of interest in Table 2 not listed in Table 4, tighter than those in Table 4, and the re- use the QC acceptance criteria developed for maining analytes (those not included in the the MS/MSD (section 8.3.3.2), or limits based 80%) must meet the acceptance criteria in on laboratory control charts. If the recov- Table 4. If an in-house lower limit for recov- eries for all analytes of interest fall within ery is lower than the lower limit in Table 4, the designated ranges, analysis of blanks and the lower limit in Table 4 must be used, and field samples may proceed. If any individual if an in-house upper limit for recovery is recovery falls outside the range, proceed ac- higher than the upper limit in Table 4, the cording to section 8.4.4. upper limit in Table 4 must be used. Many of NOTE: The large number of analytes in Ta- the analytes and surrogates do not contain bles 1 and 2 present a substantial probability acceptance criteria. The laboratory should that one or more will fail the acceptance cri- use 60–140% as interim acceptance criteria teria when all analytes are tested simulta- for recoveries of spiked analytes and surro- neously. Because a re-test is allowed in event gates that do not have recovery limits speci- of failure (sections 8.1.7 and 8.4.4), it may be fied in Table 4, and at least 80% of the surro- prudent to extract and analyze two LCSs to- gates must meet the 60–140% interim criteria gether and evaluate results of the second until in-house LCS and surrogate limits are analysis against the QC acceptance criteria developed. Alternatively, acceptance criteria only if an analyte fails the first test. for analytes that do not have recovery limits 8.4.4 Repeat the test only for those in Table 4 may be based on laboratory con- analytes that failed to meet the acceptance trol charts. In-house QC acceptance criteria criteria (P). If these analytes now pass, sys- must be updated at least every two years. tem performance is acceptable and analysis 8.5 Blank—Extract and analyze a blank of blanks and samples may proceed. Re- with each extraction batch (section 3.1) to peated failure, however, will confirm a gen- demonstrate that the reagents and equip- eral problem with the measurement system. ment used for preparation and analysis are If this occurs, repeat the test using a fresh free from contamination. LCS (section 8.2.1) or an LCS prepared with 8.5.1 Prepare the blank from reagent a fresh QC check sample concentrate (sec- water and spike it with the surrogates. The tion 8.2.1), or perform and document system volume of reagent water must be the same as repair. Subsequent to analysis of the LCS the volume used for samples, the DOC (sec- prepared with a fresh sample concentrate, or tion 8.2), the LCS (section 8.4), and the MS/ to system repair, repeat the LCS test (Sec- MSD (section 8.3). Extract, concentrate, and tion 8.4). If failure of the LCS indicates a analyze the blank using the same procedures systemic problem with samples in the batch, and reagents used for the samples, LCS, and re-extract and re-analyze the samples in the MS/MSD in the batch. Analyze the blank im- batch. See Section 8.1.7 for disposition of re- mediately after analysis of the LCS (section peated failures. 8.4) and prior to analysis of the MS/MSD and 8.4.5 After analysis of 20 LCS samples, and samples to demonstrate freedom from con- if the laboratory chooses to develop and tamination. apply optional in-house QC limits, the lab- 8.5.2 If any analyte of interest is found in oratory should calculate and apply the op- the blank at a concentration greater than tional in-house QC limits for recovery of fu- the MDL for the analyte, at a concentration

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greater than one-third the regulatory com- automatic sampling equipment. Collect 1-L pliance limit, or at a concentration greater of ambient waters, effluents, and other aque- than one-tenth the concentration in a sam- ous samples. If high concentrations of the ple in the batch (section 3.1), whichever is analytes of interest are expected (e.g., for greatest, analysis of samples must be halted untreated effluents or in-process waters), and samples in the batch must be re-ex- collect a smaller volume (e.g., 250 mL), but tracted and the extracts reanalyzed. Samples not less than 100 mL, in addition to the 1-L in a batch must be associated with an sample. Follow conventional sampling prac- uncontaminated blank before the results for tices, except do not pre-rinse the bottle with those samples may be reported or used for sample before collection. Automatic sam- permitting or regulatory compliance pur- pling equipment must be as free as possible poses. If re-testing of blanks results in re- of polyvinyl chloride or other tubing or peated failures, the laboratory should docu- ment the failures and report the problem and other potential sources of contamination. If failures with the data. needed, collect additional sample(s) for the 8.6 Surrogate recovery—The laboratory MS/MSD (section 8.3). must spike all samples with the surrogate 9.2 Ice or refrigerate the sample at ≤6 °C standard spiking solution (section 6.8.6) per from the time of collection until extraction, section 10.2.2 or 10.4.2, analyze the samples, but do not freeze. If aldrin is to be deter- and calculate the percent recovery of each mined and residual chlorine is present, add surrogate. QC acceptance criteria for surro- 80 mg/L of sodium thiosulfate but do not add gates must be developed by the laboratory excess. Any method suitable for field use (section 8.4). If any recovery fails its cri- may be employed to test for residual chlo- terion, attempt to find and correct the cause rine (Reference 14). If sodium thiosulfate of the failure, and if sufficient volume is interferes in the determination of the available, re-extract another aliquot of the analytes, an alternative preservative (e.g., affected sample; otherwise, see section 8.1.7 ascorbic acid or sodium sulfite) may be used. for disposition of repeated failures. 9.3 Extract all samples within seven days 8.7 As part of the QC program for the lab- of collection and completely analyze within oratory, it is suggested but not required that 40 days of extraction (Reference 1). If the method accuracy for wastewater samples be sample will not be extracted within 72 hours assessed and records maintained. After anal- of collection, adjust the sample pH to a ysis of five or more spiked wastewater sam- range of 5.0–9.0 with sodium hydroxide solu- ples as in Section 8.3, calculate the average tion or sulfuric acid. Record the volume of percent recovery (X) and the standard devi- acid or base used. ation of the percent recovery (sp). Express the accuracy assessment as a percent inter- 10. Sample Extraction val from X¥2sp to X+2sp. For example, if X = 90% and sp = 10%, the accuracy interval is 10.1 This section contains procedures for expressed as 70–110%. Update the accuracy separatory funnel liquid-liquid extraction assessment for each analyte on a regular (SFLLE, section 10.2), continuous liquid-liq- basis to ensure process control (e.g., after uid extraction (CLLE, section 10.4), and disk- each 5–10 new accuracy measurements). If de- based solid-phase extraction (SPE, section sired, statements of accuracy for laboratory 10.5). SFLLE is faster, but may not be as ef- performance, independent of performance on fective as CLLE for extracting polar samples, may be developed using LCSs. analytes. SFLLE is labor intensive and may 8.8 It is recommended that the laboratory result in formation of emulsions that are dif- adopt additional quality assurance practices ficult to break. CLLE is less labor intensive, for use with this method. The specific prac- avoids emulsion formation, but requires tices that are most productive depend upon more time (18–24 hours), more hood space, the needs of the laboratory and the nature of and may require more solvent. SPE can be the samples. Field duplicates may be ana- faster, unless the particulate load in an lyzed to assess the precision of environ- mental measurements. When doubt exists aqueous sample is so high that it slows the over the identification of a peak on the chro- filtration process. If an alternative extrac- matogram, confirmatory techniques such as tion scheme to those detailed in this method gas chromatography with another dissimilar is used, all QC tests must be performed and column, specific element detector, or mass all QC acceptance criteria must be met with spectrometer must be used. Whenever pos- that extraction scheme as an integral part of sible, the laboratory should analyze standard this method. reference materials and participate in rel- 10.2 Separatory funnel liquid-liquid ex- evant performance evaluation studies. traction (SFLLE). 10.2.1 The SFLLE procedure below as- 9. Sample Collection, Preservation, and sumes a sample volume of 1 L. When a dif- Handling ferent sample volume is extracted, adjust 9.1 Collect samples as grab samples in the volume of methylene chloride accord- glass bottles, or in refrigerated bottles using ingly.

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10.2.2 Mark the water meniscus on the trator tube to a 500-mL evaporative flask. side of the sample bottle for later determina- Other concentration devices or techniques tion of sample volume. Pour the entire sam- may be used in place of the K–D concen- ple into the separatory funnel. Pipet the sur- trator so long as the requirements of section rogate standard spiking solution (section 8.2 are met. 6.8.6) into the separatory funnel. If the sam- 10.3.1.2 Pour the extract through a sol- ple will be used for the LCS or MS or MSD, vent-rinsed drying column containing about pipet the appropriate QC check sample con- 10 cm of anhydrous sodium sulfate, and col- centrate (section 8.3 or 8.4) into the sepa- lect the extract in the K–D concentrator. ratory funnel. Mix well. If the sample arrives Rinse the flask and column with 20–30 mL of in a larger sample bottle, 1 L may be meas- methylene chloride to complete the quan- ured in a graduated cylinder, then added to titative transfer. the separatory funnel. 10.3.1.3 If no cleanup is to be performed on NOTE: Instances in which the sample is col- the sample, add 500 μL (0.5 mL) of isooctane lected in an oversized bottle should be re- to the extract to act as a keeper during con- ported by the laboratory to the data user. Of centration. particular concern is that fact that this 10.3.1.4 Add one or two clean boiling chips practice precludes rinsing the empty bottle and attach a three-ball Snyder column to the with solvent as described below, which could K–D evaporative flask. Pre-wet the Snyder leave hydrophobic pesticides on the wall of column by adding about 1 mL of methylene the bottle, and underestimate the actual chloride to the top. Place the K–D apparatus sample concentrations. on a hot water bath (60–65 °C) so that the 10.2.3 Add 60 mL of methylene chloride to concentrator tube is partially immersed in the sample bottle, seal, and shake for 30 sec- the hot water, and the entire lower rounded onds to rinse the inner surface. Transfer the solvent to the separatory funnel and extract surface of the flask is bathed with hot vapor. the sample by shaking the funnel for two Adjust the vertical position of the apparatus minutes with periodic venting to release ex- and the water temperature as required to cess pressure. Allow the organic layer to sep- complete the concentration in 15–20 minutes. arate from the water phase for a minimum of At the proper rate of evaporation the balls of 10 minutes. If an emulsion forms and the the column will actively chatter but the emulsion interface between the layers is chambers will not flood with condensed sol- more than one-third the volume of the sol- vent. When the apparent volume of liquid vent layer, employ mechanical techniques to reaches 1 mL or other determined amount, complete the phase separation. The optimum remove the K–D apparatus from the water technique depends upon the sample, but may bath and allow it to drain and cool for at include stirring, filtration of the emulsion least 10 minutes. through glass wool, use of phase-separation 10.3.1.5 If the extract is to be cleaned up paper, centrifugation, salting, freezing, or by sulfur removal or acid back extraction, other physical methods. Collect the meth- remove the Snyder column and rinse the ylene chloride extract in a flask. If the emul- flask and its lower joint into the concen- sion cannot be broken (recovery of less than trator tube with 1 to 2 mL of methylene 80% of the methylene chloride, corrected for chloride. A 5-mL syringe is recommended for the water of methylene chloride), this operation. Adjust the final volume to 10 transfer the sample, solvent, and emulsion mL in methylene chloride and proceed to into the extraction chamber of a continuous sulfur removal (section 11.5) or acid back ex- extractor and proceed as described in section traction (section 11.6). If the extract is to 10.4. cleaned up using one of the other cleanup 10.2.4 Add a second 60-mL volume of procedures or is to be injected into the GC, methylene chloride to the sample bottle and proceed to Kuderna-Danish micro-concentra- repeat the extraction procedure a second tion (section 10.3.2) or nitrogen evaporation time, combining the extracts in the flask. and solvent exchange (section 10.3.3). Perform a third extraction in the same man- 10.3.2 Kuderna-Danish micro concentra- ner. Proceed to macro-concentration (sec- tion—Add another one or two clean boiling tion 10.3.1). chips to the concentrator tube and attach a 10.2.5 Determine the original sample vol- two-ball micro-Snyder column. Pre-wet the ume by refilling the sample bottle to the Snyder column by adding about 0.5 mL of mark and transferring the liquid to an ap- methylene chloride to the top. Place the K- propriately sized graduated cylinder. Record D apparatus on a hot water bath (60–65 °C) so the sample volume to the nearest 5 mL. that the concentrator tube is partially im- Sample volumes may also be determined by mersed in hot water. Adjust the vertical po- weighing the container before and after ex- sition of the apparatus and the water tem- traction or filling to the mark with water. perature as required to complete the con- 10.3 Concentration. centration in 5–10 minutes. At the proper 10.3.1 Macro concentration. rate of distillation the balls of the column 10.3.1.1 Assemble a Kuderna-Danish (K–D) will actively chatter but the chambers will concentrator by attaching a 10-mL concen- not flood with condensed solvent. When the

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apparent volume of liquid reaches approxi- tion of sample volume. Transfer the sample mately 1 mL or other required amount, re- to the continuous extractor and, using a move the K–D apparatus from the water bath pipet, add surrogate standard spiking solu- and allow it to drain and cool for at least 10 tion. If the sample will be used for the LCS, minutes. Remove the Snyder column and MS, or MSD, pipet the appropriate check rinse the flask and its lower joint into the sample concentrate (section 8.2.1 or 8.3.2) concentrator tube with approximately 0.2 into the separatory funnel. Mix well. Add 60 mL of methylene chloride, and proceed to mL of methylene chloride to the sample bot- section 10.3.3 for nitrogen evaporation and tle, seal, and shake for 30 seconds to rinse solvent exchange. the inner surface. Transfer the solvent to the 10.3.3 Nitrogen evaporation and solvent extractor. exchange—Extracts to be subjected to solid- 10.4.3 Repeat the sample bottle rinse with phase cleanup (SPE) are exchanged into 1.0 two additional 50–100 mL portions of meth- mL of the SPE elution solvent (section ylene chloride and add the rinses to the ex- 6.7.2.2). Extracts to be subjected to Florisil® tractor. or alumina cleanups are exchanged into 10.4.4 Add a suitable volume of methylene hexane. Extracts that have been cleaned up chloride to the distilling flask (generally and are ready for analysis are exchanged 200–500 mL) and sufficient reagent water to into isooctane or hexane, to match the sol- ensure proper operation of the extractor, and vent used for the calibration standards. extract the sample for 18–24 hours. A shorter 10.3.3.1 Transfer the vial containing the or longer extraction time may be used if all sample extract to the nitrogen evaporation QC acceptance criteria are met. Test and, if (blowdown) device (section 5.2.5.2). Lower the necessary, adjust the pH of the water to a vial into a 50–55 °C water bath and begin con- range of 5.0–9.0 during the second or third centrating. During the solvent evaporation hour of the extraction. After extraction, process, do not allow the extract to become allow the apparatus to cool, then detach the dry. Adjust the flow of nitrogen so that the distilling flask. Dry, concentrate, solvent ex- surface of the solvent is just visibly dis- change, and transfer the extract to a vial turbed. A large vortex in the solvent may with fluoropolymer-lined cap, per Section cause analyte loss. 10.3. 10.3.3.2 Solvent exchange. 10.4.5 Determine the original sample vol- 10.3.3.2.1 When the volume of the liquid is ume by refilling the sample bottle to the approximately 500 μL, add 2 to 3 mL of the mark and transferring the liquid to an ap- desired solvent (SPE elution solvent for SPE propriately sized graduated cylinder. Record cleanup, hexane for Florisil or alumina, or the sample volume to the nearest 5 mL. isooctane for final injection into the GC) and Sample volumes may also be determined by continue concentrating to approximately 500 weighing the container before and after ex- μL. Repeat the addition of solvent and con- traction or filling to the mark with water. centrate once more. 10.5 Solid-phase extraction of aqueous 10.3.3.3.2 Adjust the volume of an extract samples. The steps in this section address to be cleaned up by SPE, Florisil®, or alu- the extraction of aqueous field samples using mina to 1.0 mL. Proceed to extract cleanup disk-based solid-phase extraction (SPE) (section 11). media, based on an ATP approved by EPA in 10.3.3.3 Extracts that have been cleaned 1995 (Reference 20). This application of SPE up and are ready for analysis—Adjust the is distinct from that used in this method for final extract volume to be consistent with the cleanup of sample extracts in section the volume extracted and the sensitivity de- 11.2. Analysts must be careful not to confuse sired. The goal is for a full-volume sample the equipment, supplies, or the procedural (e.g., 1-L) to have a final extract volume of 10 steps from these two different uses of SPE. mL, but other volumes may be used. NOTE: Changes to the extraction conditions 10.3.4 Transfer the concentrated extract described below may be made by the labora- to a vial with fluoropolymer-lined cap. Seal tory under the allowance for method flexi- the vial and label with the sample number. bility described in section 8.1, provided that Store in the dark at room temperature until the performance requirements in section 8.2 ready for GC analysis. If GC analysis will not are met. However, changes in SPE materials, be performed on the same day, store the vial formats, and solvents must meet the require- in the dark at ≤6 °C. Analyze the extract by ments in section 8.1.2 and its subsections. GC per the procedure in section 12. 10.5.1 Mark the water meniscus on the 10.4 Continuous liquid/liquid extraction side of the sample bottle for later determina- (CLLE). tion of sample volume. If the sample con- 10.4.1 Use CLLE when experience with a tains particulates, let stand to settle out the sample from a given source indicates an particulates before extraction. emulsion problem, or when an emulsion is 10.5.2 Extract the sample as follows: encountered using SFLLE. CLLE may be 10.5.2.1 Place a 90-mm standard filter ap- used for all samples, if desired. paratus on a vacuum filtration flask or 10.4.2 Mark the water meniscus on the manifold and attach to a vacuum source. The side of the sample bottle for later determina- vacuum gauge must read at least 25 in. of

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mercury when all valves are closed. Position 10.5.4 Dry, concentrate, solvent exchange, a 90-mm C18 extraction disk onto the filter and transfer the extract to a vial with screen. Wet the entire disk with methanol. fluoropolymer-lined cap, per section 10.3. To aid in filtering samples with particulates, 10.5.5 Determine the original sample vol- a 1-μm glass fiber filter or Empore® Filter ume by refilling the sample bottle to the Aid 400 can be placed on the top of the disk mark and transferring the liquid to an ap- and wetted with methanol. Install the res- propriately sized graduated cylinder. Record ervoir and clamp. Resume vacuum to dry the the sample volume to the nearest 5 mL. disk. Interrupt the vacuum. Wash the disk Sample volumes may also be determined by and reservoir with 20 mL of methylene chlo- weighing the container before and after ex- ride. Resume the vacuum briefly to pull traction or filling to the mark with water. methylene chloride through the disk. Inter- rupt the vacuum and allow the disk to soak 11. Extract Cleanup for about a minute. Resume vacuum and 11.1 Cleanup may not be necessary for rel- completely dry the disk. atively clean samples (e.g., treated effluents, 10.5.2.2 Condition the disk with 20 mL of groundwater, drinking water). If particular methanol. Apply vacuum until nearly all the circumstances require the use of a cleanup solvent has passed through the disk, inter- procedure, the laboratory may use any or all rupting it while solvent remains on the disk. of the procedures below or any other appro- Allow the disk to soak for about a minute. priate procedure (e.g., gel permeation chro- Resume vacuum to pull most of the meth- matography). However, the laboratory must anol through, but interrupting it to leave a first repeat the tests in sections 8.2, 8.3, and layer of methanol on the surface of the disk. 8.4 to demonstrate that the requirements of Do not allow disk to dry. For uniform flow those sections can be met using the cleanup and good recovery, it is critical the disk not procedure(s) as an integral part of this meth- be allowed to dry from now until the end of od. This is particularly important when the the extraction. Discard waste solvent. Rinse target analytes for the analysis include any the disk with 20 mL of deionized water. Re- of the single component pesticides in Table sume vacuum to pull most of the water 2, because some cleanups have not been opti- through, but interrupt it to leave a layer of mized for all of those analytes. water on the surface of the disk. Do not 11.1.1 The solid-phase cartridge (section allow the disk to dry. If disk does dry, recon- 11.2) removes polar organic compounds such dition with methanol as above. as phenols. 10.5.2.3 Add the water sample to the res- 11.1.2 The Florisil® column (section 11.3) ervoir and immediately apply the vacuum. If allows for selected fractionation of the particulates have settled in the sample, organochlorine analytes and will also elimi- gently decant the clear layer into the appa- nate polar interferences. ratus until most of the sample has been proc- 11.1.3 Alumina column cleanup (section essed. Then pour the remainder including the 11.4) also removes polar materials. particulates into the reservoir. Empty the 11.1.4 Elemental sulfur, which interferes sample bottle completely. When the filtra- with the electron capture gas chroma- tion is complete, dry the disk for three min- tography of some of the pesticides, may be utes. Turn off the vacuum. removed using activated copper, or TBA sul- 10.5.3 Discard sample filtrate. Insert tube fite. Sulfur removal (section 11.5) is required to collect the eluant. The tube should fit when sulfur is known or suspected to be around the drip tip of the base. Reassemble present. Some chlorinated pesticides which the apparatus. Add 5.0 mL of acetone to the also contain sulfur may be removed by this center of the disk, allowing it to spread cleanup. evenly over the disk. Turn the vacuum on 11.1.5 Acid back extraction (section 11.6) and quickly off when the filter surface nears may be useful for cleanup of PCBs and other dryness but still remains wet. Allow to soak compounds not adversely affected by sulfuric for 15 seconds. Add 20 mL of methylene chlo- acid. ride to the sample bottle, seal and shake to 11.2 Solid-phase extraction (SPE) as a rinse the inside of the bottle. Transfer the cleanup. In order to use the C18 SPE car- methylene chloride from the bottle to the tridge in section 5.5.3.5 as a cleanup proce- filter. Resume the vacuum slowly so as to dure, the sample extract must be exchanged avoid splashing. from methylene chloride to methylene chlo- Interrupt the vacuum when the filter sur- ride:acetonitrile:hexane (50:3:47). Follow the face nears dryness but still remains wet. solvent exchange steps in section 10.3.3.2 Allow disk to soak in solvent for 20 seconds. prior to attempting solid-phase cleanup. Rinse the reservoir glass and disk with 10 NOTE: This application of SPE is distinct mL of methylene chloride. Resume vacuum from that used in this method for the extrac- slowly. Interrupt vacuum when disk is cov- tion of aqueous samples in section 10.5. Ana- ered with solvent. Allow to soak for 20 sec- lysts must be careful not to confuse the onds. Resume vacuum to dry the disk. Re- equipment, supplies, or procedural steps move the sample tube. from these two different uses of SPE.

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11.2.1 Setup. the column to settle the Florisil® and add 1 11.2.1.1 Attach the VacElute Manifold to 2 cm of granular anhydrous sodium sul- (section 5.5.3.2) to a water aspirator or vacu- fate to the top. um pump with the trap and gauge installed 11.3.2 Add 60 mL of hexane to wet and between the manifold and vacuum source. rinse the sodium sulfate and Florisil®. Just 11.2.1.2 Place the SPE cartridges in the prior to exposure of the sodium sulfate layer manifold, turn on the vacuum source, and to the air, stop the elution of the hexane by adjust the vacuum to 5 to 10 psi. closing the stopcock on the chromatographic 11.2.2 Cartridge washing—Pre-elute each column. Discard the eluant. cartridge prior to use sequentially with 10- 11.3.3 Transfer the concentrated extract mL portions each of hexane, methanol, and (section 10.3.3) onto the column. Complete water using vacuum for 30 seconds after each the transfer with two 1-mL hexane rinses, eluting solvent. Follow this pre-elution with drawing the extract and rinses down to the 1 mL methylene chloride and three 10-mL level of the sodium sulfate. portions of the elution solvent (section 11.3.4 Place a clean 500-mL K–D flask and 6.7.2.2) using vacuum for 5 minutes after each concentrator tube under the column. Elute eluting solvent. Tap the cartridge lightly Fraction 1 with 200 mL of 6% (v/v) ethyl while under vacuum to dry between solvent ether in hexane at a rate of approximately 5 rinses. The three portions of elution solvent mL/min. Remove the K–D flask and set it may be collected and used as a cartridge aside for later concentration. Elute Fraction blank, if desired. Finally, elute the cartridge 2 with 200 mL of 15% (v/v) ethyl ether in with 10 mL each of methanol and water, hexane into a second K–D flask. Elute Frac- using the vacuum for 30 seconds after each tion 3 with 200 mL of 50% (v/v) ethyl ether in eluant. hexane into a third K–D flask. The elution 11.2.3 Extract cleanup. patterns for the pesticides and PCBs are 11.2.3.1 After cartridge washing (section shown in Table 6. 11.2.2), release the vacuum and place the 11.3.5 Concentrate the fractions as in Sec- rack containing the 50-mL volumetric flasks tion 10.3, except use hexane to prewet the (section 5.5.3.4) in the vacuum manifold. Re- column and set the water bath at about 85 ° establish the vacuum at 5 to 10 psi. C. When the apparatus is cool, remove the 11.2.3.2 Using a pipette or a 1-mL syringe, Snyder column and rinse the flask and its transfer 1.0 mL of extract to the SPE car- lower joint into the concentrator tube with tridge. Apply vacuum for five minutes to dry hexane. Adjust the volume of Fraction 1 to the cartridge. Tap gently to aid in drying. approximately 10 mL for sulfur removal 11.2.3.3 Elute each cartridge into its volu- (Section 11.5), if required; otherwise, adjust metric flask sequentially with three 10-mL the volume of the fractions to 10 mL, 1.0 mL, portions of the methylene chlo- or other volume needed for the sensitivity ride:acetonitrile:hexane (50:3:47) elution sol- desired. Analyze the concentrated extract by vent (section 6.7.2.2), using vacuum for five gas chromatography (Section 12). 11.4 Alumina. The sample extract must be minutes after each portion. Collect the exchanged from methylene chloride to eluants in the 50-mL volumetric flasks. 11.2.3.4 Release the vacuum and remove hexane. Follow the solvent exchange steps in the 50-mL volumetric flasks. section 10.3.3.2 prior to attempting alumina 11.2.3.5 Concentrate the eluted extracts cleanup. 11.4.1 If the chromatographic column does per Section 10.3. 11.3 Florisil®. In order to use Florisil not contain a frit at the bottom, place a cleanup, the sample extract must be ex- small plug of pre-cleaned glass wool in the changed from methylene chloride to hexane. chromatographic column (section 5.2.4) to re- Follow the solvent exchange steps in section tain the alumina. Add 10 g of alumina (sec- 10.3.3.2 prior to attempting Florisil® cleanup. tion 6.7.3) on top of the plug. Tap the column to settle the alumina. Place 1–2 g of anhy- NOTE: Alternative formats for this cleanup drous sodium sulfate on top of the alumina. may be used by the laboratory, including 11.4.2 Close the stopcock and fill the col- ® cartridges containing Florisil . If an alter- umn to just above the sodium sulfate with native format is used, consult the manufac- hexane. Add 25 mL of hexane. Open the stop- turer’s instructions and develop a formal cock and adjust the flow rate of hexane to documented procedure to replace the steps in approximately 2 mL/min. Do not allow the section 11.3 of this method and demonstrate column to go dry throughout the elutions. that the alternative meets the relevant qual- 11.4.3 When the level of the hexane is at ity control requirements of this method. the top of the column, quantitatively trans- 11.3.1 If the chromatographic column does fer the extract to the column. When the level not contain a frit at the bottom, place a of the extract is at the top of the column, small plug of pre-cleaned glass wool in the slowly add 25 mL of hexane and elute the col- column (section 5.2.4) to retain the Florisil®. umn to the level of the sodium sulfate. Dis- Place the mass of Florisil® (nominally 20 g) card the hexane. predetermined by calibration (section 7.8 and 11.4.4 Place a K–D flask (section 5.2.5.1.2) Table 6) in a chromatographic column. Tap under the column and elute the pesticides

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with approximately 150 mL of hexane:ethyl if clear crystals (precipitated sodium sulfite) ether (80:20 v/v). It may be necessary to ad- are observed, sufficient sodium sulfite is just the volume of elution solvent for slight- present. If the precipitated sodium sulfite ly different alumina activities. disappears, add more crystalline sodium sul- 11.4.5 Concentrate the extract per section fite in approximately 0.5-g portions until a 10.3. solid residue remains after repeated shaking. 11.5 Sulfur removal—Elemental sulfur 11.5.2.3 Add 5–10 mL of reagent water and will usually elute in Fraction 1 of the shake for 1–2 minutes. Centrifuge to settle Florisil® column cleanup. If Florisil® clean- the solids. up is not used, or to remove sulfur from any 11.5.2.4 Quantitatively transfer the of the Florisil® fractions, use one of the sul- hexane (top) layer through a small funnel fur removal procedures below. These proce- containing a few grams of granular anhy- dures may be applied to extracts in hexane, drous sodium sulfate to a nitrogen-evapo- ethyl ether, or methylene chloride. ration vial or tube and proceed to section NOTE: Separate procedures using copper or 10.3.3 for micro-concentration and solvent TBA sulfite are provided in this section for exchange. sulfur removal. They may be used separately 11.6 Acid back extraction (section 6.1.2). or in combination, if desired. 11.6.1 Quantitatively transfer the extract (section 10.3.1.5) to a 250-mL separatory fun- 11.5.1 Removal with copper (Reference 15). nel. NOTE: Some of the analytes in Table 2 are 11.6.2 Partition the extract against 50 mL not amenable to sulfur removal with copper of sulfuric acid solution (section 6.1.2). Dis- (e.g., atrazine and diazinon). Therefore, be- card the aqueous layer. Repeat the acid fore using copper to remove sulfur from an washing until no color is visible in the aque- extract that will be analyzed for any of the ous layer, to a maximum of four washings. non-PCB analytes in Table 2, the laboratory 11.6.3 Partition the extract against 50 mL must demonstrate that the analytes can be of sodium chloride solution (section 6.7.5). extracted from an aqueous sample matrix Discard the aqueous layer. that contains sulfur and recovered from an 11.6.4 Proceed to section 10.3.3 for micro- extract treated with copper. Acceptable per- concentration and solvent exchange. formance can be demonstrated through the preparation and analysis of a matrix spike 12. Gas Chromatography sample that meets the QC requirements for 12.1 Establish the same operating condi- recovery. tions used in section 7.1 for instrument cali- 11.5.1.1 Quantitatively transfer the ex- bration. tract to a 40- to 50-mL flask or bottle. If 12.2 If the internal standard calibration there is evidence of water in the K–D or procedure is used, add the internal standard round-bottom flask after the transfer, rinse solution (section 6.9.3) to the extract as close the flask with small portions of as possible to the time of injection to mini- hexane:acetone (40:60) and add to the flask or mize the possibility of loss by evaporation, bottle. Mark and set aside the concentration adsorption, or reaction. For example, add 1 flask for future use. μL of 10 μg/mL internal standard solution 11.5.1.2 Add 10–20 g of granular anhydrous into the extract, assuming no dilutions. Mix sodium sulfate to the flask. Swirl to dry the thoroughly. extract. 12.3 Simultaneously inject an appropriate 11.5.1.3 Add activated copper (section volume of the sample extract or standard so- 6.7.4.1.4) and allow to stand for 30–60 minutes, lution onto both columns, using split, swirling occasionally. If the copper does not splitless, solvent purge, large-volume, or on- remain bright, add more and swirl occasion- column injection. Alternatively, if using a ally for another 30–60 minutes. single-column GC configuration, inject an 11.5.1.4 After drying and sulfur removal, appropriate volume of the sample extract or quantitatively transfer the extract to a ni- standard solution onto each GC column inde- trogen-evaporation vial or tube and proceed pendently. If the sample is injected manu- to section 10.3.3 for nitrogen evaporation and ally, the solvent-flush technique should be solvent exchange, taking care to leave the used. The injection volume depends upon the sodium sulfate and copper foil in the flask. technique used and the sensitivity needed to 11.5.2 Removal with TBA sulfite. meet MDLs or reporting limits for regu- 11.5.2.1 Using small volumes of hexane, latory compliance. Injection volumes must quantitatively transfer the extract to a 40- be the same for all extracts. Record the vol- to 50-mL centrifuge tube with ume injected to the nearest 0.05 μL. fluoropolymer-lined screw cap. 12.4 Set the data system or GC control to 11.5.2.2 Add 1–2 mL of TBA sulfite reagent start the temperature program upon sample (section 6.7.4.2.4), 2–3 mL of 2-propanol, and injection, and begin data collection after the approximately 0.7 g of sodium sulfite (sec- solvent peak elutes. Set the data system to tion 6.7.4.2.2) crystals to the tube. Cap and stop data collection after the last analyte is shake for 1–2 minutes. If the sample is color- expected to elute and to return the column less or if the initial color is unchanged, and to the initial temperature.

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12.5 Perform all qualitative and quan- 13.4.1 DB–608 column—DDT and endrin titative measurements as described in Sec- aldehyde tions 14 and 15. When standards and extracts 13.4.2 DB–1701 column—alpha and gamma are not being used for analyses, store them chlordane refrigerated at <6 °C, protected from light, in screw-cap vials equipped with un-pierced NOTE: If using other GC columns or sta- fluoropolymer-lined septa. tionary phases, these resolution criteria apply to these four target analytes and any 13. System and Laboratory Performance other closely eluting analytes on those other 13.1 At the beginning of each shift during GC columns. which standards or extracts are analyzed, GC 13.5 Decomposition of DDT and endrin—If system performance and calibration must be DDT, endrin, or their breakdown products verified for all analytes and surrogates on are to be determined, this test must be per- both column/detector systems. Adjustment formed prior to calibration verification (sec- and/or recalibration (per section 7) are per- tion 13.6). DDT decomposes to DDE and DDD. formed until all performance criteria are Endrin decomposes to endrin aldehyde and met. Only after all performance criteria are endrin ketone. met may samples, blanks and other QC sam- 13.5.1 Inject 1 μL of the DDT and endrin ples, and standards be analyzed. decomposition solution (section 6.8.7). As 13.2 Inject an aliquot of the calibration noted in section 6.8.7, other injection vol- verification standard (section 6.8.4) on both umes may be used as long as the concentra- columns. Inject an aliquot of each of the multi-component standards. tions of DDT and endrin in the solution are 13.3 Retention times—The absolute reten- adjusted to introduce the masses of the two tion times of the peak maxima shall be with- analytes into the instrument that are listed in ±2 seconds of the retention times in the in section 6.8.7. calibration verification (section 7.8). 13.5.2 Measure the areas of the peaks for 13.4 GC resolution—Resolution is accept- DDT, DDE, DDD, endrin, endrin aldehyde, able if the valley height between two peaks and endrin ketone in the chromatogram and (as measured from the baseline) is less than calculate the percent breakdown as shown in 40% of the shorter of the two peaks. the equations below:

13.5.3 Both the % breakdown of DDT and based on the initial calibration data (section of endrin must be less than 20%, otherwise 7.5 or 7.6). the system is not performing acceptably for 13.6.2 For each analyte or for coeluting DDT and endrin. In this case, repair the GC analytes, compare the concentration with column system that failed and repeat the the limits for calibration verification in performance tests (sections 13.2 to 13.6) until Table 4. For coeluting analytes, use the the specification is met. coeluting analyte with the least restrictive specification (the widest range). For NOTE: DDT and endrin decomposition are analytes in Table 2 not listed in Table 4, QC usually caused by accumulations of particu- acceptance criteria must be developed by the lates in the injector and in the front end of laboratory. EPA has provided guidance for the column. Cleaning and silanizing the in- development of QC acceptance criteria (Ref- jection port liner, and breaking off a short erences 13 and 14). If the recoveries for all section of the front end of the column will analytes meet the acceptance criteria, sys- usually eliminate the decomposition prob- tem performance is acceptable and analysis lem. Either of these corrective actions may of blanks and samples may continue. If, how- affect retention times, GC resolution, and ever, any recovery falls outside the calibra- calibration linearity. tion verification range, system performance 13.6 Calibration verification. is unacceptable for that analyte. If this oc- 13.6.1 Compute the percent recovery of curs, repair the system and repeat the test each analyte and of the coeluting analytes, (section 13.6), or prepare a fresh calibration

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standard and repeat the test, or recalibrate (unique large) peaks for each multi-compo- (section 7). See Section 8.1.7 for information nent analyte. on repeated test failures. 14.2.3 Define the width of the retention 13.7 Laboratory control sample. time window as three times that standard 13.7.1 Analyze the extract of the LCS (sec- deviation. Establish the center of the reten- tion 6.8.3) extracted with each sample batch tion time window for each analyte by using (Section 8.4). See Section 8.4 for criteria ac- the absolute retention time for each analyte ceptance of the LCS. from the calibration verification standard at 13.7.2 It is suggested, but not required, the beginning of the analytical shift. For that the laboratory update statements of samples run during the same shift as an ini- data quality. Add results that pass the speci- tial calibration, use the retention time of fications in section 13.7.3 to initial (section the mid-point standard of the initial calibra- 8.7) and previous ongoing data. Update QC tion. If the calculated RT window is less charts to form a graphic representation of than 0.02 minutes, then use 0.02 minutes as continued laboratory performance. Develop a the window. statement of laboratory data quality for NOTE: Procedures for establishing reten- each analyte by calculating the average per- tion time windows from other sources may cent recovery (R) and the standard deviation be employed provided that they are clearly of percent recovery, sr. Express the accuracy documented and provide acceptable perform- as a recovery interval from R ¥ 2sr to R + ance. Such performance may be evaluated 2sr. For example, if R = 95% and sr = 5%, the using the results for the spiked QC samples accuracy is 85 to 105%. described in this method, such as laboratory 13.8 Internal standard response—If inter- control samples and matrix spike samples. nal standard calibration is used, verify that detector sensitivity has not changed by com- 14.2.4 The retention time windows must paring the response (area or height) of each be recentered when a new GC column is in- internal standard in the sample, blank, LCS, stalled or if a GC column has been shortened MS, and MSD to the response in calibration during maintenance to a degree that the re- verification (section 6.8.3). The peak area or tention times of analytes in the calibration height of the internal standard should be verification standard have shifted close to within 50% to 200% (1⁄2 to 2x) of its respective the lower limits of the established retention peak area or height in the verification stand- time windows. ard. If the area or height is not within this 14.2.5 RT windows should be checked peri- range, compute the concentration of the odically by examining the peaks in spiked analytes using the external standard method samples such as the LCS or MS/MSD to con- (section 7.5). If the analytes are affected, re- firm that peaks for known analytes are prop- prepare and reanalyze the sample, blank, erly identified. LCS, MS, or MSD, and repeat the pertinent 14.2.6 If the retention time of an analyte test. in the calibration (Section 7.4) varies by more than 5 seconds across the calibration 14. Qualitative Identification range as a function of the concentration of 14.1 Identification is accomplished by the standard, using the standard deviation of comparison of data from analysis of a sam- the retention times (section 14.2.3) to set the ple, blank, or other QC sample with data width of the retention time window may not from calibration verification (section 7.7.1 or adequately serve to identify the analyte in 13.5), and with data stored in the retention- question under routine conditions. In such time and calibration libraries (section 7.7). cases, data from additional analyses of The retention time window is determined as standards may be required to adequately described in section 14.2. Identification is model the chromatographic behavior of the confirmed when retention time agrees on analyte. both GC columns, as described below. Alter- 14.3 Identifying the analyte in a sample. natively, GC/MS identification may be used 14.3.1 In order to identify a single-compo- to provide another means of identification. nent analyte from analysis of a sample, 14.2 Establishing retention time windows. blank, or other QC sample, the peak rep- 14.2.1 Using the data from the multi-point resenting the analyte must fall within its re- initial calibration (section 7.4), determine spective retention time windows on both col- the retention time in decimal minutes (not umn/detector systems (as defined in section minutes:seconds) of each peak representing a 14.2). That identification is further supported single-component target analyte on each col- by the comparison of the numerical results umn/detector system. For the multi-compo- on both columns, as described in section 15.7. nent analytes, use the retention times of the 14.3.2 In order to identify a multi-compo- five largest peaks in the chromatograms on nent analyte, pattern matching each column/detector system. (fingerprinting) may be used, or the three to 14.2.2 Calculate the standard deviation of five exclusive (unique and largest) peaks for the retention times for each single-compo- that analyte must fall within their respec- nent analyte on each column/detector sys- tive retention time windows on both column/ tem and for the three to five exclusive detector systems (as defined in section 14.2).

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That identification is further supported by example, if the RT is very stable (i.e., varies the comparison of the numerical results on by no more than a few seconds) for the cali- both columns, as described in section 15.7. bration, calibration verification, blank, LCS, Alternatively, GC/MS identification may be and MS/MSD, the RT for the analyte of in- used. Differentiation among some of the terest in the sample should be within this Aroclors may require evaluation of more variation regardless of the window estab- than five peaks to ensure correct identifica- lished in Section 14.2. If the analyte is not tion. within this variation on both columns, it is 14.4 GC/MS confirmation. When the con- likely not present. centration of an analyte is sufficient and the 14.5.2 The possibility exists that the RT presence or identity is suspect, its presence for the analyte in a sample could shift if ex- should be confirmed by GC/MS. In order to traneous materials are present. This possi- match the sensitivity of the GC/ECD, con- bility may be able to be confirmed or refuted firmation would need to be by GC/MS–SIM, by the behavior of the surrogates in the sam- or the estimated concentration would need ple. If multiple surrogates are used that span to be 100 times higher than the GC/ECD cali- the length of the chromatographic run, the bration range. The extract may be con- RTs for the surrogates on both columns are centrated by an additional amount to allow consistent with their RTs in calibration, a further attempt at GC/MS confirmation. calibration verification, blank, LCS, and MS/ 14.5 Additional information that may aid MSD, it is unlikely that the RT for the the laboratory in the identification of an analyte of interest has shifted. analyte. The occurrence of peaks eluting 14.5.3 If the RT for the analyte is shifted near the retention time of an analyte of in- slightly later on one column and earlier on terest increases the probability of a false the other, and the surrogates have not shift- positive for the analyte. If the concentration ed, it is highly unlikely that the analyte is is insufficient for confirmation by GC/MS, present, because shifts nearly always occur the laboratory may use the cleanup proce- in the same direction on both columns. dures in this method (section 11) on a new sample aliquot to attempt to remove the 15. Quantitative Determination interferent. After attempts at cleanup are exhausted, the following steps may be help- 15.1 External standard quantitation—Cal- ful to assure that the substance that appears culate the concentration of the analyte in in the RT windows on both columns is the the extract using the calibration curve or av- analyte of interest. erage calibration factor determined in cali- 14.5.1 Determine the consistency of the bration (section 7.5.2) and the following RT data for the analyte on each column. For equation:

where: 15.2 Internal standard quantitation—Cal-

Cex = Concentration of the analyte in the ex- culate the concentration of the analyte in tract (ng/mL) the extract using the calibration curve or av- As = Peak height or area for the analyte in erage response factor determined in calibra- the standard or sample tion (section 7.6.2) and the following equa- CF = Calibration factor, as defined in Sec- tion: tion 7.5.1

where: Cis = Concentration of the internal standard (ng/mL) Cex = Concentration of the analyte in the ex- tract (ng/mL) Ais = Area of the internal standard RF = Response factor, as defined in section As = Peak height or area for the analyte in the standard or sample 7.6.1

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15.3 Calculate the concentration of the sample volume, and the dilution factor, per analyte in the sample using the concentra- the following equation: tion in the extract, the extract volume, the

where: Pollutant Discharge Elimination System (NPDES). The data reporting practices de- Cs = Concentration of the analyte in the sam- ple (μg/L) scribed here are focused on such monitoring needs and may not be relevant to other uses Vex = Final extract volume (mL) of the method. Cex = Concentration in the extract (ng/mL) 15.6.1 Report results for wastewater sam- Vs = Volume of sample (L) DF = Dilution factor ples in μg/L without correction for recovery. and the factor of 1,000 in the denominator (Other units may be used if required by in a converts the final units from ng/L to μg/L permit.) Report all QC data with the sample 15.4 If the concentration of any target results. analyte exceeds the calibration range, either 15.6.2 Reporting level. Unless specified extract and analyze a smaller sample vol- otherwise by a regulatory authority or in a ume, or dilute and analyze the diluted ex- discharge permit, results for analytes that tract. meet the identification criteria are reported 15.5 Quantitation of multi-component down to the concentration of the ML estab- analytes. lished by the laboratory through calibration 15.5.1 PCBs as Aroclors. Quantify an of the instrument (see section 7.5 or 7.6 and Aroclor by comparing the sample chromato- the glossary for the derivation of the ML). gram to that of the most similar Aroclor EPA considers the terms ‘‘reporting limit,’’ standard as indicated in section 14.3.2. Com- ‘‘quantitation limit,’’ and ‘‘minimum level’’ pare the responses of 3 to 5 major peaks in to be synonymous. the calibration standard for that Aroclor 15.6.2.1 Report the lower result from the with the peaks observed in the sample ex- two columns (see section 15.7 below) for each tract. The amount of Aroclor is calculated analyte in each sample or QC standard at or using the individual calibration factor for above the ML to 3 significant figures. Report each of the 3 to 5 characteristic peaks chosen a result for each analyte in each sample or in section 7.5.1. Determine the concentration QC standard below the ML as ‘‘

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from different analyses. Results for each would be exceeded for a re-analysis of the analyte in MS/MSD samples should be re- sample, the regulatory/control authority ported from the same GC column as used to should be consulted for disposition. report the results for that analyte in the 15.6.3 Analyze the sample by GC/MS or on unspiked sample. If the MS/MSD recoveries a third column when analytes have co-eluted and RPDs calculated in this manner do not or interfere with determination on both col- meet the acceptance criteria in Table 4, the umns. analyst may use the results from the other NOTE: Dichlone and kepone do not elute GC column to determine if the MS/MSD re- from the DB–1701 column and must be con- sults meet the acceptance criteria. If such a firmed on a DB–5 column, or by GC/MS. situation occurs, the results for the sample should be recalculated using the same GC 15.7 Quantitative information that may column data as used for the MS/MSD sam- aid in the confirmation of the presence of an ples, and reported with appropriate annota- analyte. tions that alert the data user of the issue. 15.7.1 As noted in Section 14.3, the rel- 15.6.2.4 Results from tests performed with ative agreement between the numerical re- an analytical system that is not in control sults from the two GC columns may be used (i.e., that does not meet acceptance criteria to support the identification of the target for all of QC tests in this method) must not analyte by providing evidence that co- be reported or otherwise used for permitting eluting interferences are not present at the or regulatory compliance purposes, but do retention time of the target analyte. Cal- not relieve a discharger or permittee of re- culate the percent difference (%D) between porting timely results. See section 8.1.7 for the results for the analyte from both col- dispositions of failures. If the holding time umns, as follows:

In general, if the %D of the two results is 16. Analysis of Complex Samples less than 50% (e.g., a factor of 2), then the pesticide is present. This %D is generous and 16.1 Some samples may contain high lev- μ allows for the pesticide that has the largest els (greater than 1 g/L) of the analytes of in- measurement error. terest, interfering analytes, and/or polymeric materials. Some samples may not con- NOTE: Laboratories may employ metrics centrate to 1.0 mL (section 10.3.3.3.2); others less than 50% for this comparison, including may overload the GC column and/or detec- those specified in other analytical methods tor. for these pesticides (e.g., CLP or SW–846). 16.2 When an interference is known or 15.7.2 If the amounts do not agree, and the suspected to be present, the laboratory RT data indicate the presence of the analyte should attempt to clean up the sample ex- (per Section 14), it is likely that a positive tract using the SPE cartridge (section 11.2), interference is present on the column that by Florisil® (Section 11.3), Alumina (Section yielded the higher result. That interferent 11.4), sulfur removal (section 11.5), or another may be represented by a separate peak on clean up procedure appropriate to the the other column that does not coincide with analytes of interest. If these techniques do the retention time of any of the target not remove the interference, the extract is analytes. If the interfering peak is evident diluted by a known factor and reanalyzed on the other column, report the result from (section 12). Dilution until the extract is that column and advise the data user that lightly colored is preferable. Typical dilution the interference resulted in a %D value factors are 2, 5, and 10. greater than 50%. If an interferent is not 16.3 Recovery of surrogate(s)—In most identifiable on the second column, then the samples, surrogate recoveries will be similar results must be reported as ‘‘not detected’’ to those from reagent water. If surrogate re- at the lower concentration. In this event, the covery is outside the limits developed in Sec- pesticide is not confirmed and the reporting tion 8.6, re-extract and reanalyze the sample limit is elevated. See section 8.1.7 for dis- if there is sufficient sample and if it is with- position of problem results. in the 7-day extraction holding time. If sur- NOTE: The resulting elevation of the re- rogate recovery is still outside this range, porting limit may not meet the require- extract and analyze one-tenth the volume of ments for compliance monitoring and the sample to overcome any matrix interference use of additional cleanup procedures may be problems. If a sample is highly colored or required. suspected to be high in concentration, a 1–L

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sample aliquot and a 100-mL sample aliquot threat to the environment when managed could be extracted simultaneously and still properly. Standards should be prepared in meet the holding time criteria, while pro- volumes consistent with laboratory use to viding information about a complex matrix. minimize the disposal of excess volumes of 16.4 Recovery of the matrix spike and ma- expired standards. This method utilizes sig- trix spike duplicate (MS/MSD)—In most nificant quantities of methylene chloride. samples, MS/MSD recoveries will be similar Laboratories are encouraged to recover and to those from reagent water. If either the MS recycle this and other solvents during ex- or MSD recovery is outside the range speci- tract concentration. fied in Section 8.3.3, one-tenth the volume of 18.3 For information about pollution pre- sample is spiked and analyzed. If the matrix vention that may be applied to laboratories spike recovery is still outside the range, the and research institutions, consult ‘‘Less is result for the unspiked sample may not be Better: Laboratory Chemical Management reported or used for permitting or regulatory for Waste Reduction’’ (Reference 19), avail- compliance purposes. See Section 8.1.7 for able from the American Chemical Society’s dispositions of failures. Poor matrix spike Department of Governmental Relations and recovery does not relieve a discharger or per- Science Policy, 1155 16th Street NW., Wash- mittee of reporting timely results. ington DC 20036, 202–872–4477.

17. Method Performance 19. Waste Management 17.1 This method was tested for linearity 19.1 The laboratory is responsible for of spike recovery from reagent water and has complying with all Federal, State, and local been demonstrated to be applicable over the regulations governing waste management, concentration range from 4x MDL to 1000x particularly the hazardous waste identifica- MDL with the following exceptions: tion rules and land disposal restrictions, and Chlordane recovery at 4x MDL was low to protect the air, water, and land by mini- (60%); Toxaphene recovery was demonstrated mizing and controlling all releases from linear over the range of 10x MDL to 1000x fume hoods and bench operations. Compli- MDL (Reference 3). ance is also required with any sewage dis- 17.2 The 1984 version of this method was charge permits and regulations. An overview tested by 20 laboratories using reagent of requirements can be found in Environ- water, drinking water, surface water, and mental Management Guide for Small Lab- three industrial wastewaters spiked at six oratories (EPA 233–B–98–001). concentrations (Reference 2). Concentrations 19.2 Samples at pH <2, or pH >12, are haz- used in the study ranged from 0.5 to 30 μg/L ardous and must be handled and disposed of for single-component pesticides and from 8.5 as hazardous waste, or neutralized and dis- to 400 μg/L for multi-component analytes. posed of in accordance with all federal, state, These data are for a subset of analytes de- and local regulations. It is the laboratory’s scribed in the current version of the method. responsibility to comply with all federal, 17.3 During the development of Method state, and local regulations governing waste 1656, a similar EPA procedure for the management, particularly the hazardous organochlorine pesticides, single-operator waste identification rules and land disposal precision, overall precision, and method ac- restrictions. The laboratory using this meth- curacy were found to be directly related to od has the responsibility to protect the air, the concentration of the analyte and essen- water, and land by minimizing and control- tially independent of the sample matrix. ling all releases from fume hoods and bench Linear equations to describe these relation- operations. Compliance is also required with ships are presented in Table 5. any sewage discharge permits and regula- tions. For further information on waste 18. Pollution Prevention management, see ‘‘The Waste Management 18.1 Pollution prevention encompasses Manual for Laboratory Personnel,’’ also any technique that reduces or eliminates the available from the American Chemical Soci- quantity or toxicity of waste at the point of ety at the address in section 18.3. generation. Many opportunities for pollution 19.3 Many analytes in this method decom- prevention exist in laboratory operations. pose above 500 °C. Low-level waste such as EPA has established a preferred hierarchy of absorbent paper, tissues, animal remains, environmental management techniques that and plastic gloves may be burned in an ap- places pollution prevention as the manage- propriate incinerator. Gross quantities of ment option of first choice. Whenever fea- neat or highly concentrated solutions of sible, the laboratory should use pollution toxic or hazardous chemicals should be pack- prevention techniques to address waste gen- aged securely and disposed of through com- eration. When wastes cannot be reduced at mercial or governmental channels that are the source, the Agency recommends recy- capable of handling toxic wastes. cling as the next best option. 19.4 For further information on waste 18.2 The analytes in this method are used management, consult The Waste Manage- in extremely small amounts and pose little ment Manual for Laboratory Personnel and

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Less is Better-Laboratory Chemical Manage- 1910.1450), Occupational Safety and ment for Waste Reduction, available from Health Administration, OSHA. the American Chemical Society’s Depart- 10. 40 CFR 136.6(b)(4)(j). ment of Government Relations and Science 11. Mills, P.A. ‘‘Variation of Florisil Activ- Policy, 1155 16th Street NW., Washington, DC ity: Simple Method for Measuring Ab- 20036, 202–872–4477. sorbent Capacity and Its Use in Stand- ardizing Florisil Columns,’’ Journal of 20. References the Association of Official Analytical 1. ‘‘Determination of Pesticides and PCBs in Chemists, 51:29, (1968). Industrial and Municipal Wastewaters,’’ 12. 40 CFR 136.6(b)(2)(i). EPA 600/4–82–023, National Technical In- 13. Protocol for EPA Approval of New Meth- formation Service, PB82–214222, Spring- ods for Organic and Inorganic Analytes field, Virginia 22161, April 1982. in Wastewater and Drinking Water 2. ‘‘EPA Method Study 18 Method 608- (EPA–821–B–98–003) March 1999. Organochlorine Pesticides and PCBs,’’ 14. Methods 4500 Cl F and 4500 Cl G, Standard EPA 600/4–84–061, National Technical In- Methods for the Examination of Water formation Service, PB84–211358, Spring- and Wastewater, published jointly by the field, Virginia 22161, June 1984. American Public Health Association, American Water Works Association, and 3. ‘‘Method Detection Limit and Analytical Water Environment Federation, 1015 Fif- Curve Studies, EPA Methods 606, 607, and teenth St., Washington, DC 20005, 20th 608,’’ Special letter report for EPA Con- Edition, 2000. tract 68–03–2606, U.S. Environmental Pro- 15. ‘‘Manual of Analytical Methods for the tection Agency, Environmental Moni- Analysis of Pesticides in Human and En- toring and Support Laboratory, Cin- vironmental Samples,’’ EPA–600/8–80–038, cinnati, Ohio 45268, June 1980. U.S. Environmental Protection Agency, 4. ASTM Annual Book of Standards, Part 31, Health Effects Research Laboratory, Re- D3694–78. ‘‘Standard Practice for Prepa- search Triangle Park, North Carolina. ration of Sample Containers and for 16. USEPA, 2000, Method 1656 Organo-Halide Preservation of Organic Constituents,’’ Pesticides In Wastewater, Soil, Sludge, American Society for Testing and Mate- Sediment, and Tissue by GC/HSD, EPA– rials, Philadelphia. 821–R–00–017, September 2000. 5. Giam, C.S., Chan, H.S., and Nef, G.S. 17. USEPA, 2010, Method 1668C Chlorinated ‘‘Sensitive Method for Determination of Biphenyl Congeners in Water, Soil, Sedi- Phthalate Ester Plasticizers in Open- ment, Biosolids, and Tissue by HRGC/ Ocean Biota Samples,’’ Analytical Chem- HRMS, EPA–820–R–10–005, April 2010. istry, 47:2225 (1975). 18. USEPA, 2007, Method 1699: Pesticides in 6. Giam, C.S. and Chan, H.S. ‘‘Control of Water, Soil, Sediment, Biosolids, and Blanks in the Analysis of Phthalates in Tissue by HRGC/HRMS, EPA–821–R–08– Air and Ocean Biota Samples,’’ U.S. Na- 001, December 2007. tional Bureau of Standards, Special Pub- 19. ‘‘Less is Better,’’ American Chemical So- lication 442, pp. 701–708, 1976. ciety on-line publication, http:// 7. Solutions to Analytical Chemistry Prob- www.acs.org/content/dam/acsorg/about/gov- lems with Clean Water Act Methods, ernance/committees/chemicalsafety/publica- EPA 821–R–07–002, March 2007. tions/less-is-better.pdf. 8. ‘‘Carcinogens-Working With Carcinogens,’’ 20. EPA Method 608 ATP 3M0222, An alter- Department of Health, Education, and native test procedure for the measure- Welfare, Public Health Service, Center ment of organochlorine pesticides and for Disease Control, National Institute polychlorinated biphenyls in waste for Occupational Safety and Health, Pub- water. FEDERAL REGISTER, Vol. 60, No. lication No. 77–206, August 1977. 148 August 2, 1995. 9. ‘‘Occupational Exposure to Hazardous Chemicals in Laboratories,’’ (29 CFR 21. Tables

TABLE 1—PESTICIDES 1

MDL 2 ML 3 Analyte CAS No. (ng/L) (ng/L)

Aldrin ...... 309–00–2 4 12 alpha-BHC ...... 319–84–6 3 9 beta-BHC ...... 319–85–7 6 18 delta-BHC ...... 319–86–8 9 27 gamma-BHC (Lindane) ...... 58–89–9 4 12 alpha-Chlordane 4 ...... 5103–71–9 14 42 gamma-Chlordane 4 ...... 5103–74–2 14 42 4,4′-DDD ...... 72–54–8 11 33 4,4′-DDE ...... 72–55–9 4 12 4,4′-DDT ...... 50–29–3 12 36

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TABLE 1—PESTICIDES 1—Continued

MDL 2 ML 3 Analyte CAS No. (ng/L) (ng/L)

Dieldrin ...... 60–57–1 2 6 Endosulfan I ...... 959–98–8 14 42 Endosulfan II ...... 33213–65–9 4 12 Endosulfan sulfate ...... 1031–07–8 66 198 Endrin ...... 72–20–8 6 18 Endrin aldehyde ...... 7421–93–4 23 70 Heptachlor ...... 76–44–8 3 9 Heptachlor epoxide ...... 1024–57–3 83 249 1 All analytes in this table are Priority Pollutants (40 CFR part 423, appendix A). 2 40 CFR part 136, appendix B, June 30, 1986. 3 ML = Minimum Level—see Glossary for definition and derivation, calculated as 3 times the MDL. 4 MDL based on the MDL for Chlordane.

TABLE 2—ADDITIONAL ANALYTES

MDL 3 ML 4 Analyte CAS No. (ng/L) (ng/L)

Acephate ...... 30560–19–1 Alachlor ...... 15972–60–8 Atrazine ...... 1912–24–9 Benfluralin (Benefin) ...... 1861–40–1 Bromacil ...... 314–40–9 Bromoxynil octanoate ...... 1689–99–2 Butachlor ...... 23184–66–9 Captafol ...... 2425–06–1 Captan ...... 133–06–2 Carbophenothion (Trithion) ...... 786–19–6 Chlorobenzilate ...... 510–15–6 Chloroneb (Terraneb) ...... 2675–77–6 Chloropropylate (Acaralate) ...... 5836–10–2 Chlorothalonil ...... 1897–45–6 Cyanazine ...... 21725–46–2 DCPA (Dacthal) ...... 1861–32–1 2,4′-DDD ...... 53–19–0 2,4′-DDE ...... 3424–82–6 2,4′-DDT ...... 789–02–6 Diallate (Avadex) ...... 2303–16–4 1,2-Dibromo-3-chloropropane (DBCP) ...... 96–12–8 Dichlone ...... 117–80–6 Dichloran ...... 99–30–9 Dicofol ...... 115–32–2 Endrin ketone ...... 53494–70–5 Ethalfluralin (Sonalan) ...... 55283–68–6 Etridiazole ...... 2593–15–9 Fenarimol (Rubigan) ...... 60168–88–9 Hexachlorobenzene 1 ...... 118–74–1 Hexachlorocyclopentadiene 1 ...... 77–47–4 Isodrin ...... 465–73–6 Isopropalin (Paarlan) ...... 33820–53–0 Kepone ...... 143–50–0 Methoxychlor ...... 72–43–5 ...... 51218–45–2 Metribuzin ...... 21087–64–9 Mirex ...... 2385–85–5 (TOK) ...... 1836–75–5 cis-Nonachlor ...... 5103–73–1 trans-Nonachlor ...... 39765–80–5 Norfluorazon ...... 27314–13–2 Octachlorostyrene ...... 29082–74–4 Oxychlordane ...... 27304–13–8 PCNB (Pentachloronitrobenzene) ...... 82–68–8 Pendamethalin (Prowl) ...... 40487–42–1 cis-Permethrin ...... 61949–76–6 trans-Permethrin ...... 61949–77–7 Perthane (Ethylan) ...... 72–56–0 Propachlor ...... 1918–16–7 Propanil ...... 709–98–8 Propazine ...... 139–40–2 Quintozene ...... 82–68–8 Simazine ...... 122–34–9

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TABLE 2—ADDITIONAL ANALYTES—Continued

MDL 3 ML 4 Analyte CAS No. (ng/L) (ng/L)

Strobane ...... 8001–50–1 Technazene ...... 117–18–0 Technical Chlordane 2 ...... Terbacil ...... 5902–51–2 Terbuthylazine ...... 5915–41–3 Toxaphene 1 ...... 8001–35–2 240 720 Trifluralin ...... 1582–09–8 PCB–1016 1 ...... 12674–11–2 PCB–1221 1 ...... 11104–28–2 PCB–1232 1 ...... 11141–16–5 PCB–1242 1 ...... 53469–21–9 65 95 PCB–1248 1 ...... 12672–29–6 PCB–1254 1 ...... 11097–69–1 PCB–1260 1 ...... 11096–82–5 PCB–1268 ...... 11100–14–4 1 Priority Pollutants (40 CFR part 423, appendix A). 2 Technical Chlordane may be used in cases where historical reporting has only been for this form of Chlordane. 3 40 CFR part 136, appendix B, June 30, 1986. 4 ML = Minimum Level—see Glossary for definition and derivation, calculated as 3 times the MDL.

TABLE 3—EXAMPLE RETENTION TIMES 1

Retention time 2 Analyte (min) DB–608 DB–1701

Acephate ...... 5.03 (3) Trifluralin ...... 5.16 6.79 Ethalfluralin ...... 5.28 6.49 Benfluralin ...... 5.53 6.87 Diallate-A ...... 7.15 6.23 Diallate-B ...... 7.42 6.77 alpha-BHC ...... 8.14 7.44 PCNB ...... 9.03 7.58 Simazine ...... 9.06 9.29 Atrazine ...... 9.12 9.12 Terbuthylazine ...... 9.17 9.46 gamma-BHC (Lindane) ...... 9.52 9.91 beta-BHC ...... 9.86 11.90 Heptachlor ...... 10.66 10.55 Chlorothalonil ...... 10.66 10.96 Dichlone ...... 10.80 (4) Terbacil ...... 11.11 12.63 delta-BHC ...... 11.20 12.98 Alachlor ...... 11.57 11.06 Propanil ...... 11.60 14.10 Aldrin ...... 11.84 11.46 DCPA ...... 12.18 12.09 Metribuzin ...... 12.80 11.68 Triadimefon ...... 12.99 13.57 Isopropalin ...... 13.06 13.37 Isodrin ...... 13.47 11.12 Heptachlor epoxide ...... 13.97 12.56 Pendamethalin ...... 14.21 13.46 Bromacil ...... 14.39 (3) alpha-Chlordane ...... 14.63 14.20 Butachlor ...... 15.03 15.69 gamma-Chlordane ...... 15.24 14.36 Endosulfan I ...... 15.25 13.87 4,4′-DDE ...... 16.34 14.84 Dieldrin ...... 16.41 15.25 Captan ...... 16.83 15.43 Chlorobenzilate ...... 17.58 17.28 Endrin ...... 17.80 15.86 Nitrofen (TOK) ...... 17.86 17.47 Kepone ...... 17.92 (35) 4,4′-DDD ...... 18.43 17.77 Endosulfan II ...... 18.45 18.57 Bromoxynil octanoate ...... 18.85 18.57 4,4′-DDT ...... 19.48 18.32

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TABLE 3—EXAMPLE RETENTION TIMES 1—Continued

Retention time 2 Analyte (min) DB–608 DB–1701

Carbophenothion ...... 19.65 18.21 Endrin aldehyde ...... 19.72 19.18 Endosulfan sulfate ...... 20.21 20.37 Captafol ...... 22.51 21.22 Norfluorazon ...... 20.68 22.01 Mirex ...... 22.75 19.79 Methoxychlor ...... 22.80 20.68 Endrin ketone ...... 23.00 21.79 Fenarimol ...... 24.53 23.79 cis-Permethrin ...... 25.00 23.59 trans-Permethrin ...... 25.62 23.92 PCB–1016. PCB–1221. PCB–1232. PCB–1242. PCB–1248. PCB–1254. PCB–1260 (5 peaks) ...... 15.44 14.64 15.73 15.36 16.94 16.53 17.28 18.70 19.17 19.92 Toxaphene (5 peaks) ...... 16.60 16.60 17.37 17.52 18.11 17.92 19.46 18.73 19.69 19.00 1 Data from EPA Method 1656 (Reference 16). 2 Columns: 30-m long x 0.53-mm ID fused-silica capillary; DB–608, 0.83 μm; and DB–1701, 1.0 μm. Conditions suggested to meet retention times shown: 150 °C for 0.5 minute, 150–270 °C at 5 °C/min, and 270 °C until trans- Permethrin elutes. Carrier gas flow rates approximately 7 mL/min. 3 Does not elute from DB–1701 column at level tested. 4 Not recovered from water at the levels tested. 5 Dichlone and Kepone do not elute from the DB–1701 column and should be confirmed on DB–5.

TABLE 4—QC ACCEPTANCE CRITERIA

Maximum Calibration Test X Analyte verification concentration Limit for s Range for Range for P MS/MSD μ (% SD) (%) (%) RPD (%) ( g/L) (%)

Aldrin ...... 75–125 2.0 25 54–130 42–140 35 alpha-BHC ...... 69–125 2.0 28 49–130 37–140 36 beta-BHC ...... 75–125 2.0 38 39–130 17–147 44 delta-BHC ...... 75–125 2.0 43 51–130 19–140 52 gamma-BHC ...... 75–125 2.0 29 43–130 32–140 39 alpha-Chlordane ...... 73–125 50.0 24 55–130 45–140 35 gamma-Chlordane ...... 75–125 50.0 24 55–130 45–140 35 4,4′-DDD ...... 75–125 10.0 32 48–130 31–141 39 4,4′-DDE ...... 75–125 2.0 30 54–130 30–145 35 4,4′-DDT ...... 75–125 10.0 39 46–137 25–160 42 Dieldrin ...... 48–125 2.0 42 58–130 36–146 49 Endosulfan I ...... 75–125 2.0 25 57–141 45–153 28 Endosulfan II ...... 75–125 10.0 63 22–171 D–202 53 Endosulfan sulfate ...... 70–125 10.0 32 38–132 26–144 38 Endrin ...... 5–125 10.0 42 51–130 30–147 48 Heptachlor ...... 75–125 2.0 28 43–130 34–140 43 Heptachlor epoxide ...... 75–125 2.0 22 57–132 37–142 26 Toxaphene ...... 68–134 50.0 30 56–130 41–140 41 PCB–1016 ...... 75–125 50.0 24 61–103 50–140 36 PCB–1221 ...... 75–125 50.0 50 44–150 15–178 48 PCB–1232 ...... 75–125 50.0 32 28–197 10–215 25 PCB–1242 ...... 75–125 50.0 26 50–139 39–150 29 PCB–1248 ...... 75–125 50.0 32 58–140 38–158 35 PCB–1254 ...... 75–125 50.0 34 44–130 29–140 45 PCB–1260 ...... 75–125 50.0 28 37–130 8–140 38 S = Standard deviation of four recovery measurements for the DOC (section 8.2.4). X = Average of four recovery measurements for the DOC (section 8.2.4).

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P = Recovery for the LCS (section 8.4.3). Note: These criteria were developed from data in Table 5 (Reference 2). Where necessary, limits for recovery have been broadened to assure applicability to concentrations below those in Table 5.

TABLE 5—PRECISION AND RECOVERY AS FUNCTIONS OF CONCENTRATION

Single analyst Overall Recovery, X′ ′ ′ Analyte μ precision, sr precision, S ( g/L) (μg/L) (μg/L)

Aldrin ...... 0.81C + 0.04 0.16(X) ¥ 0.04 0.20(X) ¥ 0.01 alpha-BHC ...... 0.84C + 0.03 0.13(X) + 0.04 0.23(X) ¥ 0.00 beta-BHC ...... 0.81C + 0.07 0.22(X) ¥ 0.02 0.33(X) ¥ 0.05 delta-BHC ...... 0.81C + 0.07 0.18(X) + 0.09 0.25(X) + 0.03 gamma-BHC (Lindane) ...... 0.82C ¥ 0.05 0.12(X) + 0.06 0.22(X) + 0.04 Chlordane ...... 0.82C ¥ 0.04 0.13(X) + 0.13 0.18(X) + 0.18 4,4′-DDD ...... 0.84C + 0.30 0.20(X) ¥ 0.18 0.27(X) ¥ 0.14 4,4′-DDE ...... 0.85C + 0.14 0.13(X) + 0.06 0.28(X) ¥ 0.09 4,4′-DDT ...... 0.93C ¥ 0.13 0.17(X) + 0.39 0.31(X) ¥ 0.21 Dieldrin ...... 0.90C + 0.02 0.12(X) + 0.19 0.16(X) + 0.16 Endosulfan I ...... 0.97C + 0.04 0.10(X) + 0.07 0.18(X) + 0.08 Endosulfan II ...... 0.93C + 0.34 0.41(X) ¥ 0.65 0.47(X) ¥ 0.20 Endosulfan sulfate ...... 0.89C ¥ 0.37 0.13(X) + 0.33 0.24(X) + 0.35 Endrin ...... 0.89C ¥ 0.04 0.20(X) + 0.25 0.24(X) + 0.25 Heptachlor ...... 0.69C + 0.04 0.06(X) + 0.13 0.16(X) + 0.08 Heptachlor epoxide ...... 0.89C + 0.10 0.18(X) ¥ 0.11 0.25(X) ¥ 0.08 Toxaphene ...... 0.80C + 1.74 0.09(X) + 3.20 0.20(X) + 0.22 PCB–1016 ...... 0.81C + 0.50 0.13(X) + 0.15 0.15(X) + 0.45 PCB–1221 ...... 0.96C + 0.65 0.29(X) ¥ 0.76 0.35(X) ¥ 0.62 PCB–1232 ...... 0.91C + 10.8 0.21(X) ¥ 1.93 0.31(X) + 3.50 PCB–1242 ...... 0.93C + 0.70 0.11(X) + 1.40 0.21(X) + 1.52 PCB–1248 ...... 0.97C + 1.06 0.17(X) + 0.41 0.25(X) ¥ 0.37 PCB–1254 ...... 0.76C + 2.07 0.15(X) + 1.66 0.17(X) + 3.62 PCB–1260 ...... 0.66C + 3.76 0.22(X) ¥ 2.37 0.39(X) ¥ 4.86 X′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L.

TABLE 6—DISTRIBUTION OF CHLORINATED PESTICIDES AND PCBS INTO FLORISIL® COLUMN FRACTIONS

Percent Recovery by Fraction 1 Analyte 1 2 3

Aldrin ...... 100 alpha-BHC ...... 100 beta-BHC ...... 97 delta-BHC ...... 98 gamma-BHC (Lindane) ...... 100 Chlordane ...... 100 4,4′-DDD ...... 99 4,4′-DDE ...... 98 4,4′-DDT ...... 100 Dieldrin ...... 0 100 Endosulfan I ...... 37 64 Endosulfan II ...... 0 7 91 Endosulfan sulfate ...... 0 0 106 Endrin ...... 4 96 Endrin aldehyde ...... 0 68 26 Heptachlor ...... 100 Heptachlor epoxide ...... 100 Toxaphene ...... 96 PCB–1016 ...... 97 PCB–1221 ...... 97 PCB–1232 ...... 95 4 PCB–1242 ...... 97 PCB–1248 ...... 103 PCB–1254 ...... 90 PCB–1260\. 1 Eluant composition: Fraction 1—6% ethyl ether in hexane. Fraction 2—15% ethyl ether in hexane. Fraction 3—50% ethyl ether in hexane.

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TABLE 7—SUGGESTED CALIBRATION GROUPS 1 TABLE 7—SUGGESTED CALIBRATION GROUPS 1—Continued Analyte Analyte Calibration Group 1: Acephate Endosulfan I Alachlor Mirex Atrazine Terbacil beta-BHC Terbuthylazine Bromoxynil octanoate Triadimefon Captafol Calibration Group 5: Diallate alpha-Chlordane Endosulfan sulfate Captan Endrin Chlorothalonil Isodrin ′ Pendimethalin (Prowl) 4,4 -DDD trans-Permethrin Norfluorazon Calibration Group 2: Simazine alpha-BHC Calibration Group 6: DCPA Aldrin 4,4′-DDE delta-BHC 4,4′-DDT Bromacil Dichlone Butachlor Ethalfluralin Endosulfan II Fenarimol Heptachlor Methoxychlor Kepone Metribuzin Calibration Group 7: Calibration Group 3: Carbophenothion gamma-BHC (Lindane) Chloroneb gamma-Chlordane Chloropropylate Endrin ketone DBCP Heptachlor epoxide Dicofol Isopropalin Endrin aldehyde Nitrofen (TOK) Etridiazone PCNB Perthane cis-Permethrin Propachlor Trifluralin Propanil Callibration Group 4: Propazine Benfluralin 1 The analytes may be organized in other calibration Chlorobenzilate groups, provided that there are no coelution problems and Dieldrin that all QC requirements are met.

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22. Figures

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23. Glossary 23.1 Units of weight and measure and their abbreviations. These definitions and purposes are specific 23.1.1 Symbols. to this method but have been conformed to common usage to the extent possible. °C degrees Celsius μg microgram

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μL microliter blank (section 8.5), a laboratory control sam- < less than ple (LCS, section 8.4), a matrix spike and du- ≤ less than or equal to plicate (MS/MSD; section 8.3), resulting in a > greater than minimum of five samples (1 field sample, 1 % percent blank, 1 LCS, 1 MS, and 1 MSD) and a max- 23.1.2 Abbreviations (in alphabetical imum of 24 samples (20 field samples, 1 order). blank, 1 LCS, 1 MS, and 1 MSD) for the cm centimeter batch. If greater than 20 samples are to be g gram extracted in a 24-hour shift, the samples hr hour must be separated into extraction batches of ID inside diameter 20 or fewer samples. in. inch Field Duplicates—Two samples collected L liter at the same time and place under identical M molar solution—one mole or gram molec- conditions, and treated identically through- ular weight of solute in one liter of solu- out field and laboratory procedures. Results tion of analyses the field duplicates provide an mg milligram estimate of the precision associated with min minute sample collection, preservation, and storage, mL milliliter as well as with laboratory procedures. mm millimeter Field blank—An aliquot of reagent water N Normality—one equivalent of solute in or other reference matrix that is placed in a one liter of solution sample container in the field, and treated as ng nanogram a sample in all respects, including exposure psia pounds-per-square inch absolute to sampling site conditions, storage, preser- psig pounds-per-square inch gauge vation, and all analytical procedures. The v/v volume per unit volume purpose of the field blank is to determine if w/v weight per unit volume the field or sample transporting procedures 23.2 Definitions and acronyms (in alpha- and environments have contaminated the betical order) sample. See also ‘‘Blank.’’ Analyte—A compound or mixture of com- GC—Gas chromatograph or gas chroma- pounds (e.g., PCBs) tested for by this meth- tography. od. The analytes are listed in Tables 1 and 2. Gel-permeation chromatography (GPC)—A Analytical batch—The set of samples ana- form of liquid chromatography in which the lyzed on a given instrument during a 24-hour analytes are separated based on exclusion period that begins and ends with calibration from the solid phase by size. verification (sections 7.8 and 13). See also Internal standard—A compound added to ‘‘Extraction batch.’’ an extract or standard solution in a known Blank (method blank; laboratory blank)— amount and used as a reference for quantita- An aliquot of reagent water that is treated tion of the analytes of interest and surro- exactly as a sample including exposure to all gates. Also see Internal standard quantita- glassware, equipment, solvents, reagents, in- tion. ternal standards, and surrogates that are Internal standard quantitation—A means used with samples. The blank is used to de- of determining the concentration of an termine if analytes or interferences are analyte of interest (Tables 1 and 2) by ref- present in the laboratory environment, the erence to a compound not expected to be reagents, or the apparatus. found in a sample. Calibration factor (CF)—See section 7.5.1. IDC—Initial Demonstration of Capability Calibration standard—A solution prepared (section 8.2); four aliquots of a reference ma- from stock solutions and/or a secondary trix spiked with the analytes of interest and standards and containing the analytes of in- analyzed to establish the ability of the lab- terest, surrogates, and internal standards. oratory to generate acceptable precision and This standard is used to model the response recovery. An IDC is performed prior to the of the GC instrument against analyte con- first time this method is used and any time centration. the method or instrumentation is modified. Calibration verification—The process of Laboratory Control Sample (LCS; labora- confirming that the response of the analyt- tory fortified blank; section 8.4)—An aliquot ical system remains within specified limits of reagent water spiked with known quan- of the calibration. tities of the analytes of interest and surro- Calibration verification standard—The gates. The LCS is analyzed exactly like a standard (section 6.8.4) used to verify cali- sample. Its purpose is to assure that the re- bration (sections 7.8 and 13.6). sults produced by the laboratory remain Extraction Batch—A set of up to 20 field within the limits specified in this method for samples (not including QC samples) started precision and recovery. through the extraction process in a given 24- Laboratory Fortified Sample Matrix—See hour shift. Each extraction batch of 20 or Matrix spike. fewer samples must be accompanied by a Laboratory reagent blank—See blank.

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Matrix spike (MS) and matrix spike dupli- Safety Data Sheet (SDS)—Written infor- cate (MSD) (laboratory fortified sample ma- mation on a chemical’s toxicity, health haz- trix and duplicate)—Two aliquots of an envi- ards, physical properties, fire, and reac- ronmental sample to which known quan- tivity, including storage, spill, and handling tities of the analytes of interest and surro- precautions that meet the requirements of gates are added in the laboratory. The MS/ OSHA, 29 CFR 1910.1200(g) and appendix D to MSD are prepared and analyzed exactly like § 1910.1200. United Nations Globally Har- a field sample. Their purpose is to quantify monized System of Classification and Label- any additional bias and imprecision caused ling of Chemicals (GHS), third revised edi- by the sample matrix. The background con- tion, United Nations, 2009. centrations of the analytes in the sample Should—This action, activity, or proce- matrix must be determined in a separate ali- dural step is suggested but not required. quot and the measured values in the MS/ SPE—Solid-phase extraction; a sample ex- MSD corrected for background concentra- traction or extract cleanup technique in tions. which an analyte is selectively removed May—This action, activity, or procedural from a sample or extract by passage over or step is neither required nor prohibited. through a material capable of reversibly ad- May not—This action, activity, or proce- sorbing the analyte. dural step is prohibited. Stock solution—A solution containing an Method detection limit (MDL)—A detec- analyte that is prepared using a reference tion limit determined by the procedure at 40 material traceable to EPA, the National In- CFR part 136, appendix B. The MDLs deter- stitute of Science and Technology (NIST), or mined by EPA are listed in Tables 1 and 2. As a source that will attest to the purity and noted in section 1.6, use the MDLs in Tables authenticity of the reference material. 1 and 2 in conjunction with current MDL Surrogate—A compound unlikely to be data from the laboratory actually analyzing found in a sample, which is spiked into the samples to assess the sensitivity of this pro- sample in a known amount before extrac- cedure relative to project objectives and reg- tion, and which is quantified with the same ulatory requirements (where applicable). procedures used to quantify other sample Minimum level (ML)—The term ‘‘minimum components. The purpose of the surrogate is level’’ refers to either the sample concentra- to monitor method performance with each tion equivalent to the lowest calibration sample. point in a method or a multiple of the meth- METHOD 609—NITROAROMATICS AND od detection limit (MDL), whichever is high- ISOPHORONE er. Minimum levels may be obtained in sev- eral ways: They may be published in a meth- 1. Scope and Application od; they may be based on the lowest accept- 1.1 This method covers the determination able calibration point used by a laboratory; of certain nitroaromatics and isophorone. or they may be calculated by multiplying The following parameters may be deter- the MDL in a method, or the MDL deter- mined by this method: mined by a laboratory, by a factor of 3. For the purposes of NPDES compliance moni- STORET toring, EPA considers the following terms to Parameter No. CAS No. be synonymous: ‘‘quantitation limit,’’ ‘‘re- porting limit,’’ and ‘‘minimum level.’’ 2,4-Dinitrotoluene ...... 34611 121–14–2 2,6-Dinitrotoluene ...... 34626 606–20–2 MS—Mass spectrometer or mass spectrom- Isophorone ...... 34408 78–59–1 etry. Nitrobenzene ...... 34447 98–95–3 Must—This action, activity, or procedural step is required. 1.2 This is a gas chromatographic (GC) Preparation blank—See blank. method applicable to the determination of Reagent water—Water demonstrated to be the compounds listed above in municipal and free from the analytes of interest and poten- industrial discharges as provided under 40 tially interfering substances at the MDLs for CFR 136.1. When this method is used to ana- the analytes in this method. lyze unfamiliar samples for any or all of the Regulatory compliance limit—A limit on compounds above, compound identifications the concentration or amount of a pollutant should be supported by at least one addi- or contaminant specified in a nationwide tional qualitative technique. This method standard, in a permit, or otherwise estab- describes analytical conditions for a second lished by a regulatory/control authority. gas chromatographic column that can be Relative standard deviation (RSD)—The used to confirm measurements made with standard deviation times 100 divided by the the primary column. Method 625 provides gas mean. Also termed ‘‘coefficient of vari- chromatograph/mass spectrometer (GC/MS) ation.’’ conditions appropriate for the qualitative RF—Response factor. See section 7.6.2. and quantitative confirmation of results for RPD—Relative percent difference. all of the parameters listed above, using the RSD—See relative standard deviation. extract produced by this method.

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1.3 The method detection limit (MDL, de- and rinses with tap water and distilled fined in Section 14.1) 1 for each parameter is water. The glassware should then be drained listed in Table 1. The MDL for a specific dry, and heated in a muffle furnace at 400 °C wastewater may differ from those listed, de- for 15 to 30 min. Some thermally stable ma- pending upon the nature of interferences in terials, such as PCBs, may not be eliminated the sample matrix. by this treatment. Solvent rinses with ace- 1.4 The sample extraction and concentra- tone and pesticide quality hexane may be tion steps in this method are essentially the substituted for the muffle furnace heating. same as in Methods 606, 608, 611, and 612. Thorough rinsing with such solvents usually Thus, a single sample may be extracted to eliminates PCB interference. Volumetric measure the parameters included in the ware should not be heated in a muffle fur- scope of each of these methods. When clean- nace. After drying and cooling, glassware up is required, the concentration levels must should be sealed and stored in a clean envi- be high enough to permit selecting aliquots, ronment to prevent any accumulation of as necessary, to apply appropriate cleanup dust or other contaminants. Store inverted procedures. The analyst is allowed the lati- or capped with aluminum foil. tude, under Section 12, to select 3.1.2 The use of high purity reagents and chromatographic conditions appropriate for solvents helps to minimize interference prob- the simultaneous measurement of combina- lems. Purification of solvents by distillation tions of these parameters. in all-glass systems may be required. 1.5 Any modification of this method, be- 3.2 Matrix interferences may be caused by yond those expressly permitted, shall be con- contaminants that are co-extracted from the sidered as a major modification subject to sample. The extent of matrix interferences application and approval of alternate test will vary considerably from source to source, procedures under 40 CFR 136.4 and 136.5. depending upon the nature and diversity of 1.6 This method is restricted to use by or the industrial complex or municipality being under the supervision of analysts experi- sampled. The cleanup procedure in Section enced in the use of a gas chromatograph and 11 can be used to overcome many of these in the interpretation of gas chromatograms. interferences, but unique samples may re- Each analyst must demonstrate the ability quire additional cleanup approaches to to generate acceptable results with this achieve the MDL listed in Table 1. method using the procedure described in Sec- tion 8.2. 4. Safety 4.1 The toxicity or carcinogenicity of 2. Summary of Method each reagent used in this method has not 2.1 A measured volume of sample, ap- been precisely defined; however, each chem- proximately 1–L, is extracted with meth- ical compound should be treated as a poten- ylene chloride using a separatory funnel. The tial health hazard. From this viewpoint, ex- methylene chloride extract is dried and ex- posure to these chemicals must be reduced to changed to hexane during concentration to a the lowest possible level by whatever means volume of 10 mL or less. Isophorone and available. The laboratory is responsible for nitrobenzene are measured by flame ioniza- maintaining a current awareness file of tion detector gas chromatography (FIDGC). OSHA regulations regarding the safe han- The dinitrotoluenes are measured by elec- dling of the chemicals specified in this meth- tron capture detector gas chromatography od. A reference file of material data handling (ECDGC). 2 sheets should also be made available to all 2.2 The method provides a Florisil column personnel involved in the chemical analysis. cleanup procedure to aid in the elimination Additional references to laboratory safety of interferences that may be encountered. are available and have been identified 4M6 for the information of the analyst. 3. Interferences 5. Apparatus and Materials 3.1 Method interferences may be caused by contaminants in solvents, reagents, glass- 5.1 Sampling equipment, for discrete or ware, and other sample processing hardware composite sampling. that lead to discrete artifacts and/or ele- 5.1.1 Grab sample bottle—1–L or 1-qt, vated baseliles in gas chromatograms. All of amber glass, fitted with a screw cap lined these materials must be routinely dem- with Teflon. Foil may be substituted for Tef- onstrated to be free from interferences under lon if the sample is not corrosive. If amber the conditions of the analysis by running bottles are not available, protect samples laboratory reagent blanks as described in from light. The bottle and cap liner must be Section 8.1.3. washed, rinsed with acetone or methylene 3.1.1 Glassware must be scrupulously chloride, and dried before use to minimize cleaned. 3 Clean all glassware as soon as pos- contamination. sible after use by rinsing with the last sol- 5.1.2 Automatic sampler (optional)—The vent used in it. Solvent rinsing should be fol- sampler must incorporate glass sample con- lowed by detergent washing with hot water, tainers for the collection of a minimum of

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250 mL of sample. Sample containers must be isophorone and nitrobenzene. The electron kept refrigerated at 4 °C and protected from capture detector (ECD) is used when deter- light during compositing. If the sampler uses mining the dinitrotoluenes. Both detectors a peristaltic pump, a minimum length of have proven effective in the analysis of compressible silicone rubber tubing may be wastewaters and were used in develop the used. Before use, however, the compressible method performance statements in Section tubing should be thoroughly rinsed with 14. Guidelines for the use to alternate detec- methanol, followed by repeated rinsings with tors are provided in Section 12.1. distilled water to minimize the potential for contamination of the sample. An integrating 6. Reagents flow meter is required to collect flow propor- 6.1 Reagent water—Reagent water is de- tional composites. fined as a water in which an interferent is 5.2 Glassware (All specifications are sug- not observed at the MDL of the parameters gested. Catalog numbers are included for il- of interest. lustration only.): 6.2 Sodium hydroxide solution (10 N)— 5.2.1 Separatory funnel—2–L, with Teflon Dissolve 40 g of NaOH (ACS) in reagent water stopcock. and dilute to 100 mL. 5.2.2 Drying column—Chromatographic 6.3 Sulfuric acid (1 + 1)—Slowly, add 50 column, approximately 400 mm long × 19 mm mL of H SO (ACS, sp. gr. 1.84) to 50 mL of ID, with coarse frit filter disc. 2 4 reagent water. 5.2.3 Chromatographic column—100 mm 6.4 Acetone, hexane, methanol, methylene long × 10 mm ID, with Teflon stopcock. chloride—Pesticide quality or equivalent. 5.2.4 Concentrator tube, Kuderna-Dan- 6.5 Sodium sulfate—(ACS) Granular, an- ish—10-mL, graduated (Kontes K–570050–1025 hydrous. Purify by heating at 400 °C for 4 h or equivalent). Calibration must be checked in a shallow tray. at the volumes employed in the test. Ground 6.6 Florisil—PR grade (60/100 mesh). Pur- glass stopper is used to prevent evaporation chase activated at 1250 °F and store in dark of extracts. in glass containers with ground glass stop- 5.2.5 Evaporative flask, Kuderna-Danish— pers or foil-lined screw caps. Before use, acti- 500-mL (Kontes K–570001–0500 or equivalent). vate each batch at least 16 h at 200 °C in a Attach to concentrator tube with springs. foil-covered glass container and allow to 5.2.6 Snyder column, Kuderna-Danish— cool. Three-ball macro (Kontes K–503000–0121 or 6.7 Stock standard solutions (1.00 μg/μL)— equivalent). 5.2.7 Snyder column, Kuderna-Danish— Stock standard solutions can be prepared Two-ball micro (Kontes K–569001–0219 or from pure standard materials or purchased equivalent). as certified solutions. 5.2.8 Vials—10 to 15-mL, amber glass, with 6.7.1 Prepare stock standard solutions by Teflon-lined screw cap. accurately weighing about 0.0100 g of pure 5.3 Boiling chips—Approximately 10/40 material. Dissolve the material in hexane mesh. Heat to 400 °C for 30 min or Soxhlet ex- and dilute to volume in a 10-mL volumetric tract with methylene chloride. flask. Larger volumes can be used at the con- 5.4 Water bath—Heated, with concentric venience of the analyst. When compound pu- ring cover, capable of temperature control rity is assayed to be 96% or greater, the (±2 °C). The bath should be used in a hood. weight can be used without correction to cal- 5.5 Balance—Analytical, capable of accu- culate the concentration of the stock stand- rately weighing 0.0001 g. ard. Commercially prepared stock standards 5.6 Gas chromatograph—An analytical can be used at any concentration if they are system complete with gas chromatograph certified by the manufacturer or by an inde- suitable for on-column injection and all re- pendent source. quired accessories including syringes, ana- 6.7.2 Transfer the stock standard solu- lytical columns, gases, detector, and strip- tions into Teflon-sealed screw-cap bottles. ° chart recorder. A data system is rec- Store at 4 C and protect from light. Stock ommended for measuring peak areas. standard solutions should be checked fre- 5.6.1 Column 1—1.2 m long × 2 or 4 mm ID quently for signs of degradation or evapo- glass, packed with 1.95% QF–1/1.5% OV–17 on ration, especially just prior to preparing Gas-Chrom Q (80/100 mesh) or equivalent. calibration standards from them. This column was used to develop the method 6.7.3 Stock standard solutions must be re- performance statements given in Section 14. placed after six months, or sooner if com- Guidelines for the use of alternate column parison with check standards indicates a packings are provided in Section 12.1. problem. 5.6.2 Column 2—3.0 m long × 2 or 4 mm ID 6.8 Quality control check sample con- glass, packed with 3% OV–101 on Gas-Chrom centrate—See Section 8.2.1. Q (80/100 mesh) or equivalent. 7. Calibration 5.6.3 Detectors—Flame ionization and electron capture detectors. The flame ioniza- 7.1 Establish gas chromatographic oper- tion detector (FID) is used when determining ating conditions equivalent to those given in

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Table 1. The gas chromatographic system can be calibrated using the external standard ()()ACsis technique (Section 7.2) or the internal stand- RF = ard technique (Section 7.3). ()()ACis s 7.2 External standard calibration proce- dure: where: 7.2.1 Prepare calibration standards at a As = Response for the parameter to be meas- minimum of three concentration levels for ured. each parameter of interest by adding vol- Ais = Response for the internal standard. umes of one or more stock standards to a Cis = Concentration of the internal standard volumetric flask and diluting to volume with (μg/L). hexane. One of the external standards should Cs = Concentration of the parameter to be be at a concentration near, but above, the measured (μg/L). MDL (Table 1) and the other concentrations If the RF value over the working range is should correspond to the expected range of a constant (<10% RSD), the RF can be as- concentrations found in real samples or sumed to be invariant and the average RF should define the working range of the detec- can be used for calculations. Alternatively, tor. the results can be used to plot a calibration 7.2.2 Using injections of 2 to 5 μL, analyze curve of response ratios, As/Ais, vs. RF. each calibration standard according to Sec- 7.4 The working calibration curve, cali- tion 12 and tabulate peak height or area re- bration factor, or RF must be verified on sponses against the mass injected. The re- each working day by the measurement of one sults can be used to prepare a calibration or more calibration standards. If the re- curve for each compound. Alternatively, if sponse for any parameter varies from the ± the ratio of response to amount injected predicted response by more than 15%, a new (calibration factor) is a constant over the calibration curve must be prepared for that working range (<10% relative standard devi- compound. ation, RSD) linearity through the origin can 7.5 Before using any cleanup procedure, be assumed and the average ratio or calibra- the analyst must process a series of calibra- tion factor can be used in place of a calibra- tion standards through the procedure to vali- tion curve. date elution patterns and the absence of interferences from the reagents. 7.3 Internal standard calibration proce- dure—To use this approach, the analyst must 8. Quality Control select one or more internal standards that are similar in analytical behavior to the 8.1 Each laboratory that uses this method compounds of interest. The analyst must fur- is required to operate a formal quality con- ther demonstrate that the measurement of trol program. The minimum requirements of the internal standard is not affected by this program consist of an initial demonstra- method or matrix interferences. Because of tion of laboratory capability and an ongoing these limitations, no internal standard can analysis of spiked samples to evaluate and be suggested that is applicable to all sam- document data quality. The laboratory must ples. maintain records to document the quality of data that is generated. Ongoing data quality 7.3.1 Prepare calibration standards at a checks are compared with established per- minimum of three concentration levels for formance criteria to determine if the results each parameter of interest by adding vol- of analyses meet the performance character- umes of one or more stock standards to a istics of the method. When results of sample volumetric flash. To each calibration stand- spikes indicate atypical method perform- ard, add a known constant amount of one or ance, a quality control check standard must more internal standards, and dilute to vol- be analyzed to confirm that the measure- ume with hexane. One of the standards ments were performed in an in-control mode should be at a concentration near, but above, of operation. the MDL and the other concentrations 8.1.1 The analyst must make an initial, should correspond to the expected range of one-time, demonstration of the ability to concentrations found in real samples or generate acceptable accuracy and precision should define the working range of the detec- with this method. This ability is established tor. as described in Section 8.2. 7.3.2 Using injections of 2 to 5 μL, analyze 8.1.2 In recognition of advances that are each calibration standard according to Sec- occurring in chromatography, the analyst is tion 12 and tabulate peak height or area re- permitted certain options (detailed in Sec- sponses against concentration for each com- tions 10.4, 11.1, and 12.1) to improve the sepa- pound and internal standard. Calculate re- rations or lower the cost of measurements. sponse factors (RF) for each compound using Each time such a modification is made to Equation 1. the method, the analyst is required to repeat Equation 1. the procedure in Section 8.2.

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8.1.3 Before processing any samples, the or any individual X¯ falls outside the range analyst must analyze a reagent water blank for accuracy, the system performance is un- to demonstrate that interferences from the acceptable for that parameter. Locate and analytical system and glassware are under correct the source of the problem and repeat control. Each time a set of samples is ex- the test for all parameters of interest begin- tracted or reagents are changed, a reagent ning with Section 8.2.2. water blank must be processed as a safe- 8.3 The laboratory must, on an ongoing guard against laboratory contamination. basis, spike at least 10% of the samples from 8.1.4 The laboratory must, on an ongoing each sample site being monitored to assess basis, spike and analyze a minimum of 10% accuracy. For laboratories analyzing one to of all samples to monitor and evaluate lab- ten samples per month, at least one spiked oratory data quality. This procedure is de- sample per month is required. scribed in Section 8.3. 8.3.1 The concentration of the spike in the 8.1,5 The laboratory must, on an ongoing sample should be determined as follows: basis, demonstrate through the analyses of 8.3.1.1 If, as in compliance monitoring, quality control check standards that the op- the concentration of a specific parameter in eration of the measurement system is in con- the sample is being checked against a regu- trol. This procedure is described in Section latory concentration limit, the spike should 8.4. The frequency of the check standard be at that limit or 1 to 5 times higher than analyses is equivalent to 10% of all samples the background concentration determined in analyzed but may be reduced if spike recov- Section 8.3.2, whichever concentration would eries from samples (Section 8.3) meet all be larger. specified quality control criteria. 8.3.1.2 If the concentration of a specific 8.1.6 The laboratory must maintain per- parameter in the sample is not being formance records to document the quality of checked against a limit specific to that pa- data that is generated. This procedure is de- rameter, the spike should be at the test con- scribed in Section 8.5. centration in Section 8.2.2 or 1 to 5 times 8.2 To establish the ability to generate higher than the background concentration acceptable accuracy and precision, the ana- determined in Section 8.3.2, whichever con- lyst must perform the following operations. centration would be larger. 8.2.1 A quality control (QC) check sample 8.3.1.3 If it is impractical to determile concentrate is required containing each pa- background levels before spiking (e.g., max- rameter of interest in acetone at a con- imum holding times will be exceeded), the centration of 20 μg/mL for each dinitro- spike concentration should be (1) the regu- toluene and 100 μg/mL for isophorone and latory concentration limit, if any; or, if none nitrobenzene. The QC check sample con- (2) the larger of either 5 times higher than centrate must be obtained from the U.S. En- the expected background concentration or vironmental Protection Agency, Environ- the test concentration in Section 8.2.2. mental Monitoring and Support Laboratory 8.3.2 Analyze one sample aliquot to deter- in Cincinnati, Ohio, if available. If not avail- mine the background concentration (B) of able from that source, the QC check sample each parameter. If necessary, prepare a new concentrate must be obtained from another QC check sample concentrate (Section 8.2.1) external source. If not available from either appropriate for the background concentra- source above, the QC check sample con- tions in the sample. Spike a second sample centrate must be prepared by the laboratory aliquot with 1.0 mL of the QC check sample using stock standards prepared independ- concentrate and analyze it to determine the ently from those used for calibration. concentration after spiking (A) of each pa- 8.2.2 Using a pipet, prepare QC check sam- rameter. Calculate each percent recovery (P) ples at the test concentrations shown in as 100 (A¥B)%/T, where T is the known true Table 2 by adding 1.00 mL of QC check sam- value of the spike. ple concentrate to each of four 1–L aliquots 8.3.3 Compare the percent recovery (P) for of reagent water. each parameter with the corresponding QC 8.2.3 Analyze the well-mixed QC check acceptance criteria found in Table 2. These samples according to the method beginning acceptance criteria were calculated to in- in Section 10. clude an allowance for error in measurement 8.2.4 Calculate the average recovery (X¯ ) in of both the background and spike concentra- μg/L, and the standard deviation of the re- tions, assuming a spike to background ratio covery (s) in μg/L, for each parameter using of 5:1. This error will be accounted for to the the four results. extent that the analyst’s spike to back- 8.2.5 For each parameter compare s and X¯ ground ratio approaches 5:1. 7 If spiking was with the corresponding acceptance criteria performed at a concentration lower than the for precision and accuracy, respectively, test concentration in Section 8.2.2, the ana- found in Table 2. If s and X¯ for all param- lyst must use either the QC acceptance cri- eters of interest meet the acceptance cri- teria in Table 2, or optional QC acceptance teria, the system performance is acceptable criteria calculated for the specific spike con- and analysis of actual samples can begin. If centration. To calculate optional acceptance any individual s exceeds the precision limit criteria for the recovery of a parameter: (1)

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Calculate accuracy (X′) using the equation in the samples. Field duplicates may be ana- Table 3, substituting the spike concentration lyzed to assess the precision of the environ- (T) for C; (2) calculate overall precision (S′) mental measurements. When doubt exists using the equation in Table 3, substituting X′ over the identification of a peak on the chro- for X¯ 8; (3) calculate the range for recovery at matogram, confirmatory techniques such as the spike concentration as (100 X′/T) ±2.44 gas chromatography with a dissimilar col- (100 S′/T)%. 7 umn, specific element detector, or mass 8.3.4 If any individual P falls outside the spectrometer must be used. Whenever pos- designated range for recovery, that param- sible, the laboratory should analyze standard eter has failed the acceptance criteria. A reference materials and participate in rel- check standard containing each parameter evant performance evaluation studies. that failed the criteria must be analyzed as described in Section 8.4. 9. Sample Collection, Preservation, and 8.4. If any parameter fails the acceptance Handling criteria for recovery in Section 8.3, a QC check standard containing each parameter 9.1 Grab samples must be collected in that failed must be prepared and analyzed. glass containers. Conventional sampling NOTE: The frequency for the required anal- practices 8 should be followed, except that ysis of a QC check standard will depend upon the bottle must not be prerinsed with sample the number of parameters being simulta- before collection. Composite samples should neously tested, the complexity of the sample be collected in refrigerated glass containers matrix, and the performance of the labora- in accordance with the requirements of the tory. program. Automatic sampling equipment 8.4.1 Prepare the QC check standard by must be as free as possible of Tygon tubing adding 1.0 mL of QC check sample con- and other potential sources of contamina- centrate (Section 8.2.1 or 8.3.2) to 1 L of rea- tion. gent water. The QC check standard needs 9.2 All samples must be iced or refrig- only to contain the parameters that failed erated at 4 °C from the time of collection criteria in the test in Section 8.3. until extraction. 8.4.2 Analyze the QC check standard to determine the concentration measured (A) of 9.3 All samples must be extracted within 7 each parameter. Calculate each percent re- days of collection and completely analyzed within 40 days of extraction. 2 covery (Ps) as 100 (A/T)%, where T is the true value of the standard concentration. 10. Sample Extraction 8.4.3 Compare the percent recovery (Ps) for each parameter with the corresponding 10.1 Mark the water meniscus on the side QC acceptance criteria found in Table 2. Only of the sample bottle for later determination parameters that failed the test in Section 8.3 of sample volume. Pour the entire sample need to be compared with these criteria. If into a 2–L separatory funnel. Check the pH the recovery of any such parameter falls out- of the sample with wide-range pH paper and side the designated range, the laboratory adjust to within the range of 5 to 9 with so- performance for that parameter is judged to dium hydroxide solution or sulfuric acid. be out of control, and the problem must be 10.2 Add 60 mL of methylene chloride to immediately identified and corrected. The analytical result for that parameter in the the sample bottle, seal, and shake 30 s to unspiked sample is suspect and may not be rinse the inner surface. Transfer the solvent reported for regulatory compliance purposes. to the separatory funnel and extract the 8.5 As part of QC program for the labora- sample by shaking the funnel for 2 min. with tory, method accuracy for wastewater sam- periodic venting to release excess pressure. ples must be assessed and records must be Allow the organic layer to separate from the maintained. After the analysis of five spiked water phase for a minimum of 10 min. If the wastewater samples as in Section 8.3, cal- emulsion interface between layers is more culate the average percent recovery (P¯ ) and than one-third the volume of the solvent the standard deviation of the percent recov- layer, the analyst must employ mechanical techniques to complete the phase separation. ery (sp). Express the accuracy assessment as ¯ ¯ The optimum technique depends upon the a percent recovery interval from P¥2sp to P ¯ sample, but may include stirring, filtration + 2sp. If P = 90% and sp = 10%, for example, the accuracy interval is expressed as 70– of the emulsion through glass wool, cen- 110%. Update the accuracy assessment for trifugation, or other physical methods. Col- each parameter on a regular basis (e.g. after lect the methylene chloride extract in a 250- each five to ten new accuracy measure- mL Erlenmeyer flask. ments). 10.3 Add a second 60-mL volume of meth- 8.6 It is recommended that the laboratory ylene chloride to the sample bottle and re- adopt additional quality assurance practices peat the extraction procedure a second time, for use with this method. The specific prac- combining the extracts in the Erlenmeyer tices that are most productive depend upon flask. Perform a third extraction in the same the needs of the laboratory and the nature of manner.

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10.4 Assemble a Kuderna-Danish (K-D) be stored longer than two days, it should be concentrator by attaching a 10-mL concen- transferred to a Teflon-sealed screw-cap vial. trator tube to a 500-mL evaporative flask. If the sample extract requires no further Other concentration devices or techniques cleanup, proceed with gas chromatographic may be used in place of the K-D concentrator analysis (Section 12). If the sample requires if the requirements of Section 8.2 are met. further cleanup, proceed to Section 11. 10.5 Pour the combined extract through a 10.10 Determine the original sample vol- solvent-rinsed drying column containing ume by refilling the sample bottle to the about 10 cm of anhydrous sodium sulfate, mark and transferring the liquid to a 1000- and collect the extract in the K-D concen- mL graduated cylinder. Record the sample trator. Rinse the Erlenmeyer flask and col- volume to the nearest 5 mL. umn with 20 to 30 mL of methylene chloride to complete the quantitative transfer. 11. Cleanup and Separation 10.6 Sections 10.7 and 10.8 describe a pro- 11.1 Cleanup procedures may not be nec- cedure for exchanging the methylene chlo- essary for a relatively clean sample matrix. ride solvent to hexane while concentrating If particular circumstances demand the use the extract volume to 1.0 mL. When it is not of a cleanup procedure, the analyst may use necessary to achieve the MDL in Table 2, the the procedure below or any other appropriate solvent exchange may be made by the addi- procedure. However, the analyst first must tion of 50 mL of hexane and concentration to demonstrate that the requirements of Sec- 10 mL as described in Method 606, Sections tion 8.2 can be met using the method as re- 10.7 and 10.8. vised to incorporate the cleanup procedure. 10.7 Add one or two clean boiling chips to 11.2 Florisil column cleanup: the evaporative flask and attach a three-ball 11.2.1 Prepare a slurry of 10 g of activated Snyder column. Prewet the Snyder column Florisil in methylene chloride/hexane (1 + by adding about 1 mL of methylene chloride 9)(V/V) and place the Florisil into a to the top. Place the K-D apparatus on a hot water bath (60 to 65 °C) so that the concen- chromatographic column. Tap the column to trator tube is partially immersed in the hot settle the Florisil and add 1 cm of anhydrous water, and the entire lower rounded surface sodium sulfate to the top. Adjust the elution of the flask is bathed with hot vapor. Adjust rate to about 2 mL/min. the vertical position of the apparatus and 11.2.2 Just prior to exposure of the sodium the water temperature as required to com- sulfate layer to the air, quantitatively trans- plete the concentration in 15 to 20 min. At fer the sample extract onto the column using the proper rate of distillation the balls of the an additional 2 mL of hexane to complete the column will actively chatter but the cham- transfer. Just prior to exposure of the so- bers will not flood with condensed solvent. dium sulfate layer to the air, add 30 mL of When the apparent volume of liquid reaches methylene chloride/hexane (1 + 9)(V/V) and 1 mL, remove the K-D apparatus and allow it continue the elution of the column. Discard to drain and cool for at least 10 min. the eluate. 10.8 Remove the Snyder column and rinse 11.2.3 Next, elute the column with 30 mL the flask and its lower joint into the concen- of acetone/methylene chloride (1 + 9)(V/V) trator tube with 1 to 2 mL of methylene into a 500-mL K-D flask equipped with a 10- chloride. A 5-mL syringe is recommended for mL concentrator tube. Concentrate the col- this operation. Add 1 to 2 mL of hexane and lected fraction as in Sections 10.6, 10.7, 10.8, a clean boiling chip to the concentrator tube and 10.9 including the solvent exchange to 1 and attach a two-ball micro-Snyder column. mL of hexane. This fraction should contain Prewet the column by adding about 0.5 mL of the nitroaromatics and isophorone. Analyze hexane to the top. Place the micro-K-D appa- by gas chromatography (Section 12). ratus on a hot water bath (60 to 65 °C) so that 12. Gas Chromatography the concentrator tube is partially immersed in the hot water. Adjust the vertical position 12.1 Isophorone and nitrobenzene are ana- of the apparatus and the water temperature lyzed by injection of a portion of the extract as required to complete the concentration in into an FIDGC. The dinitrotoluenes are ana- 5 to 10 min. At the proper rate of distillation lyzed by a separate injection into an ECDGC. the balls of the column will actively chatter Table 1 summarizes the recommended oper- but the chambers will not flood. When the ating conditions for the gas chromatograph. apparent volume of liquid reaches 0.5 mL, re- Included in this table are retention times move the K-D apparatus and allow it to and MDL that can be achieved under these drain and cool for at least 10 min. conditions. Examples of the separations 10.9 Remove the micro-Snyder column achieved by Column 1 are shown in Figures 1 and rinse its lower joint into the concen- and 2. Other packed or capillary (open-tubu- trator tube with a minimum amount of lar) columns, chromatographic conditions, hexane. Adjust the extract volume to 1.0 mL. or detectors may be used if the requirements Stopper the concentrator tube and store re- of Section 8.2 are met. frigerated if further processing will not be 12.2 Calibrate the system daily as de- performed immediately. If the extract will scribed in Section 7.

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12.3 If the internal standard calibration Equation 3 procedure is being used, the internal stand- ard must be added to the same extract and where: mixed thoroughly immediately before injec- As = Response for the parameter to be meas- tion into the gas chromatograph. ured. 12.4 Inject 2 to 5 μL of the sample extract Ais = Response for the internal standard. or standard into the gas chromatograph Is = Amount of internal standard added to using the solvent-flush technique. 9 Smaller each extract (μg). (1.0 μL) volumes may be injected if auto- Vo = Volume of water extracted (L). matic devices are employed. Record the vol- 13.2 Report results in μg/L without correc- ume injected to the nearest 0.05 μL, the total tion for recovery data. All QC data obtained extract volume, and the resulting peak size should be reported with the sample results. in area or peak height units. 12.5 Identify the parameters in the sample 14. Method Performance by comparing the retention times of the 14.1 The method detection limit (MDL) is peaks in the sample chromatogram with defined as the minimum concentration of a those of the peaks in standard substance that can be measured and reported chromatograms. The width of the retention with 99% confidence that the value is above time window used to make identifications zero. 1 The MDL concentrations listed in should be based upon measurements of ac- Table 1 were obtained using reagent water. 10 tual retention time variations of standards Similar results were achieved using rep- over the course of a day. Three times the resentative wastewaters. The MDL actually standard deviation of a retention time for a achieved in a given analysis will vary de- compound can be used to calculate a sug- pending on instrument sensitivity and ma- gested window size; however, the experience trix effects. of the analyst should weigh heavily in the interpretation of chromatograms. 14.2 This method has been tested for lin- 12.6 If the response for a peak exceeds the earity of spike recovery from reagent water working range of the system, dilute the ex- and has been demonstrated to be applicable × tract and reanalyze. over the concentration range from 7 MDL to 1000 × MDL. 10 12.7 If the measurement of the peak re- sponse is prevented by the presence of inter- 14.3 This method was tested by 18 labora- ferences, further cleanup is required. tories using reagent water, drinking water, surface water, and three industrial 13. Calculations wastewaters spiked at six concentrations over the range 1.0 to 515 μg/L. 11 Single oper- 13.1 Determine the concentration of indi- ator precision, overall precision, and method vidual compounds in the sample. accuracy were found to be directly related to 13.1.1 If the external standard calibration the concentration of the parameter and es- procedure is used, calculate the amount of sentially independent of the sample matrix. material injected from the peak response Linear equations to describe these relation- using the calibration curve or calibration ships are presented in Table 3. factor determined in Section 7.2.2. The con- centration in the sample can be calculated REFERENCES from Equation 2. 1. 40 CFR part 136, appendix B. ()AV() 2. ‘‘Determination of Nitroaromatic Com- Concentration (μ= g/L) t pounds and Isophorone in Industrial and Mu- ()() nicipal Wastewaters,’’ EPA 600/ 4–82–024, Na- VVis tional Technical Information Service, PB82– Equation 2 208398, Springfield, Virginia 22161, May 1982. 3. ASTM Annual Book of Standards, Part where: 31, D3694–78. ‘‘Standard Practices for Prepa- A = Amount of material injected (ng). ration of Sample Containers and for Preser- Vi = Volume of extract injected (μL). vation of Organic Constituents,’’ American Vt = Volume of total extract (μL). Society for Testing and Materials, Philadel- Vs = Volume of water extracted (mL). phia. 13.1.2 If the internal standard calibration 4. ‘‘Carcinogens—Working With Carcino- procedure is used, calculate the concentra- gens,’’ Department of Health, Education, and tion in the sample using the response factor Welfare, Public Health Service, Center for (RF) determined in Section 7.3.2 and Equa- Disease Control, National Institute for Occu- tion 3. pational Safety and Health, Publication No. 77–206, August 1977. ()()AI 5. ‘‘OSHA Safety and Health Standards, Concentration (μ= g/L) ss General Industry,’’ (29 CFR part 1910), Occu- ()ARFV()() pational Safety and Health Administration, is o OSHA 2206 (Revised, January 1976).

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6. ‘‘Safety in Academic Chemistry Labora- Aspects,’’ Journal of the Association of Official tories,’’ American Chemical Society Publica- Analytical Chemists, 48, 1037 (1965). tion, Committee on Chemical Safety, 3rd 10. ‘‘Determination of Method Detection Edition, 1979. Limit and Analytical Curve for EPA Method 7. Provost, L.P., and Elder, R.S. ‘‘Interpre- 609—Nitroaromatics and Isophorone,’’ Spe- tation of Percent Recovery Data,’’ American cial letter report for EPA Contract 68–03– 58–63 (1983). (The value 2.44 Laboratory, 15, 2624, U.S. Environmental Protection Agency, used in the equation in Section 8.3.3 is two Environmental Monitoring and Support Lab- times the value 1.22 derived in this report.) 8. ASTM Annual Book of Standards, Part oratory, Cincinnati, Ohio 45268, June 1980. 31, D3370–76. ‘‘Standard Practices for Sam- 11. ‘‘EPA Method Study 19, Method 609 pling Water,’’ American Society for Testing (Nitroaromatics and Isophorone),’’ EPA 600/ and Materials, Philadelphia. 4–84–018, National Technical Information 9. Burke, J.A. ‘‘Gas Chromatography for Service, PB84–176908, Springfield, Virginia Pesticide Residue Analysis; Some Practical 22161, March 1984.

TABLE 1—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS

Retention time (min) Method detection limit (μg/L) Parameter Col. 1 Col. 2 ECDGC FIDGC

Nitrobenzene ...... 3.31 4 .31 13 .7 3 .6 2,6-Dinitrotoluene ...... 3 .52 4 .75 0 .01 ¥ Isophorone ...... 4 .49 5 .72 15 .7 5 .7 2,4-Dinitrotoluene ...... 5 .35 6 .54 0 .02 ¥ Column 1 conditions: Gas-Chrom Q (80/100 mesh) coated with 1.95% QF–1/1.5% OV–17 packed in a 1.2 m long × 2 mm or 4 mm ID glass column. A 2 mm ID column and nitrogen carrier gas at 44 mL/min flow rate were used when determining isophorone and nitrobenzene by FIDGC. The column temperature was held isothermal at 85 °C. A 4 mm ID column and 10% methane/90% argon carrier gas at 44 mL/min flow rate were used when determining the dinitrotoluenes by ECDGC. The column temperature was held isothermal at 145 °C. Column 2 conditions: Gas-Chrom Q (80/100 mesh) coated with 3% OV–101 packed in a 3.0 m long × 2 mm or 4 mm ID glass column. A 2 mm ID column and nitrogen carrier gas at 44 mL/min flow rate were used when determining isophorone and nitrobenzene by FIDGC. The column temperature was held isothermal at 100 °C. A 4 mm ID column and 10% methane/90% argon carrier gas at 44 mL/min flow rate were used when determining the dinitrotoluenes by ECDGC. The column temperature was held isothermal at 150 °C.

TABLE 2—QC ACCEPTANCE CRITERIA—METHOD 609

¯ Parameter Test Conc. Limit for s Range for X Range for (μg/L) (μg/L) (μg/L) P, Ps (%)

2,4-Dinitrotoluene ...... 20 5 .1 3.6 –22.8 6–125 2,6-Dinitrotoluene ...... 20 4 .8 3.8 –23.0 8–126 Isophorone ...... 100 32.3 8.0 –100.0 D–117 Nitrobenzene ...... 100 33 .3 25.7 –100.0 6–118 s = Standard deviation of four recovery measurements, in μg/L (Section 8.2.4). X¯ = Average recovery for four recovery measurements, in μg/L (Section 8.2.4). P, Ps = Percent recovery measured (Section 8.3.2, Section 8.4.2). D = Detected; result must be greater than zero. NOTE: These criteria are based directly upon the method performance data in Table 3. Where necessary, the limits for recov- ery have been broadened to assure applicability of the limits to concentrations below those used to develop Table 3.

TABLE 3—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 609

Parameter Accuracy, as re- Single analyst pre- Overall precision, covery, X′ (μg/L) cision, sr′ (μg/L) S′ (μg/L)

2,4-Dinitro- toluene ...... 0.65C + 0.22 0.20X¯ + 0.08 0.37X¯ ¥0.07 2,6-Dinitro- toluene ...... 0.66C + 0.20 0.19X¯ + 0.06 0.36X¯ ¥0.00 Isophorone ...... 0.49C + 2.93 0.28X¯ + 2.77 0.46X¯ + 0.31 Nitrobenzene ...... 0.60C + 2.00 0.25X¯ + 2.53 0.37X¯ ¥0.78 X′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L. sr′ = Expected single analyst standard deviation of measurements at an average concentration found of X¯ , in μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X¯ , in μg/L. C = True value for the concentration, in μg/L. X¯ = Average recovery found for measurements of samples containing a concentration of C, in μg/L.

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COLUMN: 1.5% OV-17 /1.95% QF-1 ON GAS CHROM Q TEMPERATURE: ssoc. DETECTOR: FLAME IONIZATION

w al N zw 1%1 0 ....a: z

w z 0 a: 0 J: ~

2 6 8 10 12 RETENTION TIME, MIN. Figure 1. Gas chromatogram of nitrobenzene and isophorone.

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COLUMN: 1.5% OV-17 /1.95% QF-1 ON GAS CHROM Q TEMPERATURE: 1450C, DETECTOR: ELECTRON CAPTURE w wz w ::J z ....1 w 0 ::J t­ ....1 o 0 a: t- t- g z !::: c z .i­ 0 N tD N

2 4 6 8 RETENTION TIME, MIN. Figure 2. Gas chromatogram of dinitrotoluenes.

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METHOD 610—POLYNUCLEAR AROMATIC Thus, a single sample may be extracted to HYDROCARBONS measure the parameters included in the scope of each of these methods. When clean- 1. Scope and Application up is required, the concentration levels must 1.1 This method covers the determination be high enough to permit selecting aliquots, of certain polynuclear aromatic hydro- as necessary, to apply appropriate cleanup carbons (PAH). The following parameters procedures. Selection of the aliquots must be can be determined by this method: made prior to the solvent exchange steps of this method. The analyst is allowed the lati- Parameter STORET No. CAS No. tude, under Sections 12 and 13, to select chromatographic conditions appropriate for Acenaphthene ...... 34205 83–32–9 the simultaneous measurement of combina- Acenaphthylene ...... 34200 208–96–8 Anthracene ...... 34220 120–12–7 tions of these parameters. Benzo(a)anthracene ...... 34526 56–55–3 1.6 Any modification of this method, be- Benzo(a)pyrene ...... 34247 50–32–8 yond those expressly permitted, shall be con- Benzo(b)fluoranthene ...... 34230 205–99–2 sidered as a major modification subject to Benzo(ghi)perylene ...... 34521 191–24–2 application and approval of alternate test Benzo(k)fluoranthene ...... 34242 207–08–9 procedures under 40 CFR 136.4 and 136.5. Chrysene ...... 34320 218–01–9 Dibenzo(a,h)anthracene ...... 34556 53–70–3 1.7 This method is restricted to use by or Fluoranthene ...... 34376 206–44–0 under the supervision of analysts experi- Fluorene ...... 34381 86–73–7 enced in the use of HPLC and GC systems Indeno(1,2,3-cd)pyrene ...... 34403 193–39–5 and in the interpretation of liquid and gas Naphthalene ...... 34696 91–20–3 chromatograms. Each analyst must dem- Phenanthrene ...... 34461 85–01–8 onstrate the ability to generate acceptable Pyrene ...... 34469 129–00–0 results with this method using the procedure 1.2 This is a chromatographic method ap- described in Section 8.2. plicable to the determination of the com- 2. Summary of Method pounds listed above in municipal and indus- trial discharges as provided under 40 CFR 2.1 A measured volume of sample, ap- 136.1. When this method is used to analyze proximately 1–L, is extracted with meth- unfamiliar samples for any or all of the com- ylene chloride using a separatory funnel. The pounds above, compound identifications methylene chloride extract is dried and con- should be supported by at least one addi- centrated to a volume of 10 mL or less. The tional qualitative technique. Method 625 pro- extract is then separated by HPLC or GC. Ul- vides gas chromatograph/mass spectrometer traviolet (UV) and fluorescence detectors are (GC/MS) conditions appropriate for the qual- used with HPLC to identify and measure the itative and quantitative confirmation of re- PAHs. A flame ionization detector is used sults for many of the parameters listed with GC. 2 above, using the extract produced by this 2.2 The method provides a silica gel col- method. umn cleanup procedure to aid in the elimi- 1.3 This method provides for both high nation of interferences that may be encoun- performance liquid chromatographic (HPLC) tered. and gas chromatographic (GC) approaches 3. Interferences for the determination of PAHs. The gas chromatographic procedure does not ade- 3.1 Method interferences may be caused quately resolve the following four pairs of by contaminants in solvents, reagents, glass- compounds: Anthracene and phenanthrene; ware, and other sample processing hardward chrysene and benzo(a)anthracene; that lead to discrete artifacts and/or ele- benzo(b)fluoranthene and vated baselines in the chromatograms. All of benzo(k)fluoranthene; and dibenzo(a,h) an- these materials must be routinely dem- thracene and indeno (1,2,3-cd)pyrene. Unless onstrated to be free from interferences under the purpose for the analysis can be served by the conditions of the analysis by running reporting the sum of an unresolved pair, the laboratory reagent blanks as described in liquid chromatographic approach must be Section 8.1.3. used for these compounds. The liquid 3.1.1 Glassware must be scrupulously chromatographic method does resolve all 16 cleaned. 3 Clean all glassware as soon as pos- of the PAHs listed. sible after use by rinsing with the last sol- 1.4 The method detection limit (MDL, de- vent used in it. Solvent rinsing should be fol- fined in Section 15.1) 1 for each parameter is lowed by detergent washing with hot water, listed in Table 1. The MDL for a specific and rinses with tap water and distilled wastewater may differ from those listed, de- water. The glassware should then be drained pending upon the nature of interferences in dry, and heated in a muffle furnace at 400 °C the sample matrix. for 15 to 30 min. Some thermally stable ma- 1.5 The sample extraction and concentra- terials, such as PCBs, may not be eliminated tion steps in this method are essentially the by this treatment. Solvent rinses with ace- same as in Methods 606, 608, 609, 611, and 612. tone and pesticide quality hexane may be

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substituted for the muffle furnace heating. with Teflon. Foil may be substituted for Tef- Thorough rinsing with such solvents usually lon if the sample is not corrosive. If amber eliminates PCB interference. Volumetric bottles are not available, protect samples ware should not be heated in a muffle fur- from light. The bottle and cap liner must be nace. After drying and cooling, glassware washed, rinsed with acetone or methylene should be sealed and stored in a clean envi- chloride, and dried before use to minimize ronment to prevent any accumulation of contamination. dust or other contaminants. Store inverted 5.1.2 Automatic sampler (optional)—The or capped with aluminum foil. sampler must incorporate glass sample con- 3.1.2 The use of high purity reagents and tainers for the collection of a minimum of solvents helps to minimize interference prob- 250 mL of sample. Sample containers must be lems. Purification of solvents by distillation kept refrigerated at 4 °C and protected from in all-glass systems may be required. light during compositing. If the sampler uses 3.2 Matrix interferences may be caused by a peristaltic pump, a minimum length of contaminants that are co-extracted from the compressible silicone rubber tubing may be sample. The extent of matrix interferences used. Before use, however, the compressible will vary considerably from source to source, tubing should be thoroughly rinsed with depending upon the nature and diversity of methanol, followed by repeated rinsings with the industrial complex or municipality being distilled water to minimize the potential for sampled. The cleanup procedure in Section contamination of the sample. An integrating 11 can be used to overcome many of these flow meter is required to collect flow propor- interferences, but unique samples may re- tional composites. quire additional cleanup approaches to 5.2 Glassware (All specifications are sug- achieve the MDL listed in Table 1. gested. Catalog numbers are included for il- 3.3 The extent of interferences that may lustration only.): be encountered using liquid chromatographic 5.2.1 Separatory funnel—2–L, with Teflon techniques has not been fully assessed. Al- stopcock. though the HPLC conditions described allow 5.2.2 Drying column—Chromatographic for a unique resolution of the specific PAH column, approximately 400 mm long × 19 mm compounds covered by this method, other ID, with coarse frit filter disc. PAH compounds may interfere. 5.2.3 Concentrator tube, Kuderna-Dan- ish—10-mL, graduated (Kontes K–570050–1025 4. Safety or equivalent). Calibration must be checked at the volumes employed in the test. Ground 4.1 The toxicity or carcinogenicity of glass stopper is used to prevent evaporation each reagent used in this method have not of extracts. been precisely defined; however, each chem- 5.2.4 Evaporative flask, Kuderna-Danish— ical compound should be treated as a poten- 500-mL (Kontes K–570001–0500 or equivalent). tial health hazard. From this viewpoint, ex- Attach to concentrator tube with springs. posure to these chemicals must be reduced to 5.2.5 Snyder column, Kuderna-Danish— the lowest possible level by whatever means Three-ball macro (Kontes K–503000–0121 or available. The laboratory is responsible for equivalent). maintaining a current awareness file of 5.2.6 Snyder column, Kuderna-Danish— OSHA regulations regarding the safe han- Two-ball micro (Kontes K–569001–0219 or dling of the chemicals specified in this meth- equivalent). od. A reference file of material data handling 5.2.7 Vials—10 to 15-mL, amber glass, with sheets should also be made available to all Teflon-lined screw cap. personnel involved in the chemical analysis. 5.2.8 Chromatographic column—250 mm Additional references to laboratory safety long × 10 mm ID, with coarse frit filter disc 4M6 are available and have been identified at bottom and Teflon stopcock. for the information of the analyst. 5.3 Boiling chips—Approximately 10/40 4.2 The following parameters covered by mesh. Heat to 400 °C for 30 min or Soxhlet ex- this method have been tentatively classified tract with methylene chloride. as known or suspected, human or mamma- 5.4 Water bath—Heated, with concentric lian carcinogens: benzo(a)anthracene, ring cover, capable of temperature control benzo(a)pyrene, and dibenzo(a,h)-anthracene. (±2 °C). The bath should be used in a hood. Primary standards of these toxic compounds 5.5 Balance—Analytical, capable of accu- should be prepared in a hood. A NIOSH/ rately weighing 0.0001 g. MESA approved toxic gas respirator should 5.6 High performance liquid chro- be worn when the analyst handles high con- matograph (HPLC)—An analytical system centrations of these toxic compounds. complete with column supplies, high pres- sure syringes, detectors, and compatible 5. Apparatus and Materials strip-chart recorder. A data system is rec- 5.1 Sampling equipment, for discrete or ommended for measuring peak areas and re- composite sampling. tention times. 5.1.1 Grab sample bottle—1–L or 1-qt, 5.6.1 Gradient pumping system—Constant amber glass, fitted with a screw cap lined flow.

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5.6.2 Reverse phase column—HC-ODS Sil- 6.7.1 Prepare stock standard solutions by X, 5 micron particle diameter, in a 25 cm × accurately weighing about 0.0100 g of pure 2.6 mm ID stainless steel column (Perkin material. Dissolve the material in acetoni- Elmer No. 089–0716 or equivalent). This col- trile and dilute to volume in a 10-mL volu- umn was used to develop the method per- metric flask. Larger volumes can be used at formance statements in Section 15. Guide- the convenience of the analyst. When com- lines for the use of alternate column pound purity is assayed to be 96% or greater, packings are provided in Section 12.2. the weight can be used without correction to 5.6.3 Detectors—Fluorescence and/or UV calculate the concentration of the stock detectors. The fluorescence detector is used standard. Commercially prepared stock for excitation at 280 nm and emission greater standards can be used at any concentration than 389 nm cutoff (Corning 3–75 or equiva- if they are certified by the manufacturer or lent). Fluorometers should have dispersive by an independent source. optics for excitation and can utilize either 6.7.2 Transfer the stock standard solu- filter or dispersive optics at the emission de- tions into Teflon-sealed screw-cap bottles. tector. The UV detector is used at 254 nm Store at 4 °C and protect from light. Stock and should be coupled to the fluorescence de- standard solutions should be checked fre- tector. These detectors were used to develop quently for signs of degradation or evapo- the method performance statements in Sec- ration, especially just prior to preparing tion 15. Guidelines for the use of alternate calibration standards from them. detectors are provided in Section 12.2. 6.7.3 Stock standard solutions must be re- 5.7 Gas chromatograph—An analytical placed after six months, or sooner if com- system complete with temperature program- parison with check standards indicates a mable gas chromatograph suitable for on- problem. column or splitless injection and all required 6.8 Quality control check sample con- accessories including syringes, analytical centrate—See Section 8.2.1. columns, gases, detector, and strip-chart re- corder. A data system is recommended for 7. Calibration measuring peak areas. 7.1 Establish liquid or gas 5.7.1 Column—1.8 m long × 2 mm ID glass, chromatographic operating conditions equiv- packed with 3% OV–17 on Chromosorb W-AW- alent to those given in Table 1 or 2. The DCMS (100/120 mesh) or equivalent. This col- chromatographic system can be calibrated umn was used to develop the retention time using the external standard technique (Sec- data in Table 2. Guidelines for the use of al- tion 7.2) or the internal standard technique ternate column packings are provided in (Section 7.3). Section 13.3. 7.2 External standard calibration proce- 5.7.2 Detector—Flame ionization detector. dure: This detector has proven effective in the 7.2.1 Prepare calibration standards at a analysis of wastewaters for the parameters minimum of three concentration levels for listed in the scope (Section 1.1), excluding each parameter of interest by adding vol- the four pairs of unresolved compounds list- umes of one or more stock standards to a ed in Section 1.3. Guidelines for the use of al- volumetric flask and diluting to volume with ternate detectors are provided in Section acetonitrile. One of the external standards 13.3. should be at a concentration near, but above, the MDL (Table 1) and the other concentra- 6. Reagents tions should correspond to the expected 6.1 Reagent water—Reagent water is de- range of concentrations found in real sam- fined as a water in which an interferent is ples or should define the working range of not observed at the MDL of the parameters the detector. of interest. 7.2.2 Using injections of 5 to 25 μL for 6.2 Sodium thiosulfate—(ACS) Granular. HPLC and 2 to 5 μL for GC, analyze each cali- 6.3 Cyclohexane, methanol, acetone, bration standard according to Section 12 or methylene chloride, pentane—Pesticide qual- 13, as appropriate. Tabulate peak height or ity or equivalent. area responses against the mass injected. 6.4 Acetonitrile—HPLC quality, distilled The results can be used to prepare a calibra- in glass. tion curve for each compound. Alternatively, 6.5 Sodium sulfate—(ACS) Granular, an- if the ratio of response to amount injected hydrous. Purify by heating at 400 °C for 4 h (calibration factor) is a constant over the in a shallow tray. working range (<10% relative standard devi- 6.6 Silica gel—100/200 mesh, desiccant, ation, RSD), linearity through the origin can Davison, grade-923 or equivalent. Before use, be assumed and the average ratio or calibra- activate for at least 16 h at 130 °C in a shal- tion factor can be used in place of a calibra- low glass tray, loosely covered with foil. tion curve. 6.7 Stock standard solutions (1.00 μg/μL)— 7.3 Internal standard calibration proce- Stock standard solutions can be prepared dure—To use this approach, the analyst must from pure standard materials or purchased select one or more internal standards that as certified solutions. are similar in analytical behavior to the

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compounds of interest. The analyst must fur- 8. Quality Control ther demonstrate that the measurement of 8.1 Each laboratory that uses this method the internal standard is not affected by is required to operate a formal quality con- method or matrix interferences. Because of trol program. The minimum requirements of these limitations, no internal standard can this program consist of an initial demonstra- be suggested that is applicable to all sam- tion of laboratory capability and an ongoing ples. analysis of spiked samples to evaluate and 7.3.1 Prepare calibration standards at a document data quality. The laboratory must minimum of three concentration levels for maintain records to document the quality of each parameter of interest by adding vol- data that is generated. Ongoing data quality umes of one or more stock standards to a checks are compared with established per- volumetric flask. To each calibration stand- formance criteria to determine if the results ard, add a known constant amount of one or of analyses meet the performance character- more internal standards, and dilute to vol- istics of the method. When results of sample ume with acetonitrile. One of the standards spikes indicate atypical method perform- should be at a concentration near, but above, ance, a quality control check standard must the MDL and the other concentrations be analyzed to confirm that the measure- should correspond to the expected range of ments were performed in an in-control mode concentrations found in real samples or of operation. should define the working range of the detec- 8.1.1 The analyst must make an initial, tor. one-time, demonstration of the ability to 7.3.2 Using injections of 5 to 25 μL for generate acceptable accuracy and precision HPLC and 2 to 5 μL for GC, analyze each cali- with this method. This ability is established bration standard according to Section 12 or as described in Section 8.2. 13, as appropriate. Tabulate peak height or 8.1.2 In recognition of advances that are area responses against concentration for occurring in chromatography, the analyst is each compound and internal standard. Cal- permitted certain options (detailed in Sec- culate response factors (RF) for each com- tions 10.4, 11.1, 12.2, and 13.3) to improve the pound using Equation 1. separations or lower the cost of measure- ments. Each time such a modification is made to the method, the analyst is required ()()ACsis RF = to repeat the procedure in Section 8.2. 8.1.3 Before processing any samples the ()()ACis s analyst must analyze a reagent water blank Equation 1 to demonstrate that interferences from the analytical system and glassware are under where: control. Each time a set of samples is ex- As = Response for the parameter to be meas- tracted or reagents are changed a reagent ured. water blank must be processed as a safe- Ais = Response for the internal standard. guard against laboratory contamination. Cis = Concentration of the internal standard 8.1.4 The laboratory must, on an ongoing (μg/L). basis, spike and analyze a minimum of 10%

Cs = Concentration of the parameter to be of all samples to monitor and evaluate lab- measured (μg/L). oratory data quality. This procedure is de- scribed in Section 8.3. If the RF value over the working range is a 8.1.5 The laboratory must, on an ongoing constant (<10% RSD), the RF can be assumed basis, demonstrate through the analyses of to be invariant and the average RF can be quality control check standards that the op- used for calculations. Alternatively, the re- eration of the measurement system is in con- sults can be used to plot a calibration curve trol. This procedure is described in Section of response ratios, A /A , vs. RF. s is 8.4. The frequency of the check standard 7.4 The working calibration curve, cali- analyses is equivalent to 10% of all samples bration factor, or RF must be verified on analyzed but may be reduced if spike recov- each working day by the measurement of one eries from samples (Section 8.3) meet all or more calibration standards. If the re- specified quality control criteria. sponse for any parameter varies from the 8.1.6 The laboratory must maintain per- predicted response by more than ±15%, the formance records to document the quality of test must be repeated using a fresh calibra- data that is generated. This procedure is de- tion standard. Alternatively, a new calibra- scribed in Section 8.5. tion curve must be prepared for that com- 8.2 To establish the ability to generate pound. acceptable accuracy and precision, the ana- 7.5 Before using any cleanup procedure, lyst must perform the following operations. the analyst must process a series of calibra- 8.2.1 A quality control (QC) check sample tion standards through the procedure to vali- concentrate is required containing each pa- date elution patterns and the absence of rameter of interest at the following con- interferences from the reagents. centrations in acetonitrile: 100 μg/mL of any

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of the six early-eluting PAHs (naphthalene, 8.3.1 The concentration of the spike in the acenaphthylene, acenaphthene, fluorene, sample should be determined as follows: phenanthrene, and anthracene); 5 μg/mL of 8.3.1.1 If, as in compliance monitoring, benzo(k)fluoranthene; and 10 μg/mL of any of the concentration of a specific parameter in the other PAHs. The QC check sample con- the sample is being checked against a regu- centrate must be obtained from the U.S. En- latory concentration limit, the spike should vironmental Protection Agency, Environ- be at that limit or 1 to 5 times higher than mental Monitoring and Support Laboratory the background concentration determined in in Cincinnati, Ohio, if available. If not avail- Section 8.3.2, whichever concentration would able from that source, the QC check sample be larger. concentrate must be obtained from another 8.3.1.2 If the concentration of a specific external source. If not available from either parameter in the sample is not being source above, the QC check sample con- checked against a limit specific to that pa- centrate must be prepared by the laboratory rameter, the spike should be at the test con- using stock standards prepared independ- centration in Section 8.2.2 or 1 to 5 times ently from those used for calibration. higher than the background concentration 8.2.2 Using a pipet, prepare QC check sam- determined in Section 8.3.2, whichever con- ples at the test concentrations shown in centration would be larger. Table 3 by adding 1.00 mL of QC check sam- 8.3.1.3 If it is impractical to determine ple concentrate to each of four 1–L aliquots background levels before spiking (e.g., max- of reagent water. imum holding times will be exceeded), the 8.2.3 Analyze the well-mixed QC check spike concentration should be (1) the regu- samples according to the method beginning latory concentration limit, if any; or, if in Section 10. none, (2) the larger of either 5 times higher 8.2.4 Calculate the average recovery (X¯ ) in than the expected background concentration μg/L, and the standard deviation of the re- or the test concentration in Section 8.2.2. covery (s) in μg/L, for each parameter using 8.3.2 Analyze one sample aliquot to deter- the four results. mine the background concentration (B) of 8.2.5 For each parameter compare s and X¯ each parameter. If necessary, prepare a new with the corresponding acceptance criteria QC check sample concentrate (Section 8.2.1) for precision and accuracy, respectively, appropriate for the background concentra- found in Table 3. If s and X¯ for all param- tions in the sample. Spike a second sample eters of interest meet the acceptance cri- aliquot with 1.0 mL of the QC check sample teria, the system performance is acceptable concentrate and analyze it to determine the and analysis of actual samples can begin. If concentration after spiking (A) of each pa- any individual s exceeds the precision limit rameter. Calculate each percent recovery (P) or any individual X¯ falls outside the range as 100 (A¥B)%/T, where T is the known true for accuracy, the system performance is un- value of the spike. acceptable for that parameter. 8.3.3 Compare the percent recovery (P) for NOTE: The large number of parameters in each parameter with the corresponding QC Table 3 present a substantial probability acceptance criteria found in Table 3. These that one or more will fail at least one of the acceptance criteria were calculated to in- acceptance criteria when all parameters are clude an allowance for error in measurement analyzed. of both the background and spike concentra- 8.2.6 When one or more of the parameters tions, assuming a spike to background ratio tested fail at least one of the acceptance cri- of 5:1. This error will be accounted for to the teria, the analyst must proceed according to extent that the analyst’s spike to back- Section 8.2.6.1 or 8.2.6.2. ground ratio approaches 5:1. 7 If spiking was 8.2.6.1 Locate and correct the source of performed at a concentration lower than the the problem and repeat the test for all pa- test concentration in Section 8.2.2, the ana- rameters of interest beginning with Section lyst must use either the QC acceptance cri- 8.2.2. teria in Table 3, or optional QC acceptance 8.2.6.2 Beginning with Section 8.2.2, repeat criteria calculated for the specific spike con- the test only for those parameters that centration. To calculate optional acceptance failed to meet criteria. Repeated failure, criteria for the recovery of a parameter: (1) however, will confirm a general problem Calculate accuracy (X′) using the equation in with the measurement system. If this occurs, Table 4, substituting the spike concentration locate and correct the source of the problem (T) for C; (2) calculate overall precision (S′) and repeat the test for all compounds of in- using the equation in Table 4, substituting X′ terest beginning with Section 8.2.2. for X¯ ; (3) calculate the range for recovery at 8.3 The laboratory must, on an ongoing the spike concentration as (100 X′/T)±2.44(100 basis, spike at least 10% of the samples from S′/T)%. 7 each sample site being monitored to assess 8.3.4 If any individual P falls outside the accuracy. For laboratories analyzing one to designated range for recovery, that param- ten samples per month, at least one spiked eter has failed the acceptance criteria. A sample per month is required. check standard containing each parameter

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that failed the critiera must be analyzed as umn, specific element detector, or mass described in Section 8.4. spectrometer must be used. Whenever pos- 8.4 If any parameter fails the acceptance sible, the laboratory should analyze standard criteria for recovery in Section 8.3, a QC reference materials and participate in rel- check standard containing each parameter evant performance evaluation studies. that failed must be prepared and analyzed. NOTE: The frequency for the required anal- 9. Sample Collection, Preservation, and ysis of a QC check standard will depend upon Handling the number of parameters being simulta- neously tested, the complexity of the sample 9.1 Grab samples must be collected in matrix, and the performance of the labora- glass containers. Conventional sampling tory. If the entire list of parameters in Table practices 8 should be followed, except that 3 must be measured in the sample in Section the bottle must not be prerinsed with sample 8.3, the probability that the analysis of a QC before collection. Composite samples should check standard will be required is high. In be collected in refrigerated glass containers this case the QC check standard should be in accordance with the requirements of the routinely analyzed with the spike sample. program. Automatic sampling equipment 8.4.1 Prepare the QC check standard by must be as free as possible of Tygon tubing adding 1.0 mL of QC check sample con- and other potential sources of contamina- centrate (Section 8.2.1 or 8.3.2) to 1 L of rea- tion. gent water. The QC check standard needs 9.2 All samples must be iced or refrig- only to contain the parameters that failed erated at 4 °C from the time of collection criteria in the test in Section 8.3. until extraction. PAHs are known to be light 8.4.2 Analyze the QC check standard to sensitive; therefore, samples, extracts, and determine the concentration measured (A) of standards should be stored in amber or foil- each parameter. Calculate each percent re- wrapped bottles in order to minimize photo- covery (Ps) as 100 (A/T)%, where T is the true lytic decomposition. Fill the sample bottles value of the standard concentration. and, if residual chlorine is present, add 80 mg 8.4.3 Compare the percent recovery (Ps) of sodium thiosulfate per liter of sample and for each parameter with the corresponding mix well. EPA Methods 330.4 and 330.5 may QC acceptance criteria found in Table 3. Only be used for measurement of residual chlo- parameters that failed the test in Section 8.3 rine. 9 Field test kits are available for this need to be compared with these criteria. If purpose. the recovery of any such parameter falls out- 9.3 All samples must be extracted within 7 side the designated range, the laboratory days of collection and completely analyzed performance for that parameter is judged to within 40 days of extraction. 2 be out of control, and the problem must be immediately identified and corrected. The 10. Sample Extraction analytical result for that parameter in the unspiked sample is suspect and may not be 10.1 Mark the water meniscus on the side reported for regulatory compliance purposes. of the sample bottle for later determination 8.5 As part of the QC program for the lab- of sample volume. Pour the entire sample oratory, method accuracy for wastewater into a 2–L separatory funnel. samples must be assessed and records must 10.2 Add 60 mL of methylene chloride to be maintained. After the analysis of five the sample bottle, seal, and shake 30 s to spiked wastewater samples as in Section 8.3, rinse the inner surface. Transfer the solvent calculate the average percent recovery (P¯ ) to the separatory funnel and extract the and the standard deviation of the percent re- sample by shaking the funnel for 2 min. with covery (sp). Express the accuracy assessment periodic venting to release excess pressure. ¯ as a percent recovery interval from P–2sp to Allow the organic layer to separate from the ¯ ¯ P + 2sp. If P = 90% and sp = 10%, for example, water phase for a minimum of 10 min. If the the accuracy interval is expressed as 70– emulsion interface between layers is more 110%. Update the accuracy assessment for than one-third the volume of the solvent each parameter on a regular basis (e.g. after layer, the analyst must employ mechanical each five to ten new accuracy measure- techniques to complete the phase separation. ments). The optimum technique depends upon the 8.6 It is recommended that the laboratory sample, but may include stirring, filtration adopt additional quality assurance practices of the emulsion through glass wool, cen- for use with this method. The specific prac- trifugation, or other physical methods. Col- tices that are most productive depend upon lect the methylene chloride extract in a 250– the needs of the laboratory and the nature of mL Erlenmeyer flask. the samples. Field duplicates may be ana- 10.3 Add a second 60-mL volume of meth- lyzed to assess the precision of the environ- ylene chloride to the sample bottle and re- mental measurements. When doubt exists peat the extraction procedure a second time, over the identification of a peak on the chro- combining the extracts in the Erlenmeyer matogram, confirmatory techniques such as flask. Perform a third extraction in the same gas chromatography with a dissimilar col- manner.

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10.4 Assemble a Kuderna-Danish (K-D) concentrator tube. Add 4 mL of cyclohexane concentrator by attaching a 10-mL concen- and attach a two-ball micro-Snyder column. trator tube to a 500-mL evaporative flask. Prewet the column by adding 0.5 mL of Other concentration devices or techniques methylene chloride to the top. Place the may be used in place of the K-D concentrator micro-K-D apparatus on a boiling (100 °C) if the requirements of Section 8.2 are met. water bath so that the concentrator tube is 10.5 Pour the combined extract through a partially immersed in the hot water. Adjust solvent-rinsed drying column containing the vertical position of the apparatus and about 10 cm of anhydrous sodium sulfate, the water temperature as required to com- and collect the extract in the K-D concen- plete concentration in 5 to 10 min. At the trator. Rinse the Erlenmeyer flask and col- proper rate of distillation the balls of the umn with 20 to 30 mL of methylene chloride column will actively chatter but the cham- to complete the quantitative transfer. bers will not flood. When the apparent vol- 10.6 Add one or two clean boiling chips to ume of the liquid reaches 0.5 mL, remove the the evaporative flask and attach a three-ball K-D apparatus and allow it to drain and cool Snyder column. Prewet the Snyder column for at least 10 min. Remove the micro-Sny- by adding about 1 mL of methylene chloride der column and rinse its lower joint into the to the top. Place the K-D apparatus on a hot concentrator tube with a minimum amount water bath (60 to 65 °C) so that the concen- of cyclohexane. Adjust the extract volume to trator tube is partially immersed in the hot about 2 mL. water, and the entire lower rounded surface 11.3 Silica gel column cleanup for PAHs: of the flask is bathed with hot vapor. Adjust 11.3.1 Prepare a slurry of 10 g of activiated the vertical position of the apparatus and silica gel in methylene chloride and place the water temperature as required to com- this into a 10-mm ID chromatographic col- plete the concentration in 15 to 20 min. At umn. Tap the column to settle the silica gel the proper rate of distillation the balls of the and elute the methylene chloride. Add 1 to 2 column will actively chatter but the cham- cm of anhydrous sodium sulfate to the top of bers will not flood with condensed solvent. the silica gel. When the apparent volume of liquid reaches 11.3.2 Preelute the column with 40 mL of 1 mL, remove the K-D apparatus and allow it pentane. The rate for all elutions should be to drain and cool for at least 10 min. about 2 mL/min. Discard the eluate and just 10.7 Remove the Snyder column and rinse prior to exposure of the sodium sulfate layer the flask and its lower joint into the concen- to the air, transfer the 2-mL cyclohexane trator tube with 1 to 2 mL of methylene sample extract onto the column using an ad- chloride. A 5-mL syringe is recommended for ditional 2 mL cyclohexane to complete the this operation. Stopper the concentrator transfer. Just prior to exposure of the so- tube and store refrigerated if further proc- dium sulfate layer to the air, add 25 mL of essing will not be performed immediately. If pentane and continue the elution of the col- the extract will be stored longer than two umn. Discard this pentane eluate. days, it should be transferred to a Teflon- 11.3.3 Next, elute the column with 25 mL sealed screw-cap vial and protected from of methylene chloride/pentane (4 + 6)(V/V) light. If the sample extract requires no fur- into a 500-mL K-D flask equipped with a 10- ther cleanup, proceed with gas or liquid mL concentrator tube. Concentrate the col- chromatographic analysis (Section 12 or 13). lected fraction to less than 10 mL as in Sec- If the sample requires further cleanup, pro- tion 10.6. When the apparatus is cool, remove ceed to Section 11. the Snyder column and rinse the flask and 10.8 Determine the original sample vol- its lower joint with pentane. Proceed with ume by refilling the sample bottle to the HPLC or GC analysis. mark and transferring the liquid to a 1000- mL graduated cylinder. Record the sample 12. High Performance Liquid Chromatography volume to the nearest 5 mL. 12.1 To the extract in the concentrator tube, add 4 mL of acetonitrile and a new 11. Cleanup and Separation boiling chip, then attach a two-ball micro- 11.1 Cleanup procedures may not be nec- Snyder column. Concentrate the solvent as essary for a relatively clean sample matrix. in Section 10.6, except set the water bath at If particular circumstances demand the use 95 to 100 °C. When the apparatus is cool, re- of a cleanup procedure, the analyst may use move the micro-Snyder column and rinse its the procedure below or any other appropriate lower joint into the concentrator tube with procedure. However, the analyst first must about 0.2 mL of acetonitrile. Adjust the ex- demonstrate that the requirements of Sec- tract volume to 1.0 mL. tion 8.2 can be met using the methods as re- 12.2 Table 1 summarizes the recommended vised to incorporate the cleanup procedure. operating conditions for the HPLC. Included 11.2 Before the silica gel cleanup tech- in this table are retention times, capacity nique can be utilized, the extract solvent factors, and MDL that can be achieved under must be exchanged to cyclohexane. Add 1 to these conditions. The UV detector is rec- 10 mL of the sample extract (in methylene ommended for the determination of naph- chloride) and a boiling chip to a clean K-D thalene, acenaphthylene, acenapthene, and

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fluorene and the fluorescence detector is rec- move the K-D apparatus and allow it to ommended for the remaining PAHs. Exam- drain and cool for at least 10 min. Remove ples of the separations achieved by this the micro-Snyder column and rinse its lower HPLC column are shown in Figures 1 and 2. joint into the concentrator tube with a min- Other HPLC columns, chromatographic con- imum amount of methylene chloride. Adjust ditions, or detectors may be used if the re- the final volume to 1.0 mL and stopper the quirements of Section 8.2 are met. concentrator tube. 12.3 Calibrate the system daily as de- 13.3 Table 2 summarizes the recommended scribed in Section 7. 12.4 If the internal standard calibration operating conditions for the gas chro- procedure is being used, the internal stand- matograph. Included in this table are reten- ard must be added to the sample extract and tion times that were obtained under these mixed thoroughly immediately before injec- conditions. An example of the separations tion into the instrument. achieved by this column is shown in Figure 12.5 Inject 5 to 25 μL of the sample extract 3. Other packed or capillary (open-tubular) or standard into the HPLC using a high pres- columns, chromatographic conditions, or de- sure syringe or a constant volume sample in- tectors may be used if the requirements of jection loop. Record the volume injected to Section 8.2 are met. the nearest 0.1 μL, and the resulting peak 13.4 Calibrate the gas chromatographic size in area or peak height units. Re-equili- system daily as described in Section 7. brate the HPLC column at the initial gra- 13.5 If the internal standard calibration dient conditions for at least 10 min between procedure is being used, the internal stand- injections. ard must be added to the sample extract and 12.6 Identify the parameters in the sample by comparing the retention time of the mixed thoroughly immediately before injec- peaks in the sample chromatogram with tion into the gas chromatograph. those of the peaks in standard 13.6 Inject 2 to 5 μL of the sample extract chromatograms. The width of the retention or standard into the gas chromatograph time window used to make identifications using the solvent-flush technique. 10 Smaller should be based upon measurements of ac- (1.0 μL) volumes may be injected if auto- tual retention time variations of standards matic devices are employed. Record the vol- over the course of a day. Three times the ume injected to the nearest 0.05 μL, and the standard deviation of a retention time for a resulting peak size in area or peak height compound can be used to calculate a sug- units. gested window size; however, the experience 13.7 Identify the parameters in the sample of the analyst should weigh heavily in the by comparing the retention times of the interpretation of chromatograms. peaks in the sample chromatogram with 12.7 If the response for a peak exceeds the those of the peaks in standard working range of the system, dilute the ex- chromatograms. The width of the retention tract with acetonitrile and reanalyze. time window used to make identifications 12.8 If the measurement of the peak re- should be based upon measurements of ac- sponse is prevented by the presence of inter- ferences, further cleanup is required. tual retention time variations of standards over the course of a day. Three times the 13. Gas Chromatography standard deviation of a retention time for a compound can be used to calculate a sug- 13.1 The packed column GC procedure will gested window size; however, the experience not resolve certain isomeric pairs as indi- cated in Section 1.3 and Table 2. The liquid of the analyst should weigh heavily in the chromatographic procedure (Section 12) interpretation of chromatograms. must be used for these parameters. 13.8 If the response for a peak exceeds the 13.2 To achieve maximum sensitivity with working range of the system, dilute the ex- this method, the extract must be con- tract and reanalyze. centrated to 1.0 mL. Add a clean boiling chip 13.9 If the measurement of the peak re- to the methylene chloride extract in the con- sponse is prevented by the presence of inter- centrator tube. Attach a two-ball micro-Sny- ferences, further cleanup is required. der column. Prewet the micro-Snyder col- umn by adding about 0.5 mL of methylene 14. Calculations chloride to the top. Place the micro-K-D ap- paratus on a hot water bath (60 to 65 °C) so 14.1 Determine the concentration of indi- that the concentrator tube is partially im- vidual compounds in the sample. mersed in the hot water. Adjust the vertical 14.1.1 If the external standard calibration position of the apparatus and the water tem- procedure is used, calculate the amount of perature as required to complete the con- material injected from the peak response centration in 5 to 10 min. At the proper rate using the calibration curve or calibration of distillation the balls will actively chatter factor determined in Section 7.2.2. The con- but the chambers will not flood. When the centration in the sample can be calculated apparent volume of liquid reaches 0.5 mL, re- from Equation 2.

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Linear equations to describe these relation- ()AV() ships are presented in Table 4. Concentration (μ= g/L) t REFERENCES ()VVis() 1. 40 CFR part 136, appendix B. Equation 2 2. ‘‘Determination of Polynuclear Aro- where: matic Hydrocarbons in Industrial and Munic- A = Amount of material injected (ng). ipal Wastewaters,’’ EPA 600/4–82–025, Na- Vi = Volume of extract injected (μL). tional Technical Information Service, PB82– Vt = Volume of total extract (μL). 258799, Springfield, Virginia 22161, June 1982. Vs = Volume of water extracted (mL). 3. ASTM Annual Book of Standards, Part 13.1.2 If the internal standard calibration 31, D3694–78. ‘‘Standard Practices for Prepa- procedure is used, calculate the concentra- ration of Sample Containers and for Preser- tion in the sample using the response factor vation of Organic Constituents,’’ American (RF) determined in Section 7.3.2 and Equa- Society for Testing and Materials, Philadel- tion 3. phia. 4. ‘‘Carcinogens—Working With Carcino- ()()AI gens,’’ Department of Health, Education, and Concentration (μ= g/L) ss Welfare, Public Health Service, Center for () Disease Control, National Institute for Occu- ()ARFVis() o pational Safety and Health, Publication No. Equation 3 77–206, August 1977. 5. ‘‘OSHA Safety and Health Standards, where: General Industry,’’ (29 CFR part 1910), Occu- As = Response for the parameter to be meas- ured. pational Safety and Health Administration, A = Response for the internal standard. OSHA 2206 (Revised, January 1976). is 6. ‘‘Safety in Academic Chemistry Labora- Is = Amount of internal standard added to each extract (μg). tories,’’ American Chemical Society Publica- tion, Committee on Chemical Safety, 3rd Vo = Volume of water extracted (L). Edition, 1979. 14.2 Report results in μg/L without correc- tion for recovery data. All QC data obtained 7. Provost, L.P., and Elder, R.S. ‘‘Interpre- should be reported with the sample results. tation of Percent Recovery Data,’’ American Laboratory, 15, 58–63 (1983). (The value 2.44 15. Method Performance used in the equation in Section 8.3.3 is two times the value 1.22 derived in this report.) 15.1 The method detection limit (MDL) is 8. ASTM Annual Book of Standards, Part defined as the minimum concentration of a 31, D3370–76. ‘‘Standard Practices for Sam- substance that can be measured and reported pling Water,’’ American Society for Testing with 99% confidence that the value is above and Materials, Philadelphia. zero. 1 The MDL concentrations listed in 9. ‘‘Methods 330.4 (Titrimetric, DPD-FAS) Table 1 were obtained using reagent water. 11 Similar results were achieved using rep- and 330.5 (Spectrophotometric, DPD) for resentative wastewaters. MDL for the GC ap- Chlorine, Total Residual,’’ Methods for proach were not determined. The MDL actu- Chemical Analysis of Water and Wastes, ally achieved in a given analysis will vary EPA–600/4–79–020, U.S. Environmental Pro- depending on instrument sensitivity and ma- tection Agency, Environmental Monitoring trix effects. and Support Laboratory, Cincinnati, Ohio 15.2 This method has been tested for lin- 45268, March 1979. earity of spike recovery from reagent water 10. Burke, J.A. ‘‘Gas Chromatography for and has been demonstrated to be applicable Pesticide Residue Analysis; Some Practical over the concentration range from 8 × MDL Aspects,’’ Journal of the Association of Official to 800 × MDL 11 with the following exception: Analytical Chemists, 48, 1037 (1965). benzo(ghi)perylene recovery at 80 × and 800 × 11. Cole, T., Riggin, R., and Glaser, J. MDL were low (35% and 45%, respectively). ‘‘Evaluation of Method Detection Limits and 15.3 This method was tested by 16 labora- Analytical Curve for EPA Method 610— tories using reagent water, drinking water, PNAs,’’ International Symposium on surface water, and three industrial Polynuclear Aromatic Hydrocarbons, 5th, wastewaters spiked at six concentrations Battelle’s Columbus Laboratories, Colum- over the range 0.1 to 425 μg/L. 12 Single oper- bus, Ohio (1980). ator precision, overall precision, and method 12. ‘‘EPA Method Study 20, Method 610 accuracy were found to be directly related to (PNA’s),’’ EPA 600/4–84–063, National Tech- the concentration of the parameter and es- nical Information Service, PB84–211614, sentially independent of the sample matrix. Springfield, Virginia 22161, June 1984.

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TABLE 1—HIGH PERFORMANCE LIQUID CHROMATOGRAPHY CONDITIONS AND METHOD DETECTION LIMITS

Method Retention Column detection Parameter capacity μ time (min) factor (k′) limit ( g/ L) a

Naphthalene ...... 16.6 12 .2 1 .8 Acenaphthylene ...... 18.5 13.7 2 .3 Acenaphthene ...... 20 .5 15 .2 1 .8 Fluorene ...... 21 .2 15 .8 0 .21 Phenanthrene ...... 22 .1 16 .6 0 .64 Anthracene ...... 23.4 17.6 0 .66 Fluoranthene ...... 24 .5 18 .5 0 .21 Pyrene ...... 25 .4 19.1 0.27 Benzo(a)anthracene ...... 28.5 21 .6 0 .013 Chrysene ...... 29 .3 22 .2 0 .15 Benzo(b)fluoranthene ...... 31 .6 24 .0 0 .018 Benzo(k)fluoranthene ...... 32.9 25 .1 0 .017 Benzo(a)pyrene ...... 33.9 25.9 0 .023 Dibenzo(a,h)anthracene ...... 35.7 27 .4 0 .030 Benzo(ghi)perylene ...... 36.3 27 .8 0 .076 Indeno(1,2,3-cd)pyrene ...... 37.4 28.7 0 .043 HPLC column conditions: Reverse phase HC-ODS Sil-X, 5 micron particle size, in a 25 cm × 2.6 mm ID stainless steel col- umn. Isocratic elution for 5 min. using acetonitrile/water (4 + 6), then linear gradient elution to 100% acetonitrile over 25 min. at 0.5 mL/min flow rate. If columns having other internal diameters are used, the flow rate should be adjusted to maintain a linear velocity of 2 mm/sec. a The MDL for naphthalene, acenaphthylene, acenaphthene, and fluorene were determined using a UV detector. All others were determined using a fluorescence detector.

TABLE 2—GAS CHROMATOGRAPHIC CONDITIONS TABLE 2—GAS CHROMATOGRAPHIC CONDITIONS AND RETENTION TIMES AND RETENTION TIMES—Continued

Parameter Retention Retention time (min) Parameter time (min)

Naphthalene ...... 4.5 Benzo(k)fluoranthene ...... 28.0 Acenaphthylene ...... 10.4 Benzo(a)pyrene ...... 29.4 Acenaphthene ...... 10 .8 Dibenzo(a,h)anthracene ...... 36.2 Fluorene ...... 12 .6 Phenanthrene ...... 15 .9 Indeno(1,2,3-cd)pyrene ...... 36.2 Anthracene ...... 15.9 Benzo(ghi)perylene ...... 38.6 Fluoranthene ...... 19 .8 GC Column conditions: Chromosorb W-AW-DCMS (100/120 Pyrene ...... 20 .6 mesh) coated with 3% OV–17 packed in a 1.8 × 2 mm ID Benzo(a)anthracene ...... 24.7 glass column with nitrogen carrier gas at 40 mL/min. flow rate. Chrysene ...... 24 .7 Column temperature was held at 100 °C for 4 min., then pro- Benzo(b)fluoranthene ...... 28 .0 grammed at 8 °C/min. to a final hold at 280 °C.

TABLE 3—QC ACCEPTANCE CRITERIA—METHOD 610

¯ Parameter Test conc. Limit for s Range for X Range for (μg/L) (μg/L) (μg/L) P, Ps (%)

Acenaphthene ...... 100 40.3 D–105.7 D–124 Acenaphthylene ...... 100 45.1 22.1–112.1 D–139 Anthracene ...... 100 28.7 11.2–112.3 D–126 Benzo(a)anthracene ...... 10 4.0 3.1–11.6 12–135 Benzo(a)pyrene ...... 10 4.0 0.2–11.0 D–128 Benzo(b)fluor-anthene ...... 10 3.1 1.8–13.8 6–150 Benzo(ghi)perylene ...... 10 2.3 D–10.7 D–116 Benzo(k)fluo-ranthene ...... 5 2.5 D–7.0 D–159 Chrysene ...... 10 4.2 D–17.5 D–199 Dibenzo(a,h)an-thracene ...... 10 2.0 0.3–10.0 D–110 Fluoranthene ...... 10 3.0 2.7–11.1 14–123 Fluorene ...... 100 43.0 D–119 D–142 Indeno(1,2,3–cd)pyrene ...... 10 3.0 1.2–10.0 D–116 Naphthalene ...... 100 40.7 21.5–100.0 D–122 Phenanthrene ...... 100 37.7 8.4–133.7 D–155 Pyrene ...... 10 3.4 1.4–12.1 D–140 s = Standard deviation of four recovery measurements, in μg/L (Section 8.2.4). X¯ = Average recovery for four recovery measurements, in μg/L (Section 8.2.4). P, Ps = Percent recovery measured (Section 8.3.2, Section 8.4.2). D = Detected; result must be greater than zero. NOTE: These criteria are based directly upon the method performance data in Table 4. Where necessary, the limits for recov- ery have been broadened to assure applicability of the limits to concentrations below those used to develop Table 4.

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TABLE 4—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 610

Accuracy, as Single analyst Overall preci- Parameter recovery, X′ precision, sr′ sion, S′ (μg/L) (μg/L) (μg/L)

Acenaphthene ...... 0.52C + 0.54 0.39X¯ + 0.76 0.53X¯ + 1.32 Acenaphthylene ...... 0.69C ¥ 1.89 0.36X¯ + 0.29 0.42X¯ + 0.52 Anthracene ...... 0.63C ¥ 1.26 0.23X¯ + 1.16 0.41X¯ + 0.45 Benzo(a)anthracene ...... 0.73C + 0.05 0.28X¯ + 0.04 0.34X¯ + 0.02 Benzo(a)pyrene ...... 0.56C + 0.01 0.38X¯ ¥ 0.01 0.53X¯ ¥ 0.01 Benzo(b)fluoranthene ...... 0.78C + 0.01 0.21X¯ + 0.01 0.38X¯ ¥ 0.00 Benzo(ghi)perylene ...... 0.44C + 0.30 0.25X¯ + 0.04 0.58X¯ + 0.10 Benzo(k)fluoranthene ...... 0.59C + 0.00 0.44X¯ ¥ 0.00 0.69X¯ + 0.01 Chrysene ...... 0.77C ¥ 0.18 0.32X¯ ¥ 0.18 0.66X¯ ¥ 0.22 Dibenzo(a,h)anthracene ...... 0.41C + 0.11 0.24X¯ + 0.02 0.45X¯ + 0.03 Fluoranthene ...... 0.68C + 0.07 0.22X¯ + 0.06 0.32X¯ + 0.03 Fluorene ...... 0.56C ¥ 0.52 0.44X¯ ¥ 1.12 0.63X¯ ¥ 0.65 Indeno(1,2,3–cd)pyrene ...... 0.54C + 0.06 0.29X¯ + 0.02 0.42X¯ + 0.01 Naphthalene ...... 0.57C ¥ 0.70 0.39X¯ ¥ 0.18 0.41X¯ + 0.74 Phenanthrene ...... 0.72C ¥ 0.95 0.29X¯ + 0.05 0.47X¯ ¥ 0.25 Pyrene ...... 0.69C ¥ 0.12 0.25X¯ + 0.14 0.42X¯ ¥ 0.00 X′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L. sr′ = Expected single analyst standard deviation of measurements at an average concentration found of X¯ , in μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X¯ , in μg/L. C = True value for the concentration, in μg/L. X¯ = Average recovery found for measurements of samples containing a concentration of C, in μg/L.

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w i!i COLUMN: HC-ODS SIL-X ...J:: w z MOBILE PHASE: 40%TO 100% ACETONITRILE z :! IN WATER 0 ~z ::> w DETECTOR: FLUORESCENCE <( Ei f5 a: -a: 0 ..... ::> w 2~ z it z .!!!. w w 0 u :;; G>N <( z w a: 2 w i!i J:: .. u w ...z ~ <( z <( .. w w ...J:: z i!' ;; z w 0 <( .. N J:: w a:~ ... z Zw :ii z w Ow .. <( u =>z ~65 z <( -'w .. w ..... I:: 2 J:: .. J:: it J:: i!i .. u ...z .. <( a

8 12 RETENTION TIME. MIN. Figure 2. Liquid chromatogram of polynuclear aromatic hydrocarbons.

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METHOD 611—HALOETHERS 1. Scope and Application 1.1 This method covers the determination of certain haloethers. The following param- eters can be determined by this method:

Parameter STORET No. CAS No.

Bis(2-chloroethyl) ether ...... 34273 111–44–4 Bis(2-chloroethoxy) methane ...... 34278 111–91–1 2, 2′-oxybis (1-chloropropane) ...... 34283 108–60–1 4-Bromophenyl phenyl ether ...... 34636 101–55–3 4-Chlorophenyl phenyl ether ...... 34641 7005–72–3

1.2 This is a gas chromatographic (GC) describes analytical conditions for a second method applicable to the determination of gas chromatographic column that can be the compounds listed above in municipal and used to confirm measurements made with industrial discharges as provided under 40 the primary column. Method 625 provides gas CFR 136.1. When this method is used to ana- chromatograph/mass spectrometer (GC/MS) lyze unfamiliar samples for any or all of the conditions appropriate for the qualitative compounds above, compound identifications and quantitative confirmation of results for should be supported by at least one addi- all of the parameters listed above, using the tional qualitative technique. This method extract produced by this method.

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1.3 The method detection limit (MDL, de- dry, and heated in a muffle furnace at 400 °C fined in Section 14.1) 1 for each parameter is for 15 to 30 min. Some thermally stable ma- listed in Table 1. The MDL for a specific terials, such a PCBs, may not be eliminated wastewater may differ from those listed, de- by this treatment. Solvent rinses with ace- pending upon the nature of interferences in tone and pesticide quality hexane may be the sample matrix. substituted for the muffle furnace heating. 1.4 The sample extraction and concentra- Thorough rinsing with such solvents usually tion steps in this method are essentially the eliminates PCB interference. Volumetric same as in Methods 606, 608, 609, and 612. ware should not be heated in a muffle fur- Thus, a single sample may be extracted to nace. After drying and cooling, glassware measure the parameters included in the should be sealed and stored in a clean envi- scope of each of these methods. When clean- ronment to prevent any accumulation of up is required, the concentration levels must dust or other contaminants. Store inverted be high enough to permit selecting aliquots, or capped with aluminum foil. as necessary, to apply appropriate cleanup 3.1.2 The use of high purity reagents and procedures. The analyst is allowed the lati- solvents helps to minimize interference prob- tude, under Section 12, to select lems. Purification of solvents by distillation chromatographic conditions appropriate for in all-glass systems may be required. the simultaneous measurement of combina- 3.2 Matrix interferences may be caused by tions of these parameters. contaminants that are co-extracted from the 1.5 Any modification of this method, be- sample. The extent of matrix interferences yond those expressly permitted, shall be con- will vary considerably from source to source, sidered as a major modification subject to depending upon the nature and diversity of application and approval of alternate test the industrial complex or municipality being procedures under 40 CFR 136.4 and 136.5. sampled. The cleanup procedure in Section 1.6 This method is restricted to use by or 11 can be used to overcome many of these under the supervision of analysts experi- interferences, but unique samples may re- enced in the use of a gas chromatograph and quire additional cleanup approaches to in the interpretation of gas chromatograms. achieve the MDL listed in Table 1. Each analyst must demonstrate the ability 3.3 Dichlorobenzenes are known to to generate acceptable results with this coelute with haloethers under some gas method using the procedure described in Sec- chromatographic conditions. If these mate- tion 8.2. rials are present together in a sample, it may be necessary to analyze the extract 2. Summary of Method with two different column packings to com- 2.1 A measured volume of sample, ap- pletely resolve all of the compounds. proximately 1–L, is extracted with meth- 4. Safety ylene chloride using a separatory funnel. The methylene chloride extract is dried and ex- 4.1 The toxicity or carcinogenicity of changed to hexane during concentration to a each reagent used in this method has not volume of 10 mL or less. The extract is sepa- been precisely defined; however, each chem- rated by gas chromatography and the param- ical compound should be treated as a poten- eters are then measured with a halide spe- tial health hazard. From this viewpoint, ex- cific detector. 2 posure to these chemicals must be reduced to 2.2 The method provides a Florisil column the lowest possible level by whatever means cleanup procedure to aid in the elimination available. The laboratory is responsible for of interferences that may be encountered. maintaining a current awareness file of OSHA regulations regarding the safe han- 3. Interferences dling of the chemicals specified in this meth- 3.1 Method interferences may be caused od. A reference file of material data handling by contaminants in solvents, reagents, glass- sheets should also be made available to all ware, and other sample processing hardware personnel involved in the chemical analysis. that lead to discrete artifacts and/or ele- Additional references to laboratory safety vated baselines in gas chromatograms. All of are available and have been identified 4M6 these materials must be routinely dem- for the information of the analyst. onstrated to be free from interferences under 5. Apparatus and Materials the conditions of the analysis by running laboratory reagent blanks as described in 5.1 Sampling equipment, for discrete or Section 8.1.3. composite sampling. 3.1.1 Glassware must be scrupulously 5.1.1 Grab sample bottle—1-L or 1-qt, cleaned. 3 Clean all glassware as soon as pos- amber glass, fitted with a screw cap lined sible after use by rinsing with the last sol- with Teflon. Foil may be substituted for Tef- vent used in it. Solvent rinsing should be fol- lon if the sample is not corrosive. If amber lowed be detergent washing with hot water, bottles are not available, protect samples and rinses with tap water and distilled from light. The bottle and cap liner must be water. The glassware should then be drained washed, rinsed with acetone or methylene

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chloride, and dried before use to minimize 5.6.2 Column 2—1.8 m long × 2 mm ID contamination. glass, packed with 2,6-diphenylene oxide 5.1.2 Automatic sampler (optional)—The polymer (60/80 mesh), Tenax, or equivalent. sampler must incorporate glass sample con- 5.6.3 Detector—Halide specific detector: tainers for the collection of a minimum of electrolytic conductivity or 250 mL of sample. Sample containers must be microcoulometric. These detectors have kept refrigerated at 4 °C and protected from proven effective in the analysis of light during compositing. If the sampler uses wastewaters for the parameters listed in the a peristaltic pump, a minimum length of scope (Section 1.1). The Hall conductivity de- compressible silicone rubber tubing may be tector was used to develop the method per- used. Before use, however, the compressible formance statements in Section 14. Guide- tubing should be thoroughly rinsed with lines for the use of alternate detectors are methanol, followed by repeated rinsings with provided in Section 12.1. Although less selec- distilled water to minimize the potential for tive, an electron capture detector is an ac- contamination of the sample. An integrating ceptable alternative. flow meter is required to collect flow propor- 6. Reagents tional composites. 5.2 Glassware (All specifications are sug- 6.1 Reagent water—Reagent water is de- gested. Catalog numbers are included for il- fined as a water in which an interferent is lustration only.): not observed at the MDL of the parameters 5.2.1 Separatory funnel—2-L, with Teflon of interest. stopcock. 6.2 Sodium thiosulfate—(ACS) Granular. 5.2.2 Drying column—Chromatographic 6.3 Acetone, hexane, methanol, methylene column, approximately 400 mm long × 19 mm chloride, petroleum ether (boiling range 30– ID, with coarse frit filter disc. 60 °C)—Pesticide quality or equivalent. 5.2.3 Chromatographic column—400 mm 6.4 Sodium sulfate—(ACS) Granular, an- long × 19 mm ID, with Teflon stopcock and hydrous. Purify by heating at 400 °C for 4 h coarse frit filter disc at bottom (Kontes K– in a shallow tray. 420540–0224 or equivalent). 6.5 Florisil—PR Grade (60/100 mesh). Pur- ° 5.2.4 Concentrator tube, Kuderna-Dan- chase activated at 1250 F and store in the ish—10-mL, graduated (Kontes K–570050–1025 dark in glass containers with ground glass or equivalent). Calibration must be checked stoppers or foil-lined screw caps. Before use, ° at the volumes employed in the test. Ground activate each batch at least 16 h at 130 C in glass stopper is used to prevent evaporation a foil-covered glass container and allow to of extracts. cool. 6.6 Ethyl ether—Nanograde, redistilled in 5.2.5 Evaporative flask, Kuderna-Danish— glass if necessary. 500-mL (Kontes K–570001–0500 or equivalent). 6.6.1 Ethyl ether must be shown to be free Attach to concentrator tube with springs. of peroxides before it is used as indicated by 5.2.6 Snyder column, Kuderna-Danish— EM Laboratories Quant test strips. (Avail- Three-ball macro (Kontes K–503000–0121 or able from Scientific Products Co., Cat. No. equivalent). P1126–8, and other suppliers.) 5.2.7 Vials—10 to 15-mL, amber glass, with 6.6.2 Procedures recommended for re- Teflon-lined screw cap. moval of peroxides are provided with the test 5.3 Boiling chips—Approximately 10/40 strips. After cleanup, 20 mL of ethyl alcohol mesh. Heat to 400 °C for 30 min or Soxhlet ex- preservative must be added to each liter of tract with methylene chloride. ether. 5.4 Water bath—Heated, with concentric 6.7 Stock standard solutions (1.00 μg/μL)— ring cover, capable of temperature control Stock standard solutions can be prepared (±2 °C). The bath should be used in a hood. from pure standard materials or purchased 5.5 Balance—Analytical, capable of accu- as certified solutions. rately weighing 0.0001 g. 6.7.1 Prepare stock standard solutions by 5.6 Gas chromatograph—An analytical accurately weighing about 0.0100 g of pure system complete with temperature program- material. Dissolve the material in acetone mable gas chromatograph suitable for on- and dilute to volume in a 10-mL volumetric column injection and all required accessories flask. Larger volumes can be used at the con- including syringes, analytical columns, venience of the analyst. When compound pu- gases, detector, and strip-chart recorder. A rity is assayed to be 96% or greater, the data system is recommended for measuring weight can be used without correction to cal- peak areas. culate the concentration of the stock stand- 5.6.1 Column 1—1.8 m long × 2 mm ID ard. Commercially prepared stock standards glass, packed with 3% SP–1000 on can be used at any concentration if they are Supelcoport (100/120 mesh) or equivalent. certified by the manufacturer or by an inde- This column was used to develop the method pendent source. performance statements in Section 14. 6.7.2 Transfer the stock standard solu- Guidelines for the use of alternate column tions into Teflon-sealed screw-cap bottles. packings are provided in Section 12.1. Store at 4 °C and protect from light. Stock

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standard solutions should be checked fre- concentrations found in real samples or quently for signs of degradation or evapo- should define the working range of the detec- ration, especially just prior to preparing tor. calibration standards from them. 7.3.2 Using injections of 2 to 5 μL, analyze 6.7.3 Stock standard solutions must be re- each calibration standard according to Sec- placed after six months, or sooner if com- tion 12 and tabulate peak height or area re- parison with check standards indicates a sponses against concentration for each com- problem. pound and internal standard. Calculate re- 6.8 Quality control check sample con- sponse factors (RF) for each compound using centrate—See Section 8.2.1. Equation 1. 7. Calibration ()()ACsis 7.1 Establish gas chromatographic oper- RF = ating conditions equivalent to those given in ()()ACis s Table 1. The gas chromatographic system can be calibrated using the external standard Equation 1 technique (Section 7.2) or the internal stand- where: ard technique (Section 7.3). As = Response for the parameter to be meas- 7.2 External standard calibration proce- ured. dure: A = Response for the internal standard. 7.2.1 Prepare calibration standards at a is Cis = Concentration of the internal standard minimum of three concentration levels for (μg/L). each parameter of interest by adding vol- Cs = Concentration of the parameter to be umes of one or more stock standards to a measured (μg/L). volumetric flask and diluting to volume with hexane. One of the external standards should If the RF value over the working range is a be at a concentration near, but above, the constant (<10% RSD), the RF can be assumed MDL (Table 1) and the other concentrations to be invariant and the average RF can be should correspond to the expected range of used for calculations. Alternatively, the re- concentrations found in real samples or sults can be used to plot a calibration curve should define the working range of the detec- of response ratios, As/Ais, vs. RF. tor. 7.4 The working calibration curve, cali- 7.2.2 Using injections of 2 to 5 μL, analyze bration factor, or RF must be verified on each calibration standard according to Sec- each working day by the measurement of one tion 12 and tabulate peak height or area re- or more calibration standards. If the re- sponses against the mass injected. The re- sponse for any parameter varies from the ± sults can be used to prepare a calibration predicted response by more than 15%, a new curve for each compound. Alternatively, if calibration curve must be prepared for that the ratio of response to amount injected compound. (calibration factor) is a constant over the 7.5 The cleanup procedure in Section 11 working range (<10% relative standard devi- utilizes Florisil column chromatography. ation, RSD), linearity through the origin can Florisil from different batches or sources be assumed and the average ratio or calibra- may vary in adsorptive capacity. To stand- tion factor can be used in place of a calibra- ardize the amount of Florisil which is used, 7 tion curve. the use of lauric acid value is suggested. 7.3 Internal standard calibration proce- The referenced procedure determines the ad- dure—To use this approach, the analyst must sorption from hexane solution of lauric acid select one or more internal standards that (mg) per g of Florisil. The amount of Florisil are similar in analytical behavior to the to be used for each column is calculated by compounds of interest. The analyst must fur- dividing 110 by this ratio and multiplying by ther demonstrate that the measurement of 20 g. the internal standard is not affected by 7.6 Before using any cleanup procedure, method or matrix interferences. Because of the analyst must process a series of calibra- these limitations, no internal standard can tion standards through the procedure to vali- be suggested that is applicable to all sam- date elution patterns and the absence of ples. interferences from the reagents. 7.3.1 Prepare calibration standards at a 8. Quality Control minimum of three concentration levels for each parameter of interest by adding vol- 8.1 Each laboratory that uses this method umes of one or more stock standards to a is required to operate a formal quality con- volumetric flask. To each calibration stand- trol program. The minimum requirements of ard, add a known constant amount of one or this program consist of an initial demonstra- more internal standards, and dilute to vol- tion of laboratory capability and an ongoing ume with hexane. One of the standards analysis of spiked samples to evaluate and should be at a concentration near, but above, document data quality. The laboratory must the MDL and the other concentrations maintain records to document the quality of should correspond to the expected range of data that is generated. Ongoing data quality

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checks are compared with established per- 8.2.2 Using a pipet, prepare QC check sam- formance criteria to determine if the results ples at a concentration of 100 μg/L by adding of analyses meet the performance character- 1.00 mL of QC check sample concentrate to istics of the method. When results of sample each of four 1–L aliquots of reagent water. spikes indicate atypical method perform- 8.2.3 Analyze the well-mixed QC check ance, a quality control check standard must samples according to the method beginning be analyzed to confirm that the measure- in Section 10. ments were performed in an in-control mode 8.2.4 Calculate the average recovery (X¯ ) in of operation. μg/L, and the standard deviation of the re- 8.1.1 The analyst must make an initial, covery (s) in μg/L, for each parameter using one-time, demonstration of the ability to the four results. generate acceptable accuracy and precision 8.2.5 For each parameter compare s and X¯ with this method. This ability is established with the corresponding acceptance criteria as described in Section 8.2. for precision and accuracy, respectively, 8.1.2 In recognition of advances that are found in Table 2. If s and X¯ for all param- occurring in chromatography, the analyst is eters of interest meet the acceptance cri- permitted certain options (detailed in Sec- teria, the system performance is acceptable tions 10.4, 11.1, and 12.1) to improve the sepa- and analysis of actual samples can begin. If rations or lower the cost of measurements. any individual s exceeds the precision limit Each time such a modification is made to or any individual X¯ falls outside the range the method, the analyst is required to repeat for accuracy, the system performance is un- the procedure in Section 8.2. acceptable for that parameter. Locate and 8.1.3 Before processing any samples, the correct the source of the problem and repeat analyst must analyze a reagent water blank the test for all parameters of interest begin- to demonstrate that interferences from the ning with Section 8.2.2. analytical system and glassware are under 8.3 The laboratory must, on an ongoing control. Each time a set of samples is ex- tracted or reagents are changed, a reagent basis, spike at least 10% of the samples from water blank must be processed as a safe- each sample site being monitored to assess guard against laboratory contamination. accuracy. For laboratories analyzing one to 8.1.4 The laboratory must, on an ongoing ten samples per month, at least one spiked basis, spike and analyze a minimum of 10% sample per month is required. of all samples to monitor and evaluate lab- 8.3.1. The concentration of the spike in oratory data quality. This procedure is de- the sample should be determined as follows: scribed in Section 8.3. 8.3.1.1 If, as in compliance monitoring, 8.1.5 The laboratory must, on an ongoing the concentration of a specific parameter in basis, demonstrate through the analyses of the sample is being checked against a regu- quality control check standards that the op- latory concentration limit, the spike should eration of the measurement system is in con- be at that limit or 1 to 5 times higher than trol. This procedure is described in Section the background concentration determined in 8.4. The frequency of the check standard Section 8.3.2, whichever concentration would analyses is equivalent to 10% of all samples be larger. analyzed but may be reduced if spike recov- 8.3.1.2 If the concentration of a specific eries from samples (Section 8.3) meet all parameter in the sample is not being specified quality control criteria. checked against a limit specific to that pa- 8.1.6 The laboratory must maintain per- rameter, the spike should be at 100 μg/L or 1 formance records to document the quality of to 5 times higher than the background con- data that is generated. This procedure is de- centration determined in Section 8.3.2, scribed in Section 8.5. whichever concentration would be larger. 8.2 To establish the ability to generate 8.3.1.3 If it is impractical to determine acceptable accuracy and precision, the ana- background levels before spiking (e.g., max- lyst must perform the following operations. imum holding times will be exceeded), the 8.2.1 A quality control (QC) check sample spike concentration should be (1) the regu- concentrate is required containing each pa- latory concentration limit, if any; or, if none rameter of interest at a concentration of 100 (2) the larger of either 5 times higher than μg/mL in acetone. The QC check sample con- the expected background concentration or centrate must be obtained from the U.S. En- 100 μg/L. vironmental Protection Agency, Environ- 8.3.2 Analyze one sample aliquot to deter- mental Monitoring and Support Laboratory mine the background concentration (B) of in Cincinnati, Ohio, if available. If not avail- each parameter. If necessary, prepare a new able from that source, the QC check sample QC check sample concentrate (Section 8.2.1) concentrate must be obtained from another appropriate for the background concentra- external source. If not available from either tions in the sample. Spike a second sample source above, the QC check sample con- aliquot with 1.0 mL of the QC check sample centrate must be prepared by the laboratory concentrate and analyze it to determine the using stock standards prepared independ- concentration after spiking (A) of each pa- ently from those used for calibration. rameter. Calculate each percent recovery (P)

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as 100(A¥B)%/T, where T is the known true samples must be assessed and records must value of the spike. be maintained. After the analysis of five 8.3.3 Compare the percent recovery (P) for spiked wastewater samples as in Section 8.3, each parameter with the corresponding QC calculate the average percent recovery (P¯ ) acceptance criteria found in Table 2. These and the standard deviation of the percent re- acceptance criteria were calculated to in- covery (sp). Express the accuracy assessment ¯ clude an allowance for error in measurement as a percent recovery interval from P–2sp to ¯ ¯ of both the background and spike concentra- P + 2sp. If P = 90% and sp = 10%, for example, tions, assuming a spike to background ratio the accuracy interval is expressed as 70– of 5:1. This error will be accounted for to the 110%. Update the accuracy assessment for extent that the analyst’s spike to back- each parameter on a regular basis (e.g. after ground ratio approaches 5:1. 8 If spiking was each five to ten new accuracy measure- performed at a concentration lower than 100 ments). μg/L, the analyst must use either the QC ac- 8.6 It is recommended that the laboratory ceptance criteria in Table 2, or optional QC adopt additional quality assurance practices acceptance criteria calculated for the spe- for use with this method. The specific prac- cific spike concentration. To calculate op- tices that are most productive depend upon tional acceptance criteria for the recovery of the needs of the laboratory and the nature of a parameter: (1) Calculate accuracy (X′) the samples. Field duplicates may be ana- using the equation in Table 3, substituting lyzed to assess the precision of the environ- the spike concentration (T) for C; (2) cal- mental measurements. When doubt exists culate overall precision (S′) using the equa- over the identification of a peak on the chro- ¯ tion in Table 3, substituting X′ for X; (3) cal- matogram, confirmatory techniques such as culate the range for recovery at the spike gas chromatography with a dissimilar col- concentration as (100 X′/T)±2.44(100 S′/T)%. 8 umn, specific element detector, or mass 8.3.4 If any individual P falls outside the spectrometer must be used. Whenever pos- designated range for recovery, that param- sible, the laboratory should analyze standard eter has failed the acceptance criteria. A reference materials and participate in rel- check standard containing each parameter evant performance evaluation studies. that failed the criteria must be analyzed as described in Section 8.4. 9. Sample Collection, Preservation, and 8.4 If any parameter fails the acceptance Handling criteria for recovery in Section 8.3, a QC check standard containing each parameter 9.1 Grab samples must be collected in that failed must be prepared and analyzed. glass containers. Conventional sampling 9 NOTE: The frequency for the required anal- practices should be followed, except that ysis of a QC check standard will depend upon the bottle must not be prerinsed with sample the number of parameters being simulta- before collection. Composite samples should neously tested, the complexity of the sample be collected in refrigerated glass containers matrix, and the performance of the labora- in accordance with the requirements of the tory. program. Automatic sampling equipment 8.4.1 Prepare the QC check standard by must be as free as possible of Tygon tubing adding 1.0 m/L of QC check sample con- and other potential sources of contamina- centrate (Section 8.2.1 or 8.3.2) to 1 L of rea- tion. gent water. The QC check standard needs 9.2 All samples must be iced or refrig- only to contain the parameters that failed erated at 4 °C from the time of collection criteria in the test in Section 8.3. until extraction. Fill the sample bottles and, 8.4.2 Analyze the QC check standard to if residual chlorine is present, add 80 mg of determine the concentration measured (A) of sodium thiosulfate per liter of sample and each parameter. Calculate each percent re- mix well. EPA Methods 330.4 and 330.5 may be used for measurement of residual chlo- covery (Ps) as 100 (A/T)%, where T is the true value of the standard concentration. rine. 10 Field test kits are available for this purpose. 8.4.3 Compare the percent recovery (Ps) for each parameter with the corresponding 9.3 All samples must be extracted within 7 QC acceptance criteria found in Table 2. Only days of collection and completely analyzed parameters that failed the test in Section 8.3 within 40 days of extraction. 2 need to be compared with these criteria. If 10. Sample Extraction the recovery of any such parameter falls out- side the designated range, the laboratory 10.1 Mark the water meniscus on the side performance for that parameter is judged to of the sample bottle for later determination be out of control, and the problem must be of sample volume. Pour the entire sample immediately identified and corrected. The into a 2-L separatory funnel. analytical result for that parameter in the 10.2 Add 60 mL methylene chloride to the unspiked sample is suspect and may not be sample bottle, seal, and shake 30 s to rinse reported for regulatory compliance purposes. the inner surface. Transfer the solvent to the 8.5 As part of the QC program for the lab- separatory funnel and extract the sample by oratory, method accuracy for wastewater shaking the funnel for 2 min with periodic

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venting to release excess pressure. Allow the 10.8 Remove the Snyder column and rinse organic layer to separate from the water the flask and its lower joint into the concen- phase for a minimum of 10 min. If the emul- trator tube with 1 to 2 mL of hexane. A 5-mL sion interface between layers is more than syringe is recommended for this operation. one-third the volume of the solvent layer, Stopper the concentrator tube and store re- the analyst must employ mechanical tech- frigerated if further processing will not be niques to complete the phase separation. The performed immediately. If the extract will optimum technique depends upon the sam- be stored longer than two days, it should be ple, but may include stirring, filtration of transferred to a Teflon-sealed screw-cap vial. the emulsion through glass wool, centrifuga- If the sample extract requires no further tion, or other physical methods. Collect the cleanup, proceed with gas chromatographic methylene chloride extract in a 250-mL Er- analysis (Section 12). If the sample requires lenmeyer flask. further cleanup, proceed to Section 11. 10.3 Add a second 60-mL volume of meth- ylene chloride to the sample bottle and re- 10.9 Determine the original sample vol- peat the extraction procedure a second time, ume by refilling the sample bottle to the combining the extracts in the Erlenmeyer mark and transferring the liquid to a 1000- flask. Perform a third extraction in the same mL graduated cylinder. Record the sample manner. volume to the nearest 5 mL. 10.4 Assemble a Kuderna-Danish (K-D) concentrator by attaching a 10-mL concen- 11. Cleanup and Separation trator tube to a 500-mL evaporative flask. 11.1 Cleanup procedures may not be nec- Other concentration devices or techniques essary for a relatively clean sample matrix. may be used in place of the K-D concentrator If particular circumstances demand the use if the requirements of Section 8.2 are met. of a cleanup procedure, the analyst may use 10.5 Pour the combined extract through a the procedure below or any other appropriate solvent-rinsed drying column containing procedure. However, the analyst first must about 10 cm of anhydrous sodium sulfate, demonstrate that the requirements of Sec- and collect the extract in the K-D concen- tion 8.2 can be met using the method as re- trator. Rinse the Erlenmeyer flask and col- vised to incorporate the cleanup procedure. umn with 20 to 30 mL of methylene chloride to complete the quantitative transfer. 11.2 Florisil column cleanup for 10.6 Add one or two clean boiling chips to haloethers: the evaporative flask and attach a three-ball 11.2.1 Adjust the sample extract volume Snyder column. Prewet the Snyder column to 10 mL. by adding about 1 mL of methylene chloride 11.2.2 Place a weight of Florisil (nomi- to the top. Place the K-D apparatus on a hot nally 20 g) predetermined by calibration water bath (60 to 65 °C) so that the concen- (Section 7.5), into a chromatographic col- trator tube is partially immersed in the hot umn. Tap the column to settle the Florisil water, and the entire lower rounded surface and add 1 to 2 cm of anhydrous sodium sul- of the flask is bathed with hot vapor. Adjust fate to the top. the vertical position of the apparatus and 11.2.3 Preelute the column with 50 to 60 the water temperature as required to com- mL of petroleum ether. Discard the eluate plete the concentration in 15 to 20 min. At and just prior to exposure of the sodium sul- the proper rate of distillation the balls of the fate layer to the air, quantitatively transfer column will actively chatter but the cham- the sample extract onto the column by de- bers will not flood with condensed solvent. cantation and subsequent petroleum ether When the apparent volume of liquid reaches washings. Discard the eluate. Just prior to 1 mL, remove the K-D apparatus and allow it to drain and cool for at least 10 min. exposure of the sodium sulfate layer to the air, begin eluting the column with 300 mL of NOTE: Some of the haloethers are very volatile and significant losses will occur in ethyl ether/petroleum ether (6 + 94) (V/V). concentration steps if care is not exercised. Adjust the elution rate to approximately 5 It is important to maintain a constant mL/min and collect the eluate in a 500-mL K- gentle evaporation rate and not to allow the D flask equipped with a 10-mL concentrator liquid volume to fall below 1 to 2 mL before tube. This fraction should contain all of the removing the K-D apparatus from the hot haloethers. water bath. 11.2.4 Concentrate the fraction as in Sec- 10.7 Momentarily remove the Snyder col- tion 10.6, except use hexane to prewet the umn, add 50 mL of hexane and a new boiling column. When the apparatus is cool, remove chip, and reattach the Snyder column. Raise the Snyder column and rinse the flask and the temperature of the water bath to 85 to 90 its lower joint into the concentrator tube °C. Concentrate the extract as in Section with hexane. Adjust the volume of the 10.6, except use hexane to prewet the column. cleaned up extract to 10 mL with hexane and The elapsed time of concentration should be analyze by gas chromatography (Section 12). 5 to 10 min.

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12. Gas Chromatography A = Amount of material injected (ng). μ 12.1 Table 1 summarizes the recommended Vi = Volume of extract injected ( L). μ operating conditions for the gas chro- Vt = Volume of total extract ( L). matograph. Included in this table are reten- Vs = Volume of water extracted (mL). tion times and MDL that can be achieved 13.1.2 If the internal standard calibration under these conditions. Examples of the sep- procedure is used, calculate the concentra- arations achieved by Columns 1 and 2 are tion in the sample using the response factor shown in Figures 1 and 2, respectively. Other (RF) determined in Section 7.3.2 and Equa- packed or capillary (open-tubular) columns, tion 3. chromatographic conditions, or detectors may be used if the requirements of Section ()()AI 8.2 are met. Concentration (μ= g/L) ss 12.2 Calibrate the system daily as de- ()()() scribed in Section 7. ARFVis o 12.3 If the internal standard calibration Equation 3 procedure is being used, the internal stand- ard must be added to the sample extract and where: mixed thoroughly immediately before injec- As = Response for the parameter to be meas- tion into the gas chromatrograph. ured. μ 12.4 Inject 2 to 5 L of the sample extract A = Response for the internal standard. or standard into the gas chromatograph is I = Amount of internal standard added to using the solvent-flush technique. 11 Smaller s each extract (μg). (1.0 μL) volumes may be injected if auto- matic devices are employed. Record the vol- Vo = Volume of water extracted (L). ume injected to the nearest 0.05 μL, the total 13.2 Report results in μg/L without correc- extract volume, and the resulting peak size tion for recovery data. All QC data obtained in area or peak height units. should be reported with the sample results. 12.5 Identify the parameters in the sample by comparing the retention times of the 14. Method Performance peaks in the sample chromatogram with 14.1 The method detection limit (MDL) is those of the peaks in standard defined as the minimum concentration of a chromatograms. The width of the retention substance that can be measured and reported time window used to make identifications with 99% confidence that the value is above should be based upon measurements of ac- zero. 1 The MDL concentrations listed in tual retention time variations of standards Table 1 were obtained using reagent water. 12 over the course of a day. Three times the standard deviation of a retention time for a Similar results were achieved using rep- compound can be used to calculate a sug- resentative wastewaters. The MDL actually gested window size; however, the experience achieved in a given analysis will vary de- of the analyst should weight heavily in the pending on instrument sensitivity and ma- interpretation of chromatograms. trix effects. 12.6 If the response for a peak exceeds the 14.2 This method has been tested for lin- working range of the system, dilute the ex- earity of spike recovery from reagent water tract and reanalyze. and has been demonstrated to be applicable 12.7 If the measurement of the peak re- over the concentration range from 4 × MDL sponse is prevented by the presence of inter- to 1000 × MDL. 12 ferences, further cleanup is required. 14.3 This method was tested by 20 labora- tories using reagent water, drinking water, 13. Calculations surface water, and three industrial 13.1 Determine the concentration of indi- wastewaters spiked at six concentrations vidual compounds in the sample. over the range 1.0 to 626 μ/L. 12 Single oper- 13.1.1 If the external standard calibration ator precision, overall precision, and method procedure is used, calculate the amount of accuracy were found to be directly related to material injected from the peak response the concentration of the parameter and es- using the calibration curve or calibration sentially independent of the sample matrix. factor determined in Section 7.2.2. The con- Linear equations to describe these relation- centration in the sample can be calculated ships are presented in Table 3. from Equation 2. REFERENCES () AV()t 1. 40 CFR part 136, appendix B. Concentration (μ= g/L) 2. ‘‘Determination of Haloethers in Indus- ()VVis() trial and Municipal Wastewaters,’’ EPA 600/ 4–81–062, National Technical Information Equation 2 Service, PB81–232290, Springfield, Virginia where: 22161, July 1981.

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3. ASTM Annual Book of Standards, Part 8. Provost, L.P., and Elder, R.S. ‘‘Interpre- 31, D3694–78. ‘‘Standard Practices for Prepa- tation of Percent Recovery Data,’’ American ration of Sample Containers and for Preser- Laboratory, 15, 58–63 (1983). (The value 2.44 vation of Organic Constitutents,’’ American used in the equation in Section 8.3.3 is two Society for Testing and Materials, Philadel- times the value 1.22 derived in this report.) phia. 9. ASTM Annual Book of Standards, Part 4. ‘‘Carcinogens—Working Carcinogens, ’’ 31, D3370–76. ‘‘Standard Practices for Sam- Department of Health, Education, and Wel- pling Water,’’ American Society for Testing fare, Public Health Services, Center for Dis- and Materials, Philadelphia. ease Control, National Institute for Occupa- 10. ‘‘Methods 330.4 (Titrimetric, DPD-FAS) tional Safety and Health, Publication No. 77– and 330.5 (Spectrophotometric, DPD) for 206, August 1977. Chlorine, Total Residual,’’ Methods for 5. ‘‘OSHA Safety and Health Standards, Chemical Analysis of Water and Wastes, General Industry,’’ (29 CFR part 1910), Occu- EPA–600/4–79–020, U.S. Environmental Pro- pational Safety and Health Administration, tection Agency, Environmental Monitoring OSHA 2206 (Revised, January 1976). and Support Laboratory, Cincinnati, Ohio 6. ‘‘Safety in Academic Chemistry Labora- 45268, March 1979. tories,’’ American Chemical Society Publica- 11. Burke, J.A. ‘‘Gas Chromatography for tion, Committee on Chemical Safety, 3rd Pesticide Residue Analysis; Some Practical Edition, 1979. Aspects,’’ Journal of the Association of Official 7. Mills., P.A. ‘‘Variation of Florisil Activ- Analytical Chemists, 48, 1037 (1965). ity: Simple Method for Measuring Absorbent 12. ‘‘EPA Method Study 21, Method 611, Capacity and Its Use in Standardizing Haloethers,’’ EPA 600/4–84–052, National Florisil Columns,’’ Journal of the Association Technical Information Service, PB84–205939, of Official Analytical Chemists, 51, 29 (1968). Springfield, Virginia 22161, June 1984.

TABLE 1—CHROMATOGRAPHIC CONDITIONS AND METHODS DETECTION LIMITS

Retention time (min) Method Parameters detection Column 1 Column 2 limit (μ/L)

Bis(2-chloroisopropyl) ether ...... 8 .4 9 .7 0 .8 Bis(2-chloroethyl) ether ...... 9 .3 9 .1 0 .3 Bis(2-chloroethoxy) methane ...... 13 .1 10 .0 0 .5 4-Chlorophenyl ether ...... 19.4 15 .0 3 .9 4-Bromophenyl phenyl ether ...... 21.2 16.2 2 .3 Column 1 conditions: Supelcoport (100/120 mesh) coated with 3% SP–1000 packed in a 1.8 m long × 2 mm ID glass column with helium carrier gas at 40 mL/min. flow rate. Column temperature held at 60 °C for 2 min. after injection then programmed at 8 °C/min. to 230 °C and held for 4 min. Under these conditions the retention time for Aldrin is 22.6 min. Column 2 conditions: Tenax-GC (60/80 mesh) packed in a 1.8 m long × 2mm ID glass column with helium carrier gas at 40 mL/min. flow rate. Column temperature held at 150 °C for 4 min. after injection then programmed at 16 °C/min. to 310 °C. Under these conditions the retention time for Aldrin is 18.4 min.

TABLE 2—QC ACCEPTANCE CRITERIA—METHOD 611

Test conc. Limit for s Range for X¯ Range for Parameter μ μ μ P, Ps per- ( g/L) ( g/L) ( g/L) cent

Bis (2-chloroethyl)ether ...... 100 26.3 26.3–136.8 11–152 Bis (2-chloroethoxy)methane ...... 100 25.7 27.3–115.0 12–128 Bis (2-chloroisopropyl)ether ...... 100 32 .7 26.4–147.0 9–165 4-Bromophenyl phenyl ether ...... 100 39 .3 7.6–167.5 D–189 4-Chlorophenyl phenyl ether ...... 100 30 .7 15.4–152.5 D–170 s = Standard deviation of four recovery measurements, in μg/L (Section 8.2.4). X¯ = Average recovery for four recovery measurements, in μg/L (Section 8.2.4). P, Ps = Percent recovery measured (Section 8.3.2, Section 8.4.2). D = Detected; result must be greater than zero. NOTE: These criteria are based directly upon the method performance data in Table 3. Where necessary, the limits for recov- ery have been broadened to assure applicability of the limits to concentrations below those used to develop Table 3.

TABLE 3—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 611

Accuracy, as Single analyst ′ ′ Overall preci- Parameter recovery, X precision, sr ′ μ (μg/L) (μg/L) sion, S ( g/L)

Bis(2-chloroethyl) ether ...... 0.81C + 0.54 0.19X¯ + 0.28 0.35X¯ + 0,36 Bis(2-chloroethoxy) methane ...... 0.71C + 0.13 0.20X¯ + 0.15 0.33X¯ + 0.11 Bis(2-chloroisopropyl) ether ...... 0.85C + 1.67 0.20X¯ + 1.05 0.36X¯ + 0.79 4–Bromophenyl phenyl ether ...... 0.85C + 2.55 0.25X¯ + 0.21 0.47X¯ + 0.37

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TABLE 3—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 611— Continued

Accuracy, as Single analyst ′ ′ Overall preci- Parameter recovery, X precision, sr ′ μ (μg/L) (μg/L) sion, S ( g/L)

4–Chlorophenyl phenyl ether ...... 0.82C + 1.97 0.18X¯ + 2.13 0.41X¯ + 0.55 X′ = Expected recovery for one or more measuremelts of a sample containing a concentration of C, in μg/L. sr′ = Expected single analyst standard deviation of measurements at an average concentration found of X¯ , in μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X¯ , in μg/L. C = True value for the concentration, in μg/L. X¯ = Average recovery found for measurements of samples containing a concentration of C, in μg/L.

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METHOD 612—CHLORINATED HYDROCARBONS STORET Parameter No. CAS No. 1. Scope and Application 2-Chloronaphthalene ...... 34581 91–58–7 1.1 This method covers the determination 1,2-Dichlorobenzene ...... 34536 95–50–1 of certain chlorinated hydrocarbons. The fol- 1,3-Dichlorobenzene ...... 34566 541–73–1 lowing parameters can be determined by this 1,4-Dichlorobenzene ...... 34571 106–46–7 method: Hexachlorobenzene ...... 39700 118–74–1 Hexachlorobutadiene ...... 34391 87–68–3 Hexachlorocyclopentadiene ...... 34386 77–47–4 Hexachloroethane ...... 34396 67–72–1

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STORET 2.2 The method provides a Florisil column Parameter No. CAS No. cleanup procedure to aid in the elimination of interferences that may be encountered. 1,2,4-Trichlorobenzene ...... 34551 120–82–1 3. Interferences 1.2 This is a gas chromatographic (GC) method applicable to the determination of 3.1 Method interferences may be caused the compounds listed above in municipal and by contaminants in solvents, reagents, glass- industrial discharges as provided under 40 ware, and other sample processing hardware CFR 136.1. When this method is used to ana- that lead to discrete artifacts and/or ele- lyze unfamiliar samples for any or all of the vated baselines in gas chromatograms. All of compounds above, compound identifications these materials must be routinely dem- should be supported by at least one addi- onstrated to be free from interferences under tional qualitative technique. This method the conditions of the analysis by running describes a second gas chromatographic col- laboratory reagent blanks as described in umn that can be used to confirm measure- Section 8.1.3. ments made with the primary column. Meth- 3.1.1 Glassware must be scrupulously od 625 provides gas chromatograph/mass cleaned. 3 Clean all glassware as soon as pos- spectrometer (GC/MS) conditions appro- sible after use by rinsing with the last sol- priate for the qualitative and quantitative vent used in it. Solvent rinsing should be fol- confirmation of results for all of the param- lowed by detergent washing with hot water, eters listed above, using the extract pro- and rinses with tap water and distilled duced by this method. water. The glassware should then be drained 1.3 The method detection limit (MDL, de- dry, and heated in a muffle furnace at 400 °C fined in Section 14.1) 1 for each parameter is for 15 to 30 min. Some thermally stable ma- listed in Table 1. The MDL for a specific terials, such as PCBs, may not be eliminated wastewater may differ from those listed, de- by this treatment. Solvent rinses with ace- pending upon the nature of interferences in tone and pesticide quality hexane may be the sample matrix. substituted for the muffle furnace heating. 1.4 The sample extraction and concentra- Thorough rinsing with such solvents usually tion steps in this method are essentially the eliminates PCB interference. Volumetric same as in Methods 606, 608, 609, and 611. ware should not be heated in a muffle fur- Thus, a single sample may be extracted to nace. After drying and cooling, glassware measure the parameters included in the should be sealed and stored in a clean envi- scope of each of these methods. When clean- ronment to prevent any accumulation of up is required, the concentration levels must dust or other contaminants. Store inverted be high enough to permit selecting aliquots, or capped with aluminum foil. as necessary, to apply appropriate cleanup 3.1.2 The use of high purity reagents and procedures. The analyst is allowed the lati- solvents helps to minimize interference prob- tude, under Section 12, to select lems. Purification of solvents by distillation chromatographic conditions appropriate for in all-glass systems may be required. the simultaneous measurement of combina- 3.2 Matrix interferences may be caused by tions of these parameters. contaminants that are co-extracted from the 1.5 Any modification of this method, be- sample. The extent of matrix interferences yond those expressly permitted, shall be con- will vary considerably from source to source, sidered as a major modification subject to depending upon the nature and diversity of application and approval of alternate test the industrial complex or municipality being procedures under 40 CFR 136.4 and 136.5. sampled. The cleanup procedure in Section 1.6 This method is restricted to use by or 11 can be used to overcome many of these under the supervision of analysts experi- interferences, but unique samples may re- enced in the use of a gas chromatograph and quire additional cleanup approaches to in the interpretation of gas chromatograms. achieve the MDL listed in Table 1. Each analyst must demonstrate the ability to generate acceptable results with this 4. Safety method using the procedure described in Sec- 4.1 The toxicity or carcinogenicity of tion 8.2. each reagent used in this method has not been precisely defined; however, each chem- 2. Summary of Method ical compound should be treated as a poten- 2.1 A measured volume of sample, ap- tial health hazard. From this viewpoint, ex- proximately 1–L, is extracted with meth- posure to these chemicals must be reduced to ylene chloride using a separatory funnel. The the lowest possible level by whatever means methylene chloride extract is dried and ex- available. The laboratory is responsible for changed to hexane during concentration to a maintaining a current awareness file of volume of 10 mL or less. The extract is sepa- OSHA regulations regarding the safe han- rated by gas chromatography and the param- dling of the chemicals specified in this meth- eters are then measured with an electron od. A reference file of material data handling capture detector. 2 sheets should also be made available to all

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personnel involved in the chemical analysis. suitable for on-column injection and all re- Additional references to laboratory safety quired accessories including syringes, ana- are available and have been identified 4M6 lytical columns, gases, detector, and strip- for the information of the analyst. chart recorder. A data system is rec- ommended for measuring peak areas. 5. Apparatus and Materials 5.6.1 Column 1—1.8 m long × 2 mm ID 5.1 Sampling equipment, for discrete or glass, packed with 1% SP–1000 on composite sampling. Supelcoport (100/120 mesh) or equivalent. 5.1.1 Grab sample bottle—1cL or 1-qt, Guidelines for the use of alternate column amber glass, fitted with a screw cap lined packings are provide in Section 12.1. with Teflon. Foil may be substituted for Tef- 5.6.2 Column 2—1.8 m long × 2 mm ID lon if the sample is not corrosive. If amber glass, packed with 1.5% OV–1/2.4% OV–225 on bottles are not available, protect samples Supelcoport (80/100 mesh) or equivalent. This from light. The bottle and cap liner must be column was used to develop the method per- washed, rinsed with acetone or methylene formance statements in Section 14. chloride, and dried before use to minimize 5.6.3 Detector—Electron capture detector. contamination. This detector has proven effective in the 5.1.2 Automatic sampler (optional)—The analysis of wastewaters for the parameters sampler must incorporate glass sample con- listed in the scope (Section 1.1), and was used tainers for the collection of a minimum of to develop the method performance state- 250 mL of sample. Sample containers must be ments in Section 14. Guidelines for the use of kept refrigerated at 4 °C and protected from alternate detectors are provided in Section light during compositing. If the sampler uses 12.1. a peristaltic pump, a minimum length of 6. Reagents compressible silicone rubber tubing may be used. Before use, however, the compressible 6.1 Reagent water—Reagent water is de- tubing should be thoroughly rinsed with fined as a water in which an interferent is methanol, followed by repeated rinsings with not observed at the MDL of the parameters distilled water to minimize the potential for of interest. contamination of the sample. An integrating 6.2 Acetone, hexane, isooctane, methanol, flow meter is required to collect flow propor- methylene chloride, petroleum ether (boiling tional composites. range 30 to 60 °C)—Pesticide quality or equiv- 5.2 Glassware (All specifications are sug- alent. gested. Catalog numbers are included for il- 6.3 Sodium sulfate—(ACS) Granular, an- lustration only.): hydrous. Purify heating at 400 °C for 4 h in a 5.2.1 Separatory funnel—2–L, with Teflon shallow tray. stopcock. 6.4 Florisil—PR grade (60/100 mesh). Pur- 5.2.2 Drying column—Chromatographic chase activated at 1250 °F and store in the column, approximately 400 mm long × 19 mm dark in glass containers with ground glass ID, with coarse frit filter disc. stoppers or foil-lined screw caps. Before use, 5.2.3 Chromatographic column—300 long × activate each batch at least 16 h at 130 °C in 10 mm ID, with Teflon stopcock and coarse a foil-covered glass container and allow to frit filter disc at bottom. cool. 5.2.4 Concentrator tube, Kuderna-Dan- 6.5 Stock standard solution (1.00 μg/μL)— ish—10-mL, graduated (Kontes K–570050–1025 Stock standard solutions can be prepared or equivalent). Calibration must be checked from pure standard materials or purchased at the volumes employed in the test. Ground as certified solutions. glass stopper is used to prevent evaporation 6.5.1 Prepare stock standard solutions by of extracts. accurately weighing about 0.0100 g of pure 5.2.5 Evaporative flask, Kuderna-Danish— material. Dissolve the material in isooctane 500-mL (Kontes K–570001–0500 or equivalent). and dilute to volume in a 120-mL volumetric Attach to concentrator tube with springs. flask. Larger volumes can be used at the con- 5.2.6 Snyder column, Kuderna-Danish— venience of the analyst. When compound pu- Three-ball macro (Kontes K–503000–0121 or rity is assayed to be 96% or greater, the equivalent). weight can be used without correction to cal- 5.2.7 Vials—10 to 15-mL, amber glass, with culate the concentration of the stock stand- Teflon-lined screw cap. ard. Commercially prepared stock standards 5.3 Boiling chips—Approximately 10/40 can be used at any concentration if they are mesh. Heat to 400 °C for 30 min or Soxhlet ex- certified by the manufacturer or by an inde- tract with methylene chloride. pendent source. 5.4 Water bath—Heated, with concentric 6.5.2 Transfer the stock standard solu- ring cover, capable of temperature control tions into Teflon-sealed screw-cap bottles. (±2 °C). The bath should be used in a hood. Store at 4 °C and protect from light. Stock 5.5 Balance—Analytical, capable of accu- standard solutions should be checked fre- rately weighing 0.0001 g. quently for signs of degradation or evapo- 5.6 Gas chromatograph—An analytical ration, especially just prior to preparing system complete with gas chromatograph calibration standards from them.

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6.5.3 Stock standard solutions must be re- 7.3.2 Using injections of 2 to 5 μL, analyze placed after six months, or sooner if each calibration standard according to Sec- comparision with check standards indicates tion 12 and tabulate peak height or area re- a problem. sponses against concentration for each com- 6.6 Quality control check sample con- pound and internal standard. Calculate re- centrate—See Section 8.2.1. sponse factors (RF) for each compound using Equation 1. 7. Calibration

7.1 Establish gas chromatographic oper- ()()ACsis ating conditions equivalent to those given in RF = Table 1. The gas chromatographic system ()()ACis s can be calibrated using the external standard technique (Section 7.2) or the internal stand- Equation 1 ard technique (Section 7.3). where: 7.2 External standard calibration proce- As = Response for the parameter to be meas- dure: ured. 7.2.1 Prepare calibration standards at a A = Response for the internal standard. minimum of three concentration levels for is Cis = Concentration of the internal standard each parameter of interest by adding vol- (μg/L). umes of one or more stock standards to a Cs = Concentration of the parameter to be volumetric flask and diluting to volume with measured (μg/L). isooctane. One of the external standards should be at a concentration near, but above, If the RF value over the working range is a the MDL (Table 1) and the other concentra- constant (<10% RSD), the RF can be assumed tions should correspond to the expected to be invariant and the average RF can be range of concentrations found in real sam- used for calculations. Alternatively, the re- ples or should define the working range of sults can be used to plot a calibration curve the detector. of response ratios, As/Ais, vs. RF. 7.2.2 Using injections of 2 to 5 μL, analyze 7.4 The working calibration curve, cali- each calibration standard according to Sec- bration factor, or RF must be verified on tion 12 and tabulate peak height or area re- each working day by the measurement of one sponses against the mass injected. The re- or more calibration standards. If the re- sults can be used to prepare a calibration sponse for any parameter varies from the curve for each compound. Alternatively, if predicted response by more than ±15%, a new the ratio of response to amount injected calibration curve must be prepared for that (calibration factor) is a constant over the compound. working range (<10% relative standard devi- 7.5 Before using any cleanup procedure, ation, RSD), linearity through the origin can the analyst must process a series of calibra- be assumed and the average ratio or calibra- tion standards through the procedure to vali- tion factor can be used in place of a calibra- date elution patterns and the absence of tion curve. interferences from the reagents. 7.3 Internal standard calibration proce- 8. Quality Control dure—To use this approach, the analyst must select one or more internal standards that 8.1 Each laboratory that uses this method are similar in analytical behavior to the is required to operate a formal quality con- compounds of interest. The analyst must fur- trol program. The minimum requirements of ther demonstrate that the measurement of this program consist of an initial demonstra- the internal standard is not affected by tion of laboratory capability and an ongoing method or matrix interferences. Because of analysis of spiked samples to evaluate and these limitations, no internal standard can document data quality. The laboratory must be suggested that is applicable to all sam- maintain records to document the quality of ples. data that is generated. Ongoing data quality 7.3.1 Prepare calibration standards at a checks are compared with established per- minimum of three concentration levels for formance criteria to determine if the results each parameter of interest by adding vol- of analyses meet the performance character- umes of one or more stock standards to a istics of the method. When the results of volumetric flask. To each calibration stand- sample spikes indicate atypical method per- ard, add a known constant amount of one or formance, a quality control check standard more internal standards, and dilute to vol- must be analyzed to confirm that the meas- ume with isooctane. One of the standards urements were performed in an in-control should be at a concentration near, but above, mode of operation. the MDL and the other concentrations 8.1.1 The analyst must make an initial, should correspond to the expected range of one-time, demonstration of the ability to concentrations found in real samples or generate acceptable accuracy and precision should define the working range of the detec- with this method. This ability is established tor. as described in Section 8.2.

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8.1.2 In recognition of advances that are covery (s) in μg/L, for each parameter using occurring in chromatography, the analyst is the four results. permitted certain options (detailed in Sec- 8.2.5 For each parameter compare s and X¯ tions 10.4, 11.1, and 12.1) to improve the sepa- with the corresponding acceptance criteria rations or lower the cost of measurements. for precision and accuracy, respectively, Each time such modification is made to the found in Table 2. If s and X¯ for all param- method, the analyst is required to repeat the eters of interest meet the acceptance cri- procedure in Section 8.2. teria, the system performance is acceptable 8.1.3 Before processing any samples, the and analysis of actual samples can begin. If analyst must analyze a reagent water blank any individual s exceeds the precision limit to demonstrate that interferences from the or any individual X¯ falls outside the range analytical system and glassware are under for accuracy, the system performance is un- control. Each time a set of samples is ex- acceptable for that parameter. tracted or reagents are changed, a reagent NOTE: The large number of parameters in water blank must be processed as a safe- Table 2 presents a substantial probability guard against laboratory contamination. that one or more will fail at least one of the 8.1.4 The laboratory must, on an ongoing acceptance criteria when all parameters are basis, spike and analyze a minimum of 10% analyzed. of all samples to monitor and evaluate lab- 8.2.6 When one or more of the parameters oratory data quality. This procedure is de- tested fail at least one of the acceptance cri- scribed in Section 8.3. teria, the analyst must proceed according to 8.1.5 The laboratory must, on an ongoing Section 8.2.6.1 or 8.2.6.2. basis, demonstrate through the analyses of 8.2.6.1 Locate and correct the source of quality control check standards that the op- the problem and repeat the test for all pa- eration of the measurement system is in con- rameters of interest beginning with Section trol. This procedure is described in Section 8.2.2. 8.4. The frequency of the check standard 8.2.6.2 Beginning with Section 8.2.2, repeat analyses is equivalent to 10% of all samples the test only for those parameters that analyzed but may be reduced if spike recov- failed to meet criteria. Repeated failure, eries from samples (Section 8.3) meet all however, will confirm a general problem specified quality control criteria. with the measurement system. If this occurs, 8.1.6 The laboratory must maintain per- locate and correct the source of the problem formance records to document the quality of and repeat the test for all compounds of in- data that is generated. This procedure is de- terest beginning with Section 8.2.2. scribed in Section 8.5. 8.3 The laboratory must, on an ongoing 8.2 To establish the ability to generate basis, spike at least 10% of the samples from acceptable accuracy and precision, the ana- each sample site being monitored to assess lyst must perform the following operations. accuracy. For laboratories analyzing one to 8.2.1 A quality control (QC) check sample ten samples per month, at least one spike concentrate is required containing each pa- sample per month is required. rameter of interest at the following con- 8.3.1 The concentration of the spike in the centrations in acetone: Hexachloro-sub- sample should be determined as follows: stituted parameters, 10 μg/mL; any other 8.3.1.1 If, as in compliance monitoring, chlorinated hydrocarbon, 100 μg/mL. The QC the concentration of a specific parameter in check sample concentrate must be obtained the sample is being checked against a regu- from the U.S. Environmental Protection latory concentration limit, the spike should Agency, Environmental Monitoring and Sup- be at that limit or 1 to 5 times higher than port Laboratory in Cincinnati, Ohio, if avail- the background concentration determined in able. If not available from that source, the Section 8.3.2, whichever concentration would QC check sample concentrate must be ob- be larger. tained from another external source. If not 8.3.1.2 If the concentration of a specific available from either source above, the QC parameter in the sample is not being check sample concentrate must be prepared checked against a limit specific to that pa- by the laboratory using stock standards pre- rameter, the spike should be at the test con- pared independently from those used for cali- centration in Section 8.2.2 or 1 to 5 times bration. higher than the background concentration 8.2.2 Using a pipet, prepare QC check sam- determined in Section 8.3.2, whichever con- ples at the test concentrations shown in centration would be larger. Table 2 by adding 1.00 mL of QC check sam- 8.3.1.3 If it is impractical to determine ple concentrate to each of four 1–L aliquots background levels before spiking (e.g., max- of reagent water. imum holding times will be exceeded), the 8.2.3 Analyze the well-mixed QC check spike concentration should be (1) the regu- samples according to the method beginning latory concentration limit, if any; or, if none in Section 10. by (2) the larger of either 5 times higher than 8.2.4 Calculate the average recovery (X¯ ) in the expected background concentration or μg/L, and the standard deviation of the re- the test concentration in Section 8.2.2.

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8.3.2 Analyze one sample aliquot to deter- need to be compared with these criteria. If mine the background concentration (B) of the recovery of any such parameter falls out- each parameter. In necessary, prepare a new side the designated range, the laboratory QC check sample concentrate (Section 8.2.1) performance for that parameter is judged to appropriate for the background concentra- be out of control, and the problem must be tions in the sample. Spike a second sample immediately identified and corrected. The aliquot with 1.0 mL of the QC check sample analytical result for that parameter in the concentrate and analyze it to determine the unspiked sample is suspect and may not be concentration after spiking (A) of each pa- reported for regulatory compliance purposes. rameter. Calculate each percent recovery (P) 8.5 As part of the QC program for the lab- as 100 (A¥B)%/T, where T is the known true oratory, method accuracy for wastewater value of the spike. samples must be assessed and records must 8.3.3 Compare the percent recovery (P) for be maintained. After the analysis of five each parameter with the corresponding QC spiked wastewater samples as in Section 8.3, acceptance criteria found in Table 2. These calculate the average percent recovery (P¯ ) acceptance criteria were calculated to in- and the standard deviation of the percent re- clude an allowance for error in measurement covery (sp). Express the accuracy assessment of both the background and spike concentra- as a percent recovery interval from P¥2sp to tions, assuming a spike to background ratio P + 2sp. If P = 90% and sp = 10%, for example, of 5:1. This error will be accounted for to the the accuracy interval is expressed as 70– extent that the analyst’s spike to back- 110%. Update the accuracy assessment for ground ratio approaches 5:1. 7 If spiking was each parameter on a regular basis (e.g. after performed at a concentration lower than the each five to ten new accuracy measure- test concentration in Section 8.2.2, the ana- ments). lyst must use either the QC acceptance cri- 8.6 It is recommended that the laboratory teria in Table 2, or optional QC acceptance adopt additional quality assurance practices criteria calculated for the specific spike con- for use with this method. The specific prac- centration. To calculate optional acceptance tices that are most productive depend upon criteria for the recovery of a parameter: (1) the needs of the laboratory and the nature of ′ Calculate accuracy (X ) using the equation in the samples. Field duplicates may be ana- Table 3, substituting the spike concentration lyzed to assess the precision of the environ- ′ (T) for C; (2) calculate overall precision (S ) mental measurements. When doubt exists using the equation in Table 3, substituting X′ ¯ over the identification of a peak on the chro- for X; (3) calculate the range for recovery at matogram, confirmatory techniques such as ′ ± the spike concentration as (100 X /T) 2.44 gas chromatography with a dissimilar col- ′ 7 (100 S /T)%. umn, specific element detector, or mass 8.3.4 If any individual P falls outside the spectrometer must be used. Whenever pos- designated range for recovery, that param- sible, the laboratory should analyze standard eter has failed the acceptance criteria. A reference materials and participate in check standard containing each parameter relevent performance evaluation studies. that failed the criteria must be analyzed as described in Section 8.4. 9. Sample Collection, Preservation, and 8.4. If any parameter fails the acceptance Handling criteria for recovery in Section 8.3, a QC check standard containing each parameter 9.1 Grab samples must be collected in that failed must be prepared and analyzed. glass containers. Conventional sampling 8 NOTE: The frequency for the required anal- practices should be followed, except that ysis of a QC check standard will depend upon the bottle must not be prerinsed with sample the number of parameters being simulta- before collection. Composite samples should neously tested, the complexity of the sample be collected in refrigerated glass containers matrix, and the performance of the labora- in accordance with the requirements of the tory. program. Automatic sampling equipment 8.4.1 Prepare the QC check standard by must be as free as possible of Tygon tubing adding 1.0 mL of QC check sample con- and other potential sources of contamina- centrate (Sections 8.2.1 or 8.3.2) to 1 L of rea- tion. gent water. The QC check standard needs 9.2 All samples must be iced or refrig- only to contain the parameters that failed erated at 4 °C from the time of collection criteria in the test in Section 8.3. until extraction. 8.4.2 Analyze the QC check standard to 9.3 All samples must be extracted within 7 determine the concentration measured (A) of days of collection and completely analyzed each parameter. Calculate each percent re- within 40 days of extraction. 2 covery (P ) as 100 (A/T)%, where T is the true s 10. Sample Extraction value of the standard concentration. 8.4.3 Compare the percent recovery (Ps) 10.1 Mark the water meniscus on the side for each parameter with the corresponding of the sample bottle for later determination QC acceptance criteria found in Table 2. Only of sample volume. Pour the entire sample parameters that failed the test in Section 8.3 into a 2–L separatory funnel.

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10.2 Add 60 mL of methylele chloride to °C. Concentrate the extract as in Section the sample bottle, seal, and shake 30 s to 10.6, except use hexane to prewet the column. rinse the inner surface. Transfer the solvent The elapsed time of concentration should be to the separatory funnel and extract the 5 to 10 min. sample by shaking the funnel for 2 min with 10.8 Romove the Snyder column and rinse periodic venting to release excess pressure. the flask and its lower joint into the concen- Allow the organic layer to separate from the trator tube with 1 to 2 mL of hexane. A 5-mL water phase for a minimum of 10 min. If the syringe is recommended for this operation. emulsion interface between layers is more than one-third the volume of the solvent Stopper the concentrator tube and store re- layer, the analyst must employ mechanical frigerated if further processing will not be techniques to complete the phase separation. performed immediately. If the extract will The optimum technique depends upon the be stored longer than two days, it should be sample, but may include stirring, filtration transferred to a Teflon-sealed screw-cap vial. of the emulsion through glass wool, cen- If the sample extract requires no further trifugation, or other physical methods. Col- cleanup, proceed with gas chromatographic lect the methylene chloride extract in a 250- analysis (Section 12). If the sample requires mL Erlenmeyer flask. further cleanup, proceed to Section 11. 10.3 Add a second 60-mL volume of meth- 10.9 Determine the original sample vol- ylene chloride to the sample bottle and re- ume by refilling the sample bottle to the peat the extraction procedure a second time, mark and transferring the liquid to a 1000- combining the extracts in the Erlenmeyer mL graduated cylinder. Record the sample flask. Perform a third extraction in the same volume to the nearest 5 mL. manner. 10.4 Assemble a Kuderna-Danish (K-D) 11. Cleanup and Separation concentrator by attaching a 10-mL concen- trator tube to a 500-mL evaporative flask. 11.1 Cleanup procedures may not be nec- Other concentration devices or techniques essary for a relatively clean sample matrix. may be used in place of the K-D concentrator If particular circumstances demand the use if the requirements of Section 8.2 are met. of a cleanup procedure, the analyst may use 10.5 Pour the combined extract through a the procedure below or any other appropriate solvent-rinsed drying column containing procedure. However, the analyst first must about 10 cm of anhydrous sodium sulfate, demonstrate that the requirements of Sec- and collect the extract in the K-D concen- tion 8.2 can be met using the method as re- trator. Rinse the Erlenmeyer flask and col- vised to incorporate the cleanup procedure. umn with 20 to 30 mL of methylene chloride to complete the quantitative transfer. 11.2 Florisil column cleanup for 10.6 Add one or two clean boiling chips to chlorinated hydrocarbons: the evaporative flask and attach a three-ball 11.2.1 Adjust the sample extract to 10 mL Snyder column. Prewet the Snyder column with hexane. by adding about 1 mL of methylene chloride 11.2.2 Place 12 g of Florisil into a to the top. Place the K-D apparatus on a hot chromatographic column. Tap the column to water bath (60 to 65 °C) so that the concen- settle the Florisil and add 1 to 2 cm of anhy- trator tube is partially immersed in the hot drous sodium sulfate to the top. water, and the entire lower rounded surface 11.2.3 Preelute the column with 100 mL of of the flask is bathed with hot vapor. Adjust petroleum ether. Discard the eluate and just the vertical position of the apparatus and prior to exposure of the sodium sulfate layer the water temperature as required to com- to the air, quantitatively transfer the sam- plete the concentration in 15 to 20 min. At ple extract onto the column by decantation the proper rate of distillation the balls of the and subsequent petroleum ether washings. column will actively chatter but the cham- Discard the eluate. Just prior to exposure of bers will not flood with condensed solvent. When the apparent volume of liquid reaches the sodium sulfate layer to the air, begin 1 to 2 mL, remove the K-D apparatus and eluting the column with 200 mL of petroleum allow it to drain and cool for at least 10 min. ether and collect the eluate in a 500-mL K-D NOTE: The dichloribenzenes have a suffi- flask equipped with a 10-mL concentrator ciently high volatility that significant losses tube. This fraction should contain all of the may occur in concentration steps if care is chlorinated hydrocarbons. not exercised. It is important to maintain a 11.2.4 Concentrate the fraction as in Sec- constant gentle evaporation rate and not to tion 10.6, except use hexane to prewet the allow the liquid volume to fall below 1 to 2 column. When the apparatus is cool, remove mL before removing the K-D apparatus from the Snyder column and rinse the flask and the hot water bath. its lower joint into the concentrator tube 10.7 Momentarily remove the Snyder col- with hexane. Analyze by gas chroma- umn, add 50 mL of hexane and a new boiling tography (Section 12). chip, and reattach the Snyder column. Raise the tempeature of the water bath to 85 to 90

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12. Gas Chromatography Vi = Volume of extract injected (μL). V = Volume of total extract (μL). 12.1 Table 1 summarizes the recommended t V = Volume of water extracted (mL). operating conditions for the gas chro- s matograph. Included in this table are reten- 13.1.2 If the internal standard calibration tion times and MDL that can be achieved procedure is used, calculate the concentra- under these conditions. Examples of the sep- tion in the sample using the response factor arations achieved by Columl 2 are shown in (RF) determined in Section 7.3.2 and Equa- Figures 1 and 2. Other packed or capillary tion 3. (open-tubular) columns, chromatographic conditions, or detectors may be used if the ()()AI μ= ss requirements of Section 8.2 are met. Concentration ( g/L) () 12.2 Calibrate the system daily as de- ()ARFVis() o scribed in Section 7. 12.3 If the internal standard calibration Equation 3 procedure is being used, the internal stand- where: ard must be added to the sample extract and As = Response for the parameter to be meas- mixed throughly immediately before injec- ured. tion into the gas chromatograph. Ais = Response for the internal standard. 12.4 Inject 2 to 5 μL of the sample extract Is = Amount of internal standard added to or standard into the gas chromatograph each extract (μg). using the solvent-flush techlique. 9 Smaller Vo = Volume of water extracted (L). (1.0 μL) volumes may be injected if auto- 13.2 Report results in μg/L without correc- matic devices are employed. Record the vol- tion for recovery data. All QC data obtained ume injected to the nearest 0.05 μL, the total should be reported with the sample results. extract volume, and the resulting peak size in area or peak height units. 14. Method Performance 12.5 Identify the parameters in the sample by comparing the retention times of the 14.1 The method detection limit (MDL) is peaks in the sample chromatogram with defined as the minimum concentration of a those of the peaks in standard substance that can be measured and reported chromatograms. The width of the retention with 99% confidence that the value is above time window used to make identifications zero. 1 The MDL concentrations listed in should be based upon measurements of ac- Table 1 were obtained using reagent water. 10 tual retention time variations of standards Similar results were achieved using rep- over the course of a day. Three times the resentative wastewaters. The MDL actually standard deviation of a retention time for a achieved in a given analysis will vary de- compound can be used to calculate a sug- pending on instrument sensitivity and ma- gested window size; however, the experience trix effects. of the analyst should weigh heavily in the 14.2 This method has been tested for lin- interpretation of chromatograms. earity of spike recovery from reagent water 12.6 If the response for a peak exceeds the and has been demonstrated to be applicable × working range of the system, dilute the ex- over the concentration range from 4 MDL × 10 tract and reanalyze. to 1000 MDL. 12.7 If the measurement of the peak re- 14.3 This method was tested by 20 labora- sponse is prevented by the presence of inter- tories using reagent water, drinking water, ferences, further cleanup is required. surface water, and three industrial wastewaters spiked at six concentrations 13. Calculations over the range 1.0 to 356 μg/L. 11 Single oper- ator precision, overall precision, and method 13.1 Determine the concentration of indi- accuracy were found to be directly related to vidual compounds in the sample. the concentration of the parameter and es- 13.1.1 If the external standard calibration sentially independent of the sample matrix. procedure is used, calculate the amount of Linear equations to describe these relation- material injected from the peak response ships are presented in Table 3. using the calibration curve or calibration factor determined in Section 7.2.2. The con- REFERENCES centration in the sample can be calculated from Equation 2. 1. 40 CFR part 136, appendix B. 2. ‘‘Determination of Chlorinated Hydro- ()AV() carbons In Industrial and Municipal μ= t Wastewaters, ‘‘EPA 6090/4–84–ABC, National Concentration ( g/L) ()() Technical Information Service, PBXYZ, VVis Springfield, Virginia, 22161 November 1984. 3. ASTM Annual Book of Standards, Part Equation 2 31, D3694–78. ‘‘Standard Practices for Prepa- where: ration of Sample Containers and for Preser- A = Amount of material injected (ng). vation of Organic Constituents,’’ American

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Society for Testing and Materials, Philadel- pling Water,’’ American Society for Testing phia. and Materials, Philadelphia. 4. ‘‘Carcinogens—Working With Carcino- 9. Burke, J.A. ‘‘Gas Chromatography for gens,’’ Department of Health, Education, and Pesticide Residue Analysis; Some Practical Welfare, Public Health Service, Center for Aspects,’’ Journal of the Association of Official Disease Control, National Institute for Occu- Analytical Chemists, 48, 1037 (1965). pational Safety and Health, Publication No. 10. ‘‘Development of Detection Limits, 77–206, August 1977. EPA Method 612, Chlorinated Hydro- 5. ‘‘OSHA Safety and Health Standards, carbons,’’ Special letter report for EPA Con- General Industry,’’ (29 CFR part 1910), Occu- tract 68–03–2625, U.S. Environmental Protec- pational Safety and Health Administration, tion Agency, Environmental Monitoring and OSHA 2206 (Revised, January 1976). Support Laboratory, Cincinnati, Ohio 45268. 6. ‘‘Safety in Academic Chemistry Labora- 11. ‘‘EPA Method Study Method 612— tories,’’ American Chemical Society Publica- Chlorinated Hydrocarbons,’’ EPA 600/4–84– tion, Committee on Chemical Safety, 3rd 039, National Technical Information Service, Edition, 1979. PB84–187772, Springfield, Virginia 22161, May 7. Provost, L.P., and Elder, R.S. ‘‘Interpre- 1984. tation of Percent Recovery Data,’’American 12. ‘‘Method Performance for Laboratory, 15, 58–63 (1983). (The value 2.44 Hexachlorocyclopentadiene by Method 612,’’ used in the equation in Section 8.3.3 is two Memorandum from R. Slater, U.S. Environ- times the value 1.22 derived in this report.) mental Protection Agency, Environmental 8. ASTM Annual Book of Standards, Part Monitoring and Support Laboratory, Cin- 31, D3370–76. ‘‘Standard Practices for Sam- cinnati, Ohio 45268, December 7, 1983.

TABLE 1—CHROMATOGRAPHIC CONDITIONS AND METHOD DETECTION LIMITS

Retention time (min) Method de- Parameter tection limit Column 1 Column 2 (μg/L)

1,3-Dichlorobenzene ...... 4.5 6.8 1.19 Hexachloroethane ...... 4.9 8.3 0.03 1,4-Dichlorobenzene ...... 5.2 7.6 1.34 1,2-Dichlorobenzene ...... 6.6 9.3 1.14 Hexachlorobutadiene ...... 7.7 20.0 0.34 1,2,4-Trichlorobenzene ...... 15.5 22.3 0.05 Hexachlorocyclopentadiene ...... nd c 16.5 0.40 2-Chloronaphthalene ...... a 2.7 b 3.6 0.94 Hexachlorobenzene ...... a 5.6 b 10.1 0.05 Column 1 conditions: Supelcoport (100/120 mesh) coated with 1% SP–1000 packed in a 1.8 m × 2 mm ID glass column with 5% methane/95% argon carrier gas at 25 mL/min. flow rate. Column temperature held isothermal at 65 °C, except where other- wise indicated. Column 2 conditions: Supelcoport (80/100 mesh) coated with 1.5% OV–1/2.4% OV–225 packed in a 1.8 m × 2 mm ID glass column with 5% methane/95% argon carrier gas at 25 mL/min. flow rate. Column temperature held isothermal at 75 °C, except where otherwise indicated. nd = Not determined. a 150 °C column temperature. b 165 °C column temperature. c 100 °C column temperature.

TABLE 2—QC ACCEPTANCE CRITERIA—METHOD 612

Test ¯ Range for μ Limit for s Range for X Parameter conc. ( g/ μ μ P, Ps L) ( g/L) ( g/L) (percent)

2-Chloronaphthalene ...... 100 37.3 29.5–126.9 9–148 1,2-Dichlorobenzene ...... 100 28.3 23.5–145.1 9–160 1,3-Dichlorobenzene ...... 100 26.4 7.2–138.6 D–150 1,4-Dichlorobenzene ...... 100 20.8 22.7–126.9 13–137 Hexachlorobenzene ...... 10 2.4 2.6–14.8 15–159 Hexachlorobutadiene ...... 10 2.2 D–12.7 D–139 Hexachlorocyclopentadiene ...... 10 2.5 D–10.4 D–111 Hexachloroethane ...... 10 3.3 2.4–12.3 8–139 1,2,4-Trichlorobenzene ...... 100 31.6 20.2–133.7 5–149 s = Standard deviation of four recovery measurements, in μg/L (Section 8.2.4). X¯ = Average recovery for four recovery measurements, in μg/L (Section 8.2.4). P, Ps = Percent recovery measured (Section 8.3.2, Section 8.4.2). D = Detected; result must be greater than zero. NOTE: These criteria are based directly upon the method performance data in Table 3. Where necessary, the limits for recov- ery have been broadened to assure applicability of the limits to concentrations below those used to develop Table 3.

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TABLE 3—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 612

Acccuracy, as Single analyst ′ ′ Overall preci- Parameter recovery, X precision, sr ′ μ (μg/L) (μg/L) sion, S ( g/L)

2-Chloronaphthalene ...... 0.75C + 3.21 0.28X¯ ¥1.17 0.38X¯ ¥1.39 1,2-Dichlorobenzene ...... 0.85C¥0.70 0.22X¯ ¥2.95 0.41X¯ ¥3.92 1,3-Dichlorobenzene ...... 0.72C + 0.87 0.21X¯ ¥1.03 0.49X¯ ¥3.98 1,4-Dichlorobenzene ...... 0.72C + 2.80 0.16X¯ ¥0.48 0.35X¯ ¥0.57 Hexachlorobenzene ...... 0.87C¥0.02 0.14X¯ + 0.07 0.36X¯ ¥0.19 Hexachlorobutadiene ...... 0.61C + 0.03 0.18X¯ + 0.08 0.53X¯ ¥0.12 Hexachlorocyclopentadiene a ...... 0.47C 0.24X¯ 0.50X¯ Hexachloroethane ...... 0.74C¥0.02 0.23X¯ + 0.07 0.36X¯ ¥0.00 1,2,4-Trichlorobenzene ...... 0.76C + 0.98 0.23X¯ ¥0.44 0.40X¯ ¥1.37 X′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L. sr′ = Expected single analyst standard deviation of measurements at an average concentration found of X¯ , in μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X¯ , in μg/L. C = True value for the concentration, in μg/L. X¯ = Average recovery found for measurements of samples containing a concentration of C, in μg/L. a Estimates based upon the performance in a single laboratory. 12

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COLUMN: 1.5% OV-1 /2.4% OV-225 ON SUPELGOPORT TEMPERATURE: 75 -c DETECTOR: aECTRON CAPTURE

w 2w 25 < ....::;:, = 0 a: w 0 _, 2w :X: N w (J w 2 < Cj w2 < >C = 2wwN :X: w 0 N2 .... ::1: a: zw w 0 w= 0 ...1 CICIO a: ::1: o= 0 (J ceO _,w cc 0-1 ::1:2 ...... J::I: Uw I

0 4 8 12 16 20 24 RETENTION TIME. MIN.

Figure 1. Gas chromatogram of chlorinated hydrocarbons.

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COLUMN: 1.5% OV-1 /2.4% OV-225 ON SUPELCOPORT TEMPERATURE: 165•c DETECTOR: ELECTRON CAPTURE

w ....as w c:c ifj iE N ::c a.c:c ~ 2 i g i ::c g u ::c < u >C I w N ::c

0 4 8 12 RETENTION TIME, MIN.

Figure 2. Gas chromatogram of chlorinated hydrocarbons.

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METHOD 613—2,3,7,8-TETRACHLORODIBENZO-P- 3. Interferences DIOXIN 3.1 Method interferences may be caused 1. Scope and Application by contaminants in solvents, reagents, glass- ware, and other sample processing hardware 1.1 This method covers the determination that lead to discrete artifacts and/or ele- of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8- vated backgrounds at the masses (m/z) mon- TCDD). The following parameter may be de- itored. All of these materials must be rou- termined by this method: tinely demonstrated to be free from inter- ferences under the conditions of the analysis Parameter STORET GAS No. by running laboratory reagent blanks as de- No. scribed in Section 8.1.3. 2,3,7,8-TCDD ...... 34675 1746–01–6 3.1.1 Glassware must be scrupulously cleaned. 4 Clean all glassware as soon as pos- 1.2 This is a gas chromatographic/mass sible after use by rinsing with the last sol- spectrometer (GC/MS) method applicable to vent used in it. Solvent rinsing should be fol- lowed by detergent washing with hot water, the determination of 2,3,7,8–TCDD in munic- and rinses with tap water and distilled ipal and industrial discharges as provided water. The glassware should then be drained under 40 CFR 136.1. Method 625 may be used dry, and heated in a muffle furnace at 400 °C to screen samples for 2,3,7,8–TCDD. When the for 15 to 30 min. Some thermally stable ma- screening test is positive, the final quali- terials, such as PCBs, may not be eliminated tative confirmation and quantification must by the treatment. Solvent rinses with ace- be made using Method 613. tone and pesticide quality hexane may be 1.3 The method detection limit (MDL, de- substituted for the muffle furnace heating. fined in Section 14.1) 1 for 2,3,7,8–TCDD is Thorough rinsing with such solvents usually listed in Table 1. The MDL for a specific eliminates PCB interference. Volumetric wastewater may be different from that list- ware should not be heated in a muffle fur- ed, depending upon the nature of inter- nace. After drying and cooling, glassware ferences in the sample matrix. should be sealed and stored in a clean envi- 1.4 Because of the extreme toxicity of this ronment to prevent any accumulation of compound, the analyst must prevent expo- dust or other contaminants. Store inverted sure to himself, of to others, by materials or capped with aluminum foil. knows or believed to contain 2,3,7,8–TCDD. 3.1.2 The use of high purity reagents and Section 4 of this method contains guidelines solvents helps to mininmize interference and protocols that serve as minimum safe- problems. Purification of solvents by dis- handling standards in a limited-access lab- tillation in all-glass systems may be re- oratory. quired. 1.5 Any modification of this method, be- 3.2 Matrix interferences may be caused by yond those expressly permitted, shall be con- contaminants that are coextracted from the sidered as a major modification subject to sample. The extent of matrix interferences application and approval of alternate test will vary considerably from source to source, procedures under 40 CFR 136.4 and 136.5. depending upon the nature and diversity of 1.6 This method is restricted to use by or the industrial complex or municipality being under the supervision of analysts experi- sampled. 2,3,7,8–TCDD is often associated enced in the use of a gas chromatograph/ with other interfering chlorinated com- mass spectrometer and in the interpretation pounds which are at concentrations several of mass spectra. Each analyst must dem- magnitudes higher than that of 2,3,7,8–TCDD. onstrate the ability to generate acceptable The cleanup producers in Section 11 can be results with this method using the procedure used to overcome many of these inter- described in Section 8.2. ferences, but unique samples may require ad- ditional cleanup approaches 1 5M7 to elimi- 2. Summary of Method nate false positives and achieve the MDL 2.1 A measured volume of sample, ap- listed in Table 1. proximately 1–L, is spiked with an internal 3.3 The primary column, SP–2330 or equiv- standard of labeled 2,3,7,8–TCDD and ex- alent, resolves 2,3,7,8–TCDD from the other tracted with methylene chloride using a 21 TCDD insomers. Positive results using separatory funnel. The methylene chloride any other gas chromatographic column must extract is exchanged to hexane during con- be confirmed using the primary column. centration to a volume of 1.0 mL or less. The extract is then analyzed by capillary column 4. Safety GC/MS to separate and measure 2,3,7,8– 4.1 The toxicity or carcinogenicity of TCDD. 23 each reagent used in this method has not 2.2 The method provides selected column been precisely defined; however, each chem- chromatographic cleanup proceudres to aid ical compound should be treated as a poten- in the elimination of interferences that may tial health hazard. From this viewpoint, ex- be encountered. posure to these chemicals must be reduced to

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the lowest possible level by whatever means 4.3.1.3 Personal hygiene—Thorough wash- available. The laboratory is responsible for ing of hands and forearms after each manipu- maintaining a current awareness file of lation and before breaks (coffee, lunch, and OSHA regulations regarding the safe han- shift). dling of the chemicals specified in this meth- 4.3.1.4 Confinement—Isolated work area, od. A reference file of material data handling posted with signs, segregated glassware and sheets should also be made available to all tools, plastic-backed absorbent paper on personnel involved in the chemical analysis. benchtops. Additional references to laboratory safety 4.3.1.5 Waste—Good technique includes 8M10 are available and have been identified minimizing contaminated waste. Plastic bag for the information of the analyst. Benzene liners should be used in waste cans. Janitors and 2,3,7,8–TCDD have been identified as sus- must be trained in the safe handling of pected human or mammalian carcinogens. waste. 4.2 Each laboratory must develop a strict safety program for handling 2,3,7,8–TCDD. 4.3.1.6 Disposal of wastes—2,3,7,8–TCDD ° The following laboratory practices are rec- decomposes above 800 C. Low-level waste ommended: such as absorbent paper, tissues, animal re- 4.2.1 Contamination of the laboratory will mains, and plastic gloves may be burned in a be minimized by conducting all manipula- good incinerator. Gross quantities (milli- tions in a hood. grams) should be packaged securely and dis- 4.2.2 The effluents of sample splitters for posed through commercial or governmental the gas chromatograph and roughing pumps channels which are capable of handling high- on the GC/MS should pass through either a level radioactive wastes or extremely toxic column of activated charcoal or be bubbled wastes. Liquids should be allowed to evapo- through a trap containing oil or high-boiling rate in a good hood and in a disposable con- alcohols. tainer. Residues may then be handled as 4.2.3 Liquid waste should be dissolved in above. methanol or ethanol and irradiated with ul- 4.3.1.7 Decontamination—For personal de- traviolet light with a wavelength greater contamination, use any mild soap with plen- than 290 nm for several days. (Use F 40 BL ty of scrubbing action. For decontamination lamps or equivalent). Analyze liquid wastes of glassware, tools, and surfaces, and dispose of the solutions when 2,3,7,8– Chlorothene NU Solvent (Trademark of the TCDD can no longer be detected. Dow Chemical Company) is the least toxic 4.3 Dow Chemical U.S.A. has issued the solvent shown to be effective. Satisfactory following precautimns (revised November cleaning may be accomplished by rinsing 1978) for safe handling of 2,3,7,8–TCDD in the with Chlorothene, then washing with any de- laboratory: tergent and water. Dishwater may be dis- 4.3.1 The following statements on safe posed to the sewer. It is prudent to minimize handling are as complete as possible on the solvent wastes because they may require spe- basis of available toxicological information. cial disposal through commercial sources The precautions for safe handling and use which are expensive. are necessarily general in nature since de- 4.3.1.8 Laundry—Clothing known to be tailed, specific recommendations can be contaminated should be disposed with the made only for the particular exposure and precautions described under Section 4.3.1.6. circumstances of each individual use. Inquir- Lab coats or other clothing worn in 2,3,7,8– ies about specific operations or uses may be TCDD work areas may be laundered. addressed to the Dow Chemical Company. Assistance in evaluating the health hazards Clothing should be collected in plastic of particular plant conditions may be ob- bags. Persons who convey the bags and laun- tained from certain consulting laboratories der the clothing should be advised of the haz- and from State Departments of Health or of ard and trained in proper handling. The Labor, many of which have an industrial clothing may be put into a washer without health service. 2,3,7,8–TCDD is extremely contact if the launderer knows the problem. toxic to laboratory animals. However, it has The washer should be run through a cycle be- been handled for years without injury in an- fore being used again for other clothing. alytical and biological laboratories. Tech- 4.3.1.9 Wipe tests—A useful method of de- niques used in handling radioactive and in- termining cleanliness of work surfaces and fectious materials are applicable to 2,3,7,8,– tools is to wipe the surface with a piece of TCDD. filter paper. Extraction and analysis by gas 4.3.1.1 Protective equipment—Throw- chromatography can achieve a limit of sensi- away plastic gloves, apron or lab coat, safety tivity of 0.1 μg per wipe. Less than 1 μg of glasses, and a lab hood adequate for radio- 2,3,7,8–TCDD per sample indicates acceptable active work. cleanliness; anything higher warrants fur- 4.3.1.2 Training—Workers must be trained ther cleaning. More than 10 μg on a wipe in the proper method of removing contami- sample constitutes an acute hazard and re- nated gloves and clothing without con- quires prompt cleaning before further use of tacting the exterior surfaces. the equipment or work space. A high (>10 μg)

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2,3,7,8–TCDD level indicates that unaccept- 5.2.6 Vials—10 to 15–mL, amber glass, with able work practices have been employed in Teflon-lined screw cap. the past. 5.2.7 Chromatographic column—300 mm 4.3.1.10 Inhalation—Any procedure that long × 10 mm ID, with Teflon stopcock and may produce airborne contamination must coarse frit filter disc at bottom. be done with good ventilation. Gross losses 5.2.8 Chromatographic column—400 mm to a ventilation system must not be allowed. long × 11 mm ID, with Teflon stopcock and Handling of the dilute solutions normally coarse frit filter disc at bottom. used in analytical and animal work presents 5.3 Boiling chips—Approximately 10/40 no inhalation hazards except in the case of mesh. Heat to 400 °C for 30 min or Soxhlet ex- an accident. tract with methylene chloride. 4.3.1.11 Accidents—Remove contaminated 5.4 Water bath—Heated, with concentric clothing immediately, taking precautions ring cover, capable of temperature control not to contaminate skin or other articles. (±2 °C). The bath should be used in a hood. Wash exposed skin vigorously and repeatedly 5.5 GC/MS system: until medical attention is obtained. 5.5.1 Gas chromatograph—An analytical system complete with a temperature pro- 5. Apparatus and Materials grammable gas chromatograph and all re- 5.1 Sampling equipment, for discrete or quired accessories including syringes, ana- composite sampling. lytical columns, and gases. The injection 5.1.1 Grab sample bottle—1–L or 1-qt, port must be designed for capillary columns. amber glass, fitted with a screw cap lined Either split, splitless, or on-column injection with Teflon. Foil may be substituted for Tef- techniques may be employed, as long as the lon if the sample is not corrosive. If amber requirements of Section 7.1.1 are achieved. bottles are not available, protect samples 5.5.2 Column—60 m long × 0.25 mm ID from light. The bottle and cap liner must be glass or fused silica, coated with SP–2330 (or washed, rinsed with acetone or methylene equivalent) with a film thickness of 0.2 μm. chloride, and dried before use to minimize Any equivalent column must resolve 2, 3, 7, contamination. 8–TCDD from the other 21 TCDD isomers. 16 5.1.2 Automatic sampler (optional)—The 5.5.3 Mass spectrometer—Either a low res- sampler must incorporate glass sample con- olution mass spectrometer (LRMS) or a high tainers for the collection of a minimum of resolution mass spectrometer (HRMS) may 250 mL of sample. Sample containers must be be used. The mass spectrometer must be kept refrigerated at 4 °C and protected from equipped with a 70 V (nominal) ion source light during compositing. If the sampler uses and be capable of aquiring m/z abundance a peristaltic pump, a minimum length of data in real time selected ion monitoring compressible silicone rubber tubing may be (SIM) for groups of four or more masses. used. Before use, however, the compressible 5.5.4 GC/MS interface—Any GC to MS tubing should be thoroughly rinsed with interface can be used that achieves the re- methanol, followed by repeated rinsings with quirements of Section 7.1.1. GC to MS inter- distilled water to minimize the potential for faces constructed of all glass or glass-lined contamination of the sample. An integrating materials are recommended. Glass surfaces flow meter is required to collect flow propor- can be deactivated by silanizing with tional composites. dichlorodimethylsilane. To achieve max- 5.1.3 Clearly label all samples as ‘‘POI- imum sensitivity, the exit end of the cap- SON’’ and ship according to U.S. Department illary column should be placed in the ion of Transportation regulations. source. A short piece of fused silica capillary 5.2 Glassware (All specifications are sug- can be used as the interface to overcome gested. Catalog numbers are included for il- problems associated with straightening the lustration only.): exit end of glass capillary columns. 5.2.1 Separatory funnels—2–L and 125-mL, 5.5.5 The SIM data acquired during the with Teflon stopcock. chromatographic program is defined as the 5.2.2 Concentrator tube, Kuderna-Dan- Selected Ion Current Profile (SICP). The ish—10-mL, graduated (Kontes K–570050–1025 SICP can be acquired under computer con- or equivalent). Calibration must be checked trol or as a real time analog output. If com- at the volumes employed in the test. Ground puter control is used, there must be software glass stopper is used to prevent evaporation available to plot the SICP and report peak of extracts. height or area data for any m/z in the SICP 5.2.3 Evaporative flask, Kuderna-Danish— between specified time or scan number lim- 500–mL (Kontes K–570001–0500 or equivalent). its. Attach to concentrator tube with springs. 5.6 Balance—Analytical, capable of accu- 5.2.4 Snyder column, Kuderna-Danish— rately weighing 0.0001 g. Three-ball macro (Kontes K–503000–0121 or 6. Reagents equivalent). 5.2.5 Snyder column, Kuderna-Danish— 6.1 Reagent water—Reagent water is de- Two-ball micro (Kontes K–569001–0219 or fined as a water in which an interferent is equivalent). not observed at the MDL of 2, 3, 7, 8–TCDD.

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6.2 Sodium hydroxide solution (10 N)— 7. Calibration Dissolve 40 g of NaOH (ACS) in reagent water 7.1 Establish gas chromatograhic oper- and dilute to 100 mL. Wash the solution with ating conditions equivalent to those given in methylene chloride and hexane before use. Table 1 and SIM conditions for the mass 6.3 Sodium thiosulfate—(ACS) Granular. spectrometer as described in Section 12.2 The 6.4 Sulfuric acid—Concentrated (ACS, sp. GC/MS system must be calibrated using the gr. 1.84). internal standard technique. 6.5 Acetone, methylene chloride, hexane, 7.1.1 Using stock standards, prepare cali- benzene, ortho-xylene, tetradecane—Pes- bration standards that will allow measure- ticide quality or equivalent. ment of relative response factors of at least 6.6 Sodium sulfate—(ACS) Granular, an- three concentration ratios of 2,3,7,8–TCDD to ° hydrous. Purify by heating at 400 C for 4 h internal standard. Each calibration standard in a shallow tray. must be prepared to contain the internal 6.7 Alumina—Neutral, 80/200 mesh (Fisher standard at a concentration of 25 ng/mL. If Scientific Co., No. A–540 or equivalent). Be- any interferences are contributed by the in- fore use, activate for 24 h at 130 °C in a foil- ternal standard at m/z 320 and 322, its con- covered glass container. centration may be reduced in the calibration 6.8 Silica gel—High purity grade, 100/120 standards and in the internal standard spik- mesh (Fisher Scientific Co., No. S–679 or ing solution (Section 6.10). One of the cali- equivalent). bration standards should contain 2,3,7,8– 6.9 Stock standard solutions (1.00 μg/μL)— TCDD at a concentration near, but above, Stock standard solutimns can be prepared the MDL and the other 2,3,7,8–TCDD con- from pure standard materials or purchased centrations should correspond to the ex- as certified solutions. Acetone should be pected range of concentrations found in real used as the solvent for spiking solutions; samples or should define the working range ortho-xylene is recommended for calibration of the GC/MS system. standards for split injectors; and tetradecane 7.1.2 Using injections of 2 to 5 μL, analyze is recommended for splitless or on-colum each calibration standardaccording to Sec- injectors. Analyze stock internal standards tion 12 and tabulate peak height or area re- to verify the absence of native 2,3,7,8–TCDD. sponse against the concentration of 2,3,7,8– 6.9.1 Prepare stock standard solutions of TCDD and internal standard. Calculate re- 37 2,3,7,8–TCDD (mol wt 320) and either C14 sponse factors (RF) for 2,3,7,8–TCDD using 13 2,3,7,8–TCDD (mol wt 328) or C112 2,3,7,8– Equation 1. TCDD (mol wt 332) in an isolated area by ac- curately weighing about 0.0100 g of pure ma- ()()AC terial. Dissolve the material in pesticide RF = sis quality solvent and dilute to volume in a 10- ()()ACis s mL volumetric flask. When compound purity is assayed to be 96% or greater, the weight Equation 1 can be used without correction to calculate the concentration of the stock standard. where: Commercially prepared stock standards can As = SIM response for 2,3,7,8–TCDD m/z 320. be used at any concentration if they are cer- Ais = SIM response for the internal standard, 13 tified by the manufacturer or by an inde- m/z 332 for C12 2,3,7,8–TCDD m/z 328 for 37 pendent source. Cl4 2,3,7,8–TCDD. 6.9.2 Transfer the stock standard solu- Cis = Concentration of the internal standard tions into Teflon-sealed screw-cap bottles. (μg/L). Store in an isolated refrigerator protected Cs = Concentration of 2,3,7,8–TCDD (μg/L). from light. Stock standard solutions should If the RF value over the working range is a be checked frequently for signs of degrada- constant (<10% relative standard deviation, tion or evaporation, especially just prior to RSD), the RF can be assumed to be invariant preparing calibration standards or spiking and the average RF can be used for calcula- solutions from them. tions. Alternatively, the results can be used 6.9.3 Stock standard solutions must be re- to plot a calibration curve of response ratios, placed after six months, or sooner if com- As/Ais, vs. RF. parison with check standards indicates a 7.1.3 The working calibration curve or RF problem. must be verified on each working day by the 6.10 Internal standard spiking solution (25 measurement of one or more 2,3,7,8–TCDD ng/mL)—Using stock standard solution, pre- calibration standards. If the response for pare a spiking solution in acetone of either 2,3,7,8–TCDD varies from the predicted re- 13 37 Cl12 or Cl4 2,3,7,8–TCDD at a concentra- sponse by more than ±15%, the test must be tion of 25 ng/mL. (See Section 10.2) repeated using a fresh calibration standard. 6.11 Quality control check sample con- Alternatively, a new calibration curve must centrate—See Section 8.2.1. be prepared.

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7.2 Before using any cleanup procedure, 8.2 To establish the ability to generate the analyst must process a series of calibra- acceptable accuracy and precision, the ana- tion standards through the procedure to vali- lyst must perform the following operations. date elution patterns and the absence of 8.2.1 A quality control (QC) check sample interferences from the reagents. concentrate is required containing 2,3,7,8– TCDD at a concentration of 0.100 μg/mL in 8. Quality Control acetone. The QC check sample concentrate 8.1 Each laboratory that uses this method must be obtained from the U.S. Environ- is required to operate a formal quality con- mental Protection Agency, Environmental trol program. The minimum requirements of Monitoring and Support Laboratory in Cin- this program consist of an initial demonstra- cinnati, Ohio, if available. If not available from that source, the QC check sample con- tion of laboratory capability and an ongoing centrate must be obtained from another ex- analysis of spiked samples to evaluate and ternal source. If not available from either document data quality. The laboratory must source above, the QC check sample con- maintain records to document the quality of centrate must be prepared by the laboratory data that is generated. Ongoing data quality using stock standards prepared independ- checks are compared with established per- ently from those used for calibration. formance criteria to determine if the results 8.2.2 Using a pipet, prepare QC check sam- of analyses meet the performance character- ples at a concentration of 0.100 μg/L (100 ng/ istics of the method. When results of sample L) by adding 1.00 mL of QC check sample spikes indicate atypical method perform- concentrate to each of four 1–L aliquots of ance, a quality control check standard must reagent water. be analyzed to confirm that the measure- 8.2.3 Analyze the well-mixed QC check ments were performed in an in-control mode samples according to the method beginning of operation. in Section 10. 8.1.1 The analyst must make an initial, 8.2.4 Calculate the average recovery (X¯ ) in one-time, demonstration of the ability to μg/L, and the standard deviation of the re- generate acceptable accuracy and precision covery (s) in μg/L, for 2,3,7,8–TCDD using the with this method. This ability is established four results. as described in Section 8.2. 8.2.5 Compare s and (X¯ ) with the cor- 8.1.2 In recognition of advances that are responding acceptance criteria for precision occurring in chromatography, the analyst is and accuracy, respectively, found in Table 2. permitted certain options (detailed in Sec- If s and X¯ meet the acceptance criteria, the tions 10.5, 11.1, and 12.1) to improve the sepa- system performance is acceptable and anal- rations or lower the cost of measurements. ysis of actual samples can begin. If s exceeds Each time such a modification is made to the precision limit or X¯ falls outside the the method, the analyst is required to repeat range for accuracy, the system performance the procedure in Section 8.2 is unacceptable for 2,3,7,8–TCDD. Locate and 8.1.3 Before processing any samples, the correct the source of the problem and repeat analyst must analyze a reagent water blank the test beginning with Section 8.2.2. to demonstrate that interferences from the 8.3 The laboratory must, on an ongoing analytical system and glassware are under basis, spike at least 10% of the samples from control. Each time a set of samples is ex- each sample site being monitored to assess tracted or reagents are changed, a reagent accuracy. For laboratories analyzing one to water blank must be processed as a safe- ten samples per month, at least one spiked guard against laboratory contamination. sample per month is required. 8.1.4 The laboratory must, on an ongoing 8.3.1 The concentration of the spike in the basis, spike and analyze a minimum of 10% sample should be determined as follows: of all samples with native 2,3,7,8–TCDD to 8.3.1.1 If, as in compliance monitoring, monitor and evaluate laboratory data qual- the concentration of 2,3,7,8–TCDD in the ity. This procedure is described in Section sample is being checked against a regulatory 8.3. concentration limit, the spike should be at 8.1.5 The laboratory must, on an ongoing that limit or 1 to 5 times higher than the basis, demonstrate through the analyses of background concentration determined in quality control check standards that the op- Section 8.3.2, whichever concentration would eration of the measurement system is in con- be larger. trol. This procedure is described in Section 8.3.1.2 If the concentration of 2,3,7,8–TCDD 8.4. The frequency of the check standard in the sample is not being checked against a analyses is equivalent to 10% of all samples limit specific to that parameter, the spike analyzed but may be reduced if spike recov- should be at 0.100 μg/L or 1 to 5 times higher eries from samples (Section 8.3) meet all than the background concentration deter- specified quality control criteria. mined in Section 8.3.2, whichever concentra- 8.1.6 The laboratory must maintain per- tion would be larger. formance records to document the quality of 8.3.1.3 If it is impractical to determine data that is generated. This procedure is de- background levels before spiking (e.g., max- scribed in Section 8.5. imum holding times will be exceeded), the

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spike concentration should be (1) the regu- be immediately identified and corrected. The latory concentration limit, if any; or, if none analytical result for 2,3,7,8–TCDD in the (2) the larger of either 5 times higher than unspiked sample is suspect and may not be the expected background concentration or reported for regulatory compliance purposes. 0.100 μg/L. 8.5 As part of the QC program for the lab- 8.3.2 Analyze one sample aliquot to deter- oratory, method accuracy for wastewater mine the background concentration (B) of samples must be assessed and records must 2,3,7,8–TCDD. If necessary, prepare a new QC be maintained. After the analysis of five check sample concentrate (Section 8.2.1) ap- spiked wastewater samples as in Section 8.3, propriate for the background concentration calculate the average percent recovery (P¯ ) in the sample. Spike a second sample aliquot and the spandard deviation of the percent re- with 1.0 mL of the QC check sample con- covery (sp). Express the accuracy assessment centrate and analyze it to determine the ¯ as a percent recovery interval from P¥2sp to concentration after spiking (A) of 2,3,7,8– ¯ ¯ P + 2sp. If P = 90% and sp = 10%, for example, TCDD. Calculate percent recovery (P) as the accuracy interval is expressed as 70– 100(A¥B)%T, where T is the known true 110%. Update the accuracy assessment on a value of the spike. regular basis (e.g. after each five to ten new 8.3.3 Compare the percent recovery (P) for accuracy measurements). 2,3,7,8–TCDD with the corresponding QC ac- 8.6 It is recommended that the laboratory ceptance criteria found in Table 2. These ac- adopt additional quality assurance practices ceptance criteria were calculated to include for use with this method. The specific prac- an allowance for error in measurement of tices that are most productive depend upon both the background and spike concentra- the needs of the laboratory and the nature of tions, assuming a spike to background ratio the samples. Field duplicates may be ana- of 5:1. This error will be accounted for to the lyzed to assess the precision of the environ- extent that the analyst’s spike to back- mental measurements. Whenever possible, ground ratio approaches 5:1. 11 If spiking was performed at a concentration lower than the laboratory should analyze standard ref- 0.100 μg/L, the analyst must use either the erence materials and participate in relevant QC acceptance criteria in Table 2, or op- performance evaluation studies. tional QC acceptance criteria calculated for 9. Sample Collection, Preservation, and the specific spike concentration. To cal- Handling culate optional acceptance criteria for the recovery of 2,3,7,8–TCDD: (1) Calculate accu- 9.1 Grab samples must be collected in racy (X′) using the equation in Table 3, sub- glass containers. Conventional sampling stituting the spike concentration (T) for C; practices 12 should be followed, except that (2) calculate overall precision (S′) using the the bottle must not be prerinsed with sample equation in Table 3, substituting X′ for X; (3) before collection. Composite samples should calculate the range for recovery at the spike be collected in refrigerated glass containers concentration as (100 X′/T)±2.44(100 S′/T)%. 11 in accordance with the requirements of the 8.3.4 If the recovery of 2,3,7,8–TCDD falls program. Automatic sampling equipment outside the designated range for recovery, a must be as free as possible of Tygon tubing check standard must be analyzed as de- and other potential sources of contamina- scribed in Section 8.4. tion. 8.4 If the recovery of 2,3,7,8–TCDD fails 9.2 All samples must be iced or refrig- the acceptance criteria for recovery in Sec- erated at 4 °C and protected from light from tion 8.3, a QC check standard must be pre- the time of collection until extraction. Fill pared and analyzed. the sample bottles and, if residual chlorine is NOTE: The frequency for the required anal- present, add 80 mg of sodium thiosulfate per ysis of a QC check standard will depend upon liter of sample and mix well. EPA Methods the complexity of the sample matrix and the 330.4 and 330.5 may be used for measurement performance of the laboratory. of residual chlorine. 13 Field test kits are 8.4.1 Prepare the QC check standard by available for this purpose. adding 1.0 mL of QC check sample con- 9.3 Label all samples and containers centrate (Section 8.2.1 or 8.3.2) to 1 L of rea- ‘‘’’ and ship according to applicable gent water. U.S. Department of Transportation regula- 8.4.2 Analyze the QC check standard to tions. determine the concentration measured (A) of 9.4 All samples must be extracted within 7 2,3,7,8–TCDD. Calculate the percent recovery days of collection and completely analyzed (P ) as 100 (A/T)%, where T is the true value s within 40 days of extraction. 2 of the standard concentration. 8.4.3 Compare the percent recovery (Ps) 10. Sample Extraction with the corresponding QC acceptance cri- teria found in Table 2. If the recovery of CAUTION: When using this method to ana- 2,3,7,8–TCDD falls outside the designated lyze for 2,3,7,8–TCDD, all of the following op- range, the laboratory performance is judged erations must be performed in a limited-ac- to be out of control, and the problem must cess laboratory with the analyst wearing full

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protective covering for all exposed skin sur- 10.8 Momentarily remove the Snyder col- faces. See Section 4.2. umn, add 50 mL of hexane and a new boiling 10.1 Mark the water meniscus on the side chip, and reattach the Snyder column. Raise of the sample bottle for later determination the temperature of the water bath to 85 to 90 of sample volume. Pour the entire sample °C. Concentrate the extract as in Section into a 2–L separatory funnel. 10.7, except use hexane to prewet the column. 10.2 Add 1.00 mL of internal standard Remove the Snyder column and rinse the spiking solution to the sample in the sepa- flask and its lower joint into the concen- ratory funnel. If the final extract will be trator tube with 1 to 2 mL of hexane. A 5-mL concentrated to a fixed volume below 1.00 syringe is recommended for this operation. mL (Section 12.3), only that volume of spik- Set aside the K-D glassware for reuse in Sec- ing solution should be added to the sample so tion 10.14. that the final extract will contain 25 ng/mL 10.9 Pour the hexane extract from the of internal standard at the time of analysis. concentrator tube into a 125-mL separatory 10.3 Add 60 mL of methylene chloride to funnel. Rinse the concentrator tube four the sample bottle, seal, and shake 30 s to times with 10-mL aliquots of hexane. Com- rinse the inner surface. Transfer the solvent bine all rinses in the 125-mL separatory fun- to the separatory funnel and extract the nel. sample by shaking the funnel for 2 min. with 10.10 Add 50 mL of sodium hydroxide solu- periodic venting to release excess pressure. tion to the funnel and shake for 30 to 60 s. Allow the organic layer to separate from the Discard the aqueous phase. water phase for a minimum of 10 min. If the 10.11 Perform a second wash of the or- emulsion interface between layers is more ganic layer with 50 mL of reagent water. Dis- than one-third the vmlume of the solvent card the aqueous phase. layer, the analyst must employ mechanical 10.12 Wash the hexane layer with a least techniques to complete the phase separation. two 50-mL aliquots of concentrated sulfuric The optimum technique depends upon the acid. Continue washing the hexane layer sample, but may include stirring, filtration with 50-mL aliquots of concentrated sulfuric of the emulsion through glass wool, cen- acid until the acid layer remains colorless. trifugation, or other physical methods. Col- Discard all acid fractions. lect the methylene chloride extract in a 250- 10.13 Wash the hexane layer with two 50- mL Erlenmeyer flask. mL aliquots of reagent water. Discard the 10.4 Add a second 60-mL volume of meth- aqueous phases. ylene chloride to the sample bottle and re- 10.14 Transfer the hexane extract into a peat the extraction procedure a second time, 125-mL Erlenmeyer flask containing 1 to 2 g combining the extracts in the Erlenmeyer of anhydrous sodium sulfate. Swirl the flask flask. Perform a third extraction in the same for 30 s and decant the hexane extract into manner. the reassembled K-D apparatus. Complete 10.5 Assemble a Kuderna-Danish (K-D) the quantitative transfer with two 10-mL concentrator by attaching a 10-mL concen- hexane rinses of the Erlenmeyer flask. trator tube to a 500-mL evaporative flask. 10.15 Replace the one or two clean boiling Other concentration devices or techniques chips and concentrate the extract to 6 to 10 may be used in place of the K-D concentrator mL as in Section 10.8. if the requirements of Section 8.2 are met. 10.16 Add a clean boiling chip to the con- 10.6 Pour the combined extract into the centrator tube and attach a two-ball micro- K-D concentrator. Rinse the Erlenmeyer Snyder column. Prewet the column by add- flask with 20 to 30 mL of methylele chloride ing about 1 mL of hexane to the top. Place to complete the quantitative transfer. the micro-K-D apparatus on the water bath 10.7 Add one or two clean boiling chips to so that the concentrator tube is partially the evaporative flask and attach a three-ball immersed in the hot water. Adjust the Snyder column. Prewet the Snyder column vertical position of the apparatus and the by adding about 1 mL of methylene chloride water temperature as required to complete to the top. Place the K-D apparatus on a hot the concentration in 5 to 10 min. At the prop- water bath (60 to 65 °C) so that the concen- er rate of distillation the balls of the column trator tube is partially immersed in the hot will actively chatter but the chambers will water, and the entire lower rounded surface not flood. When the apparent volume of liq- of the flask is bathed with hot vapor. Adjust uid reaches about 0.5 mL, remove the K-D the vertical position of the apparatus and apparatus and allow it to drain and cool for the water temperature as required to com- at least 10 min. Remove the micro-Snyder plete the concentration in 15 to 20 min. At column and rinse its lower joint into the the proper rate of distillation the balls of the concentrator tube with 0.2 mL of hexane. column will actively chatter but the cham- Adjust the extract volume to 1.0 mL with bers will not flood with condensed solvent. hexane. Stopper the concentrator tube and When the apparent volume of liquid reaches store refrigerated and protected from light if 1 mL, remove the K-D apparatus and allow it further processing will not be performed im- to drain and cool for at least 10 min. mediately. If the extract will be stored

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longer than two days, it should be trans- using two 2-mL portions of 20% benzene/80% ferred to a Teflon-sealed screw-cap vial. If hexane to complete the transfer. the sample extract requires no further clean- 11.3.3 Just prior to exposure of the sodium up, proceed with GC/MS analysis (Section sulfate layer to the air, add 40 mL of 20% 12). If the sample requires further cleanup, benzene/80% hexane to the column. Collect proceed to Section 11. the eluate in a clean 500-mL K-D flask 10.17 Determine the original sample vol- equipped with a 10-mL concentrator tube. ume by refilling the sample bottle to the Concentrate the collected fraction to 1.0 mL mark and transferring the liquid to a 1000- as in Section 10.16 and analyze by GC/MS. mL graduated cylinder. Record the sample volume to the nearest 5 mL. 12. GC/MS Analysis 12.1 Table 1 summarizes the recommended 11. Cleanup and Separation operating conditions for the gas chro- 11.1 Cleanup procedures may not be nec- matograph. Included in this table are reten- essary for a relatively clean sample matrix. tion times and MDL that can be achieved If particular circumstances demand the use under these conditions. Other capillary col- of a cleanup procedure, the analyst may use umns or chromatographic conditions may be either procedure below or any other appro- used if the requirements of Sections 5.5.2 and priate procedure. 1 5M7 However, the analyst 8.2 are met. first must demonstrate that the require- 12.2 Analyze standards and samples with ments of Section 8.2 can be met using the the mass spectrometer operating in the se- method as revised to incorporate the cleanup lected ion monitoring (SIM) mode using a procedure. Two cleanup column options are dwell time to give at least seven points per offered to the analyst in this section. The peak. For LRMS, use masses at m/z 320, 322, alumina column should be used first to over- and 257 for 2,3,7,8–TCDD and either m/z 328 37 13 come interferences. If background problems for Cl4 2,3,7,8–TCDD or m/z 332 for C12 are still encountered, the silica gel column 2,3,7,8–TCDD. For HRMS, use masses at m/z may be helpful. 319.8965 and 321.8936 for 2,3,7,8–TCDD and ei- 37 11.2 Alumina column cleanup for 2,3,7,8– ther m/z 327.8847 for Cl4 2,3,7,8–TCDD or m/ 13 TCDD: z 331.9367 for C12 2,3,7,8–TCDD. 11.2.1 Fill a 300 mm long × 10 mm ID 12.3 If lower detection limits are required, chromatographic column with activated alu- the extract may be carefully evaporated to mina to the 150 mm level. Tap the column dryness under a gentle stream of nitrogen gently to settle the alumina and add 10 mm with the concentrator tube in a water bath of anhydrous sodium sulfate to the top. at about 40 °C. Conduct this operation imme- 11.2.2 Preelute the column with 50 mL of diately before GC/MS analysis. Redissolve hexane. Adjust the elution rate to 1 mL/min. the extract in the desired final volume of Discard the eluate and just prior to exposure ortho-xylene or tetradecane. of the sodium sulfate layer to the air, quan- 12.4 Calibrate the system daily as de- titatively transfer the 1.0-mL sample extract scribed in Section 7. onto the column using two 2-mL portions of 12.5 Inject 2 to 5 μL of the sample extract hexane to complete the transfer. into the gas chromatograph. The volume of 11.2.3 Just prior to exposure of the sodium calibration standard injected must be meas- sulfate layer to the air, add 50 mL of 3% ured, or be the same as all sample injection methylene chloride/95% hexane (V/V) and volumes. continue the elution of the column. Discard 12.6 The presence of 2,3,7,8–TCDD is quali- the eluate. tatively confirmed if all of the following cri- 11.2.4 Next, elute the column with 50 mL teria are achieved: of 20% methylene chloride/80% hexane (V/V) 12.6.1 The gas chromatographic column into a 500-mL K-D flask equipped with a 10- must resolve 2,3,7,8–TCDD from the other 21 mL concentrator tube. Concentrate the col- TCDD isomers. lected fraction to 1.0 mL as in Section 10.16 12.6.2 The masses for native 2,3,7,8–TCDD and analyze by GC/MS (Section 12). (LRMS-m/z 320, 322, and 257 and HRMS-m/z 11.3 Silica gel column cleanup for 2,3,7,8– 320 and 322) and labeled 2,3,7,8–TCDD (m/z 328 TCDD: or 332) must exhibit a simultaneous max- 11.3.1 Fill a 400 mm long × 11 mm ID imum at a retention time that matches that chromatmgraphic column with silica gel to of native 2,3,7,8–TCDD in the calibration the 300 mm level. Tap the column gently to standard, with the performance specifica- settle the silica gel and add 10 mm of anhy- tions of the analytical system. drous sodium sulfate to the top. 12.6.3 The chlorine isotope ratio at m/z 320 11.3.2 Preelute the column with 50 mL of and m/z 322 must agree to within±10% of that 20% benzene/80% hexane (V/V). Adjust the in the calibration standard. elution rate to 1 mL/min. Discard the eluate 12.6.4 The signal of all peaks must be and just prior to exposure of the sodium sul- greater than 2.5 times the noise level. fate layer to the air, quantitatively transfer 12.7 For quantitation, measure the re- the 1.0-mL sample extract onto the column sponse of the m/z 320 peak for 2,3,7,8–TCDD

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13 and the m/z 332 peak for C12 2,3,7,8–TCDD or 14. Method Performance the m/z 328 peak for 37Cl 2,3,7,8–TCDD. 4 14.1 The method detection limit (MDL) is 12.8 Co-eluting impurities are suspected if defined as the minimum concentration of a all criteria are achieved except those in Sec- substance that can be measured and reported tion 12.6.3. In this case, another SIM analysis with 99% confidence that the value is above using masses at m/z 257, 259, 320 and either m/ zero. 1 The MDL concentration listed in a 328 or m/z 322 can be performed. The masses Table 1 was obtained using reagent water. 14 at m/z 257 and m/z 259 are indicative of the loss of one chlorine and one The MDL actually achieved in a given anal- from 2,3,7,8–TCDD. If masses m/z 257 and m/z ysis will vary depending on instrument sen- 259 give a chlorine isotope ratio that agrees sitivity and matrix effects. to within ±10% of the same cluster in the 14.2 This method was tested by 11 labora- calibration standards, then the presence of tories using reagent water, drinking water, TCDD can be confirmed. Co-eluting DDD, surface water, and three industrial DDE, and PCB residues can be confirmed, wastewaters spiked at six concentrations but will require another injection using the over the range 0.02 to 0.20 μg/L. 15 Single oper- appropriate SIM masses or full repetitive ator precision, overall precision, and method 37 accuracy were found to be directly related to mass scans. If the response for Cl4 2,3,7,8– TCDD at m/z 328 is too large, PCB contami- the concentration of the parameter and es- nation is suspected and can be confirmed by sentially independent of the sample matrix. examining the response at both m/z 326 and Linear equations to describe these relation- 37 m/z 328. The Cl4 2,3,7,8–TCDD internal ships are presented in Table 3. standard gives negligible response at m/z 326. These pesticide residues can be removed REFERENCES using the alumina column cleanup proce- 1. 40 CFR part 136, appendix B. dure. 2. ‘‘Determination of TCDD in Industrial 12.9 If broad background interference re- and Municipal Wastewaters,’’ EPA 600/4–82– stricts the sensitivity of the GC/MS analysis, 028, National Technical Information Service, the analyst should employ additional clean- PB82–196882, Springfield, Virginia 22161, April up procedures and reanalyze by GC/MS. 1982. 12.10 In those circumstances where these 3. Buser, H.R., and Rappe, C. ‘‘High Resolu- procedures do not yield a definitive conclu- tion Gas Chromatography of the 22 sion, the use of high resolution mass spec- Tetrachlorodibenzo-p-dioxin Isomers,’’ Ana- 5 trometry is suggested. lytical Chemistry, 52, 2257 (1980). 13. Calculations 4. ASTM Annual Book of Standards, Part 31, D3694–78. ‘‘Standard Practices for Prepa- 13.1 Calculate the concentration of 2,3,7,8– ration of Sample Containers and for Preser- TCDD in the sample using the response fac- vation of Organic Constituents,’’ American tor (RF) determined in Section 7.1.2 and Society for Testing and Materials, Philadel- Equation 2. phia. 5. Harless, R. L., Oswald, E. O., and () Wilkinson, M. K. ‘‘Sample Preparation and AV()t Concentration (μ= g/L) Gas Chromatography/Mass Spectrometry De- ()VVis() termination of 2,3,7,8-Tetrachlorodibenzo-p- dioxin,’’ Analytical Chemistry, 52, 1239 (1980). Equation 2 6. Lamparski, L. L., and Nestrick, T. J. ‘‘Determination of Tetra-, Hepta-, and where: Octachlorodibenzo-p-dioxin Isomers in Par- As = SIM response for 2,3,7,8–TCDD at m/z ticulate Samples at Parts per Trillion Lev- 320. els,’’ Analytical Chemistry, 52, 2045 (1980). Ais = SIM response for the internal standard 7. Longhorst, M. L., and Shadoff, L. A. at m/z 328 or 332. ‘‘Determination of Parts-per-Trillion Con- Is = Amount of internal standard added to centrations of Tetra-, Hexa-, and μ each extract ( g). Octachlorodibenzo-p-dioxins in Human Vo = Volume of water extracted (L). Milk,’’ Analytical Chemistry, 52, 2037 (1980). 13.2 For each sample, calculate the per- 8. ‘‘Carcinogens—Working with Carcino- cent recovery of the internal standard by gens,’’ Department of Health, Education, and comparing the area of the m/z peak measured Welfare, Public Health Service, Center for in the sample to the area of the same peak Disease Control, National Institute for Occu- in the calibration standard. If the recovery is pational Safety and Health, Publication No. below 50%, the analyst should review all as- 77–206, August 1977. pects of his analytical technique. 9. ‘‘OSHA Safety and Health Standards, 13.3 Report results in μg/L without correc- General Industry,’’ (29 CFR part 1910), tion for recovery data. All QC data obtained Occuptional Safety and Health Administra- should be reported with the sample results. tion, OSHA 2206 (Revised, January 1976).

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10. ‘‘Safety in Academic Chemistry Labora- Service, PB84–188879, Springfield, Virginia tories,’’ American Chemical Society Publica- 22161, May 1984. tion, Committee on Chemical Safety, 3rd Edition, 1979. TABLE 1—CHROMATOGRAPHIC CONDITIONS AND 11. Provost, L. P., and Elder, R. S., ‘‘Inter- METHOD DETECTION LIMIT pretation of Percent Recovery Data,’’ Amer- Method ican Laboratory, 15, 58–63 (1983). (The value Retention detection 2.44 used in the equation in Section 8.3.3 is Parameter time (min) limit (μg/ two times the value 1.22 derived in this re- L) port.) 2,3,7,8–TCDD ...... 13.1 0.002 12. ASTM Annual Book of Standards, Part 31, D3370–76, ‘‘Standard Practices for Sam- Column conditions: SP–2330 coated on a 60 m long × 0.25 mm ID glass column with hydrogen carrier gas at 40 cm/sec pling Water,’’ American Society for Testing linear velocity, splitless injection using tetradecane. Column and Materials, Philadelphia. temperature held isothermal at 200 °C for 1 min, then pro- grammed at 8 °C/min to 250 °C and held. Use of helium car- 13. ‘‘Methods, 330.4 (Titrimetric, DPD-FAS) rier gas will approximately double the retention time. and 330.5 (Spectrophotometric DPD) for Chlorine, Total Residual,’’ Methods for TABLE 2—QC ACCEPTANCE CRITERIA—METHOD Chemical Analysis of Water and Wastes, 613 EPA–600/4–79–020, U.S. Environmental Pro- tection Agency, Environmental Monitoring Test Limit Range for X Range and Support Laboratory, Cincinnati, Ohio Parameter conc. for s μ for P, (μg/L) (μg/L) ( g/L) P (%) 45268, March 1979. s 14. Wong, A.S. et al. ‘‘The Determination 2,3,7,8–TCDD 0.100 0.0276 0.0523–0.1226 45–129 of 2,3,7,8–TCDD in Industrial and Municipal s = Standard deviation of four recovery measurements, in Wastewaters, Method 613, Part 1—Develop- μg/L (Section 8.2.4). ment and Detection Limits,’’ G. Choudhay, X¯ = Average recovery for four recovery measurements, in μg/L (Section 8.2.4). L. Keith, and C. Ruppe, ed., Butterworth P, Ps = Percent recovery measured (Section 8.3.2, Section Inc., (1983). 8.4.2). 15. ‘‘EPA Method Study 26, Method 613: NOTE: These criteria are based directly upon the method performance data in Table 3. Where necessary, the limits for 2,3,7,8–Tetrachlorodibenzo-p-dioxin,’’ EPA recovery have been broadened to assure applicability of the 600/4–84–037, National Technical Information limits to concentrations below those used to develop Table 3.

TABLE 3—METHOD ACCURACY AND PRECISION AS FUNCTIONS OF CONCENTRATION—METHOD 613

Parameter Accuracy, as recov- Single analyst, pre- Overall precision, ery, X″ (μg/L) cision, sr″ (μ/L) S″ (μ/g/L)

2,3,7,8-TCDD ...... 0.86C + 0.00145 0.13X¯ + 0.00129 0.19X¯ + 0.00028 X′ = Expected recovery for one or more measurements. of a sample containing a concentration of C, in μg/L. sr′ = Expected single analyst standard deviation of measurements at an average concentration found of X¯ , in μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X¯ , in μg/L. C = True value for the concentration, in μg/L. X¯ = Average recovery found for measurements of samples containing a concentration of C, in μg/L.

METHOD 624.1—PURGEABLES BY GC/MS may be extended to determine the analytes listed in Table 2; however, poor purging effi- 1. Scope and Application ciency or gas chromatography of some of 1.1 This method is for determination of these analytes may make quantitative deter- purgeable organic pollutants in industrial mination difficult. For example, an elevated discharges and other environmental samples temperature may be required to purge some by gas chromatography combined with mass analytes from water. If an elevated tempera- spectrometry (GC/MS), as provided under 40 ture is used, calibration and all quality con- CFR 136.1. This revision is based on previous trol (QC) tests must be performed at the ele- protocols (References 1—3), on the revision vated temperature. EPA encourages the use promulgated October 26, 1984, and on an of this method to determine additional com- interlaboratory method validation study pounds amenable to purge-and-trap GC/MS. (Reference 4). Although this method was 1.3 The large number of analytes in Ta- validated through an interlaboratory study bles 1 and 2 of this method makes testing dif- conducted in the early 1980s, the funda- ficult if all analytes are determined simulta- mental chemistry principles used in this neously. Therefore, it is necessary to deter- method remain sound and continue to apply. mine and perform QC tests for ‘‘analytes of 1.2 The analytes that may be quali- interest’’ only. Analytes of interest are those tatively and quantitatively determined required to be determined by a regulatory/ using this method and their CAS Registry control authority or in a permit, or by a cli- numbers are listed in Table 1. The method ent. If a list of analytes is not specified, the

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analytes in Table 1 must be determined, at a 1.7 Terms and units of measure used in minimum, and QC testing must be performed this method are given in the glossary at the for these analytes. The analytes in Table 1 end of the method. and some of the analytes in Table 2 have been identified as Toxic Pollutants (40 CFR 2. Summary of Method 401.15), expanded to a list of Priority Pollut- 2.1 A gas is bubbled through a measured ants (40 CFR part 423, appendix A). volume of water in a specially-designed purg- 1.4 Method detection limits (MDLs; Ref- ing chamber. The purgeables are efficiently erence 5) for the analytes in Table 1 are list- transferred from the aqueous phase to the ed in that table. These MDLs were deter- vapor phase. The vapor is swept through a mined in reagent water (Reference 6). Ad- sorbent trap where the purgeables are vances in analytical technology, particularly trapped. After purging is completed, the trap the use of capillary (open-tubular) columns, is heated and backflushed with the gas to allowed laboratories to routinely achieve desorb the purgeables onto a gas MDLs for the analytes in this method that chromatographic column. The column is are 2–10 times lower than those in the temperature programmed to separate the version promulgated in 1984. The MDL for a purgeables which are then detected with a specific wastewater may differ from those mass spectrometer. listed, depending on the nature of inter- 2.2 Different sample sizes in the range of ferences in the sample matrix. 5–25 mL are allowed in order to meet dif- 1.4.1 EPA has promulgated this method at fering sensitivity requirements. Calibration 40 CFR part 136 for use in wastewater com- and QC samples must have the same volume pliance monitoring under the National Pol- as field samples. lutant Discharge Elimination System (NPDES). The data reporting practices de- 3. Interferences scribed in section 13.2 are focused on such 3.1 Impurities in the purge gas, organic monitoring needs and may not be relevant to compounds outgassing from the plumbing other uses of the method. ahead of the trap, and solvent vapors in the 1.4.2 This method includes ‘‘reporting lim- laboratory account for the majority of con- its’’ based on EPA’s ‘‘minimum level’’ (ML) tamination problems. The analytical system concept (see the glossary in section 20). must be demonstrated to be free from con- Table 1 contains MDL values and ML values tamination under the conditions of the anal- for many of the analytes. The MDL for an ysis by analyzing blanks initially and with analyte in a specific wastewater may differ each analytical batch (samples analyzed on a from that listed in Table 1, depending upon given 12-hour shift, to a maximum of 20 sam- the nature of interferences in the sample ma- ples), as described in Section 8.5. trix. Fluoropolymer tubing, fittings, and thread 1.5 This method is performance-based. It sealant should be used to avoid contamina- may be modified to improve performance tion. (e.g., to overcome interferences or improve 3.2 Samples can be contaminated by diffu- the accuracy of results) provided all per- sion of volatile organics (particularly fluoro- formance requirements are met. carbons and methylene chloride) through the 1.5.1 Examples of allowed method modi- septum seal into the sample during shipment fications are described at 40 CFR 136.6. Other and storage. Protect samples from sources of examples of allowed modifications specific to volatiles during collection, shipment, and this method are described in section 8.1.2. storage. A reagent water field blank carried 1.5.2 Any modification beyond those ex- through sampling and analysis can serve as a pressly allowed at 40 CFR 136.6 or in section check on such contamination. 8.1.2 of this method shall be considered a 3.3 Contamination by carry-over can major modification that is subject to appli- occur whenever high level and low level sam- cation and approval of an alternate test pro- ples are analyzed sequentially. To reduce the cedure under 40 CFR 136.4 and 136.5. potential for carry-over, the purging device 1.5.3 For regulatory compliance, any and sample syringe must be rinsed with rea- modification must be demonstrated to gent water between sample analyses. When- produce results equivalent or superior to re- ever an unusually concentrated sample is en- sults produced by this method when applied countered, it should be followed by an anal- to relevant wastewaters (section 8.3). ysis of a blank to check for cross contamina- 1.6 This method is restricted to use by or tion. For samples containing large amounts under the supervision of analysts experi- of water-soluble materials, suspended solids, enced in the operation of a purge-and-trap high boiling compounds or high purgeable system and a gas chromatograph/mass spec- levels, it may be necessary to wash the purg- trometer and in the interpretation of mass ing device with a detergent solution, rinse it spectra. Each analyst must demonstrate the with distilled water, and then dry it in a 105 ability to generate acceptable results with °C oven between analyses. The trap and this method using the procedure in section other parts of the system are also subject to 8.2. contamination; therefore, frequent bakeout

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and purging of the entire system may be re- 5.1.2 Septum—Fluoropolymer-faced sili- quired. Screening samples at high dilution cone (Fisher #12722 or equivalent). Unless may prevent introduction of contaminants pre-cleaned, detergent wash, rinse with tap into the system. and reagent water, and dry at 105 ± 5 °C for one hour before use. 4. Safety 5.2 Purge-and-trap system—The purge- and-trap system consists of three separate 4.1 The toxicity or carcinogenicity of pieces of equipment: A purging device, trap, each reagent used in this method has not and desorber. Several complete systems are been precisely defined; however, each chem- commercially available with autosamplers. ical compound should be treated as a poten- Any system that meets the performance re- tial health hazard. From this viewpoint, ex- quirements in this method may be used. posure to these chemicals must be reduced to 5.2.1 The purging device should accept 5- the lowest possible level. The laboratory is to 25-mL samples with a water column at responsible for maintaining a current aware- least 3 cm deep. The purge gas must pass ness file of OSHA regulations regarding the though the water column as finely divided safe handling of the chemicals specified in bubbles. The purge gas must be introduced this method. A reference file of safety data no more than 5 mm from the base of the sheets (SDSs, OSHA, 29 CFR 1910.1200(g)) water column. Purge devices of a different should also be made available to all per- volume may be used so long as the perform- sonnel involved in sample handling and ance requirements in this method are met. chemical analysis. Additional references to 5.2.2 The trap should be at least 25 cm laboratory safety are available and have long and have an inside diameter of at least been identified (References 7–9) for the infor- 0.105 in. The trap should be packed to con- mation of the analyst. tain the following minimum lengths of ad- 4.2. The following analytes covered by sorbents: 1.0 cm of methyl silicone coated this method have been tentatively classified packing (section 6.3.2), 15 cm of 2,6- as known or suspected human or mammalian diphenylene oxide polymer (section 6.3.1), carcinogens: Benzene; carbon tetrachloride; and 8 cm of silica gel (section 6.3.3). A trap chloroform; 1,4-dichlorobenzene; 1,2- with different dimensions and packing mate- dichloroethane; 1,2-dichloropropane; meth- rials is acceptable so long as the perform- ylene chloride; tetrachloroethylene; tri- ance requirements in this method are met. chloroethylene; and vinyl chloride. Primary 5.2.3 The desorber should be capable of standards of these toxic compounds should rapidly heating the trap to the temperature be prepared in a chemical fume hood, and a necessary to desorb the analytes of interest, NIOSH/MESA approved toxic gas respirator and of maintaining this temperature during should be worn when handling high con- desorption. The trap should not be heated centrations of these compounds. higher than the maximum temperature rec- 4.3 This method allows the use of hydro- ommended by the manufacturer. gen as a carrier gas in place of helium (Sec- 5.2.4 The purge-and-trap system may be tion 5.3.1.2). The laboratory should take the assembled as a separate unit or coupled to a necessary precautions in dealing with hydro- gas chromatograph. gen, and should limit hydrogen flow at the 5.3 GC/MS system. source to prevent buildup of an explosive 5.3.1 Gas chromatograph (GC)—An analyt- mixture of hydrogen in air. ical system complete with a temperature programmable gas chromatograph and all re- 5. Apparatus and Materials quired accessories, including syringes and analytical columns. Autosamplers designed NOTE: Brand names, suppliers, and part for purge-and-trap analysis of volatiles also numbers are cited for illustration purposes may be used. only. No endorsement is implied. Equivalent 5.3.1.1 Injection port—Volatiles interface, performance may be achieved using equip- split, splitless, temperature programmable ment and materials other than those speci- split/splitless (PTV), large volume, on-col- fied here. Demonstration of equivalent per- umn, backflushed, or other. formance that meets the requirements of 5.3.1.2 Carrier gas—Data in the tables in this method is the responsibility of the lab- this method were obtained using helium car- oratory. Suppliers for equipment and mate- rier gas. If another carrier gas is used, ana- rials in this method may be found through lytical conditions may need to be adjusted an on-line search. for optimum performance, and calibration 5.1 Sampling equipment for discrete sam- and all QC tests must be performed with the pling. alternative carrier gas. See Section 4.3 for 5.1.1 Vial—25- or 40-mL capacity, or larg- precautions regarding the use of hydrogen as er, with screw cap with a hole in the center a carrier gas. (Fisher #13075 or equivalent). Unless pre- 5.3.2 GC column—See the footnote to cleaned, detergent wash, rinse with tap and Table 3. Other columns or column systems reagent water, and dry at 105 ± 5 °C before may be used provided all requirements in use. this method are met.

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5.3.3 Mass spectrometer—Capable of re- 6.3.3 Silica gel—35/60 mesh, Davison, petitively scanning from 35–260 Daltons Grade-15 or equivalent. (amu) every 2 seconds or less, utilizing a 70 6.3.4 Other trap materials are acceptable eV (nominal) electron energy in the electron if performance requirements in this method impact ionization mode, and producing a are met. mass spectrum which meets all criteria in 6.4 Methanol—Demonstrated to be free Table 4 when 50 ng or less of 4- from the target analytes and potentially bromofluorobenzene (BFB) is injected interfering compounds. through the GC inlet. If acrolein, acrylo- 6.5 Stock standard solutions—Stock , chloromethane, and vinyl chloride standard solutions may be prepared from are to be determined, it may be necessary to pure materials, or purchased as certified so- scan from below 25 Daltons to measure the lutions. Traceability must be to the National peaks in the 26–35 Dalton range for reliable Institute of Standards and Technology identification. (NIST) or other national or international 5.3.4 GC/MS interface—Any GC to MS standard, when available. Stock solution interface that meets all performance re- concentrations alternative to those below quirements in this method may be used. may be used. Prepare stock standard solu- 5.3.5 Data system—A computer system tions in methanol using assayed liquids or must be interfaced to the mass spectrometer gases as appropriate. Because some of the that allows continuous acquisition and stor- compounds in this method are known to be age of mass spectra throughout the toxic, primary dilutions should be prepared chromatographic program. The computer in a hood, and a NIOSH/MESA approved must have software that allows searching toxic gas respirator should be worn when any GC/MS data file for specific m/z’s high concentrations of neat materials are (masses) and plotting m/z abundances versus handled. The following procedure may be time or scan number. This type of plot is de- used to prepare standards from neat mate- fined as an extracted ion current profile rials: (EICP). Software must also be available that 6.5.1 Place about 9.8 mL of methanol in a allows integrating the abundance at any 10-mL ground-glass-stoppered volumetric EICP between specified time or scan number flask. Allow the flask to stand, unstoppered, for about 10 minutes or until all alcohol limits. wetted surfaces have dried. Weigh the flask 5.4 Syringes—Graduated, 5–25 mL, glass to the nearest 0.1 mg. hypodermic with Luerlok tip, compatible 6.5.2 Add the assayed reference material. with the purging device. 6.5.2.1 Liquids—Using a 100 μL syringe, 5.5 Micro syringes—Graduated, 25–1000 μL, immediately add two or more drops of as- with 0.006 in. ID needle. sayed reference material to the flask. Be 5.6 Syringe valve—Two-way, with Luer sure that the drops fall directly into the al- ends. cohol without contacting the neck of the 5.7 Syringe—5 mL, gas-tight with shut-off flask. Reweigh, dilute to volume, stopper, valve. then mix by inverting the flask several 5.8 Bottle—15 mL, screw-cap, with Teflon times. Calculate the concentration in μg/μL cap liner. from the net gain in weight. 5.9 Balance—Analytical, capable of accu- 6.5.2.2 Gases—To prepare standards for rately weighing 0.0001 g. any of compounds that boil below 30 °C, fill 6. Reagents a 5-mL valved gas-tight syringe with ref- erence standard vapor to the 5.0 mL mark. 6.1 Reagent water—Reagent water is de- Lower the needle to 5 mm above the meth- fined as water in which the analytes of inter- anol meniscus. Slowly introduce the vapor est and interfering compounds are not de- above the surface of the liquid (the vapor tected at the MDLs of the analytes of inter- will rapidly dissolve in the methanol). Re- est. It may be generated by passing deionized weigh, dilute to volume, stopper, then mix water, distilled water, or tap water through by inverting the flask several times. Cal- a carbon bed, passing the water through a culate the concentration in μg/μL from the water purifier, or heating the water to be- net gain in weight. tween 90 and 100 °C while bubbling contami- 6.5.3 When compound purity is assayed to nant-free gas through it for approximately 1 be 96% or greater, the weight may be used hour. While still hot, transfer the water to without correction to calculate the con- screw-cap bottles and seal with a centration of the stock standard. Commer- fluoropolymer-lined cap. cially prepared stock standards may be used 6.2 Sodium thiosulfate—(ACS) Granular. at any concentration if they are certified by 6.3 Trap materials. the manufacturer or by an independent 6.3.1 2,6-Diphenylene oxide polymer— source. Tenax, 60/80 mesh, chromatographic grade, or 6.5.4 Prepare fresh standards weekly for equivalent. the gases and 2-chloroethylvinyl ether. Un- 6.3.2 Methyl silicone packing—3% OV–1 on less stated otherwise in this method, store Chromosorb-W, 60/80 mesh, or equivalent. non-aqueous standards in fluoropolymer-

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lined screw-cap, or heat-sealed, glass con- 7. Calibration tainers, in the dark at ¥20 to ¥10 °C. Store 7.1 Assemble a purge-and-trap system aqueous standards; e.g., the aqueous LCS that meets the specifications in Section 5.2. (section 8.4.1) in the dark at ≤6 °C (but do not Prior to first use, condition the trap over- freeze) with zero headspace; e.g., in VOA night at 180 °C by backflushing with gas at a vials (section 5.1.1). Standards prepared by flow rate of at least 20 mL/min. Condition the laboratory may be stored for up to one the trap after each analysis at a temperature month, except when comparison with QC and time sufficient to prevent detectable check standards indicates that a standard concentrations of the analytes or contami- has degraded or become more concentrated nants in successive analyses. due to evaporation, or unless the laboratory 7.2 Connect the purge-and-trap system to has data on file to prove stability for a the gas chromatograph. The gas chro- longer period. Commercially prepared stand- matograph should be operated using tem- ards may be stored until the expiration date perature and flow rate conditions equivalent provided by the vendor, except when com- to those given in the footnotes to Table 3. parison with QC check standards indicates Alternative temperature and flow rate condi- that a standard has degraded or become tions may be used provided that performance more concentrated due to evaporation, or requirements in this method are met. unless the laboratory has data from the ven- 7.3 Internal standard calibration. dor on file to prove stability for a longer pe- 7.3.1 Internal standards. riod. 7.3.1.1 Select three or more internal NOTE: 2-Chloroethylvinyl ether has been standards similar in chromatographic behav- shown to be stable for as long as one month ior to the compounds of interest. Suggested if prepared as a separate standard, and the internal standards are listed in Table 5. Use other analytes have been shown to be stable the base peak m/z as the primary m/z for for as long as 2 months if stored at less than quantification of the standards. If inter- ¥10 °C with minimal headspace in sealed, ferences are found at the base peak, use one miniature inert-valved vials. of the next two most intense m/z’s for quan- titation. Demonstrate that measurements of 6.6 Secondary dilution standards—Using the internal standards are not affected by stock solutions, prepare secondary dilution method or matrix interferences. standards in methanol that contain the com- 7.3.1.2 To assure accurate analyte identi- pounds of interest, either singly or mixed. fication, particularly when selected ion mon- Secondary dilution standards should be pre- itoring (SIM) is used, it may be advan- pared at concentrations such that the aque- tageous to include more internal standards ous calibration standards prepared in section than those suggested in Section 7.3.1.1. An 7.3.2 will bracket the working range of the analyte will be located most accurately if its analytical system. retention time relative to an internal stand- 6.7 Surrogate standard spiking solution— ard is in the range of 0.8 to 1.2. Select a minimum of three surrogate com- 7.3.1.3 Prepare a stock standard solution pounds from Table 5. The surrogates selected for each internal standard in methanol as de- should match the purging characteristics of scribed in Section 6.5, and prepare a solution the analytes of interest as closely as pos- for spiking the internal standards into all sible. Prepare a stock standard solution for blanks, LCSs, and MS/MSDs. Prepare the each surrogate in methanol as described in spiking solution such that spiking a small section 6.5, and prepare a solution for spik- volume will result in a constant concentra- ing the surrogates into all blanks, LCSs, and tion of the internal standards. For example, MS/MSDs. Prepare the spiking solution such add 10 μL of a spiking solution containing that spiking a small volume will result in a the internal standards at a concentration of constant concentration of the surrogates. 15 μg/mL in methanol to a 5-mL aliquot of For example, add 10 μL of a spiking solution water to produce a concentration of 30 μg/L containing the surrogates at a concentration for each internal standard. Other concentra- of 15 μg/mL in methanol to a 5-mL aliquot of tions may be used. The internal standard so- water to produce a concentration of 30 μg/L lution and the surrogate standard spiking so- for each surrogate. Other surrogate con- lution (Section 6.7) may be combined, if de- centrations may be used. Store per section sired. Store per section 6.5.4. 6.5.4. 7.3.2 Calibration. 6.8 BFB standard—Prepare a solution of 7.3.2.1 Calibration standards. BFB in methanol as described in Sections 6.5 7.3.2.1.1 Prepare calibration standards at and 6.6. The solution should be prepared such a minimum of five concentration levels for that an injection or purging from water will each analyte of interest by adding appro- result in introduction of ≤ 50 ng into the GC. priate volumes of one or more stock stand- BFB may be included in a mixture with the ards to a fixed volume (e.g., 40 mL) of rea- internal standards and/or surrogates. gent water in volumetric glassware. Fewer 6.9 Quality control check sample con- levels may be necessary for some analytes centrate—See Section 8.2.1. based on the sensitivity of the MS, but no

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fewer than 3 levels may be used, and only the as the spectrum is not distorted to meet the highest or lowest point(s) may be dropped criteria in Table 4. from the calibration. One of the calibration 7.3.2.3 Analyze the mid-point standard standards should be at a concentration at or and enter or review the retention time, rel- below the ML or as specified by a regulatory/ ative retention time, mass spectrum, and control authority or in a permit. The ML quantitation m/z in the data system for each value may be rounded to a whole number analyte of interest, surrogate, and internal that is more convenient for preparing the standard. If additional analytes (Table 2) are standard, but must not exceed the ML values to be quantified, include these analytes in listed in Table 1 for those analytes which list the standard. The mass spectrum for each ML values. Alternatively, the laboratory analyte must be comprised of a minimum of may establish the ML for each analyte based 2 m/z’s; 3 to 5 m/z’s assure more reliable on the concentration of the lowest calibra- analyte identification. Suggested quantita- tion standard in a series of standards pro- tion m/z’s are shown in Table 6 as the pri- duced in the laboratory or obtained from a mary m/z. For analytes in Table 6 that do commercial vendor, again, provided that the not have a secondary m/z, acquire a mass ML value does not exceed the MLs in Table spectrum and enter one or more secondary 1, and provided that the resulting calibration m/z’s for more reliable identification. If an meets the acceptance criteria in Section interference occurs at the primary m/z, use 7.3.4, based on the RSD, RSE, or R2. The con- one of the secondary m/z’s or an alternative centrations of the higher standards should m/z. A single m/z only is required for quan- correspond to the expected range of con- titation. centrations found in real samples, or should 7.3.2.4 For SIM operation, determine the define the working range of the GC/MS sys- analytes in each descriptor, the quantitation tem for full-scan and/or SIM operation, as m/z for each analyte (the quantitation m/z appropriate. A minimum of six concentra- can be the same as for full-scan operation; tion levels is required for a second order, Section 7.3.2.3), the dwell time on each m/z non-linear (e.g., quadratic; ax2 + bx + c = 0) for each analyte, and the beginning and end- calibration. Calibrations higher than second ing retention time for each descriptor. Ana- order are not allowed. lyze the verification standard in scan mode to verify m/z’s and establish retention times 7.3.2.1.2 To each calibration standard or for the analytes. There must be a minimum standard mixture, add a known constant vol- of two m/z’s for each analyte to assure ume of the internal standard spiking solu- analyte identification. To maintain sensi- tion (section 7.3.1.3) and surrogate standard tivity, the number of m/z’s in a descriptor spiking solution (section 6.7) or the com- should be limited. For example, for a bined internal standard solution and surro- descriptor with 10 m/z’s and a gate spiking solution (section 7.3.1.3). Aque- chromatographic peak width of 5 sec, a dwell ous standards may be stored up to 24 hours, time of 100 ms at each m/z would result in a if held in sealed vials with zero headspace. If scan time of 1 second and provide 5 scans not so stored, they must be discarded after across the GC peak. The quantitation m/z one hour. will usually be the most intense peak in the 7.3.2.2 Prior to analysis of the calibration mass spectrum. The quantitation m/z and standards, analyze the BFB standard (sec- dwell time may be optimized for each tion 6.8) and adjust the scan rate of the MS analyte. The acquisition table used for SIM to produce a minimum of 5 mass spectra must take into account the mass defect (usu- across the BFB GC peak, but do not exceed ally less than 0.2 Dalton) that can occur at 2 seconds per scan. Adjust instrument condi- each m/z monitored. Refer to the footnotes tions until the BFB criteria in Table 4 are to Table 3 for establishing operating condi- met. Once the scan conditions are estab- tions and to section 7.3.2.2 for establishing lished, they must be used for analyses of all scan conditions. standards, blanks, and samples. 7.3.2.5 For combined scan and SIM oper- NOTE: The BFB spectrum may be evaluated ation, set up the scan segments and by summing the intensities of the m/z’s descriptors to meet requirements in sections across the GC peak, subtracting the back- 7.3.2.2–7.3.2.4. Analyze unfamiliar samples in ground at each m/z in a region of the chro- the scan mode to assure that the analytes of matogram within 20 scans of but not includ- interest are determined. ing any part of the BFB peak. The BFB spec- 7.3.3 Analyze each calibration standard trum may also be evaluated by fitting a according to Section 10 and tabulate the area Gaussian to each m/z and using the intensity at the quantitation m/z against concentra- at the maximum for each Gaussian, or by in- tion for each analyte of interest, surrogate, tegrating the area at each m/z and using the and internal standard. Calculate the re- integrated areas. Other means may be used sponse factor (RF) for each compound at for evaluation of the BFB spectrum so long each concentration using Equation 1.

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Where: erated. Results of ongoing performance tests are compared with established QC accept- As = Area of the characteristic m/z for the analyte to be measured. ance criteria to determine if the results of Ais = Area of the characteristic m/z for the analyses meet performance requirements of internal standard. this method. When results of spiked samples Cis = Concentration of the internal standard do not meet the QC acceptance criteria in (μg/L). this method, a quality control check sample Cs = Concentration of the analyte to be (laboratory control sample; LCS) must be measured (μg/L). analyzed to confirm that the measurements 7.3.4 Calculate the mean (average) and were performed in an in-control mode of op- relative standard deviation (RSD) of the re- eration. A laboratory may develop its own sponse factors. If the RSD is less than 35%, performance criteria (as QC acceptance cri- the RF can be assumed to be invariant and teria), provided such criteria are as or more the average RF can be used for calculations. restrictive than the criteria in this method. Alternatively, the results can be used to fit 8.1.1 The laboratory must make an initial a linear or quadratic regression of response demonstration of capability (DOC) to gen- ratios, As/Ais, vs. concentration ratios Cs/Cis. erate acceptable precision and recovery with If used, the regression must be weighted in- this method. This demonstration is detailed versely proportional to concentration (1/C). in Section 8.2. On a continuing basis, the lab- The coefficient of determination (R2) of the oratory must repeat demonstration of capa- weighted regression must be greater than bility (DOC) at least annually. 0.920 (this value roughly corresponds to the 8.1.2 In recognition of advances that are RSD limit of 35%). Alternatively, the rel- occurring in analytical technology, and to ative standard error (Reference 10) may be overcome matrix interferences, the labora- used as an acceptance criterion. As with the tory is permitted certain options (section 1.5 RSD, the RSE must be less than 35%. If an and 40 CFR 136.6(b)) to improve separations RSE less than 35% cannot be achieved for a or lower the costs of measurements. These quadratic regression, system performance is options may include an alternative purge- unacceptable, and the system must be ad- and-trap device, and changes in both column justed and re-calibrated. and type of mass spectrometer (see 40 CFR NOTE: Using capillary columns and current 136.6(b)(4)(xvi)). Alternative determinative instrumentation, it is quite likely that a lab- techniques, such as substitution of oratory can calibrate the target analytes in spectroscopic or immunoassay techniques, this method and achieve a linearity metric and changes that degrade method perform- (either RSD or RSE) well below 35%. There- ance, are not allowed. If an analytical tech- fore, laboratories are permitted to use more nique other than GC/MS is used, that tech- stringent acceptance criteria for calibration nique must have a specificity equal to or than described here, for example, to har- greater than the specificity of GC/MS for the monize their application of this method with analytes of interest. The laboratory is also those from other sources. encouraged to participate in inter-compari- 7.4 Calibration verification—Because the son and performance evaluation studies (see analytical system is calibrated by purge of section 8.8). the analytes from water, calibration 8.1.2.1 Each time a modification is made verification is performed using the labora- to this method, the laboratory is required to tory control sample (LCS). See section 8.4 for repeat the procedure in section 8.2. If the de- requirements for calibration verification tection limit of the method will be affected using the LCS, and the Glossary for further by the change, the laboratory must dem- definition. onstrate that the MDLs (40 CFR part 136, ap- pendix B) are lower than one-third the regu- 8. Quality Control latory compliance limit or the MDLs in this 8.1 Each laboratory that uses this method method, whichever are greater. If calibration is required to operate a formal quality assur- will be affected by the change, the instru- ance program. The minimum requirements ment must be recalibrated per section 7. of this program consist of an initial dem- Once the modification is demonstrated to onstration of laboratory capability and on- produce results equivalent or superior to re- going analysis of spiked samples and blanks sults produced by this method, that modi- to evaluate and document data quality (40 fication may be used routinely thereafter, so CFR 136.7). The laboratory must maintain long as the other requirements in this meth- records to document the quality of data gen- od are met (e.g., matrix spike/matrix spike

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duplicate recovery and relative percent dif- formed the analyses and modification, and of ference). the quality control officer that witnessed 8.1.2.1.1 If a modification is to be applied and will verify the analyses and modifica- to a specific discharge, the laboratory must tions. prepare and analyze matrix spike/matrix 8.1.2.2.2 A list of analytes, by name and spike duplicate (MS/MSD) samples (Section CAS Registry Number. 8.3) and LCS samples (section 8.4). The lab- 8.1.2.2.3 A narrative stating reason(s) for oratory must include internal standards and the modifications. surrogates (section 8.7) in each of the sam- 8.1.2.2.4 Results from all quality control ples. The MS/MSD and LCS samples must be (QC) tests comparing the modified method to fortified with the analytes of interest (sec- this method, including: tion 1.3.). If the modification is for nation- (a) Calibration (section 7). wide use, MS/MSD samples must be prepared (b) Calibration verification/LCS (section from a minimum of nine different discharges 8.4). (See section 8.1.2.1.2), and all QC acceptance (c) Initial demonstration of capability (sec- criteria in this method must be met. This tion 8.2). evaluation only needs to be performed once, (d) Analysis of blanks (section 8.5). other than for the routine QC required by (e) Matrix spike/matrix spike duplicate this method (for example it could be per- analysis (section 8.3). formed by the vendor of the alternative ma- (f) Laboratory control sample analysis terials) but any laboratory using that spe- (section 8.4). cific material must have the results of the 8.1.2.2.5 Data that will allow an inde- study available. This includes a full data pendent reviewer to validate each deter- package with the raw data that will allow an mination by tracing the instrument output independent reviewer to verify each deter- (peak height, area, or other signal) to the mination and calculation performed by the final result. These data are to include: laboratory (see section 8.1.2.2.5, items (a)– (a) Sample numbers and other identifiers. (l)). (b) Analysis dates and times. 8.1.2.1.2 Sample matrices on which MS/ (c) Analysis sequence/run chronology. MSD tests must be performed for nationwide (d) Sample volume (Section 10). use of an allowed modification: (e) Sample dilution (Section 13.2). (a) Effluent from a publicly owned treat- (f) Instrument and operating conditions. ment works (POTW). (b) ASTM D5905 Standard Specification for (g) Column (dimensions, material, etc). Substitute Wastewater. (h) Operating conditions (temperature pro- (c) Sewage sludge, if sewage sludge will be gram, flow rate, etc). in the permit. (i) Detector (type, operating conditions, (d) ASTM D1141 Standard Specification for etc). Substitute Ocean Water, if ocean water will (j) Chromatograms, mass spectra, and be in the permit. other recordings of raw data. (e) Untreated and treated wastewaters up (k) Quantitation reports, data system out- to a total of nine matrix types (see https:// puts, and other data to link the raw data to www.epa.gov/eg/industrial-effluent-guidelines the results reported. for a list of industrial categories with exist- (l) A written Standard Operating Proce- ing effluent guidelines). dure (SOP). (i) At least one of the above wastewater 8.1.2.2.6 Each individual laboratory wish- matrix types must have at least one of the ing to use a given modification must perform following characteristics: the start-up tests in section 8.1.2 (e.g., DOC, (A) Total suspended solids greater than 40 MDL), with the modification as an integral mg/L. part of this method prior to applying the (B) Total dissolved solids greater than 100 modification to specific discharges. Results mg/L. of the DOC must meet the QC acceptance cri- (C) Oil and grease greater than 20 mg/L. teria in Table 7 for the analytes of interest (D) NaCl greater than 120 mg/L. (section 1.3), and the MDLs must be equal to (E) CaCO3 greater than 140 mg/L. or lower than the MDLs in Table3 for the (ii) Results of MS/MSD tests must meet QC analytes of interest acceptance criteria in section 8.3. 8.1.3 Before analyzing samples, the lab- (f) A proficiency testing (PT) sample from oratory must analyze a blank to dem- a recognized provider, in addition to tests of onstrate that interferences from the analyt- the nine matrices (section 8.1.2.1.1). ical system, labware, and reagents are under 8.1.2.2 The laboratory is required to main- control. Each time a batch of samples is ana- tain records of modifications made to this lyzed or reagents are changed, a blank must method. These records include the following, be analyzed as a safeguard against labora- at a minimum: tory contamination. Requirements for the 8.1.2.2.1 The names, titles, and business blank are given in section 8.5. street addresses, telephone numbers, and 8.1.4 The laboratory must, on an ongoing email addresses of the analyst(s) that per- basis, spike and analyze samples to monitor

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and evaluate method and laboratory per- 8.2.2 Using a pipet or micro-syringe, pre- formance on the sample matrix. The proce- pare four LCSs by adding an appropriate vol- dure for spiking and analysis is given in sec- ume of the concentrate to each of four tion 8.3. aliquots of reagent water. The volume of rea- 8.1.5 The laboratory must, on an ongoing gent water must be the same as the volume basis, demonstrate through analysis of a that will be used for the sample, blank (sec- quality control check sample (laboratory tion 8.5), and MS/MSD (section 8.3). A volume control sample, LCS; on-going precision and of 5 mL and a concentration of 20 μg/L were recovery sample, OPR) that the measure- used to develop the QC acceptance criteria in ment system is in control. This procedure is Table 7. An alternative volume and sample given in section 8.4. concentration may be used, provided that all 8.1.6 The laboratory must maintain per- QC tests are performed and all QC accept- formance records to document the quality of ance criteria in this method are met. Also data that is generated. This procedure is add an aliquot of the surrogate spiking solu- given in section 8.8. tion (section 6.7) and internal standard spik- 8.1.7 The large number of analytes tested ing solution (section 7.3.1.3) to the reagent- in performance tests in this method present water aliquots. a substantial probability that one or more 8.2.3 Analyze the four LCSs according to will fail acceptance criteria when many the method beginning in section 10. analytes are tested simultaneously, and a re- 8.2.4 Calculate the average percent recov- test is allowed if this situation should occur. ery (X) and the standard deviation of the per- If, however, continued re-testing results in cent recovery (s) for each analyte using the further repeated failures, the laboratory four results. must document and report the failures (e.g., 8.2.5 For each analyte, compare s and X as qualifiers on results), unless the failures with the corresponding acceptance criteria are not required to be reported as deter- for precision and recovery in Table 7. For mined by the regulatory/control authority. analytes in Tables 1 and 2 not listed in Table Results associated with a QC failure for an 7, DOC QC acceptance criteria must be devel- analyte regulated in a discharge cannot be oped by the laboratory. EPA has provided used to demonstrate regulatory compliance. guidance for development of QC acceptance QC failures do not relieve a discharger or criteria (References 11 and 12). Alternatively, permittee of reporting timely results. acceptance criteria for analytes not listed in 8.2 Initial demonstration of capability Table 7 may be based on laboratory control (DOC)—To establish the ability to generate charts. If s and X for all analytes of interest acceptable recovery and precision, the lab- meet the acceptance criteria, system per- oratory must perform the DOC in sections formance is acceptable and analysis of 8.2.1 through 8.2.6 for the analytes of inter- blanks and samples may begin. If any indi- est. The laboratory must also establish vidual s exceeds the precision limit or any MDLs for the analytes of interest using the individual X falls outside the range for recov- MDL procedure at 40 CFR part 136, appendix ery, system performance is unacceptable for B. The laboratory’s MDLs must be equal to that analyte. or lower than those listed in Table 1 for NOTE: The large number of analytes in Ta- those analytes which list MDL values, or bles 1 and 2 present a substantial probability lower than one-third the regulatory compli- that one or more will fail at least one of the ance limit, whichever is greater. For MDLs acceptance criteria when many or all not listed in Table 1, the laboratory must de- analytes are determined simultaneously. termine the MDLs using the MDL procedure Therefore, the analyst is permitted to con- at 40 CFR part 136, appendix B under the duct a ‘‘re-test’’ as described in section 8.2.6. same conditions used to determine the MDLs for the analytes listed in Table 1. All proce- 8.2.6 When one or more of the analytes dures used in the analysis must be included tested fail at least one of the acceptance cri- in the DOC. teria, repeat the test for only the analytes 8.2.1 For the DOC, a QC check sample con- that failed. If results for these analytes pass, centrate (LCS concentrate) containing each system performance is acceptable and anal- analyte of interest (section 1.3) is prepared in ysis of samples and blanks may proceed. If methanol. The QC check sample concentrate one or more of the analytes again fail, sys- must be prepared independently from those tem performance is unacceptable for the used for calibration, but may be from the analytes that failed the acceptance criteria. same source as the second-source standard Correct the problem and repeat the test (sec- used for calibration verification/LCS (sec- tion 8.2). See section 8.1.7 for disposition of tions 7.4 and 8.4). The concentrate should repeated failures. produce concentrations of the analytes of in- NOTE: To maintain the validity of the test terest in water at the mid-point of the cali- and re-test, system maintenance and/or ad- bration range, and may be at the same con- justment is not permitted between this pair centration as the LCS (section 8.4). of tests. NOTE: QC check sample concentrates are 8.3 Matrix spike and matrix spike dupli- no longer available from EPA. cate (MS/MSD)—The purpose of the MS/MSD

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requirement is to provide data that dem- error in measurement of both the back- onstrate the effectiveness of the method as ground and spike concentrations, assuming a applied to the samples in question by a given spike to background ratio of 5:1. This error laboratory, and both the data user (dis- will be accounted for to the extent that the charger, permittee, regulated entity, regu- spike to background ratio approaches 5:1 latory/control authority, customer, other) (Reference 13) and is applied to spike con- and the laboratory share responsibility for centrations of 20 μg/L and higher. If spiking provision of such data. The data user should is performed at a concentration lower than identify the sample and the analytes of in- 20 μg/L, the laboratory must use the QC ac- terest (section 1.3) to be spiked and provide ceptance criteria in Table 7, the optional QC sufficient sample volume to perform MS/ acceptance criteria calculated for the spe- MSD analyses. The laboratory must, on an cific spike concentration in Table 8, or op- ongoing basis, spike at least 5% of the sam- tional in-house criteria (Section 8.3.4). To ples in duplicate from each discharge being use the acceptance criteria in Table 8: (1) monitored to assess accuracy (recovery and Calculate recovery (X’) using the equation in precision). If direction cannot be obtained Table 8, substituting the spike concentration from the data user, the laboratory must (T) for C; (2) Calculate overall precision (S’) spike at least one sample in duplicate per ex- using the equation in Table 8, substituting traction batch of up to 20 samples with the X’ for X; (3) Calculate the range for recovery analytes in Table 1. Spiked sample results at the spike concentration as (100 X’/T) ± should be reported only to the data user 2.44(100 S’/T)% (Reference 4). For analytes of whose sample was spiked, or as requested or interest in Tables 1 and 2 not listed in Table required by a regulatory/control authority, 7, QC acceptance criteria must be developed or in a permit. by the laboratory. EPA has provided guid- 8.3.1 If, as in compliance monitoring, the ance for development of QC acceptance cri- concentration of a specific analyte will be teria (References 11 and 12). Alternatively, checked against a regulatory concentration acceptance criteria may be based on labora- limit, the concentration of the spike should tory control charts. In-house LCS QC accept- be at that limit; otherwise, the concentra- ance criteria must be updated at least every tion of the spike should be one to five times two years. higher than the background concentration 8.3.4 After analysis of a minimum of 20 determined in section 8.3.2, at or near the MS/MSD samples for each target analyte and mid-point of the calibration range, or at the surrogate, and if the laboratory chooses to concentration in the LCS (section 8.4) which- develop and apply in-house QC limits, the ever concentration would be larger. laboratory should calculate and apply in- 8.3.2 Analyze one sample aliquot to deter- house QC limits for recovery and RPD of fu- mine the background concentration (B) of ture MS/MSD samples (section 8.3). The QC the each analyte of interest. If necessary, limits for recovery are calculated as the prepare a new check sample concentrate mean observed recovery ± 3 standard devi- (section 8.2.1) appropriate for the background ations, and the upper QC limit for RPD is concentration. Spike and analyze two addi- calculated as the mean RPD plus 3 standard tional sample aliquots, and determine the deviations of the RPDs. The in-house QC lim- concentration after spiking (A1 and A2) of its must be updated at least every two years each analyte. Calculate the percent recov- and re-established after any major change in eries (P1 and P2) as 100 (A1¥B)/T and 100 the analytical instrumentation or process. If (A2¥B)/T, where T is the known true value of in-house QC limits are developed, at least the spike. Also calculate the relative percent 80% of the analytes tested in the MS/MSD difference (RPD) between the concentrations must have in-house QC acceptance criteria (A1 and A2) as 200 √A1¥A2√/(A1 + A2). If nec- that are tighter than those in Table 7 and essary, adjust the concentrations used to the remaining analytes (those other than the calculate the RPD to account for differences analytes included in the 80%) must meet the in the volumes of the spiked aliquots. acceptance criteria in Table 7. If an in-house 8.3.3 Compare the percent recoveries (P1 QC limit for the RPD is greater than the and P2) and the RPD for each analyte in the limit in Table 7, then the limit in Table 7 MS/MSD aliquots with the corresponding QC must be used. Similarly, if an in-house lower acceptance criteria in Table 7. A laboratory limit for recovery is below the lower limit in may develop and apply QC acceptance cri- Table 7, then the lower limit in Table 7 must teria more restrictive than the criteria in be used, and if an in-house upper limit for re- Table 7, if desired. covery is above the upper limit in Table 7, 8.3.3.1 If any individual P falls outside the then the upper limit in Table 7 must be used. designated range for recovery in either ali- 8.4 Calibration verification/laboratory quot, or the RPD limit is exceeded, the re- control sample (LCS)—The working calibra- sult for the analyte in the unspiked sample tion curve or RF must be verified imme- is suspect. See Section 8.1.7 for disposition of diately after calibration and at the begin- failures. ning of each 12-hour shift by the measure- 8.3.3.2 The acceptance criteria in Table 7 ment of an LCS. The LCS must be from a were calculated to include an allowance for source different from the source used for

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calibration (section 7.3.2.1), but may be the NOTE: To maintain the validity of the test same as the sample prepared for the DOC and re-test, system maintenance and/or ad- (section 8.2.1). justment is not permitted between the pair NOTE: The 12-hour shift begins after anal- of tests. ysis of BFB, the LCS, and the blank, and 8.4.5 After analysis of 20 LCS samples, and ends 12 hours later. BFB, the LCS, and blank if the laboratory chooses to develop and are outside of the 12-hour shift (Section 11.4). apply in-house QC limits, the laboratory The MS and MSD are treated as samples and should calculate and apply in-house QC lim- are analyzed within the 12-hour shift. its for recovery to future LCS samples (sec- tion 8.4). Limits for recovery in the LCS cal- 8.4.1 Prepare the LCS by adding QC check culated as the mean recovery ±3 standard de- sample concentrate (section 8.2.1) to reagent viations. A minimum of 80% of the analytes water. Include all analytes of interest (Sec- tested for in the LCS must have QC accept- tion 1.3) in the LCS. The volume of reagent ance criteria tighter than those in Table 7, water must be the same as the volume used and the remaining analytes (those other for the sample, blank (Section 8.5), and MS/ than the analytes included in the 80%) must MSD (section 8.3). Also add an aliquot of the meet the acceptance criteria in Table 7. If an surrogate solution (Section 6.7) and internal in-house lower limit for recovery is lower standard solution (section 7.3.1.3). The con- than the lower limit in Table 7, the lower centration of the analytes in reagent water limit in Table 7 must be used, and if an in- should be the same as the concentration in house upper limit for recovery is higher than the DOC (section 8.2.2). the upper limit in Table 7, the upper limit in 8.4.2 Analyze the LCS prior to analysis of Table 7 must be used. Many of the analytes field samples in the batch of samples ana- and surrogates do not have acceptance cri- lyzed during the 12-hour shift (see the NOTE teria. The laboratory should use 60–140% as at section 8.4). Determine the concentration interim acceptance criteria for recoveries of (A) of each analyte. Calculate the percent re- spiked analytes that do not have recovery covery (Q) as 100 (A/T) %, where T is the true limits specified in Table 7, and least 80% of value of the concentration in the LCS. the analytes should meet the 60–140% in- 8.4.3 Compare the percent recovery (Q) for terim criteria until in-house LCS limits are each analyte with its corresponding QC ac- developed. Alternatively, acceptance criteria ceptance criterion in Table 7. For analytes of for analytes that do not have recovery limits interest in Tables 1 and 2 not listed in Table in Table 7 may be based on laboratory con- 7, use the QC acceptance criteria developed trol charts. In-house QC acceptance criteria for the LCS (section 8.4.5). If the recoveries must be updated at least every two years. for all analytes of interest fall within their 8.5 Blank—A blank must be analyzed respective QC acceptance criteria, analysis prior to each 12-hour shift to demonstrate of blanks and field samples may proceed. If freedom from contamination. A blank must any individual Q falls outside the range, pro- also be analyzed after a sample containing a ceed according to section 8.4.4. high concentration of an analyte or poten- NOTE: The large number of analytes in Ta- tially interfering compound to demonstrate bles 1—2 present a substantial probability freedom from carry-over. that one or more will fail the acceptance cri- 8.5.1 Spike the internal standards and sur- teria when all analytes are tested simulta- rogates into the blank. Analyze the blank neously. Because a re-test is allowed in event immediately after analysis of the LCS (Sec- of failure (sections 8.1.7 and 8.4.3), it may be tion 8.4) and prior to analysis of the MS/MSD prudent to analyze two LCSs together and and samples to demonstrate freedom from evaluate results of the second analysis contamination. against the QC acceptance criteria only if an 8.5.2 If any analyte of interest is found in analyte fails the first test. the blank: At a concentration greater than 8.4.4 Repeat the test only for those the MDL for the analyte, at a concentration analytes that failed to meet the acceptance greater than one-third the regulatory com- criteria (Q). If these analytes now pass, sys- pliance limit, or at a concentration greater tem performance is acceptable and analysis than one-tenth the concentration in a sam- of blanks and samples may proceed. Re- ple analyzed during the 12-hour shift (section peated failure, however, will confirm a gen- 8.4), whichever is greater; analysis of samples eral problem with the measurement system. must be halted and samples affected by the If this occurs, repeat the test (section 8.4.2). blank must be re-analyzed. If, however, con- using a fresh LCS (section 8.2.2) or an LCS tinued re-testing results in repeated blank prepared with a fresh QC check sample con- contamination, the laboratory must docu- centrate (section 8.2.1), or perform and docu- ment and report the failures (e.g., as quali- ment system repair. Subsequent to repair, fiers on results), unless the failures are not repeat the calibration verification/LCS test required to be reported as determined by the (section 8.4). If the acceptance criteria for Q regulatory/control authority. Results associ- cannot be met, re-calibrate the instrument ated with blank contamination for an (section 7). See section 8.1.7 for disposition of analyte regulated in a discharge cannot be repeated failures. used to demonstrate regulatory compliance.

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QC failures do not relieve a discharger or or report it if required by the regulatory/con- permittee of reporting timely results. trol authority. 8.6 Surrogate recoveries—The laboratory 8.8 As part of the QC program for the lab- must evaluate surrogate recovery data in oratory, control charts or statements of ac- each sample against its in-house surrogate curacy for wastewater samples must be as- recovery limits for surrogates that do not sessed and records maintained periodically have acceptance criteria in Table 7. The lab- (see 40 CFR 136.7(c)(1)(viii)). After analysis of oratory may use 60–140% as interim accept- five or more spiked wastewater samples as in ance criteria for recoveries for surrogates section 8.3, calculate the average percent re- not listed in Table 5. At least 80% of the sur- covery (PX) and the standard deviation of the rogates must meet the 60–140% interim cri- percent recovery (sp). Express the accuracy teria until in-house limits are developed. Al- assessment as a percent interval from PX¥2sp ternatively, surrogate recovery limits may to PX + 2sp. For example, if PX = 90% and sp be developed from laboratory control charts. = 10%, the accuracy interval is expressed as 8.6.1 Spike the surrogates into all sam- 70–110%. Update the accuracy assessment for ples, blanks, LCSs, and MS/MSDs. Compare each analyte on a regular basis (e.g., after surrogate recoveries against the QC accept- each 5–10 new accuracy measurements). If de- ance criteria in Table 7. For surrogates in sired, statements of accuracy for laboratory Table 5 without QC acceptance criteria in performance, independent of performance on Table 7, and for other surrogates that may be samples, may be developed using LCSs. used by the laboratory, limits must be devel- 8.9 It is recommended that the laboratory oped by the laboratory. EPA has provided adopt additional quality assurance practices guidance for development of QC acceptance for use with this method. The specific prac- criteria (References 11 and 12). Alternatively, tices that are most productive depend upon surrogate recovery limits may be developed the needs of the laboratory and the nature of from laboratory control charts. In-house QC the samples. Field duplicates may be ana- acceptance criteria must be updated at least lyzed to assess the precision of environ- every two years. mental measurements. Whenever possible, 8.6.2 If any recovery fails its criteria, at- the laboratory should analyze standard ref- tempt to find and correct the cause of the erence materials and participate in relevant failure. See section 8.1.7 for disposition of performance evaluation studies. failures. 8.7 Internal standard responses. 9. Sample Collection, Preservation, and 8.7.1 Calibration verification/LCS—The Handling responses (GC peak heights or areas) of the 9.1 Collect the sample as a grab sample in internal standards in the calibration a glass container having a total volume of at verification/LCS must be within 50% to 200% least 25 mL. Fill the sample bottle just to (1/2 to 2×) of their respective responses in the overflowing in such a manner that no air mid-point calibration standard. If they are bubbles pass through the sample as the bot- not, repeat the LCS test using a fresh QC tle is being filled. Seal the bottle so that no check sample (section 8.4.1) or perform and air bubbles are entrapped in it. If needed, document system repair. Subsequent to re- collect additional sample(s) for the MS/MSD pair, repeat the calibration verification/LCS (section 8.3). test (section 8.4). If the responses are still 9.2 Ice or refrigerate samples at ≤6 °C not within 50% to 200%, re-calibrate the in- from the time of collection until analysis, strument (section 7) and repeat the calibra- but do not freeze. If residual chlorine is tion verification/LCS test. present, add sodium thiosulfate preservative 8.7.2 Samples, blanks, and MS/MSDs—The (10 mg/40 mL is sufficient for up to 5 ppm Cl2) responses (GC peak heights or areas) of each to the empty sample bottle just prior to internal standard in each sample, blank, and shipping to the sampling site. Any method MS/MSD must be within 50% to 200% (1/2 to suitable for field use may be employed to 2×) of its respective response in the mid- test for residual chlorine (Reference 14). point calibration standard. If, as a group, all Field test kits are also available for this pur- internal standards are not within this range, pose. If sodium thiosulfate interferes in the perform and document system repair, repeat determination of the analytes, an alter- the calibration verification/LCS test (section native preservative (e.g., ascorbic acid or so- 8.4), and re-analyze the affected samples. If a dium sulfite) may be used. If preservative single internal standard is not within the has been added, shake the sample vigorously 50% to 200% range, use an alternative inter- for one minute. Maintain the hermetic seal nal standard for quantitation of the analyte on the sample bottle until time of analysis. referenced to the affected internal standard. 9.3 If acrolein is to be determined, analyze It may be necessary to use the data system the sample within 3 days. To extend the to calculate a new response factor from cali- holding time to 14 days, acidify a separate bration data for the alternative internal sample to pH 4–5 with HCl using the proce- standard/analyte pair. If an internal stand- dure in section 9.7. ard fails the 50–200% criteria and no analytes 9.4 Experimental evidence indicates that are detected in the sample, ignore the failure some aromatic compounds, notably benzene,

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toluene, and ethyl benzene are susceptible to just short of overflowing. Replace the sy- rapid biological degradation under certain ringe plunger and compress the sample. Open environmental conditions (Reference 3). Re- the syringe valve and vent any residual air frigeration alone may not be adequate to while adjusting the sample volume. Since preserve these compounds in wastewaters for this process of taking an aliquot destroys more than seven days. To extend the holding the validity of the sample for future anal- time for aromatic compounds to 14 days, ysis, the analyst should fill a second syringe acidify the sample to approximately pH 2 at this time to protect against possible loss using the procedure in section 9.7. of data. Add the surrogate spiking solution 9.5 If halocarbons are to be determined, (section 6.7) and internal standard spiking either use the acidified aromatics sample in solution (section 7.3.1.3) through the valve section 9.4 or acidify a separate sample to a bore, then close the valve. The surrogate and pH of about 2 using the procedure in section internal standards may be mixed and added 9.7. as a single spiking solution. Autosamplers 9.6 The ethers listed in Table 2 are prone designed for purge-and-trap analysis of to at pH 2 when a heated purge is volatiles also may be used. used. Aqueous samples should not be acid 10.4 Attach the syringe valve assembly to preserved if these ethers are of interest, or if the syringe valve on the purging device. the alcohols they would form upon hydrol- Open the syringe valve and inject the sample ysis are of interest and the ethers are antici- into the purging chamber. pated to present. 10.5 Close both valves and purge the sam- 9.7 Sample acidification—Collect about ple at a temperature, flow rate, and duration 500 mL of sample in a clean container and sufficient to purge the less-volatile analytes adjust the pH of the sample to 4–5 for acro- onto the trap, yet short enough to prevent lein (section 9.3), or to about 2 for the aro- blowing the more-volatile analytes through matic compounds (section 9.4) by adding 1+1 the trap. The temperature, flow rate, and HCl while swirling or stirring. Check the pH time should be determined by test. The same with narrow range pH paper. Fill a sample purge temperature, flow rate, and purge time container as described in section 9.1. Alter- must be used for all calibration, QC, and natively, fill a precleaned vial (section 5.1.1) field samples. that contains approximately 0.25 mL of 1+1 10.6 After the purge, set the purge-and- HCl with sample as in section 9.1. If pre- trap system to the desorb mode, and begin served using this alternative procedure, the the temperature program of the gas chro- pH of the sample can be verified to be <2 matograph. Introduce the trapped materials after some of the sample is removed for anal- to the GC column by rapidly heating the ysis. Acidification will destroy 2- trap to the desorb temperature while chloroethylvinyl ether; therefore, determine backflushing the trap with carrier gas at the 2-chloroethylvinyl ether from the flow rate and for the time necessary to unacidified sample. desorb the analytes of interest. The optimum 9.8 All samples must be analyzed within temperature, flow rate, and time should be 14 days of collection (Reference 3), unless determined by test. The same temperature, specified otherwise in sections 9.3–9.7. desorb time, and flow rate must be used for all calibration, QC, and field samples. If 10. Sample Purging and Gas Chromatography heating of the trap does not result in sharp 10.1 The footnote to Table 3 gives the sug- peaks for the early eluting analytes, the GC gested GC column and operating conditions column may be used as a secondary trap by MDLs and MLs for many of the analytes are cooling to an ambient or subambient tem- given in Table 1. Retention times for many perature. To avoid carry-over and inter- of the analytes are given in Table 3. Sections ferences, maintain the trap at the desorb 10.2 through 10.7 suggest procedures that temperature and flow rate until the may be used with a manual purge-and-trap analytes, interfering compounds, and excess system. Auto-samplers and other columns or water are desorbed. The optimum conditions chromatographic conditions may be used if should be determined by test. requirements in this method are met. Prior 10.7 Start MS data acquisition at the to performing analyses, and between anal- start of the desorb cycle and stop data col- yses, it may be necessary to bake the purge- lection when the analytes of interest, poten- and-trap and GC systems (section 3.3). tially interfering compounds, and water have 10.2 Attach the trap inlet to the purging eluted (see the footnote to Table 3 for condi- device, and set the purge-and-trap system to tions). purge. Open the syringe valve located on the 10.8 Cool the trap to the purge tempera- purging device sample introduction needle. ture and return the trap to the purge mode. 10.3 Allow the sample to come to ambient When the trap is cool, the next sample can temperature prior to pouring an aliquot into be analyzed. the syringe. Remove the plunger from a sy- 11. Performance Tests ringe and attach a closed syringe valve. Open the sample bottle (or standard) and carefully 11.1 At the beginning of each 12-hour shift pour the sample into the syringe barrel to during which standards or samples will be

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analyzed, perform the tests in sections 11.2– and the LCS or other standard. When signifi- 11.3 to verify system performance. Use the cant retention time drifts are observed, dilu- instrument operating conditions in the foot- tions or spiked samples may help the analyst notes to Table 3 for these performance tests. determine the effects of the matrix on Alternative conditions may be used so as elution of the target analytes and to assist long as all QC requirements are met. in qualitative identification. 11.2 BFB—Inject 50 ng of BFB solution di- 12.1.3 Either the background corrected rectly on the column. Alternatively, add EICP areas, or the corrected relative inten- BFB to reagent water or an aqueous stand- sities of the mass spectral peaks at the GC ard such that 50 ng or less of BFB will be in- peak maximum, must agree within 50% to troduced into the GC. Analyze according to 200% (1⁄2 to 2 times) for the quantitation and section 10. Confirm that all criteria in sec- secondary m/z’s in the reference mass spec- tion 7.3.2.2 and Table 4 are met. If all criteria trum stored in the data system (section are not met, perform system repair, retune 7.3.2.3), or from a reference library. For ex- the mass spectrometer, and repeat the test ample, if a peak has an intensity of 20% rel- until all criteria are met. ative to the base peak, the analyte is identi- 11.3 Verify calibration with the LCS (sec- fied if the intensity of the peak in the sam- tion 8.4) after the criteria for BFB are met ple is in the range of 10% to 40% of the base (Reference 15) and prior to analysis of a peak. blank or sample. After verification, analyze 12.1.4 If the acquired mass spectrum is a blank (section 8.5) to demonstrate freedom contaminated, or if identification is ambig- from contamination and carry-over at the uous, an experienced spectrometrist (section MDL. Tests for BFB, the LCS, and the blank 1.6) must determine the presence or absence are outside of the 12-hour shift, and the 12- of the compound. hour shift includes samples and matrix spikes and matrix spike duplicates (section 12.2 Structural isomers that produce very 8.4). The total time for analysis of BFB, the similar mass spectra should be identified as LCS, the blank, and the 12-hour shift must individual isomers if they have sufficiently not exceed 14 hours. different gas chromatographic retention times. Sufficient gas chromatographic reso- 12. Qualitative Identification lution is achieved if the height of the valley between two isomer peaks is less than 50% of 12.1 Identification is accomplished by the average of the two peak heights. Other- comparison of results from analysis of a wise, structural isomers are identified as iso- sample or blank with data stored in the GC/ meric pairs. The resolution should be MS data system (section 7.3.2.3). Identifica- verified on the mid-point concentration of tion of an analyte is confirmed per sections the initial calibration as well as the labora- 12.1.1 through 12.1.4. tory designated continuing calibration 12.1.1 The signals for the quantitation and verification level if closely eluting isomers secondary m/z’s stored in the data system are to be reported. (section 7.3.2.3) for each analyte of interest must be present and must maximize within 13. Calculations the same two consecutive scans. 12.1.2 The retention time for the analyte 13.1 When an analyte has been identified, should be within ± 10 seconds of the analyte quantitation of that analyte is based on the in the LCS run at the beginning of the shift integrated abundance from the EICP of the (section 8.4). primary characteristic m/z in Table 5 or 6. NOTE: Retention time windows other than Calculate the concentration using the re- ± 10 seconds may be appropriate depending sponse factor (RF) determined in section on the performance of the gas chro- 7.3.3 and Equation 2. If a calibration curve matograph or observed retention time drifts was used, calculate the concentration using due to certain types of matrix effects. Rel- the regression equation for the curve. If the ative retention time (RRT) may be used as concentration of an analyte exceeds the cali- an alternative to absolute retention times if bration range, dilute the sample by the min- retention time drift is a concern. RRT is a imum amount to bring the concentration unitless quantity (see section 20.2), although into the calibration range, and re-analyze. some procedures refer to ‘‘RRT units’’ in Determine a dilution factor (DF) from the providing the specification for the agree- amount of the dilution. For example, if the ment between the RRT values in the sample extract is diluted by a factor of 2, DF = 2.

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Where: (i.e., that does not meet acceptance criteria for any of the QC test in this method) must Cs = Concentration of the analyte in the sam- ple, and the other terms are as defined in be documented and reported (e.g., as a quali- Section 7.3.3. fier on results), unless the failure is not re- 13.2 Reporting of results quired to be reported as determined by the As noted in section 1.4.1, EPA has promul- regulatory/control authority. Results associ- gated this method at 40 CFR part 136 for use ated with a QC failure cannot be used to in wastewater compliance monitoring under demonstrate regulatory compliance. QC fail- the National Pollutant Discharge Elimi- ures do not relieve a discharger or permittee nation System (NPDES). The data reporting of reporting timely results. If the holding practices described here are focused on such time would be exceeded for a re-analysis of monitoring needs and may not be relevant to the sample, the regulatory/control authority other uses of this method. should be consulted for disposition. 13.2.1 Report results for wastewater sam- ples in μg/L without correction for recovery. 14. Method Performance (Other units may be used if required by a 14.1 This method was tested by 15 labora- permit.) Report all QC data with the sample tories using reagent water, drinking water, results. surface water, and industrial wastewaters 13.2.2 Reporting level. Unless otherwise spiked at six concentrations over the range specified in by a regulatory authority or in a 5–600 μg/L (References 4 and 16). Single-oper- discharge permit, results for analytes that ator precision, overall precision, and method meet the identification criteria are reported accuracy were found to be directly related to down to the concentration of the ML estab- the concentration of the analyte and essen- lished by the laboratory through calibration tially independent of the sample matrix. of the instrument (see section 7.3.2 and the Linear equations to describe these relation- glossary for the derivation of the ML). EPA considers the terms ‘‘reporting limit,’’ ships are presented in Table 8. ‘‘limit of quantitation,’’ ‘‘quantitation 14.2 As noted in section 1.1, this method limit,’’ and ‘‘minimum level’’ to be synony- was validated through an interlaboratory mous. study conducted in the early 1980s. However, 13.2.2.1 Report a result for each analyte in the fundamental chemistry principles used each field sample or QC standard at or above in this method remain sound and continue to the ML to 3 significant figures. Report a re- apply. sult for each analyte found in each field sam- ple or QC standard below the ML as ‘‘

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16. Waste Management Pollutants,’’ U.S. Environmental Protection Agency, Environmental Monitoring and Sup- 16.1 The laboratory is responsible for port Laboratory, Cincinnati, Ohio 45268, complying with all Federal, State, and local March 1977, Revised April 1977. regulations governing waste management, 3. Bellar, T.A. and Lichtenberg, J.J. particularly the hazardous waste identifica- ‘‘Semi-Automated Headspace Analysis of tion rules and land disposal restrictions, and Drinking Waters and Industrial Waters for to protect the air, water, and land by mini- Purgeable Volatile Organic Compounds,’’ mizing and controlling all releases from Measurement of Organic Pollutants in Water fume hoods and bench operations. Compli- and Wastewater, C.E. Van Hall, editor, ance is also required with any sewage dis- American Society for Testing and Materials, charge permits and regulations. An overview Philadelphia, PA. Special Technical Publica- of requirements can be found in Environ- tion 686, 1978. mental Management Guide for Small Lab- 4. ‘‘EPA Method Study 29 EPA Method 624- oratories (EPA 233–B–98–001). Purgeables,’’ EPA 600/4–84–054, National 16.2 Samples at pH <2, or pH >12, are haz- Technical Information Service, PB84–209915, ardous and must be handled and disposed of Springfield, Virginia 22161, June 1984. as hazardous waste, or neutralized and dis- 5. 40 CFR part 136, appendix B. posed of in accordance with all federal, state, 6. ‘‘Method Detection Limit for Methods and local regulations. It is the laboratory’s 624 and 625,’’ Olynyk, P., Budde, W.L., and responsibility to comply with all federal, Eichelberger, J.W. Unpublished report, May state, and local regulations governing waste 14, 1980. management, particularly the hazardous 7. ‘‘Carcinogens-Working With Carcino- waste identification rules and land disposal gens,’’ Department of Health, Education, and restrictions. The laboratory using this meth- Welfare, Public Health Service, Center for od has the responsibility to protect the air, Disease Control, National Institute for Occu- water, and land by minimizing and control- pational Safety and Health, Publication No. ling all releases from fume hoods and bench 77–206, August 1977. operations. Compliance is also required with 8. ‘‘OSHA Safety and Health Standards, any sewage discharge permits and regula- General Industry,’’ (29 CFR part 1910), Occu- tions. For further information on waste pational Safety and Health Administration, management, see ‘‘The Waste Management OSHA 2206 (Revised, January 1976). Manual for Laboratory Personnel,’’ also 9. ‘‘Safety in Academic Chemistry Labora- available from the American Chemical Soci- tories,’’ American Chemical Society Publica- ety at the address in Section 15.3. tion, Committee on Chemical Safety, 7th 16.3 Many analytes in this method decom- Edition, 2003. pose above 500 °C. Low-level waste such as 10. 40 CFR 136.6(b)(5)(x). absorbent paper, tissues, and plastic gloves 11. 40 CFR 136.6(b)(2)(i). may be burned in an appropriate incinerator. 12. Protocol for EPA Approval of New Gross quantities of neat or highly con- Methods for Organic and Inorganic Analytes centrated solutions of toxic or hazardous in Wastewater and Drinking Water (EPA– chemicals should be packaged securely and 821–B–98–003) March 1999. disposed of through commercial or govern- 13. Provost, L.P. and Elder, R.S. ‘‘Inter- mental channels that are capable of handling pretation of Percent Recovery Data,’’ Amer- these types of wastes. ican Laboratory, 15, 58–63 (1983). 16.4 For further information on waste 14. 40 CFR 136.3(a), Table IB, Chlorine— management, consult ‘‘Waste Management Total residual. Manual for Laboratory Personnel and Less is 15. Budde, W.L. and Eichelberger, J.W. Better-Laboratory Chemical Management ‘‘Performance Tests for the Evaluation of for Waste Reduction,’’ available from the Computerized Gas Chromatography/Mass American Chemical Society’s Department of Spectrometry Equipment and Laboratories,’’ Government Relations and Science Policy, EPA–600/4–80–025, U.S. Environmental Pro- 1155 16th Street NW., Washington, DC 20036, tection Agency, Environmental Monitoring 202–872–4477. and Support Laboratory, Cincinnati, Ohio 17. References 45268, April 1980. 16. ‘‘Method Performance Data for Method 1. Bellar, T.A. and Lichtenberg, J.J. ‘‘De- 624,’’ Memorandum from R. Slater and T. termining Volatile Organics at Microgram- Pressley, U.S. Environmental Protection per-Litre Levels by Gas Chromatography,’’ Agency, Environmental Monitoring and Sup- Journal American Water Works Association, port Laboratory, Cincinnati, Ohio 45268, Jan- 66: 739 (1974). uary 17, 1984. 2. ‘‘Sampling and Analysis Procedures for Screening of Industrial Effluents for Priority 18. Tables

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TABLE 1—PURGEABLES 1

CAS Registry μ 2 μ 3 Analyte No. MDL ( g/L) ML ( g/L)

Acrolein ...... 107–02–8 Acrylonitrile ...... 107–13–1 Benzene ...... 71–43–2 4.4 13.2 Bromodichloromethane ...... 75–27–4 2.2 6.6 Bromoform ...... 75–25–2 4.7 14.1 Bromomethane ...... 74–83–9 Carbon tetrachloride ...... 56–23–5 2.8 8.4 Chlorobenzene ...... 108–90–7 6.0 18.0 Chloroethane ...... 75–00–3 2-Chloroethylvinyl ether ...... 110–75–8 Chloroform ...... 67–66–3 1.6 4.8 Chloromethane ...... 74–87–3 Dibromochloromethane ...... 124–48–1 3.1 9.3 1,2-Dichlorobenzene ...... 95–50–1 1,3-Dichlorobenzene ...... 541–73–1 1,4-Dichlorobenzene ...... 106–46–7 1,1-Dichloroethane ...... 75–34–3 4.7 14.1 1,2-Dichloroethane ...... 107–06–2 2.8 8.4 1,1-Dichloroethene ...... 75–35–4 2.8 8.4 trans-1,2-Dichloroethene ...... 156–60–5 1.6 4.8 1,2-Dichloropropane ...... 78–87–5 6.0 18.0 cis-1,3-Dichloropropene ...... 10061–01–5 5.0 15.0 trans-1,3-Dichloropropene ...... 10061–02–6 Ethyl benzene ...... 100–41–4 7.2 21.6 Methylene chloride ...... 75–09–2 2.8 8.4 1,1,2,2-Tetrachloroethane ...... 79–34–5 6.9 20.7 Tetrachloroethene ...... 127–18–4 4.1 12.3 Toluene ...... 108–88–3 6.0 18.0 1,1,1-Trichloroethane ...... 71–55–6 3.8 11.4 1,1,2-Trichloroethane ...... 79–00–5 5.0 15.0 Trichloroethene ...... 79–01–6 1.9 5.7 Vinyl chloride ...... 75–01–4 1 All the analytes in this table are Priority Pollutants (40 CFR part 423, appendix A). 2 MDL values from the 1984 promulgated version of Method 624. 3 ML = Minimum Level—see Glossary for definition and derivation.

TABLE 2—ADDITIONAL PURGEABLES TABLE 2—ADDITIONAL PURGEABLES—Continued

Analyte CAS Registry Analyte CAS Registry

Acetone 1 ...... 67–64–1 2-Chlorotoluene ...... 95–49–8 Acetonitrile 2 ...... 75–05–8 4-Chlorotoluene ...... 106–43–4 Acrolein ...... 107–02–8 Crotonaldehyde 12 ...... 123–73–9 Acrylonitrile ...... 107–13–1 Cyclohexanone ...... 108–94–1 Allyl alcohol 1 ...... 107–18–6 1,2-Dibromo-3-chloropropane ...... 96–12–8 Allyl chloride ...... 107–05–1 1,2-Dibromoethane ...... 106–93–4 t-Amyl ethyl ether (TAEE) ...... 919–94–8 Dibromomethane ...... 74–95–3 t-Amyl methyl ether (TAME) ...... 994–058 cis-1,4-Dichloro-2-butene ...... 1476–11–5 Benzyl chloride ...... 100–44–7 trans-1,4-Dichloro-2-butene ...... 110–57–6 Bromoacetone 2 ...... 598–31–2 cis-1,2-Dichloroethene ...... 156–59–2 Bromobenzene ...... 108–86–1 Dichlorodifluoromethane ...... 75–71–8 Bromochloromethane ...... 74–97–5 1,3-Dichloropropane ...... 142–28–9 1,3-Butadiene ...... 106–99–0 2,2-Dichloropropane ...... 590–20–7 n-Butanol 1 ...... 71–36–3 1,3-Dichloro-2-propanol 2 ...... 96–23–1 2-Butanone (MEK) 12 ...... 78–93–3 1,1-Dichloropropene ...... 563–58–6 t-Butyl alcohol (TBA) ...... 75–65–0 cis-1,3-Dichloropropene ...... 10061–01–5 n-Butylbenzene ...... 104–51–8 1:2,3:4-Diepoxybutane ...... 1464–53–5 sec-Butylbenzene ...... 135–98–8 ...... 60–29–7 t-Butylbenzene ...... 98–06–6 Diisopropyl ether (DIPE) ...... 108–20–3 t-Butyl ethyl ether (ETBE) ...... 637–92–3 1,4-Dioxane 2 ...... 123–91–1 ...... 75–15–0 Epichlorohydrin 2 ...... 106–89–8 Chloral hydrate 2 ...... 302–17–0 Ethanol 2 ...... 64–17–5 Chloroacetonitrile 1 ...... 107–14–2 Ethyl acetate 2 ...... 141–78–6 1-Chlorobutane ...... 109–69–3 Ethyl methacrylate ...... 97–63–2 Chlorodifluoromethane ...... 75–45–6 Ethylene oxide 2 ...... 75–21–8 2-Chloroethanol 2 ...... 107–07–3 Hexachlorobutadiene ...... 87–63–3 bis (2-Chloroethyl) sulfide 2 ...... 505–60–2 Hexachloroethane ...... 67–72–1 1-Chlorohexanone ...... 20261–68–1 2-Hexanone 2 ...... 591–78–6 Chloroprene (2-chloro-1,3-butadiene) ...... 126–99–8 Iodomethane ...... 74–88–4 3-Chloropropene ...... 107–05–1 Isobutyl alcohol 1 ...... 78–83–1 3-Chloropropionitrile ...... 542–76–7 Isopropylbenzene ...... 98–82–8

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TABLE 2—ADDITIONAL PURGEABLES—Continued TABLE 3—EXAMPLE RETENTION TIMES— Continued Analyte CAS Registry Retention time p-Isopropyltoluene ...... 99–87–6 Analyte (min) Methacrylonitrile 2 ...... 126–98–7 Methanol 2 ...... 67–56–1 1,1-Dichloroethane ...... 8.45 2 Malonitrile ...... 109–77–3 Vinyl acetate ...... 8.55 Methyl acetate ...... 79–20–9 Allyl alcohol ...... 8.58 Methyl acrylate ...... 96–33–3 Methyl cyclohexane ...... 108–87–2 2-Chloro-1,3-butadiene ...... 8.65 Methyl iodide ...... 74–88–4 Methyl ethyl ketone ...... 9.50 Methyl methacrylate ...... 78–83–1 cis-1,2-Dichloroethene ...... 9.50 4-Methyl-2-pentanone (MIBK) 2 ...... 108–10–1 Ethyl cyanide ...... 9.57 Methyl-t-butyl ether (MTBE) ...... 1634–04–4 Methacrylonitrile ...... 9.83 Naphthalene ...... 91–20–3 Chloroform ...... 10.05 Nitrobenzene ...... 98–95–3 1,1,1-Trichloroethane ...... 10.37 N-Nitroso-di-n-butylamine 2 ...... 924–16–3 Carbon tetrachloride ...... 10.70 2-Nitropropane ...... 79–46–9 Isobutanol ...... 10.77 Paraldehyde 2 ...... 123–63–7 Benzene ...... 10.98 Pentachloroethane 2 ...... 76–01–7 Pentafluorobenzene ...... 363–72–4 1,2-Dichloroethane ...... 11.00 2-Pentanone 2 ...... 107–19–7 Crotonaldehyde ...... 11.45 2-Picoline 2 ...... 109–06–8 Trichloroethene ...... 12.08 1-Propanol 1 ...... 71–23–8 1,2-Dichloropropane ...... 12.37 2-Propanol 1 ...... 67–63–0 Methyl methacrylate ...... 12.55 Propargyl alcohol 2 ...... 107–19–7 p-Dioxane ...... 12.63 beta-Propiolactone 2 ...... 57–58–8 Dibromomethane ...... 12.65 Propionitrile (ethyl cyanide) 1 ...... 107–12–0 Bromodichloromethane ...... 12.95 n-Propylamine ...... 107–10–8 Chloroacetonitrile ...... 13.27 n-Propylbenzene ...... 103–65–1 2-Chloroethylvinyl ether ...... 13.45 2 ...... 110–86–1 cis-1,3-Dichloropropene ...... 13.65 Styrene ...... 100–42–5 1,1,1,2-Tetrachloroethane ...... 630–20–6 4-Methyl-2-pentanone ...... 13.83 Tetrahydrofuran ...... 109–99–9 Toluene ...... 14.18 o-Toluidine 2 ...... 95–53–4 trans-1,3-Dichloropropene ...... 14.57 1,2,3-Trichlorobenzene ...... 87–61–6 Ethyl methacrylate ...... 14.70 Trichlorofluoromethane ...... 75–69–4 1,1,2-Trichloroethane ...... 14.93 1,2,3-Trichloropropane ...... 96–18–4 1,3-Dichloropropane ...... 15.18 1,2,3-Trimethylbenzene ...... 526–73–8 Tetrachloroethene ...... 15.22 1,2,4-Trimethylbenzene ...... 95–63–6 2-Hexanone ...... 15.30 1,3,5-Trimethylbenzene ...... 108–67–8 Dibromochloromethane ...... 15.68 Vinyl acetate ...... 108–05–4 1,2-Dibromoethane ...... 15.90 m-Xylene 3 ...... 108–38–3 Chlorobenzene ...... 16.78 3 o-Xylene ...... 95–47–6 Ethylbenzene ...... 16.82 3 p-Xylene ...... 106–42–3 1,1,1,2-Tetrachloroethane ...... 16.87 m+o-Xylene 3 ...... 179601–22–0 m+p-Xylene ...... 17.08 m+p-Xylene 3 ...... 179601–23–1 o-Xylene ...... 17.82 o+p-Xylene 3 ...... 136777–61–2 Bromoform ...... 18.27 1 Determined at a purge temperature of 80 °C. Bromofluorobenzene ...... 18.80 2 ° May be detectable at a purge temperature of 80 C. 1,1,2,2-Tetrachloroethane ...... 18.98 3 Determined in combination separated by GC column. Most GC columns will resolve o-xylene from m+p-xylene. Re- 1,2,3-Trichloropropane ...... 19.08 port using the CAS number for the individual xylene or the trans-1,4-Dichloro-2-butene ...... 19.12 combination, as determined. Column: 75 m x 0.53 mm ID x 3.0 μm wide-bore DB–624 Conditions: 40 °C for 4 min, 9 °C/min to 200 °C, 20 °C/min TABLE 3—EXAMPLE RETENTION TIMES (or higher) to 250 °C, hold for 20 min at 250 °C to remove water. Carrier gas flow rate: 6–7 mL/min at 40 °C. Analyte Retention time (min) Inlet split ratio: 3:1. Interface split ratio: 7:2. Chloromethane ...... 3.68 Vinyl chloride ...... 3.92 TABLE 4—BFB KEY m/z ABUNDANCE CRITERIA 1 Bromomethane ...... 4.50 Chloroethane ...... 4.65 m/z Abundance criteria Trichlorofluoromethane ...... 5.25 Diethyl ether ...... 5.88 50 ...... 15–40% of m/z 95. Acrolein ...... 6.12 75 ...... 30–60% of m/z 95. 1,1-Dichloroethene ...... 6.30 Acetone ...... 6.40 95 ...... Base Peak, 100% Relative Iodomethane ...... 6.58 Abundance. Carbon disulfide ...... 6.72 96 ...... 5–9% of m/z 95. 3-Chloropropene ...... 6.98 173 ...... <2% of m/z 174. Methylene chloride ...... 7.22 174 ...... >50% of m/z 95. Acrylonitrile ...... 7.63 175 ...... 5–9% of m/z 174. trans-1,2-Dichloroethene ...... 7.73 176 ...... >95% but <101% of m/z 174.

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TABLE 4—BFB KEY m/z ABUNDANCE CRITERIA 1—Continued

m/z Abundance criteria

177 ...... 5–9% of m/z 176. 1 Abundance criteria are for a quadrupole mass spectrom- eter. Alternative tuning criteria from other published EPA ref- erence methods may be used, provided method performance is not adversely affected. Alternative tuning criteria specified by an instrument manufacturer may also be used for another type of mass spectrometer, or for an alternative carrier gas, provided method performance is not adversely affected.

TABLE 5—SUGGESTED SURROGATE AND INTERNAL STANDARDS

Analyte Retention time Primary m/z Secondary (min) 1 m/z’s

Benzene-d6 ...... 10.95 84 4-Bromofluorobenzene ...... 18.80 95 174, 176 Bromochloromethane ...... 9.88 128 49, 130, 51 2-Bromo-1-chloropropane ...... 14.80 77 79, 156 2-Butanone-d5 ...... 9.33 77 Chloroethane-d5 ...... 4.63 71 Chloroform-13C ...... 10.00 86 1,2-Dichlorobenzene-d4 ...... 152 1,4-Dichlorobutane ...... 18.57 55 90, 92 1,2-Dichloroethane-d4 ...... 10.88 102 1,1-Dichloroethene-d2 ...... 6.30 65 1,2-Dichloropropane-d6 ...... 12.27 67 trans-1,3-Dichloropropene-d4 ...... 14.50 79 1,4-Difluorobenzene ...... 114 63, 88 Ethylbenzene-d10 ...... 16.77 98 Fluorobenzene ...... 96 70 2-Hexanone-d5 ...... 15.30 63 Pentafluorobenzene ...... 168 1,1,2,2-Tetrachloroethane-d2 ...... 18.93 84 Toluene-d8 ...... 14.13 100 Vinyl chloride-d3 ...... 3.87 65 1 For chromatographic conditions, see the footnote to Table 3.

TABLE 6—CHARACTERISTIC m/z’s FOR PURGEABLE ORGANICS

Analyte Primary m/z Secondary m/z’s

Acrolein ...... 56 55 and 58. Acrylonitrile ...... 53 52 and 51. Chloromethane ...... 50 52. Bromomethane ...... 94 96. Vinyl chloride ...... 62 64. Chloroethane ...... 64 66. Methylene chloride ...... 84 49, 51, and 86. Trichlorofluoromethane ...... 101 103. 1,1-Dichloroethene ...... 96 61 and 98. 1,1-Dichloroethane ...... 63 65, 83, 85, 98, and 100. trans-1,2-Dichloroethene ...... 96 61 and 98. Chloroform ...... 83 85. 1,2-Dichloroethane ...... 98 62, 64, and 100. 1,1,1-Trichloroethane ...... 97 99, 117, and 119. Carbon tetrachloride ...... 117 119 and 121. Bromodichloromethane ...... 83 127, 85, and 129. 1,2-Dichloropropane ...... 63 112, 65, and 114. trans-1,3-Dichloropropene ...... 75 77. Trichloroethene ...... 130 95, 97, and 132. Benzene ...... 78 Dibromochloromethane ...... 127 129, 208, and 206. 1,1,2-Trichloroethane ...... 97 83, 85, 99, 132, and 134. cis-1,3-Dichloropropene ...... 75 77. 2-Chloroethylvinyl ether ...... 106 63 and 65. Bromoform ...... 173 171, 175, 250, 252, 254, and 256. 1,1,2,2-Tetrachloroethane ...... 168 83, 85, 131, 133, and 166. Tetrachloroethene ...... 164 129, 131, and 166. Toluene ...... 92 91. Chlorobenzene ...... 112 114.

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TABLE 6—CHARACTERISTIC m/z’s FOR PURGEABLE ORGANICS—Continued

Analyte Primary m/z Secondary m/z’s

Ethyl benzene ...... 106 91. 1,3-Dichlorobenzene ...... 146 148 and 111. 1,2-Dichlorobenzene ...... 146 148 and 111. 1,4-Dichlorobenzene ...... 146 148 and 111.

TABLE 7—LCS (Q), DOC (S AND X), AND MS/MSD (P AND RPD) ACCEPTANCE CRITERIA 1

Range for Q Limit for s Range for X Range for P1, Analyte P2 Limit for RPD (%) (%) (%) (%)

Acrolein ...... 60–140 30 50–150 40–160 60 Acrylonitrile ...... 60–140 30 50–150 40–160 60 Benzene ...... 65–135 33 75–125 37–151 61

Benzene-d6 ...... Bromodichloromethane ...... 65–135 34 50–140 35–155 56 Bromoform ...... 70–130 25 57–156 45–169 42 Bromomethane ...... 15–185 90 D–206 D–242 61

2-Butanone-d5 ...... Carbon tetrachloride...... 70–130 26 65–125 70–140 41 Chlorobenzene ...... 65–135 29 82–137 37–160 53 Chloroethane ...... 40–160 47 42–202 14–230 78

Chloroethane-d5 ...... 2-Chloroethylvinyl ether ...... D–225 130 D–252 D–305 71 Chloroform ...... 70–135 32 68–121 51–138 54 Chloroform-13C ...... Chloromethane ...... D–205 472 D–230 D–273 60 Dibromochloromethane ...... 70–135 30 69–133 53–149 50 1,2-Dichlorobenzene ...... 65–135 31 59–174 18–190 57

1,2-Dichlorobenzene-d4 ...... 1,3-Dichlorobenzene ...... 70–130 24 75–144 59–156 43 1,4-Dichlorobenzene ...... 65–135 31 59–174 18–190 57 1,1-Dichloroethane ...... 70–130 24 71–143 59–155 40 1,2-Dichloroethane ...... 70–130 29 72–137 49–155 49

1,2-Dichloroethane-d4 ...... 1,1-Dichloroethene ...... 50–150 40 19–212 D–234 32

1,1-Dichloroethene-d2 ...... trans-1,2-Dichloroethene ...... 70–130 27 68–143 54–156 45 1,2-Dichloropropane ...... 35–165 69 19–181 D–210 55

1,2-Dichloropropane-d6 ...... cis-1,3-Dichloropropene ...... 25–175 79 5–195 D–227 58 trans-1,3-Dichloropropene ...... 50–150 52 38–162 17–183 86

trans-1,3-Dichloropropene-d4 ...... Ethyl benzene ...... 60–140 34 75–134 37–162 63

2-Hexanone-d5 ...... Methylene chloride ...... 60–140 192 D–205 D–221 28 1,1,2,2-Tetrachloroethane ...... 60–140 36 68–136 46–157 61

1,1,2,2-Tetrachloroethane-d2 ...... Tetrachloroethene ...... 70–130 23 65–133 64–148 39 Toluene ...... 70–130 22 75–134 47–150 41

Toluene-d8 ...... 1,1,1-Trichloroethane ...... 70–130 21 69–151 52–162 36 1,1,2-Trichloroethane ...... 70–130 27 75–136 52–150 45 Trichloroethene ...... 65–135 29 75–138 70–157 48 Trichlorofluoromethane ...... 50–150 50 45–158 17–181 84 Vinyl chloride ...... 5–195 100 D–218 D–251 66

Vinyl chloride-d3. 1 Criteria were calculated using an LCS concentration of 20 μg/L. Q = Percent recovery in calibration verification/LCS (section 8.4). s = Standard deviation of percent recovery for four recovery measurements (section 8.2.4). X = Average percent recovery for four recovery measurements (section 8.2.4). P = Percent recovery for the MS or MSD (section 8.3.3). D = Detected; result must be greater than zero. Notes: 1. Criteria for pollutants are based upon the method performance data in Reference 4. Where necessary, limits have been broadened to assure applicability to concentrations below those used to develop Table 7. 2. Criteria for surrogates are from EPA CLP SOM01.2D.

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TABLE 8—RECOVERY AND PRECISION AS FUNCTIONS OF CONCENTRATION

Single analyst Overall Recovery, X′ ′ ′ Analyte μ precision, sr precision, S ( g/L) (μg/L) (μg/L)

Benzene ...... 0.93C+2.00 20.26 X¥1.74 0.25 X¥1.33 Bromodichloromethane ...... 1.03C¥1.58 0.15 X+0.59 0.20 X+1.13 Bromoform ...... 1.18C¥2.35 0.12 X+0.36 0.17 X+1.38 Bromomethane a ...... 1.00C 0.43 X 0.58 X Carbon tetrachloride ...... 1.10C¥1.68 0.12 X+0.25 0.11 X+0.37 Chlorobenzene ...... 0.98C+2.28 0.16 X¥0.09 0.26 X¥1.92 Chloroethane ...... 1.18C+0.81 0.14 X+2.78 0.29 X+1.75 2-Chloroethylvinyl ether a ...... 1.00C 0.62 X 0.84 X Chloroform ...... 0.93C+0.33 0.16 X+0.22 0.18 X+0.16 Chloromethane ...... 1.03C+0.81 0.37 X+2.14 0.58 X+0.43 Dibromochloromethane ...... 1.01C¥0.03 0.17 X¥0.18 0.17 X+0.49 1,2-Dichlorobenzene b ...... 0.94C+4.47 0.22 X¥1.45 0.30 X¥1.20 1,3-Dichlorobenzene ...... 1.06C+1.68 0.14 X¥0.48 0.18 X¥0.82 1,4-Dichlorobenzene b ...... 0.94C+4.47 0.22 X¥1.45 0.30 X¥1.20 1,1-Dichloroethane ...... 1.05C+0.36 0.13 X¥0.05 0.16 X+0.47 1,2-Dichloroethane ...... 1.02C+0.45 0.17 X¥0.32 0.21 X¥0.38 1,1-Dichloroethene ...... 1.12C+0.61 0.17 X+1.06 0.43 X¥0.22 trans-1,2,-Dichloroethene ...... 1.05C+0.03 0.14 X¥+0.09 0.19 X¥+0.17 1,2-Dichloropropane a ...... 1.00C 0.33 X 0.45 X cis-1,3-Dichloropropene a ...... 1.00C 0.38 X 0.52 X trans-1,3-Dichloropropene a ...... 1.00C 0.25 X 0.34 X Ethyl benzene ...... 0.98C+2.48 0.14 X+1.00 0.26 X¥1.72 Methylene chloride ...... 0.87C+1.88 0.15 X+1.07 0.32 X+4.00 1,1,2,2-Tetrachloroethane ...... 0.93C+1.76 0.16 X+0.69 0.20 X+0.41 Tetrachloroethene ...... 1.06C+0.60 0.13 X¥0.18 0.16 X¥0.45 Toluene ...... 0.98C+2.03 0.15 X¥0.71 0.22 X¥1.71 1,1,1-Trichloroethane ...... 1.06C+0.73 0.12 X¥0.15 0.21 X¥0.39 1,1,2-Trichloroethane ...... 0.95C+1.71 0.14 X+0.02 0.18 X+0.00 Trichloroethene ...... 1.04C+2.27 0.13 X+0.36 0.12 X+0.59 Trichlorofluoromethane ...... 0.99C+0.39 0.33 X¥1.48 0.34 X¥0.39 Vinyl chloride ...... 1.00C 0.48 X 0.65 X X′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L. Sr′ = Expected single analyst standard deviation of measurements at an average concentration found of X, in μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X, in μg/L. C = True value for the concentration, in μg/L. X = Average recovery found for measurements of samples containing a concentration of C, in μg/L. a Estimates based upon the performance in a single laboratory (References 4 and 16). b Due to coelutions, performance statements for these isomers are based upon the sums of their concentrations.

19. Glossary ms millisecond m/z mass-to-charge ratio These definitions and purposes are specific N normal; gram molecular weight of solute to this method, but have been conformed to divided by hydrogen equivalent of solute, common usage to the extent possible. 19.1 Units of weight and measure and per liter of solution their abbreviations. ng nanogram 19.1.1 Symbols. pg picogram °C degrees Celsius ppb part-per-billion μg microgram ppm part-per-million μL microliter ppt part-per-trillion < less than psig pounds-per-square inch gauge > greater than v/v volume per unit volume % percent w/v weight per unit volume 19.1.2 Abbreviations (in alphabetical 19.2 Definitions and acronyms (in alpha- order). betical order). cm centimeter Analyte—A compound tested for by this g gram method. The analytes are listed in Tables 1 h hour and 2. ID inside diameter Analyte of interest—An analyte of interest in. inch is an analyte required to be determined by a L liter regulatory/control authority or in a permit, m mass or by a client. mg milligram Analytical batch—The set of samples ana- min minute lyzed on a given instrument during a 12-hour mL milliliter period that begins with analysis of a calibra- mm millimeter tion verification/LCS. See section 8.4.

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Blank—An aliquot of reagent water that is and analyzed to establish the ability of the treated exactly as a sample including expo- laboratory to generate acceptable precision sure to all glassware, equipment, solvents, and recovery. A DOC is performed prior to reagents, internal standards, and surrogates the first time this method is used and any that are used with samples. The blank is time the method or instrumentation is modi- used to determine if analytes or inter- fied. ferences are present in the laboratory envi- Laboratory control sample (LCS; labora- ronment, the reagents, or the apparatus. See tory fortified blank (LFB); on-going preci- section 8.5. sion and recovery sample; OPR)—An aliquot Calibration—The process of determining of reagent water spiked with known quan- the relationship between the output or re- tities of the analytes of interest and surro- sponse of a measuring instrument and the gates. The LCS is analyzed exactly like a value of an input standard. Historically, sample. Its purpose is to assure that the re- EPA has referred to a multi-point calibra- sults produced by the laboratory remain tion as the ‘‘initial calibration,’’ to differen- within the limits specified in this method for tiate it from a single-point calibration precision and recovery. In this method, the verification. LCS is synonymous with a calibration Calibration standard—A solution prepared verification sample (See sections 7.4 and 8.4). from stock solutions and/or a secondary Laboratory fortified sample matrix—See standards and containing the analytes of in- Matrix spike. terest, surrogates, and internal standards. Laboratory reagent blank—See Blank. The calibration standard is used to calibrate Matrix spike (MS) and matrix spike dupli- the response of the GC/MS instrument cate (MSD) (laboratory fortified sample ma- against analyte concentration. trix and duplicate)—Two aliquots of an envi- Calibration verification standard—The lab- ronmental sample to which known quan- oratory control sample (LCS) used to verify tities of the analytes of interest and surro- calibration. See Section 8.4. gates are added in the laboratory. The MS/ Descriptor—In SIM, the beginning and end- MSD are prepared and analyzed exactly like ing retention times for the RT window, the a field sample. Their purpose is to quantify m/z’s sampled in the RT window, and the any additional bias and imprecision caused dwell time at each m/z. by the sample matrix. The background con- Extracted ion current profile (EICP)—The centrations of the analytes in the sample line described by the signal at a given m/z. matrix must be determined in a separate ali- Field duplicates—Two samples collected at quot and the measured values in the MS/ the same time and place under identical con- MSD corrected for background concentra- ditions, and treated identically throughout tions. field and laboratory procedures. Results of May—This action, activity, or procedural analyses of field duplicates provide an esti- step is neither required nor prohibited. mate of the precision associated with sample May not—This action, activity, or proce- collection, preservation, and storage, as well dural step is prohibited. as with laboratory procedures. Method blank (laboratory reagent blank)— Field blank—An aliquot of reagent water See Blank. or other reference matrix that is placed in a Method detection limit (MDL)—A detec- sample container in the field, and treated as tion limit determined by the procedure at 40 a sample in all respects, including exposure CFR part 136, appendix B. The MDLs deter- to sampling site conditions, storage, preser- mined by EPA in the original version of the vation, and all analytical procedures. The method are listed in Table 1. As noted in Sec. purpose of the field blank is to determine if 1.4, use the MDLs in Table 1 in conjunction the field or sample transporting procedures with current MDL data from the laboratory and environments have contaminated the actually analyzing samples to assess the sen- sample. sitivity of this procedure relative to project GC—Gas chromatograph or gas chroma- objectives and regulatory requirements tography. (where applicable). Internal standard—A compound added to a Minimum level (ML)—The term ‘‘minimum sample in a known amount and used as a ref- level’’ refers to either the sample concentra- erence for quantitation of the analytes of in- tion equivalent to the lowest calibration terest and surrogates. Internal standards are point in a method or a multiple of the meth- listed in Table 5. Also see Internal standard od detection limit (MDL), whichever is high- quantitation. er. Minimum levels may be obtained in sev- Internal standard quantitation—A means eral ways: They may be published in a meth- of determining the concentration of an od; they may be based on the lowest accept- analyte of interest (Tables 1 and 2) by ref- able calibration point used by a laboratory; erence to a compound added to a sample and or they may be calculated by multiplying not expected to be found in the sample. the MDL in a method, or the MDL deter- DOC—Initial demonstration of capability mined by a laboratory, by a factor of 3. For (DOC; section 8.2); four aliquots of reagent the purposes of NPDES compliance moni- water spiked with the analytes of interest toring, EPA considers the following terms to

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be synonymous: ‘‘quantitation limit,’’ ‘‘re- age) of the noise to the peak maximum di- porting limit,’’ and ‘‘minimum level.’’ vided by the width of the noise. MS—Mass spectrometer or mass spectrom- SIM—See Selection Ion Monitoring. etry. Should—This action, activity, or proce- Must—This action, activity, or procedural dural step is suggested but not required. step is required. Stock solution—A solution containing an m/z—The ratio of the mass of an ion (m) analyte that is prepared using a reference detected in the mass spectrometer to the material traceable to EPA, the National In- charge (z) of that ion. stitute of Science and Technology (NIST), or Quality control sample (QCS)—A sample a source that will attest to the purity and containing analytes of interest at known authenticity of the reference material. concentrations. The QCS is obtained from a Surrogate—A compound unlikely to be source external to the laboratory or is pre- found in a sample, and which is spiked into pared from standards obtained from a dif- sample in a known amount before purge-and- ferent source than the calibration standards. trap. The surrogate is quantitated with the The purpose is to check laboratory per- same procedures used to quantitate the formance using test materials that have analytes of interest. The purpose of the sur- been prepared independent of the normal rogate is to monitor method performance preparation process. with each sample. VOA—Volatile organic analysis: e.g., the Reagent water—Water demonstrated to be analysis performed by this method. free from the analytes of interest and poten- tially interfering substances at the MDLs for METHOD 625.1—BASE/NEUTRALS AND ACIDS BY the analytes in this method. GC/MS Regulatory compliance limit (or regu- latory concentration limit)—A limit on the 1. Scope and Application concentration or amount of a pollutant or 1.1 This method is for determination of contaminant specified in a nationwide stand- semivolatile organic pollutants in industrial ard, in a permit, or otherwise established by discharges and other environmental samples a regulatory/control authority. by gas chromatography combined with mass Relative retention time (RRT)—The ratio spectrometry (GC/MS), as provided under 40 of the retention time of an analyte to the re- CFR 136.1. This revision is based on a pre- tention time of its associated internal stand- vious protocol (Reference 1), on the basic re- ard. RRT compensates for small changes in vision promulgated October 26, 1984, and on the GC temperature program that can affect an interlaboratory method validation study the absolute retention times of the analyte (Reference 2). Although this method was and internal standard. RRT is a unitless validated through an interlaboratory study quantity. conducted in the early 1980s, the funda- Relative standard deviation (RSD)—The mental chemistry principles used in this standard deviation times 100 divided by the method remain sound and continue to apply. mean. Also termed ‘‘coefficient of vari- 1.2 The analytes that may be quali- ation.’’ tatively and quantitatively determined RF—Response factor. See section 7.3.3. using this method and their CAS Registry RSD—See relative standard deviation. numbers are listed in Tables 1 and 2. The Safety Data Sheet (SDS)—Written infor- method may be extended to determine the mation on a chemical’s toxicity, health haz- analytes listed in Table 3; however, extrac- ards, physical properties, fire, and reac- tion or gas chromatography of some of these tivity, including storage, spill, and handling analytes may make quantitative determina- precautions that meet the requirements of tion difficult. For example, benzidine is sub- OSHA, 29 CFR 1910.1200(g) and appendix D to ject to oxidative losses during extraction § 1910.1200. United Nations Globally Har- and/or solvent concentration. Under the al- monized System of Classification and Label- kaline conditions of the extraction, alpha- ling of Chemicals (GHS), third revised edi- BHC, gamma-BHC, endosulfan I and II, and tion, United Nations, 2009. endrin are subject to decomposition. Selected Ion Monitoring (SIM)—An MS Hexachlorocyclopentadiene is subject to technique in which a few m/z’s are mon- thermal decomposition in the inlet of the gas itored. When used with gas chromatography, chromatograph, chemical reaction in ace- the m/z’s monitored are usually changed pe- tone solution, and photochemical decomposi- riodically throughout the chromatographic tion. N-nitrosodiphenylamine and other run to correlate with the characteristic m/z’s nitrosoamines may decompose in the gas for the analytes, surrogates, and internal chromatographic inlet. The sample may be standards as they elute from the extracted at neutral pH if necessary to over- chromatographic column. The technique is come these or other decomposition problems often used to increase sensitivity and mini- that could occur at alkaline or acidic pH. mize interferences. EPA also has provided other methods (e.g., Signal-to-noise ratio (S/N)—The height of Method 607—Nitrosamines) that may be used the signal as measured from the mean (aver- for determination of some of these analytes.

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EPA encourages use of Method 625.1 to deter- differ from those listed, depending upon the mine additional compounds amenable to ex- nature of interferences in the sample matrix. traction and GC/MS. 1.5.1 EPA has promulgated this method at 1.3 The large number of analytes in Ta- 40 CFR part 136 for use in wastewater com- bles 1–3 of this method makes testing dif- pliance monitoring under the National Pol- ficult if all analytes are determined simulta- lutant Discharge Elimination System neously. Therefore, it is necessary to deter- (NPDES). The data reporting practices de- mine and perform quality control (QC) tests scribed in section 15.2 are focused on such for the ‘‘analytes of interest’’ only. Analytes monitoring needs and may not be relevant to of interest are those required to be deter- other uses of the method. mined by a regulatory/control authority or 1.5.2 This method includes ‘‘reporting lim- in a permit, or by a client. If a list of its’’ based on EPA’s ‘‘minimum level’’ (ML) analytes is not specified, the analytes in Ta- concept (see the glossary in section 22). Ta- bles 1 and 2 must be determined, at a min- bles 1, 2, and 3 contain MDL values and ML imum, and QC testing must be performed for values for many of the analytes. these analytes. The analytes in Tables 1 and 1.6 This method is performance-based. It 2, and some of the analytes in Table 3 have may be modified to improve performance been identified as Toxic Pollutants (40 CFR (e.g., to overcome interferences or improve 401.15), expanded to a list of Priority Pollut- the accuracy of results) provided all per- ants (40 CFR part 423, appendix A). formance requirements are met. 1.4 In this revision to Method 625, the pes- 1.6.1 Examples of allowed method modi- ticides and polychlorinated biphenyls (PCBs) fications are described at 40 CFR 136.6. Other have been moved from Table 1 to Table 3 examples of allowed modifications specific to (Additional Analytes) to distinguish these this method, including solid-phase extrac- analytes from the analytes required in qual- tion (SPE) are described in section 8.1.2. ity control tests (Tables 1 and 2). QC accept- 1.6.2 Any modification beyond those ex- ance criteria for pesticides and PCBs have pressly permitted at 40 CFR 136.6 or in sec- been retained in Table 6 and may continue to tion 8.1.2 of this method shall be considered be applied if desired, or if requested or re- a major modification subject to application quired by a regulatory/control authority or and approval of an alternate test procedure in a permit. Method 608.3 should be used for under 40 CFR 136.4 and 136.5. determination of pesticides and PCBs. How- 1.6.3 For regulatory compliance, any ever, if pesticides and/or PCBs are to be de- modification must be demonstrated to termined, an additional sample must be col- produce results equivalent or superior to re- lected and extracted using the pH adjust- sults produced by this method when applied ment and extraction procedures specified in to relevant wastewaters (section 8.3). Method 608.3. Method 1668C may be useful for 1.7 This method is restricted to use by or determination of PCBs as individual under the supervision of analysts experi- chlorinated biphenyl congeners, and Method enced in the use of a gas chromatograph/ 1699 may be useful for determination of pes- mass spectrometer and in the interpretation ticides. At the time of writing of this revi- of mass spectra. Each laboratory that uses sion, Methods 1668C and 1699 had not been ap- this method must demonstrate the ability to proved for use at 40 CFR part 136. The screen- generate acceptable results using the proce- ing procedure for 2,3,7,8-tetrachlorodibenzo- dure in Section 8.2. p-dioxin (2,3,7,8-TCDD) contained in the 1.8 Terms and units of measure used in version of Method 625 promulgated October this method are given in the glossary at the 26, 1984 has been replaced with procedures for end of the method. selected ion monitoring (SIM), and 2,3,7,8- TCDD may be determined using the SIM pro- 2. Summary of Method cedures. However, EPA Method 613 or 1613B should be used for analyte-specific deter- 2.1 A measured volume of sample, suffi- mination of 2,3,7,8-TCDD because of the focus cient to meet an MDL or reporting limit, is of these methods on this compound. Methods serially extracted with methylene chloride 613 and 1613B are approved for use at 40 CFR at pH 11–13 and again at a pH less than 2 part 136. using a separatory funnel or continuous liq- 1.5 Method detection limits (MDLs; Ref- uid/liquid extractor. erence 3) for the analytes in Tables 1, 2, and 2.2 The extract is concentrated to a vol- 3 are listed in those tables. These MDLs were ume necessary to meet the required compli- determined in reagent water (Reference 4). ance or detection limit, and analyzed by GC/ Advances in analytical technology, particu- MS. Qualitative identification of an analyte larly the use of capillary (open-tubular) col- in the extract is performed using the reten- umns, allowed laboratories to routinely tion time and the relative abundance of two achieve MDLs for the analytes in this meth- or more characteristic masses (m/z’s). Quan- od that are 2–10 times lower than those in titative analysis is performed using the in- the version promulgated in 1984. The MDL ternal standard technique with a single char- for an analyte in a specific wastewater may acteristic m/z.

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3. Contamination and Interferences etry or as a primary method for identifica- tion and quantification. While the use of 3.1 Solvents, reagents, glassware, and these enhanced techniques is encouraged, it other sample processing labware may yield is not required. artifacts, elevated baselines, or matrix inter- ferences causing misinterpretation of 4. Safety chromatograms and mass spectra. All mate- rials used in the analysis must be dem- 4.1 Hazards associated with each reagent onstrated to be free from contamination and used in this method have not been precisely interferences by analyzing blanks initially defined; however, each chemical compound and with each extraction batch (samples should be treated as a potential health haz- started through the extraction process in a ard. From this viewpoint, exposure to these given 24-hour period, to a maximum of 20 chemicals must be reduced to the lowest pos- samples—see Glossary for detailed defini- sible level by whatever means available. The tion), as described in Section 8.5. Specific se- laboratory is responsible for maintaining a lection of reagents and purification of sol- current awareness file of OSHA regulations vents by distillation in all-glass systems regarding the safe handling of the chemicals may be required. Where possible, labware is specified in this method. A reference file of cleaned by extraction or solvent rinse, or safety data sheets (SDSs, OSHA, 29 CFR baking in a kiln or oven. 1910.1200(g)) should also be made available to 3.2 Glassware must be scrupulously all personnel involved in sample handling cleaned (Reference 5). Clean all glassware as and chemical analysis. Additional references soon as possible after use by rinsing with the to laboratory safety are available and have last solvent used in it. Solvent rinsing been identified (References 7–9) for the infor- should be followed by detergent washing mation of the analyst. with hot water, and rinses with tap water 4.2 The following analytes covered by this and reagent water. The glassware should method have been tentatively classified as then be drained dry, and heated at 400 °C for known or suspected human or mammalian 15–30 minutes. Some thermally stable mate- carcinogens: Benzo(a)anthracene, benzidine, rials, such as PCBs, may require higher tem- 3,3′-dichlorobenzidine, benzo(a)pyrene, alpha- peratures and longer baking times for re- BHC, beta-BHC, delta-BHC, gamma-BHC, moval. Solvent rinses with pesticide quality Dibenz(a,h)-anthracene, N- acetone, hexane, or other solvents may be nitrosodimethylamine, 4,4′-DDT, and PCBs. substituted for heating. Do not heat volu- Other compounds in Table 3 may also be metric labware above 90 °C. After drying and toxic. Primary standards of toxic compounds cooling, store inverted or capped with sol- should be prepared in a chemical fume hood, vent-rinsed or baked aluminum foil in a and a NIOSH/MESA approved toxic gas res- clean environment to prevent accumulation pirator should be worn when handling high of dust or other contaminants. concentrations of these compounds. 3.3 Matrix interferences may be caused by 4.3 This method allows the use of hydro- contaminants co-extracted from the sample. gen as a carrier gas in place of helium (sec- The extent of matrix interferences will vary tion 5.6.1.2). The laboratory should take the considerably from source to source, depend- necessary precautions in dealing with hydro- ing upon the nature and diversity of the in- gen, and should limit hydrogen flow at the dustrial complex or municipality being sam- source to prevent buildup of an explosive pled. Interferences extracted from samples mixture of hydrogen in air. high in total organic carbon (TOC) may re- 5. Apparatus and Materials sult in elevated baselines, or by enhancing or suppressing a signal at or near the retention NOTE: Brand names, suppliers, and part time of an analyte of interest. Analyses of numbers are for illustration purposes only. the matrix spike and duplicate (section 8.3) No endorsement is implied. Equivalent per- may be useful in identifying matrix inter- formance may be achieved using equipment ferences, and gel permeation chroma- and materials other than those specified tography (GPC; Section 11.1) and sulfur re- here. Demonstrating that the equipment and moval (section 11.2) may aid in eliminating supplies used in the laboratory achieves the these interferences. EPA has provided guid- required performance is the responsibility of ance that may aid in overcoming matrix the laboratory. Suppliers for equipment and interferences (Reference 6). materials in this method may be found 3.4 In samples that contain an inordinate through an on-line search. Please do not con- number of interferences, the use of chemical tact EPA for supplier information. ionization (CI) or triple quadrupole (MRM) 5.1 Sampling equipment, for discrete or mass spectrometry may make identification composite sampling. easier. Tables 4 and 5 give characteristic CI 5.1.1 Grab sample bottle—amber glass bot- and MRM m/z’s for many of the analytes cov- tle large enough to contain the necessary ered by this method. The use of CI or MRM sample volume, fitted with a fluoropolymer- mass spectrometry may be utilized to sup- lined screw cap. Foil may be substituted for port electron ionization (EI) mass spectrom- fluoropolymer if the sample is not corrosive.

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If amber bottles are not available, protect 5.5 Balances. samples from light. Unless pre-cleaned, the 5.5.1 Analytical, capable of accurately bottle and cap liner must be washed, rinsed weighing 0.1 mg. with acetone or methylene chloride, and 5.5.2 Top loading, capable of accurately dried before use to minimize contamination. weighing 10 mg. 5.1.2 Automatic sampler (optional)—the 5.6 GC/MS system. sampler must incorporate a pre-cleaned glass 5.6.1 Gas chromatograph (GC)—An analyt- sample container. Samples must be kept re- ical system complete with a temperature frigerated at ≤6 °C and protected from light programmable gas chromatograph and all re- during compositing. If the sampler uses a quired accessories, including syringes and peristaltic pump, a minimum length of com- analytical columns. pressible silicone rubber tubing may be used. 5.6.1.1 Injection port—Can be split, Before use, however, rinse the compressible splitless, temperature programmable vapor- tubing with methanol, followed by repeated ization split/splitless (PTV), solvent-purge, rinsing with reagent water, to minimize the large-volume, on-column, backflushed, or potential for sample contamination. An inte- other. An autosampler is highly rec- grating flow meter is required to collect ommended because it injects volumes more flow-proportioned composites. precisely than volumes injected manually. 5.2 Glassware. 5.6.1.2 Carrier gas—Helium or hydrogen. 5.2.1 Separatory funnel—Size appropriate Data in the tables in this method were ob- to hold sample volume and extraction sol- tained using helium carrier gas. If hydrogen vent volume, and equipped with is used, analytical conditions may need to be fluoropolymer stopcock. adjusted for optimum performance, and cali- 5.2.2 Drying column—Chromatographic bration and all QC tests must be performed column, approximately 400 mm long by 19 mm ID, with coarse frit, or equivalent, suffi- with hydrogen carrier gas. See Section 4.3 cient to hold 15 g of anhydrous sodium sul- for precautions regarding the use of hydro- fate. gen as a carrier gas. 5.2.3 Concentrator tube, Kuderna-Dan- 5.6.2 GC column—See the footnotes to Ta- ish—10 mL, graduated (Kontes 570050–1025 or bles 4 and 5. Other columns or column sys- equivalent). Calibration must be checked at tems may be used provided all requirements the volumes employed in the test. A ground in this method are met. glass stopper is used to prevent evaporation 5.6.3 Mass spectrometer—Capable of re- of extracts. petitively scanning from 35–450 Daltons 5.2.4 Evaporative flask, Kuderna-Danish— (amu) every two seconds or less, utilizing a 500 mL (Kontes 57001–0500 or equivalent). At- 70 eV (nominal) electron energy in the elec- tach to concentrator tube with springs. tron impact ionization mode, and producing NOTE: Use of a solvent recovery system a mass spectrum which meets all the criteria with the K–D or other solvent evaporation in Table 9A or 9B when 50 ng or less of apparatus is strongly recommended. decafluorotriphenyl phosphine (DFTPP; CAS 5.2.5 Snyder column, Kuderna-Danish— 5074–71–5; bis(pentafluorophenyl) phenyl Three-ball macro (Kontes 503000–0121 or phosphine) is injected into the GC. equivalent). 5.6.4 GC/MS interface—Any GC to MS 5.2.6 Snyder column, Kuderna-Danish— interface that meets all performance re- Two-ball micro (Kontes 569001–0219 or equiva- quirements in this method may be used. lent). 5.6.5 Data system—A computer system 5.2.7 Vials—10–15 mL, amber glass, with must be interfaced to the mass spectrometer Teflon-lined screw cap. that allows the continuous acquisition and 5.2.8 Continuous liquid-liquid extractor— storage of mass spectra acquired throughout Equipped with fluoropolymer or glass con- the chromatographic program. The computer necting joints and stopcocks requiring no lu- must have software that allows searching brication. (Hershberg-Wolf Extractor, Ace any GC/MS data file for specific m/z’s Glass Company, Vineland, NJ, P/N 6848–20, or (masses) and plotting m/z abundances versus equivalent.) time or scan number. This type of plot is de- 5.2.9 In addition to the glassware listed fined as an extracted ion current profile above, the laboratory should be equipped (EICP). Software must also be available that with all necessary pipets, volumetric flasks, allows integrating the abundance at any beakers, and other glassware listed in this EICP between specified time or scan number method and necessary to perform analyses limits. successfully. 5.7 Automated gel permeation chro- 5.3 Boiling chips—Approximately 10/40 matograph (GPC). mesh, glass, silicon , or equivalent. 5.7.1 GPC column—150–700 mm long × 21–25 Heat to 400 °C for 30 minutes, or solvent rinse mm ID, packed with 70 g of SX–3 Biobeads; or Soxhlet extract with methylene chloride. Bio-Rad Labs, or equivalent. 5.4 Water bath—Heated, with concentric 5.7.2 Pump, injection valve, UV detector, ring cover, capable of temperature control and other apparatus necessary to meet the (±2 °C). The bath should be used in a hood. requirements in this method.

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5.8 Nitrogen evaporation device— fluoropolymer-lined screw-cap, or heat- Equipped with a water bath than can be sealed, glass containers, in the dark at ¥20 maintained at 30–45 °C; N-Evap, to ¥10 °C. Store aqueous standards; e.g., the Organomation Associates, or equivalent. aqueous LCS (section 8.4.1), in the dark at ≤ 5.9 Muffle furnace or kiln—Capable of 6 °C, but do not freeze. Standards prepared baking glassware or sodium sulfate in the by the laboratory may be stored for up to range of 400–450 °C. one year, except when comparison with QC check standards indicates that a standard 6. Reagents has degraded or become more concentrated 6.1 Reagent water—Reagent water is de- due to evaporation, or unless the laboratory fined as water in which the analytes of inter- has data on file to prove stability for a est and interfering compounds are not de- longer period. Commercially prepared stand- tected at the MDLs of the analytes of inter- ards may be stored until the expiration date est. provided by the vendor, except when com- 6.2 Sodium hydroxide solution (10 N)— parison with QC check standards indicates Dissolve 40 g of NaOH (ACS) in reagent water that a standard has degraded or become and dilute to 100 mL. more concentrated due to evaporation, or 6.3 Sodium thiosulfate—(ACS) granular. unless the laboratory has data from the ven- 6.4 Sulfuric acid (1+1)—Slowly add 50 mL dor on file to prove stability for a longer pe- of H2SO4 (ACS, sp. gr. 1.84) to 50 mL of rea- riod. gent water. 6.8 Surrogate standard spiking solution. 6.5 Acetone, methanol, methylene chlo- 6.8.1 Select a minimum of three surrogate ride, 2-propanol—High purity pesticide qual- compounds from Table 8 that most closely ity, or equivalent, demonstrated to be free of match the recovery of the analytes of inter- the analytes of interest and interferences est. For example, if all analytes tested are (Section 3). Purification of solvents by dis- considered acids, use surrogates that have tillation in all-glass systems may be re- similar chemical attributes. Other com- quired. pounds may be used as surrogates so long as 6.6 Sodium sulfate—(ACS) granular, anhy- they do not interfere in the analysis. If only drous, rinsed or Soxhlet extracted with one or two analytes are determined, one or methylene chloride (20 mL/g), baked in a two surrogates may be used. ° shallow tray at 450 C for one hour minimum, 6.8.2 Prepare a solution containing each cooled in a desiccator, and stored in a pre- selected surrogate such that the concentra- cleaned glass bottle with screw cap that pre- tion in the sample would match the con- vents moisture from entering. centration in the mid-point calibration 6.7 Stock standard solutions (1.00 μg/μL)— standard. For example, if the midpoint of the Stock standard solutions may be prepared calibration is 100 μg/L, prepare the spiking from pure materials, or purchased as cer- solution at a concentration of 100 μg/mL in tified solutions. Traceability must be to the methanol. Addition of 1.00 mL of this solu- National Institute of Standards and Tech- tion to 1000 mL of sample will produce a con- nology (NIST) or other national or inter- centration of 100 μg/L of the surrogate. Alter- national standard, when available. Stock so- lution concentrations alternate to those nate volumes and concentrations appropriate below may be used. Because of the toxicity to the response of the GC/MS instrument or of some of the compounds, primary dilutions for selective ion monitoring (SIM) may be should be prepared in a hood, and a NIOSH/ used, if desired. Store per section 6.7.2. MESA approved toxic gas respirator should 6.9 Internal standard spiking solution. be worn when high concentrations of neat 6.9.1 Select three or more internal stand- materials are handled. The following proce- ards similar in chromatographic behavior to dure may be used to prepare standards from the analytes of interest. Internal standards neat materials. are listed in Table 8. Suggested internal 6.7.1 Prepare stock standard solutions by standards are: 1,4-dichlorobenzene-d4; naph- accurately weighing about 0.0100 g of pure thalene-d8; acenaphthene-d10; phenanthrene- material. Dissolve the material in pesticide d10; chrysene-d12; and perylene-d12. The lab- quality methanol or other suitable solvent oratory must demonstrate that measure- and dilute to volume in a 10-mL volumetric ment of the internal standards is not af- flask. Larger volumes may be used at the fected by method or matrix interferences convenience of the laboratory. When com- (see also section 7.3.4). pound purity is assayed to be 96% or greater, 6.9.2 Prepare the internal standards at a the weight may be used without correction concentration of 10 mg/mL in methylene to calculate the concentration of the stock chloride or other suitable solvent. When 10 standard. Commercially prepared stock μL of this solution is spiked into a 1-mL ex- standards may be used at any concentration tract, the concentration of the internal if they are certified by the manufacturer or standards will be 100 μg/mL. A lower con- by an independent source. centration appropriate to the response of the 6.7.2 Unless stated otherwise in this meth- GC/MS instrument or for SIM may be used, if od, store non-aqueous standards in desired. Store per section 6.7.3.

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6.9.3 To assure accurate analyte identi- all traces of acid, then 3 times with acetone, fication, particularly when SIM is used, it then 3 times with hexane. may be advantageous to include more inter- 6.13.1.4 For copper foil, cover with hexane nal standards than those suggested in sec- after the final rinse. Store in a stoppered tion 6.9.1. An analyte will be located most flask under nitrogen until used. For the pow- accurately if its retention time relative to der, dry on a rotary evaporator or under a an internal standard is in the range of 0.8 to stream of nitrogen. Store in a stoppered 1.2. flask under nitrogen until used. Inspect the 6.10 DFTPP standard—Prepare a solution copper foil or powder before each use. It of DFTPP in methanol or other suitable sol- must have a bright, non-oxidized appearance vent such that 50 ng or less will be injected to be effective. Copper foil or powder that (see section 13.2). An alternative concentra- has oxidized may be reactivated using the tion may be used to compensate for specific procedure described above. injection volumes or to assure that the oper- 6.13.2 Tetrabutylammonium sodium sul- ating range of the instrument is not exceed- fite (TBA sodium sulfite). ed, so long as the total injected is 50 ng or 6.13.2.1 Tetrabutylammonium hydrogen less. Include benzidine and sulfate, [CH3(CH2)3]4NHSO4. pentachlorophenol in this solution such that 6.13.2.2 Sodium sulfite, Na2SO3. ≤100 ng of benzidine and ≤50 ng of 6.13.2.3 Dissolve approximately 3 g pentachlorophenol will be injected. tetrabutylammonium hydrogen sulfate in 100 6.11 Quality control check sample con- mL of reagent water in an amber bottle with centrate—See section 8.2.1. fluoropolymer-lined screw cap. Extract with three 20-mL portions of hexane and discard 6.12 GPC calibration solution. the hexane extracts. 6.12.1 Prepare a methylene chloride solu- 6.13.2.4 Add 25 g sodium sulfite to produce tion to contain corn oil, bis(2-ethylhexyl) a saturated solution. Store at room tempera- phthalate (BEHP), perylene, and sulfur at ture. Replace after 1 month. the concentrations in section 6.12.2, or at 6.14 DDT and endrin decomposition concentrations appropriate to the response (breakdown) solution—Prepare a solution of the detector. containing endrin at a concentration of 1 μg/ NOTE: Sulfur does not readily dissolve in mL and 4,4′-DDT at a concentration of 2 μg/ methylene chloride, but is soluble in warm mL, in isooctane or hexane. A 1-μL injection corn oil. The following procedure is sug- of this standard will contain 1 nanogram (ng) gested for preparation of the solution. of endrin and 2 ng of DDT. The concentration 6.12.2 Weigh 8 mg sulfur and 2.5 g corn oil of the solution may be adjusted by the lab- into a 100-mL volumetric flask and warm to oratory to accommodate other injection vol- dissolve the sulfur. Separately weigh 100 mg umes such that the same masses of the two BEHP, 20 mg pentachlorophenol, and 2 mg analytes are introduced into the instrument. perylene and add to flask. Bring to volume with methylene chloride and mix thor- 7. Calibration oughly. 7.1 Establish operating conditions equiva- 6.12.3 Store the solution in an amber glass lent to those in the footnote to Table 4 or 5 bottle with a fluoropolymer-lined screw cap for the base/neutral or acid fraction, respec- ° at 0–6 C. Protect from light. Refrigeration tively. If a combined base/neutral/acid frac- may cause the corn oil to precipitate. Before tion will be analyzed, use the conditions in use, allow the solution to stand at room tem- the footnote to Table 4. Alternative tempera- perature until the corn oil dissolves, or ture program and flow rate conditions may warm slightly to aid in dissolution. Replace be used. It is necessary to calibrate the GC/ the solution every year, or more frequently MS for the analytes of interest (Section 1.3) if the response of a component changes. only. 6.13 Sulfur removal—Copper foil or pow- 7.2 Internal standard calibration. der (bright, non-oxidized), or 7.2.1 Prepare calibration standards for the tetrabutylammonium sulfite (TBA sulfite). analytes of interest and surrogates at a min- 6.13.1 Copper foil, or powder—Fisher, Alfa imum of five concentration levels by adding Aesar 42455–18, 625 mesh, or equivalent. Cut appropriate volumes of one or more stock copper foil into approximately 1-cm squares. standards to volumetric flasks. One of the Copper must be activated before it may be calibration standards should be at a con- used, as described below: centration at or below the ML specified in 6.13.1.1 Place the quantity of copper need- Table 1, 2, or 3, or as specified by a regu- ed for sulfur removal (section 11.2.1.3) in a latory/control authority or in a permit. The ground-glass-stoppered Erlenmeyer flask or ML value may be rounded to a whole number bottle. Cover the foil or powder with meth- that is more convenient for preparing the anol. standard, but must not exceed the ML in 6.13.1.2 Add HCl dropwise (0.5–1.0 mL) Table 1, 2, or 3 for those analytes which list while swirling, until the copper brightens. ML values. Alternatively, the laboratory 6.13.1.3 Pour off the methanol/HCl and may establish a laboratory ML for each rinse 3 times with reagent water to remove analyte based on the concentration in a

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nominal whole-volume sample that is equiv- 7.2.1.2 Analyze the mid-point combined alent to the concentration of the lowest cali- base/neutral and acid calibration standard bration standard in a series of standards pro- and enter or review the retention time, rel- duced in the laboratory or obtained from a ative retention time, mass spectrum, and commercial vendor. The laboratory’s ML quantitation m/z in the data system for each must not exceed the ML in Table 1, 2, or 3, analyte of interest, surrogate, and internal and the resulting calibration must meet the standard. If additional analytes (Table 3) are acceptance criteria in Section 7.2.3, based on to be quantified, include these analytes in the RSD, RSE, or R2. The concentrations of the standard. The mass spectrum for each the other calibration standards should cor- analyte must be comprised of a minimum of respond to the expected range of concentra- 2 m/z’s (Tables 4 and 5); 3 to 5 m/z’s assure tions found in real samples or should define more reliable analyte identification. Sug- the working range of the GC/MS system for gested quantitation m/z’s are shown in Ta- full-scan and/or SIM operation, as appro- bles 4 and 5 as the primary m/z. If an inter- priate. A minimum of six concentration lev- ference occurs at the primary m/z, use one of els is required for a second order, non-linear the secondary m/z’s or an alternate m/z. A (e.g., quadratic; ax2 + bx + c = 0) calibration single m/z only is required for quantitation. (section 7.2.3). Calibrations higher than sec- 7.2.1.3 For SIM operation, determine the ond order are not allowed. To each calibra- analytes in each descriptor, the quantitation tion standard or standard mixture, add a m/z for each analyte (the quantitation m/z known constant volume of the internal can be the same as for full-scan operation; standard solution (section 6.9), and dilute to section 7.2.1.2), the dwell time on each m/z volume with methylene chloride. for each analyte, and the beginning and end- ing retention time for each descriptor. Ana- NOTE: The large number of analytes in Ta- bles 1 through 3 may not be soluble or stable lyze the verification standard in scan mode in a single solution; multiple solutions may to verify m/z’s and establish retention times be required if a large number of analytes are for the analytes. There must be a minimum to be determined simultaneously. of two m/z’s for each analyte to assure analyte identification. To maintain sensi- 7.2.1.1 Prior to analysis of the calibration tivity, the number of m/z’s in a descriptor standards, inject the DFTPP standard (Sec- should be limited. For example, for a tion 6.10) and adjust the scan rate of the descriptor with 10 m/z’s and a mass spectrometer to produce a minimum of chromatographic peak width of 5 sec, a dwell 5 mass spectra across the DFTPP GC peak. time of 100 ms at each m/z would result in a Adjust instrument conditions until the scan time of 1 second and provide 5 scans DFTPP criteria in Table 9A or 9B are met. across the GC peak. The quantitation m/z Calculate peak tailing factors for benzidine will usually be the most intense peak in the and pentachlorophenol. Calculation of the mass spectrum. The quantitation m/z and tailing factor is illustrated in Figure 1. The dwell time may be optimized for each tailing factor for benzidine and analyte. The acquisition table used for SIM pentachlorophenol must be <2; otherwise, ad- must take into account the mass defect (usu- just instrument conditions and either re- ally less than 0.2 Dalton) that can occur at place the column or break off a short section each m/z monitored. Refer to the footnotes of the front end of the column, and repeat to Table 4 or 5 for establishing operating the test. Once the scan conditions are estab- conditions and to section 7.2.1.1 for estab- lished, they must be used for analyses of all lishing scan conditions. standards, blanks, and samples. 7.2.1.4 For combined scan and SIM oper- NOTE: The DFTPP spectrum may be evalu- ation, set up the scan segments and ated by summing the intensities of the m/z’s descriptors to meet requirements in sections across the GC peak, subtracting the back- 7.2.1.1–7.2.1.3. Analyze unfamiliar samples in ground at each m/z in a region of the chro- the scan mode to assure that the analytes of matogram within 20 scans of but not includ- interest are determined. ing any part of, the DFTPP peak. The 7.2.2 Analyze each calibration standard DFTPP spectrum may also be evaluated by according to section 12 and tabulate the area fitting a Gaussian to each m/z and using the at the quantitation m/z against concentra- intensity at the maximum for each Gaussian tion for each analyte of interest, surrogate, or by integrating the area at each m/z and and internal standard. If an interference is using the integrated areas. Other means may encountered, use a secondary m/z (Table 4 or be used for evaluation of the DFTPP spec- 5) for quantitation. Calculate a response fac- trum so long as the spectrum is not distorted tor (RF) for each analyte of interest at each to meet the criteria in Table 9A or 9B. concentration using Equation 1.

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where: calibration verification tests are outside of the 12-hour shift. As = Area of the characteristic m/z for the analyte of interest or surrogate. 7.3.1 Analyze the calibration verification standard(s) beginning in section 12. Calculate Ais = Area of the characteristic m/z for the internal standard. the percent recovery of each analyte. Com- pare the recoveries for the analytes of inter- Cis = Concentration of the internal standard (μg/mL). est against the acceptance criteria for recov- ery (Q) in Table 6, and the recoveries for the Cs = Concentration of the analyte of interest or surrogate (μg/mL). surrogates against the acceptance criteria in Table 8. If recovery of the analytes of inter- 7.2. Calculate the mean (average) and rel- est and surrogates meet acceptance criteria, ative standard deviation (RSD) of the re- system performance is acceptable and anal- sponses factors. If the RSD is less than 35%, the RF can be assumed to be invariant and ysis of samples may continue. If any indi- the average RF can be used for calculations. vidual recovery is outside its limit, system Alternatively, the results can be used to fit performance is unacceptable for that a linear or quadratic regression of response analyte. ratios, As/Ais, vs. concentration ratios Cs/ NOTE: The large number of analytes in Ta- Cis. If used, the regression must be weighted bles 6 and 8 present a substantial probability inversely proportional to concentration. The that one or more will fail acceptance criteria coefficient of determination (R2; Reference when all analytes are tested simultaneously. 10) of the weighted regression must be great- 7.3.2 When one or more analytes fail ac- er than 0.920 (this value roughly corresponds ceptance criteria, analyze a second aliquot of to the RSD limit of 35%). Alternatively, the the calibration verification standard and relative standard error (Reference 11) may be compare ONLY those analytes that failed used as an acceptance criterion. As with the the first test (section 7.3.1) with their respec- RSD, the RSE must be less than 35%. If an tive acceptance criteria. If these analytes RSE less than 35% cannot be achieved for a now pass, system performance is acceptable quadratic regression, system performance is and analysis of samples may continue. A re- unacceptable and the system must be ad- peat failure of any analyte that failed the justed and re-calibrated. first test, however, will confirm a general NOTE: Using capillary columns and current problem with the measurement system. If instrumentation, it is quite likely that a lab- this occurs, repair the system (section oratory can calibrate the target analytes in 7.2.1.1) and repeat the test (section 7.3.1), or this method and achieve a linearity metric prepare a fresh calibration standard and re- (either RSD or RSE) well below 35%. There- peat the test. If calibration cannot be fore, laboratories are permitted to use more verified after maintenance or injection of stringent acceptance criteria for calibration the fresh calibration standard, re-calibrate than described here, for example, to har- the instrument. monize their application of this method with NOTE: If it is necessary to perform a repeat those from other sources. verification test frequently; i.e., perform two 7.3 Calibration verification—The RF or tests in order to pass, it may be prudent to calibration curve must be verified imme- perform two injections in succession and re- diately after calibration and at the begin- view the results, rather than perform one in- ning of each 12-hour shift, by analysis of a jection, review the results, then perform the standard at or near the concentration of the second injection if results from the first in- mid-point calibration standard (section jection fail. To maintain the validity of the 7.2.1). The standard(s) must be obtained from test and re-test, system maintenance and/or a second manufacturer or a manufacturer’s adjustment is not permitted between the in- batch prepared independently from the batch jections. used for calibration. Traceability must be to 7.3.3 Many of the analytes in Table 3 do a national standard, when available. Include not have QC acceptance criteria in Table 6, the surrogates (section 6.8) in this solution. and some of the surrogates in Table 8 do not It is necessary to verify calibration for the have acceptance criteria. If calibration is to analytes of interest (section 1.3) only. be verified and other QC tests are to be per- NOTE: The 12-hour shift begins after the formed for these analytes, acceptance cri- DFTPP (section 13.1) and DDT/endrin tests teria must be developed and applied. EPA (if DDT and endrin are to be determined), has provided guidance for development of QC and after analysis of the calibration acceptance criteria (References 12 and 13). verification standard. The 12-hour shift ends Alternatively, analytes that do not have ac- 12 hours later. The DFTPP, DDT/endrin, and ceptance criteria in Table 6 or Table 8 may

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be based on laboratory control charts, or 60 degrade method performance, are not al- to 140% may be used. lowed. If an analytical technique other than 7.3.4 Internal standard responses—Verify GC/MS is used, that technique must have a that detector sensitivity has not changed by specificity equal to or greater than the speci- comparing the response of each internal ficity of GC/MS for the analytes of interest. standard in the calibration verification The laboratory is also encouraged to partici- standard (section 7.3) to the response of the pate in inter-comparison and performance respective internal standard in the midpoint evaluation studies (see section 8.10). calibration standard (section 7.2.1). The peak 8.1.2.1 Each time a modification is made areas or heights of the internal standards in to this method, the laboratory is required to the calibration verification standard must be repeat the procedure in section 8.2. If the de- within 50% to 200% (1/2 to 2x) of their respec- tection limit of the method will be affected tive peak areas or heights in the mid-point by the change, the laboratory must dem- calibration standard. If not, repeat the cali- onstrate that the MDLs (40 CFR part 136, ap- bration verification test using a fresh cali- pendix B) are lower than one-third the regu- bration verification standard (7.3), or per- latory compliance limit or the MDLs in this form and document system repair. Subse- method, whichever are greater. If calibration quent to repair, repeat the calibration will be affected by the change, the instru- verification test (section 7.3.1). If the re- ment must be recalibrated per section 7. sponses are still not within 50% to 200%, re- Once the modification is demonstrated to calibrate the instrument (section 7.2.2) and produce results equivalent or superior to re- repeat the calibration verification test. sults produced by this method, that modi- fication may be used routinely thereafter, so 8. Quality Control long as the other requirements in this meth- 8.1 Each laboratory that uses this method od are met (e.g., matrix spike/matrix spike is required to operate a formal quality assur- duplicate recovery and relative percent dif- ance program. The minimum requirements ference). of this program consist of an initial dem- 8.1.2.1.1 If SPE, or another allowed meth- onstration of laboratory capability and on- od modification, is to be applied to a specific going analysis of spiked samples and blanks discharge, the laboratory must prepare and to evaluate and document data quality (40 analyze matrix spike/matrix spike duplicate CFR 136.7). The laboratory must maintain (MS/MSD) samples (section 8.3) and LCS records to document the quality of data gen- samples (section 8.4). The laboratory must erated. Results of ongoing performance tests include surrogates (section 8.7) in each of the are compared with established QC accept- samples. The MS/MSD and LCS samples ance criteria to determine if the results of must be fortified with the analytes of inter- analyses meet performance requirements of est (Section 1.3). If the modification is for this method. When results of spiked samples nationwide use, MS/MSD samples must be do not meet the QC acceptance criteria in prepared from a minimum of nine different this method, a quality control check sample discharges (See section 8.1.2.1.2), and all QC (laboratory control sample; LCS) must be acceptance criteria in this method must be analyzed to confirm that the measurements met. This evaluation only needs to be per- were performed in an in-control mode of op- formed once other than for the routine QC eration. A laboratory may develop its own required by this method (for example it performance criteria (as QC acceptance cri- could be performed by the vendor of the SPE teria), provided such criteria are as or more materials) but any laboratory using that restrictive than the criteria in this method. specific material must have the results of 8.1.1 The laboratory must make an initial the study available. This includes a full data demonstration of capability (DOC) to gen- package with the raw data that will allow an erate acceptable precision and recovery with independent reviewer to verify each deter- this method. This demonstration is detailed mination and calculation performed by the in Section 8.2. laboratory (see section 8.1.2.2.5, items (a)– 8.1.2 In recognition of advances that are (q)). occurring in analytical technology, and to 8.1.2.1.2 Sample matrices on which MS/ overcome matrix interferences, the labora- MSD tests must be performed for nationwide tory is permitted certain options (section 1.6 use of an allowed modification: and 40 CFR 136.6(b)) to improve separations (a) Effluent from a POTW. or lower the costs of measurements. These (b) ASTM D5905 Standard Specification for options may include alternate extraction, Substitute Wastewater. concentration, and cleanup procedures (e.g., (c) Sewage sludge, if sewage sludge will be solid-phase extraction; rotary-evaporator in the permit. concentration; column chromatography (d) ASTM D1141 Standard Specification for cleanup), changes in column and type of Substitute Ocean Water, if ocean water will mass spectrometer (40 CFR 136.6(b)(4)(xvi)). be in the permit. Alternate determinative techniques, such as (e) Untreated and treated wastewaters up substitution of spectroscopic or to a total of nine matrix types (see https:// immunoassay techniques, and changes that www.epa.gov/eg/industrial-effluent-guidelines

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for a list of industrial categories with exist- (n) Detector (type, operating conditions, ing effluent guidelines). etc). (i) At least one of the above wastewater (o) Chromatograms, mass spectra, and matrix types must have at least one of the other recordings of raw data. following characteristics: (p) Quantitation reports, data system out- (A) Total suspended solids greater than 40 puts, and other data to link the raw data to mg/L. the results reported. (B) Total dissolved solids greater than 100 (q) A written Standard Operating Proce- mg/L. dure (SOP). (C) Oil and grease greater than 20 mg/L. 8.1.2.2.6 Each individual laboratory wish- (D) NaCl greater than 120 mg/L. ing to use a given modification must perform (E) CaCO3 greater than 140 mg/L. the start-up tests in section 8.1.2 (e.g., DOC, (ii) Results of MS/MSD tests must meet QC MDL), with the modification as an integral acceptance criteria in Section 8.3. part of this method prior to applying the (f) A proficiency testing (PT) sample from modification to specific discharges. Results a recognized provider, in addition to tests of of the DOC must meet the QC acceptance cri- the nine matrices (section 8.1.2.1.1). teria in Table 6 for the analytes of interest 8.1.2.2 The laboratory is required to main- (section 1.3), and the MDLs must be equal to tain records of modifications made to this or lower than the MDLs in Tables 1, 2, or 3 method. These records include the following, for the analytes of interest. at a minimum: 8.1.3 Before analyzing samples, the lab- 8.1.2.2.1 The names, titles, and business oratory must analyze a blank to dem- street addresses, telephone numbers, and onstrate that interferences from the analyt- email addresses, of the analyst(s) that per- ical system, labware, and reagents, are under formed the analyses and modification, and of control. Each time a batch of samples is ex- the quality control officer that witnessed tracted or reagents are changed, a blank and will verify the analyses and modifica- must be extracted and analyzed as a safe- tions. guard against laboratory contamination. Re- 8.1.2.2.2 A list of analytes, by name and quirements for the blank are given in section CAS Registry Number. 8.5. 8.1.2.2.3 A narrative stating reason(s) for 8.1.4 The laboratory must, on an ongoing the modifications. basis, spike and analyze to monitor and 8.1.2.2.4 Results from all quality control evaluate method and laboratory performance (QC) tests comparing the modified method to on the sample matrix. The procedure for this method, including: spiking and analysis is given in section 8.3. (a) Calibration (section 7). 8.1.5 The laboratory must, on an ongoing (b) Calibration verification (section 7). basis, demonstrate through analysis of a (c) Initial demonstration of capability (sec- quality control check sample (laboratory tion 8.2). control sample, LCS; on-going precision and (d) Analysis of blanks (section 8.5). recovery sample, OPR) that the measure- (e) Matrix spike/matrix spike duplicate ment system is in control. This procedure is analysis (section 8.3). given in section 8.4. (f) Laboratory control sample analysis 8.1.6 The laboratory must maintain per- (section 8.4). formance records to document the quality of 8.1.2.2.5 Data that will allow an inde- data that is generated. This procedure is pendent reviewer to validate each deter- given in section 8.9. mination by tracing the instrument output 8.1.7 The large number of analytes tested (peak height, area, or other signal) to the in performance tests in this method present final result. These data are to include: a substantial probability that one or more (a) Sample numbers and other identifiers. will fail acceptance criteria when many (b) Extraction dates. analytes are tested simultaneously, and a re- (c) Analysis dates and times. test is allowed if this situation should occur. (d) Analysis sequence/run chronology. If, however, continued re-testing results in (e) Sample weight or volume (ssection 10). further repeated failures, the laboratory (f) Extract volume prior to each cleanup must document and report the failures (e.g., step (sections 10 and 11). as qualifiers on results), unless the failures (g) Extract volume after each cleanup step are not required to be reported as deter- (section 11). mined by the regulatory/control authority. (h) Final extract volume prior to injection Results associated with a QC failure for an (sections 10 and 12). analyte regulated in a discharge cannot be (i) Injection volume (section 12.2.3). used to demonstrate regulatory compliance. (j) Sample or extract dilution (section QC failures do not relieve a discharger or 12.2.3.2). permittee of reporting timely results. (k) Instrument and operating conditions. 8.2 Initial demonstration of capability (l) Column (dimensions, material, etc). (DOC)—To establish the ability to generate (m) Operating conditions (temperature acceptable recovery and precision, the lab- program, flow rate, etc). oratory must perform the DOC in sections

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8.2.1 through 8.2.6 for the analytes of inter- outside the range for recovery, system per- est. The laboratory must also establish formance is unacceptable for that analyte. MDLs for the analytes of interest using the NOTE: The large number of analytes in Ta- MDL procedure at 40 CFR part 136, appendix bles 1–3 present a substantial probability B. The laboratory’s MDLs must be equal to that one or more will fail at least one of the or lower than those listed in Tables 1, 2, or acceptance criteria when many or all 3 or lower than one third the regulatory analytes are determined simultaneously. compliance limit, whichever is greater. For Therefore, the analyst is permitted to con- MDLs not listed in Tables 4 and 5, the lab- duct a ‘‘re-test’’ as described in section 8.2.6. oratory must determine the MDLs using the 8.2.6 When one or more of the analytes MDL procedure at 40 CFR part 136, appendix tested fail at least one of the acceptance cri- B under the same conditions used to deter- teria, repeat the test for only the analytes mine the MDLs for the analytes listed in Ta- that failed. If results for these analytes pass, bles 1, 2, and 3. All procedures used in the system performance is acceptable and anal- analysis, including cleanup procedures, must ysis of samples and blanks may proceed. If be included in the DOC. one or more of the analytes again fail, sys- 8.2.1 For the DOC, a QC check sample con- tem performance is unacceptable for the centrate (LCS concentrate) containing each analytes that failed the acceptance criteria. analyte of interest (section 1.3) is prepared in Correct the problem and repeat the test (sec- a water-miscible solvent. The QC check sam- tion 8.2). See section 8.1.7 for disposition of ple concentrate must be prepared independ- repeated failures. ently from those used for calibration, but NOTE: To maintain the validity of the test may be from the same source as the second- and re-test, system maintenance and/or ad- source standard used for calibration justment is not permitted between this pair verification (Section 7.3). The concentrate of tests. should produce concentrations of the 8.3 Matrix spike and matrix spike dupli- cate (MS/MSD)—The purpose of the MS/MSD analytes of interest in water at the mid- requirement is to provide data that dem- point of the calibration range, and may be at onstrate the effectiveness of the method as the same concentration as the LCS (section applied to the samples in question by a given 8.4). Multiple solutions may be required. laboratory, and both the data user (dis- OTE: QC check sample concentrates are N charger, permittee, regulated entity, regu- no longer available from EPA. latory/control authority, customer, other) 8.2.2 Using a pipet or micro-syringe, pre- and the laboratory share responsibility for pare four LCSs by adding an appropriate vol- provision of such data. The data user should ume of the concentrate to each of four identify the sample and the analytes of in- aliquots of reagent water, and mix well. The terest (section 1.3) to be spiked and provide volume of reagent water must be the same as sufficient sample volume to perform MS/ the volume that will be used for the sample, MSD analyses. The laboratory must, on an blank (section 8.5), and MS/MSD (section 8.3). ongoing basis, spike at least 5% of the sam- A volume of 1–L and a concentration of 100 ples in duplicate from each discharge being μg/L were used to develop the QC acceptance monitored to assess accuracy (recovery and criteria in Table 6. Also add an aliquot of the precision). If direction cannot be obtained surrogate spiking solution (section 6.8) to from the data user, the laboratory must the reagent-water aliquots. spike at least one sample in duplicate per ex- 8.2.3 Extract and analyze the four LCSs traction batch of up to 20 samples with the according to the method beginning in Sec- analytes in Table 1. Spiked sample results tion 10. should be reported only to the data user 8.2.4 Calculate the average percent recov- whose sample was spiked, or as requested or ery (X) and the standard deviation of the per- required by a regulatory/control authority, cent recovery (s) for each analyte using the or in a permit. four results. 8.3.1 If, as in compliance monitoring, the 8.2.5 For each analyte, compare s and (X) concentration of a specific analyte will be with the corresponding acceptance criteria checked against a regulatory concentration for precision and recovery in Table 6. For limit, the concentration of the spike should analytes in Table 3 not listed in Table 6, DOC be at that limit; otherwise, the concentra- QC acceptance criteria must be developed by tion of the spike should be one to five times the laboratory. EPA has provided guidance higher than the background concentration for development of QC acceptance criteria determined in section 8.3.2, at or near the (References 12 and 13). Alternatively, accept- midpoint of the calibration range, or at the ance criteria for analytes not listed in Table concentration in the LCS (section 8.4) which- 6 may be based on laboratory control charts. ever concentration would be larger. If s and (X) for all analytes of interest meet 8.3.2 Analyze one sample aliquot to deter- the acceptance criteria, system performance mine the background concentration (B) of is acceptable and analysis of blanks and sam- the each analyte of interest. If necessary, ples may begin. If any individual s exceeds prepare a new check sample concentrate the precision limit or any individual (X) falls (section 8.2.1) appropriate for the background

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concentration. Spike and analyze two addi- updated at least every two years and re-es- tional sample aliquots, and determine the tablished after any major change in the ana- concentration after spiking (A1 and A2) of lytical instrumentation or process. If in- each analyte. Calculate the percent recov- house QC limits are developed, at least 80% eries (P1 and P2) as 100 (A1 ¥ B)/T and 100 (A2 of the analytes tested in the MS/MSD must ¥ B)/T, where T is the known true value of have in-house QC acceptance criteria that the spike. Also calculate the relative percent are tighter than those in Table 6, and the re- difference (RPD) between the concentrations maining analytes (those other than the (A1 and A2) as 200 |A1 ¥ A2|/(A1 + A2). If nec- analytes included in the 80%) must meet the essary, adjust the concentrations used to acceptance criteria in Table 6. If an in-house calculate the RPD to account for differences QC limit for the RPD is greater than the in the volumes of the spiked aliquots. limit in Table 6, then the limit in Table 6 8.3.3 Compare the percent recoveries (P1 must be used. Similarly, if an in-house lower and P2) and the RPD for each analyte in the limit for recovery is below the lower limit in MS/MSD aliquots with the corresponding QC Table 6, then the lower limit in Table 6 must acceptance criteria in Table 6. A laboratory be used, and if an in-house upper limit for re- may develop and apply QC acceptance cri- covery is above the upper limit in Table 6, teria more restrictive than the criteria in then the upper limit in Table 6 must be used. Table 6, if desired. 8.4 Laboratory control sample (LCS)—A 8.3.3.1 If any individual P falls outside the QC check sample (laboratory control sample, designated range for recovery in either ali- LCS; on-going precision and recovery sam- quot, or the RPD limit is exceeded, the re- ple, OPR) containing each analyte of interest sult for the analyte in the unspiked sample (Section 1.3) and surrogate must be prepared is suspect. See Section 8.1.7 for disposition of and analyzed with each extraction batch of failures. up to 20 samples to demonstrate acceptable 8.3.3.2 The acceptance criteria in Table 6 recovery of the analytes of interest from a were calculated to include an allowance for clean sample matrix. error in measurement of both the back- 8.4.1 Prepare the LCS by adding QC check ground and spike concentrations, assuming a sample concentrate (section 8.2.1) to reagent spike to background ratio of 5:1. This error will be accounted for to the extent that the water. Include all analytes of interest (sec- spike to background ratio approaches 5:1 tion 1.3) in the LCS. The LCS may be the (Reference 14) and is applied to spike con- same sample prepared for the DOC (section centrations of 100 μg/L and higher. If spiking 8.2.1). The volume of reagent water must be is performed at a concentration lower than the same as the volume used for the sample, 100 μg/L, the laboratory must use the QC ac- blank (section 8.5), and MS/MSD (Section ceptance criteria in Table 6, the optional QC 8.3). Also add an aliquot of the surrogate acceptance criteria calculated for the spe- spiking solution (section 6.8). The concentra- cific spike concentration in Table 7, or op- tion of the analytes in reagent water should tional in-house criteria (section 8.3.4). To use be the same as the concentration in the DOC the acceptance criteria in Table 7: (1) Cal- (section 8.2.2). culate recovery (X′) using the equation in 8.4.2 Analyze the LCS prior to analysis of Table 7, substituting the spike concentration field samples in the extraction batch. Deter- (T) for C; (2) Calculate overall precision (S′) mine the concentration (A) of each analyte. using the equation in Table 7, substituting X′ Calculate the percent recovery (PS) as 100 for X; (3) Calculate the range for recovery at (A/T)%, where T is the true value of the con- the spike concentration as (100 X′/T) ± centration in the LCS. 2.44(100 S′/T)% (Reference 14). For analytes in 8.4.3 Compare the percent recovery (PS) Table 3 not listed in Table 6, QC acceptance for each analyte with its corresponding QC criteria must be developed by the laboratory. acceptance criterion in Table 6. For analytes EPA has provided guidance for development of interest in Table 3 not listed in Table 6, of QC acceptance criteria (References 12 and use the QC acceptance criteria developed for 13). Alternatively, acceptance criteria may the LCS (section 8.4.5), or limits based on be based on laboratory control charts. laboratory control charts. If the recoveries 8.3.4 After analysis of a minimum of 20 for all analytes of interest fall within their MS/MSD samples for each target analyte and respective QC acceptance criteria, analysis surrogate, and if the laboratory chooses to of blanks and field samples may proceed. If develop and apply the optional in-house QC any individual PS falls outside the range, limits (Section 8.3.3), the laboratory should proceed according to section 8.4.4. calculate and apply the optional in-house QC NOTE: The large number of analytes in Ta- limits for recovery and RPD of future MS/ bles 1–3 present a substantial probability MSD samples (Section 8.3). The QC limits for that one or more will fail the acceptance cri- recovery are calculated as the mean ob- teria when all analytes are tested simulta- served recovery ±3 standard deviations, and neously. Because a re-test is allowed in event the upper QC limit for RPD is calculated as of failure (sections 8.1.7 and 8.4.3), it may be the mean RPD plus 3 standard deviations of prudent to extract and analyze two LCSs to- the RPDs. The in-house QC limits must be gether and evaluate results of the second

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analysis against the QC acceptance criteria ysis of the LCS (section 8.4) and prior to only if an analyte fails the first test. analysis of the MS/MSD and samples to dem- 8.4.4 Repeat the test only for those onstrate freedom from contamination. analytes that failed to meet the acceptance 8.5.2 If an analyte of interest is found in criteria (PS). If these analytes now pass, sys- the blank: At a concentration greater than tem performance is acceptable and analysis the MDL for the analyte, at a concentration of blanks and samples may proceed. Re- greater than one-third the regulatory com- peated failure, however, will confirm a gen- pliance limit, or at a concentration greater eral problem with the measurement system. than one-tenth the concentration in a sam- If this occurs, repeat the test using a fresh ple in the extraction batch, whichever is LCS (section 8.2.2) or an LCS prepared with greater, analysis of samples must be halted, a fresh QC check sample concentrate (sec- and the problem corrected. If the contamina- tion 8.2.1), or perform and document system tion is traceable to the extraction batch, repair. Subsequent to analysis of the LCS samples affected by the blank must be re-ex- prepared with a fresh sample concentrate, or tracted and the extracts re-analyzed. If, how- to system repair, repeat the LCS test (sec- ever, continued re-testing results in repeated tion 8.4). If failure of the LCS indicates a blank contamination, the laboratory must systemic problem with samples in the batch, document and report the failures (e.g., as re-extract and re-analyze the samples in the qualifiers on results), unless the failures are batch. See section 8.1.7 for disposition of re- not required to be reported as determined by peated failures. the regulatory/control authority. Results as- NOTE: To maintain the validity of the test sociated with blank contamination for an and re-test, system maintenance and/or ad- analyte regulated in a discharge cannot be justment is not permitted between the pair used to demonstrate regulatory compliance. of tests. QC failures do not relieve a discharger or 8.4.5 After analysis of 20 LCS samples, and permittee of reporting timely results. if the laboratory chooses to develop and 8.6 Internal standards responses. apply in-house QC limits, the laboratory should calculate and apply in-house QC lim- 8.6.1 Calibration verification—The re- its for recovery to future LCS samples (sec- sponses (GC peak heights or areas) of the in- tion 8.4). Limits for recovery in the LCS ternal standards in the calibration should be calculated as the mean recovery ±3 verification must be within 50% to 200% (1/2 standard deviations. A minimum of 80% of to 2x) of their respective responses in the the analytes tested for in the LCS must have mid-point calibration standard. If they are QC acceptance criteria tighter than those in not, repeat the calibration verification (Sec- Table 6, and the remaining analytes (those tion 7.4) test or perform and document sys- other than the analytes included in the 80%) tem repair. Subsequent to repair, repeat the must meet the acceptance criteria in Table calibration verification. If the responses are 6. If an in-house lower limit for recovery is still not within 50% to 200%, re-calibrate the lower than the lower limit in Table 6, the instrument (Section 7) and repeat the cali- lower limit in Table 6 must be used, and if an bration verification test. in-house upper limit for recovery is higher 8.6.2 Samples, blanks, LCSs, and MS/ than the upper limit in Table 6, the upper MSDs—The responses (GC peak heights or limit in Table 6 must be used. Many of the areas) of each internal standard in each sam- analytes and surrogates do not contain ac- ple, blank, and MS/MSD must be within 50% ceptance criteria. The laboratory should use to 200% (1/2 to 2x) of its respective response 60–140% as interim acceptance criteria for re- in the LCS for the extraction batch. If, as a coveries of spiked analytes and surrogates group, all internal standards are not within that do not have recovery limits specified in this range, perform and document system re- Table 8, and at least 80% of the surrogates pair, repeat the calibration verification (sec- must meet the 60–140% interim criteria until tion 8.4), and re-analyze the affected sam- in-house LCS and surrogate limits are devel- ples. If a single internal standard is not oped. Alternatively, acceptance criteria for within the 50% to 200% range, use an alter- analytes that do not have recovery limits in nate internal standard for quantitation of Table 6 may be based on laboratory control the analyte referenced to the affected inter- charts. In-house QC acceptance criteria must nal standard. It may be necessary to use the be updated at least every two years. data system to calculate a new response fac- 8.5 Blank—A blank must be extracted and tor from calibration data for the alternate analyzed with each extraction batch to dem- internal standard/analyte pair. If an internal onstrate that the reagents and equipment standard fails the 50–200% criteria and no used for preparation and analysis are free analytes are detected in the sample, ignore from contamination. the failure or report it if required by the reg- 8.5.1 Spike the surrogates into the blank. ulatory/control authority. Extract and concentrate the blank using the 8.7 Surrogate recoveries—The laboratory same procedures and reagents used for the must evaluate surrogate recovery data in samples, LCS, and MS/MSD in the batch. each sample against its in-house surrogate Analyze the blank immediately after anal- recovery limits. The laboratory may use 60–

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140% as interim acceptance criteria for re- collection. Automatic sampling equipment coveries for surrogates not listed in Table 8. must be as free as possible of polyvinyl chlo- At least 80% of the surrogates must meet the ride or other tubing or other potential 60–140% interim criteria until in-house limits sources of contamination. If needed, collect are developed. Alternatively, surrogate re- additional sample(s) for the MS/MSD (sec- covery limits may be developed from labora- tion 8.3). tory control charts, but such limits must be 9.2 Ice or refrigerate samples at ≤6 °C at least as restrictive as those in Table 8. from the time of collection until extraction, Spike the surrogates into all samples, but do not freeze. If residual chlorine is blanks, LCSs, and MS/MSDs. Compare surro- present, add 80 mg of sodium thiosulfate per gate recoveries against the QC acceptance liter of sample and mix well. Any method criteria in Table 8 and/or those developed in suitable for field use may be employed to section 7.3.3 or 8.4.5. If any recovery fails its test for residual chlorine (Reference 16). Add criteria, attempt to find and correct the more sodium sulfate if 80 mg/L is insufficient cause of the failure. See section 8.1.7 for dis- but do not add excess sodium thiosulfate. If position of failures. sodium thiosulfate interferes in the deter- 8.8 DDT and endrin decomposition (break- mination of the analytes, an alternate pre- down)—If DDT and/or endrin are to be ana- servative (e.g., ascorbic acid or sodium sul- lyzed using this method, the DDT/endrin de- fite) may be used. If preservative has been composition test in section 13.8 must be per- added, shake the sample vigorously for one formed to reliably quantify these two pes- minute. Maintain the hermetic seal on the ticides. sample bottle until time of analysis. 8.9 As part of the QC program for the lab- 9.3 All samples must be extracted within 7 oratory, control charts or statements of ac- days of collection and sample extracts must curacy for wastewater samples must be as- be analyzed within 40 days of extraction. sessed and records maintained (40 CFR 136.7(c)(1)(viii)). After analysis of five or 10. Extraction more spiked wastewater samples as in sec- tion 8.3, calculate the average percent recov- 10.1 This section contains procedures for separatory funnel liquid-liquid extraction ery (PX) and the standard deviation of the percent recovery (sp). Express the accuracy (SFLLE) and continuous liquid-liquid ex- traction (CLLE). SFLLE is faster, but may assessment as a percent interval from PX not be as effective as CLLE for recovery of ¥2sp to PX +2sp. For example, if PX = 90% and sp = 10%, the accuracy interval is ex- polar analytes such as phenol. SFLLE is pressed as 70–110%. Update the accuracy as- labor intensive and may result in formation sessment for each analyte on a regular basis of emulsions that are difficult to break. (e.g., after each 5–10 new accuracy measure- CLLE is less labor intensive, avoids emulsion ments). If desired, statements of accuracy formation, but requires more time (18–24 for laboratory performance, independent of hours) and more hood space, and may require performance on samples, may be developed more solvent. The procedures assume base- using LCSs. neutral extraction followed by acid extrac- 8.10 It is recommended that the labora- tion. For some matrices and analytes of in- tory adopt additional quality assurance terest, improved results may be obtained by practices for use with this method. The spe- acid-neutral extraction followed by base ex- cific practices that are most productive de- traction. A single acid or base extraction pend upon the needs of the laboratory and may also be performed. If an extraction the nature of the samples. Field duplicates scheme alternate to base-neutral followed by may be analyzed to assess the precision of acid extraction is used, all QC tests must be environmental measurements. Whenever performed and all QC acceptance criteria possible, the laboratory should analyze must be met with that extraction scheme as standard reference materials and participate an integral part of this method. Solid-phase in relevant performance evaluation studies. extraction (SPE) may be used provided re- quirements in section 8.1.2 are met. 9. Sample Collection, Preservation, and 10.2 Separatory funnel liquid-liquid ex- Handling traction (SFLLE) and extract concentration. 9.1 Collect samples as grab samples in 10.2.1 The SFLLE procedure below as- amber or clear glass bottles, or in refrig- sumes a sample volume of 1 L. When a dif- erated bottles using automatic sampling ferent sample volume is extracted, adjust equipment. If clear glass is used, protect the volume of methylene chloride accord- samples from light. Collect 1–L of ambient ingly. waters, effluents, and other aqueous samples. 10.2.2 Mark the water meniscus on the If the sensitivity of the analytical system is side of the sample bottle for later determina- sufficient, a smaller volume (e.g., 250 mL), tion of sample volume. Pour the entire sam- but no less than 100 mL, may be used. Con- ple into the separatory funnel. Pipet the sur- ventional sampling practices (Reference 15) rogate standard spiking solution (section 6.8) should be followed, except that the bottle into the separatory funnel. If the sample will must not be pre-rinsed with sample before be used for the LCS or MS or MSD, pipet the

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appropriate check sample concentrate (sec- lect the extract in the K–D concentrator. tion 8.2.1 or 8.3.2) into the separatory funnel. Rinse the Erlenmeyer flask and column with Mix well. Check the pH of the sample with 20–30 mL of methylene chloride to complete wide-range pH paper and adjust to pH 11–13 the quantitative transfer. with sodium hydroxide solution. 10.2.8 Add one or two clean boiling chips 10.2.3 Add 60 mL of methylene chloride to and attach a three-ball Snyder column to the the sample bottle, seal, and shake for ap- evaporative flask for each fraction (section proximately 30 seconds to rinse the inner 10.2.7). Pre-wet the Snyder column by adding surface. Transfer the solvent to the sepa- about 1 mL of methylene chloride to the top. ratory funnel and extract the sample by Place the K–D apparatus on a hot water bath shaking the funnel for two minutes with (60–65 °C) so that the concentrator tube is periodic venting to release excess pressure. partially immersed in the hot water, and the Allow the organic layer to separate from the entire lower rounded surface of the flask is water phase for a minimum of 10 minutes. If bathed with hot vapor. Adjust the vertical the emulsion interface between layers is position of the apparatus and the water tem- more than one-third the volume of the sol- perature as required to complete the con- vent layer, the analyst must employ me- centration in 15–20 minutes. At the proper chanical techniques to complete the phase rate of distillation, the balls of the column separation. The optimum technique depends will actively chatter but the chambers will upon the sample, but may include stirring, not flood with condensed solvent. When the filtration of the emulsion through glass wool apparent volume of liquid reaches 1 mL or or phase-separation paper, salting, cen- trifugation, or other physical methods. Col- other determined amount, remove the K–D lect the methylene chloride extract in a apparatus from the water bath and allow to flask. If the emulsion cannot be broken (re- drain and cool for at least 10 minutes. Re- covery of <80% of the methylene chloride), move the Snyder column and rinse the flask transfer the sample, solvent, and emulsion and its lower joint into the concentrator into a continuous extractor and proceed as tube with 1–2 mL of methylene chloride. A 5- described in section 10.3. mL syringe is recommended for this oper- 10.2.4 Add a second 60-mL volume of ation. If the sample will be cleaned up, re- methylene chloride to the sample bottle and serve the K–D apparatus for concentration of repeat the extraction procedure a second the cleaned up extract. Adjust the volume to time, combining the extracts in the Erlen- 5 mL with methylene chloride and proceed to meyer flask. Perform a third extraction in section 11 for cleanup; otherwise, further the same manner. concentrate the extract for GC/MS analysis 10.2.5 Adjust the pH of the aqueous phase per section 10.2.9 or 10.2.10. to less than 2 using sulfuric acid. Serially ex- 10.2.9 Micro Kuderna-Danish concentra- tract the acidified aqueous phase three times tion—Add another one or two clean boiling with 60 mL aliquots of methylene chloride. chips to the concentrator tube for each frac- Collect and combine the extracts in a flask tion and attach a two-ball micro-Snyder col- in the same manner as the base/neutral ex- umn. Pre-wet the Snyder column by adding tracts. about 0.5 mL of methylene chloride to the NOTE: Base/neutral and acid extracts may top. Place the K–D apparatus on a hot water be combined for concentration and analysis bath (60–65 °C) so that the concentrator tube provided all QC tests are performed and all is partially immersed in hot water. Adjust QC acceptance criteria met for the analytes the vertical position of the apparatus and of interest with the combined extract as an the water temperature as required to com- integral part of this method, and provided plete the concentration in 5–10 minutes. At that the analytes of interest are as reliably the proper rate of distillation the balls of the identified and quantified as when the ex- column will actively chatter but the cham- tracts are analyzed separately. If doubt ex- bers will not flood with condensed solvent. ists as to whether identification and quan- When the apparent volume of liquid reaches titation will be affected by use of a combined about 1 mL or other determined amount, re- extract, the fractions must be analyzed sepa- move the K–D apparatus from the water bath rately. and allow it to drain and cool for at least 10 10.2.6 For each fraction or the combined minutes. Remove the Snyder column and fractions, assemble a Kuderna-Danish (K–D) rinse the flask and its lower joint into the concentrator by attaching a 10-mL concen- concentrator tube with approximately 0.2 trator tube to a 500-mL evaporative flask. mL of or methylene chloride. Adjust the Other concentration devices or techniques final volume to 1.0 mL or a volume appro- may be used in place of the K–D concen- priate to the sensitivity desired (e.g., to trator so long as the requirements in section meet lower MDLs or for selected ion moni- 8.2 are met. toring). Record the volume, stopper the con- 10.2.7 For each fraction or the combined centrator tube and store refrigerated if fur- fractions, pour the extract through a sol- ther processing will not be performed imme- vent-rinsed drying column containing about diately. If the extracts will be stored longer 10 cm of anhydrous sodium sulfate, and col- than two days, they should be transferred to

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fluoropolymer-lined screw-cap vials and la- the vial and label with the sample number. beled base/neutral or acid fraction as appro- Store in the dark at room temperature until priate. Mark the level of the extract on the ready for GC analysis. If GC analysis will not vial so that solvent loss can be detected. be performed on the same day, store the vial 10.2.10 Nitrogen evaporation and solvent in the dark at ≤6 °C. Analyze the extract by exchange—Extracts may be concentrated for GC/MS per the procedure in section 12. analysis using nitrogen evaporation in place 10.2.12 Determine the original sample vol- of micro K–D concentration (section 10.2.9). ume by refilling the sample bottle to the Extracts that have been cleaned up using mark and transferring the liquid to an ap- sulfur removal (section 11.2) and are ready propriately sized graduated cylinder. For for analysis are exchanged into methylene sample volumes on the order of 1000 mL, chloride. record the sample volume to the nearest 10 10.2.10.1 Transfer the vial containing the mL; for sample volumes on the order of 100 sample extract to the nitrogen evaporation mL, record the volume to the nearest 1 mL. (blowdown) device (section 5.8). Lower the Sample volumes may also be determined by vial into the water bath and begin concen- weighing the container before and after fill- trating. If the more volatile analytes (sec- ing to the mark with water. tion 1.2) are to be concentrated, use room 10.3 Continuous liquid/liquid extraction temperature for concentration; otherwise, a (CLLE). slightly elevated (e.g., 30–45 °C) may be used. NOTE: With CLLE, phenol, 2,4-dimethyl During the solvent evaporation process, keep phenol, and some other analytes may be the solvent level below the water level of the preferentially extracted into the base-neu- bath and do not allow the extract to become tral fraction. Determine an analyte in the dry. Adjust the flow of nitrogen so that the fraction in which it is identified and quan- surface of the solvent is just visibly dis- tified most reliably. Also, the short-chain turbed. A large vortex in the solvent may phthalate esters (e.g., dimethyl phthalate, cause analyte loss. diethyl phthalate) and some other com- 10.2.10.2 Extracts to be solvent ex- pounds may hydrolyze during prolonged ex- changed—When the volume of the liquid is posure to basic conditions required for con- approximately 200 μL, add 2 to 3 mL of meth- tinuous extraction, resulting in low recovery ylene chloride and continue concentrating to of these analytes. When these analytes are of approximately 100 μL. Repeat the addition of interest, their recovery may be improved by solvent and concentrate once more. Adjust performing the acid extraction first. the final extract volume to be consistent 10.3.1 Use CLLE when experience with a with the volume extracted and the sensi- sample from a given source indicates an tivity desired. emulsion problem, or when an emulsion is 10.2.10.3 For extracts that have been encountered during SFLLE. CLLE may be cleaned up by GPC and that are to be con- used for all samples, if desired. centrated to a nominal volume of 1 mL, ad- 10.3.2 Mark the water meniscus on the just the final volume to compensate the GPC side of the sample bottle for later determina- loss. For a 50% GPC loss, concentrate the ex- tion of sample volume. Check the pH of the tract to 1/2000 of the volume extracted. For sample with wide-range pH paper and adjust example, if the volume extracted is 950 mL, to pH 11–13 with sodium hydroxide solution. adjust the final volume to 0.48 mL. For ex- Transfer the sample to the continuous ex- tracts that have not been cleaned up by GPC tractor. Pipet surrogate standard spiking so- and are to be concentrated to a nominal vol- lution (section 6.8) into the sample. If the ume of 1.0 mL, adjust the final extract vol- sample will be used for the LCS or MS or ume to 1/1000 of the volume extracted. For MSD, pipet the appropriate check sample example, if the volume extracted is 950 mL, concentrate (section 8.2.1 or 8.3.2) into the adjust the final extract volume to 0.95 mL. extractor. Mix well. Add 60 mL of methylene Alternative means of compensating the loss chloride to the sample bottle, seal, and during GPC are acceptable so long as they shake for 30 seconds to rinse the inner sur- produce results as accurate as results pro- face. Transfer the solvent to the extractor. duced using the procedure detailed in this 10.3.3 Repeat the sample bottle rinse with Section. An alternative final volume may be an additional 50–100 mL portion of methylene used, if desired, and the calculations ad- chloride and add the rinse to the extractor. justed accordingly. 10.3.4 Add a suitable volume of methylene NOTE: The difference in the volume frac- chloride to the distilling flask (generally tion for an extract cleaned up by GPC ac- 200–500 mL), add sufficient reagent water to counts for the loss in GPC cleanup. Also, by ensure proper operation, and extract for 18– preserving the ratio between the volume ex- 24 hours. A shorter or longer extraction time tracted and the final extract volume, the may be used if all QC acceptance criteria are concentrations and detection limits do not met. Test and, if necessary, adjust the pH of need to be adjusted for differences in the vol- the water during the second or third hour of ume extracted and the extract volume. the extraction. After extraction, allow the 10.2.11 Transfer the concentrated extract apparatus to cool, then detach the distilling to a vial with fluoropolymer-lined cap. Seal flask. Dry, concentrate, and seal the extract

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per sections 10.2.6 through 10.2.11. See the 11.1.2.1 Filter the extract or load through note at section 10.2.5 regarding combining the filter holder to remove particulates. extracts of the base/neutral and acid frac- Load the extract into the sample loop. The tions. maximum capacity of the column is 0.5–1.0 g. 10.3.5 Charge the distilling flask with If necessary, split the extract into multiple methylene chloride and attach it to the con- aliquots to prevent column overload. tinuous extractor. Carefully, while stirring, 11.1.2.2 Elute the extract using the cali- adjust the pH of the aqueous phase to less bration data determined in Section 11.1.1. than 2 using sulfuric acid. Extract for 18–24 Collect the eluate in the K–D apparatus re- hours. A shorter or longer extraction time served in section 10.2.8. may be used if all QC acceptance criteria are 11.1.3 Concentrate the cleaned up extract met. Test and, if necessary, adjust the pH of per sections 10.2.8 and 10.2.9 or 10.2.10. the water during the second or third hour of 11.1.4 Rinse the sample loading tube thor- the extraction. After extraction, allow the oughly with methylene chloride between ex- apparatus to cool, then detach the distilling tracts to prepare for the next sample. flask. Dry, concentrate, and seal the extract 11.1.5 If a particularly dirty extract is en- per sections 10.2.6 through 10.2.11. Determine countered, run a methylene chloride blank the sample volume per section 10.2.12. through the system to check for carry-over. 11.2 Sulfur removal. 11. Extract Cleanup NOTE: Separate procedures using copper or NOTE: Cleanup may not be necessary for TBA sulfite are provided in this section for relatively clean samples (e.g., treated sulfur removal. They may be used separately effluents, groundwater, drinking water). If or in combination, if desired. particular circumstances require the use of a 11.2.1 Removal with copper (Reference 17). cleanup procedure, the laboratory may use NOTE: If an additional compound (Table 3) any or all of the procedures below or any is to be determined; sulfur is to be removed; other appropriate procedure. Before using a copper will be used for sulfur removal; and a cleanup procedure, the laboratory must dem- sulfur matrix is known or suspected to be onstrate that the requirements of section present, the laboratory must demonstrate 8.1.2 can be met using the cleanup procedure that the additional compound can be suc- as an integral part of this method. cessfully extracted and treated with copper 11.1 Gel permeation chromatography in the sulfur matrix. Some of the additional (GPC). compounds (Table 3) are known not to be 11.1.1 Calibration. amenable to sulfur removal with copper (e.g., 11.1.1.1 Load the calibration solution (sec- Atrazine and Diazinon). tion 6.12) into the sample loop. 11.2.1.1 Quantitatively transfer the ex- 11.1.1.2 Inject the calibration solution and tract from section 10.2.8 to a 40- to 50-mL record the signal from the detector. The flask or bottle. If there is evidence of water elution pattern will be corn oil, bis(2- in the concentrator tube after the transfer, ethylhexyl) phthalate, pentachlorophenol, rinse the tube with small portions of perylene, and sulfur. hexane:acetone (40:60) and add to the flask or 11.1.1.3 Set the ‘‘dump time’’ to allow bottle. Mark and set aside the concentrator >85% removal of the corn oil and >85% col- tube for use in re-concentrating the extract. lection of the phthalate. 11.2.1.2 Add 10–20 g of granular anhydrous 11.1.1.4 Set the ‘‘collect time’’ to the peak sodium sulfate to the flask. Swirl to dry the minimum between perylene and sulfur. extract. 11.1.1.5 Verify calibration with the cali- 11.2.1.3 Add activated copper (section bration solution after every 20 or fewer ex- 6.13.1.4) and allow to stand for 30—60 min- tracts. Calibration is verified if the recovery utes, swirling occasionally. If the copper of the pentachlorophenol is greater than does not remain bright, add more and swirl 85%. If calibration is not verified, recalibrate occasionally for another 30–60 minutes. using the calibration solution, and re-extract 11.2.1.4 After drying and sulfur removal, and clean up the preceding extracts using quantitatively transfer the extract to a ni- the calibrated GPC system. trogen-evaporation vial or tube and proceed 11.1.2 Extract cleanup—GPC requires that to section 10.2.10 for nitrogen evaporation the column not be overloaded. The column and solvent exchange, taking care to leave specified in this method is designed to han- the sodium sulfate and copper in the flask. dle a maximum of 0.5 g of high molecular 11.2.2 Removal with TBA sulfite. weight material in a 5-mL extract. If the ex- 11.2.2.1 Using small volumes of hexane, tract is known or expected to contain more quantitatively transfer the extract to a 40- than 0.5 g, the extract is split into fractions to 50-mL centrifuge tube with for GPC and the fractions are combined after fluoropolymer-lined screw cap. elution from the column. The solids content 11.2.2.2 Add 1–2 mL of TBA sulfite reagent of the extract may be obtained gravimetri- (section 6.13.2.4), 2–3 mL of 2-propanol, and cally by evaporating the solvent from a 50-μL approximately 0.7 g of sodium sulfite (sec- aliquot. tion 6.13.2.2) crystals to the tube. Cap and

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shake for 1–2 minutes. If the sample is color- sample volume, or dilute and analyze the di- less or if the initial color is unchanged, and luted extract after bringing the concentra- if clear crystals (precipitated sodium sulfite) tions of the internal standards to the levels are observed, sufficient sodium sulfite is in the undiluted extract. present. If the precipitated sodium sulfite 12.2.4 Perform all qualitative and quan- disappears, add more crystalline sodium sul- titative measurements as described in Sec- fite in approximately 0.5 g portions until a tions 14 and 15. When standards and extracts solid residue remains after repeated shaking. are not being used for analyses, store them 11.2.2.3 Add 5–10 mL of reagent water and refrigerated at ≤6 °C protected from light in shake for 1–2 minutes. Centrifuge to settle screw-cap vials equipped with un-pierced the solids. fluoropolymer-lined septa. 11.2.2.4 Quantitatively transfer the hexane (top) layer through a small funnel 13. Performance Tests containing a few grams of granular anhy- 13.1 At the beginning of each 12-hour shift drous sodium sulfate to a nitrogen-evapo- during which standards or extracts will be ration vial or tube and proceed to section analyzed, perform the tests in sections 13.2– 10.2.10 for nitrogen evaporation and solvent 13.4 to verify system performance. If an ex- exchange. tract is concentrated for greater sensitivity (e.g., by SIM), all tests must be performed at 12. Gas Chromatography/Mass Spectrometry levels consistent with the reduced extract 12.1 Establish the operating conditions in volume. Table 4 or 5 for analysis of a base/neutral or 13.2 DFTPP—Inject the DFTPP standard acid extract, respectively. For analysis of a (section 6.10) and verify that the criteria for combined extract (section 10.2.5, note), use DFTPP in section 7.2.1.1 and Table 9A (Ref- the operating conditions in Table 4 MDLs erence 18) for a quadrupole MS, or Table 9B and MLs for the analytes are given in Tables (Reference 19) for a time-of-flight MS, are 1, 2, and 3. Retention times for many of the met. analytes are given in Tables 4 and 5. Exam- 13.3 GC resolution—The resolution should ples of the separations achieved are shown in be verified on the mid-point concentration of Figure 2 for the combined extract. Alter- the initial calibration as well as the labora- native columns or chromatographic condi- tory designated continuing calibration tions may be used if the requirements of sec- verification level if closely eluting isomers tion 8.2 are met. Verify system performance are to be reported (e.g., benzo(b)fluoranthene per section 13. and benzo(k)fluoranthene). Sufficient gas 12.2 Analysis of a standard or extract. chromatographic resolution is achieved if 12.2.1 Bring the standard or concentrated the height of the valley between two isomer extract (section 10.2.9 or 10.2.11) to room peaks is less than 50% of the average of the temperature and verify that any precipitate two peak heights. has redissolved. Verify the level on the ex- 13.4 Calibration verification—Verify cali- tract and bring to the mark with solvent if bration per sections 7.3 and Table 6. required. 13.5 Peak tailing—Verify the tailing fac- 12.2.2 Add the internal standard solution tor specifications are met per Section 7.2.1.1. (section 6.9) to the extract. Mix thoroughly. 13.6 Laboratory control sample and 12.2.3 Inject an appropriate volume of the blank—Analyze the extracts of the LCS and sample extract or standard solution using blank at the beginning of analyses of sam- split, splitless, solvent purge, large-volume, ples in the extraction batch (section 3.1). The or on-column injection. If the sample is in- LCS must meet the requirements in section jected manually the solvent-flush technique 8.4, and the blank must meet the require- should be used. The injection volume de- ments in section 8.5 before sample extracts pends upon the technique used and the abil- may be analyzed. ity to meet MDLs or reporting limits for reg- 13.7 Analysis of DFTPP, the DDT/Endrin ulatory compliance. Injected volumes must decomposition test (if used), the LCS, and be the same for standards and sample ex- the blank are outside of the 12-hour analysis tracts. Record the volume injected to two shift (section 3.1). The total time for DFTPP, significant figures. DDT/Endrin, the LCS, the blank, and the 12- 12.2.3.1 Start the GC column oven pro- hour shift must not exceed 15 hours. gram upon injection. Start MS data collec- 13.8 Decomposition of DDT and endrin—If tion after the solvent peak elutes. Stop data DDT and/or endrin are to be determined, this collection after benzo(ghi)perylene elutes for test must be performed prior to calibration the base/neutral or combined fractions, or verification (section 13.4). The QC acceptance after pentachlorophenol elutes for the acid criteria (section 13.8.3) must be met before fraction. Return the column to the initial analyzing samples for DDE and/or Endrin. temperature for analysis of the next stand- DDT decomposes to DDE and DDD. Endrin ard solution or extract. decomposes to endrin aldehyde and endrin 12.2.3.2 If the concentration of any ketone. analyte of interest exceeds the calibration 13.8.1 Inject 1 μL of the DDT and endrin range, either extract and analyze a smaller decomposition solution (section 6.14). As

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noted in section 6.14, other injection volumes 13.8.2 Measure the areas of the peaks for may be used as long as the concentrations of DDT, DDE, DDD, Endrin, Endrin aldehyde, DDT and endrin in the solution are adjusted and Endrin ketone. Calculate the percent to introduce the masses of the two analytes breakdown as shown in the equations below: into the instrument that are listed in section 6.14.

13.8.3 Both the % breakdown of DDT and some procedures refer to ‘‘RRT units’’ in of Endrin must be less than 20%, otherwise providing the specification for the agree- the system is not performing acceptably for ment between the RRT values in the sample DDT and endrin. In this case, repair the GC and the calibration verification or other column system that failed and repeat the standard. When significant retention time performance tests (sections 13.2 to 13.6) until drifts are observed, dilutions or spiked sam- the specification is met. ples may help the analyst determine the ef- NOTE: DDT and endrin decomposition are fects of the matrix on elution of the target usually caused by accumulation of particu- analytes and to assist in qualitative identi- lates in the injector and in the front end of fication. the column. Cleaning and silanizing the in- 14.1.3 Either the background corrected jection port liner, and breaking off a short EICP areas, or the corrected relative inten- section of the front end of the column will sities of the mass spectral peaks at the GC usually eliminate the decomposition prob- peak maximum, must agree within 50% to lem. Either of these corrective actions may 200% (1/2 to 2 times) for the quantitation and affect retention times, GC resolution, and secondary m/z’s in the reference mass spec- calibration linearity. trum stored in the data system (section 7.2.1.2), or from a reference library. For ex- 14. Qualitative Identification ample, if a peak has an intensity of 20% rel- 14.1 Identification is accomplished by ative to the base peak, the analyte is identi- comparison of data from analysis of a sample fied if the intensity of the peak in the sam- or blank with data stored in the GC/MS data ple is in the range of 10% to 40% of the base system (sections 5.6.5 and 7.2.1.2). Identifica- peak. If identification is ambiguous, an expe- tion of an analyte is confirmed per sections rienced spectrometrist (section 1.7) must de- 14.1.1 through 14.1.4. termine the presence or absence of the com- 14.1.1 The signals for the quantitation and pound. secondary m/z’s stored in the data system for 14.2 Structural isomers that produce very each analyte of interest must be present and similar mass spectra should be identified as must maximize within the same two con- individual isomers if they have sufficiently secutive scans. different gas chromatographic retention 14.1.2 The retention time for the analyte times. Sufficient gas chromatographic reso- should be within ± 10 seconds of the analyte lution is achieved if the height of the valley in the calibration verification run at the be- between two isomer peaks is less than 50% of ginning of the shift (section 7.3 or 13.4). the average of the two peak heights. Other- NOTE: Retention time windows other than wise, structural isomers are identified as iso- ± 10 seconds may be appropriate depending meric pairs. on the performance of the gas chro- 15. Calculations matograph or observed retention time drifts due to certain types of matrix effects. Rel- 15.1 When an analyte has been identified, ative retention time (RRT) may be used as quantitation of that analyte is based on the an alternative to absolute retention times if integrated abundance from the EICP of the retention time drift is a concern. RRT is a primary characteristic m/z in Table 4 or 5. unitless quantity (see Sec. 22.2), although Calculate the concentration in the extract

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using the response factor (RF) determined in into the calibration range, and re-analyze Section 7.2.2 and Equation 2. If the con- the extract. Determine a dilution factor (DF) centration of an analyte exceeds the calibra- from the amount of the dilution. For exam- tion range, dilute the extract by the min- ple, if the extract is diluted by a factor of 2, imum amount to bring the concentration DF = 2.

where: Calculate the concentration of the analyte in the sample using the concentration in the Cex = Concentration of the analyte in the ex- tract, in μg/mL, and the other terms are extract, the extract volume, the sample vol- as defined in section 7.2.2. ume, and the dilution factor, per Equation 3:

where: where ML is the concentration of the analyte at the ML, or as required by the reg- Csamp = Concentration of the analyte in the sample ulatory/control authority or permit. Report a result for each analyte in a blank at or Cex = Concentration of the analyte in the ex- tract, in μg/mL above the MDL to 2 significant figures. Re- port a result for each analyte found in a V = Volume of extract (mL) ex blank below the MDL as ‘‘MDL,’’ where MDL V = Volume of sample (L) s is the concentration of the analyte at the DF = Dilution factor MDL, or as required by the regulatory/con- 15.2 Reporting of results. As noted in sec- trol authority or permit. tion 1.4.1, EPA has promulgated this method 15.2.2.2 In addition to reporting results for at 40 CFR part 136 for use in wastewater samples and blanks separately, the con- compliance monitoring under the National centration of each analyte in a blank associ- Pollutant Discharge Elimination System ated with the sample may be subtracted (NPDES). The data reporting practices de- from the result for that sample, but only if scribed here are focused on such monitoring requested or required by a regulatory au- needs and may not be relevant to other uses thority or in a permit. In this case, both the of the method. sample result and the blank results must be 15.2.1 Report results for wastewater sam- reported together. ples in μg/L without correction for recovery. 15.2.2.3 Report a result for an analyte (Other units may be used if required by in a found in a sample or extract that has been permit.) Report all QC data with the sample diluted at the least dilute level at which the results. area at the quantitation m/z is within the 15.2.2 Reporting level. Unless specified calibration range (i.e., above the ML for the otherwise by a regulatory authority or in a analyte) and the MS/MSD recovery and RPD discharge permit, results for analytes that are within their respective QC acceptance meet the identification criteria are reported criteria (Table 6). This may require reporting down to the concentration of the ML estab- results for some analytes from different lished by the laboratory through calibration analyses. of the instrument (see section 7.3.2 and the 15.2.3 Results from tests performed with glossary for the derivation of the ML). EPA an analytical system that is not in control considers the terms ‘‘reporting limit,’’ (i.e., that does not meet acceptance criteria ‘‘quantitation limit,’’ ‘‘limit of quantita- for any QC test in this method) must be doc- tion,’’ and ‘‘minimum level’’ to be synony- umented and reported (e.g., as a qualifier on mous. results), unless the failure is not required to 15.2.2.1 Report a result for each analyte in be reported as determined by the regulatory/ each field sample or QC standard at or above control authority. Results associated with a the ML to 3 significant figures. Report a re- QC failure cannot be used to demonstrate sult for each analyte found in each field sam- regulatory compliance. QC failures do not re- ple or QC standard below the ML as ‘‘ML’’ lieve a discharger or permittee of reporting

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timely results. If the holding time would be regulations governing waste management, exceeded for a re-analysis of the sample, the particularly the hazardous waste identifica- regulatory/control authority should be con- tion rules and land disposal restrictions, and sulted for disposition. to protect the air, water, and land by mini- mizing and controlling all releases from 16. Method Performance fume hoods and bench operations. Compli- 16.1 The basic version of this method was ance is also required with any sewage dis- tested by 15 laboratories using reagent charge permits and regulations. An overview water, drinking water, surface water, and in- of requirements can be found in Environ- dustrial wastewaters spiked at six con- mental Management Guide for Small Lab- centrations over the range 5–1300 μg/L (Ref- oratories (EPA 233–B–98–001). erence 2). Single operator precision, overall 18.2 Samples at pH <2, or pH >12, are haz- precision, and method accuracy were found ardous and must be handled and disposed of to be directly related to the concentration of as hazardous waste, or neutralized and dis- the analyte and essentially independent of posed of in accordance with all federal, state, the sample matrix. Linear equations to de- and local regulations. It is the laboratory’s scribe these relationships are presented in responsibility to comply with all federal, Table 7. state, and local regulations governing waste 16.2 As noted in section 1.1, this method management, particularly the hazardous was validated through an interlaboratory waste identification rules and land disposal study in the early 1980s. However, the funda- restrictions. The laboratory using this meth- mental chemistry principles used in this od has the responsibility to protect the air, method remain sound and continue to apply. water, and land by minimizing and control- 16.3 A chromatogram of the combined ling all releases from fume hoods and bench acid/base/neutral calibration standard is operations. Compliance is also required with shown in Figure 2. any sewage discharge permits and regula- 17. Pollution Prevention tions. For further information on waste management, see ‘‘The Waste Management 17.1 Pollution prevention encompasses Manual for Laboratory Personnel,’’ also any technique that reduces or eliminates the available from the American Chemical Soci- quantity or toxicity of waste at the point of ety at the address in section 17.3. generation. Many opportunities for pollution 18.3 Many analytes in this method decom- prevention exist in laboratory operations. pose above 500 ßC. Low-level waste such as EPA has established a preferred hierarchy of absorbent paper, tissues, and plastic gloves environmental management techniques that may be burned in an appropriate incinerator. places pollution prevention as the manage- Gross quantities of neat or highly con- ment option of first choice. Whenever fea- centrated solutions of toxic or hazardous sible, the laboratory should use pollution chemicals should be packaged securely and prevention techniques to address waste gen- disposed of through commercial or govern- eration. When wastes cannot be reduced at mental channels that are capable of handling the source, the Agency recommends recy- these types of wastes. cling as the next best option. 18.4 For further information on waste 17.2 The analytes in this method are used management, consult The Waste Manage- in extremely small amounts and pose little ment Manual for Laboratory Personnel and threat to the environment when managed Less is Better-Laboratory Chemical Manage- properly. Standards should be prepared in ment for Waste Reduction, available from volumes consistent with laboratory use to the American Chemical Society’s Depart- minimize the disposal of excess volumes of ment of Government Relations and Science expired standards. This method utilizes sig- Policy, 1155 16th Street NW., Washington, DC nificant quantities of methylene chloride. 20036, 202–872–4477. Laboratories are encouraged to recover and recycle this and other solvents during ex- 19. References tract concentration. 17.3 For information about pollution pre- 1. ‘‘Sampling and Analysis Procedures for vention that may be applied to laboratories Screening of Industrial Effluents for Pri- and research institutions, consult Less is ority Pollutants,’’ U.S. Environmental Better: Laboratory Chemical Management Protection Agency, Environmental Moni- for Waste Reduction, available from the toring and Support Laboratory, Cin- American Chemical Society’s Department of cinnati, Ohio 45268, March 1977, Revised Governmental Relations and Science Policy, April 1977. 1155 16th Street NW., Washington DC 20036, 2. ‘‘EPA Method Study 30, Method 625, Base/ 202–872–4477. Neutrals, Acids, and Pesticides,’’ EPA 600/4–84–053, National Technical Informa- 18. Waste Management tion Service, PB84–206572, Springfield, 18.1 The laboratory is responsible for Virginia 22161, June 1984. complying with all Federal, State, and local 3. 40 CFR part 136, appendix B.

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4. Olynyk, P., Budde, W.L. and Eichelberger, in Wastewater and Drinking Water J.W. ‘‘Method Detection Limit for Meth- (EPA–821–B–98–003) March 1999. ods 624 and 625,’’ Unpublished report, 14. Provost, L.P. and Elder, R.S. ‘‘Interpre- May 14, 1980. tation of Percent Recovery Data,’’ Amer- 5. Annual Book of ASTM Standards, Volume ican Laboratory, 15, 58–63 (1983). (The 11.02, D3694–96, ‘‘Standard Practices for value 2.44 used in the equation in section Preparation of Sample Containers and 8.3.3 is two times the value 1.22 derived in for Preservation of Organic Constitu- this report.) ents,’’ American Society for Testing and Materials, Philadelphia. 15. ASTM Annual Book of Standards, Part 31, 6. Solutions to Analytical Chemistry Prob- D3370–76. ‘‘Standard Practices for Sam- lems with Clean Water Act Methods, pling Water,’’ American Society for Test- EPA 821–R–07–002, March 2007. ing and Materials, Philadelphia. 7. ‘‘Carcinogens-Working With Carcinogens,’’ 16. 40 CFR 136.3(a), Table IB, Chlorine—Total Department of Health, Education, and Residual. Welfare, Public Health Service, Center 17. ‘‘Manual of Analytical Methods for the for Disease Control, National Institute Analysis of Pesticides in Human and En- for Occupational Safety and Health, Pub- vironmental Samples,’’ EPA–600/8–80–038, lication No. 77–206, August 1977. U.S. Environmental Protection Agency, 8. ‘‘OSHA Safety and Health Standards, Gen- Health Effects Research Laboratory, Re- eral Industry,’’ (29 CFR part 1910), Occu- search Triangle Park, North Carolina. pational Safety and Health Administra- tion, OSHA 2206 (Revised, January 1976). 18. Eichelberger, J.W., Harris, L.E., and 9. ‘‘Safety in Academic Chemistry Labora- Budde, W.L. ‘‘Reference Compound to tories,’’ American Chemical Society Pub- Calibrate Ion Abundance Measurement in lication, Committee on Chemical Safety, Gas Chromatography-Mass Spectrom- 7th Edition, 2003. etry,’’ Analytical Chemistry, 47, 995 10. Johnson, R.A., and Wichern, D.W., ‘‘Ap- (1975). plied Multivariate Statistical Analysis,’’ 19. Letter of approval of acceptance criteria 3rd edition, Prentice Hall, Englewood for DFTPP for time-of-flight mass spec- Cliffs, NJ, 1992. trometers from William A. Telliard and 11. 40 CFR 136.6(b)(4)(x). Herb Brass of EPA to Jack Cochran of 12. 40 CFR 136.6(b)(2)(i). LECO Corporation, February 9, 2005. 13. Protocol for EPA Approval of New Meth- ods for Organic and Inorganic Analytes 20. Tables

TABLE 1—NON PESTICIDE/PCB BASE/NEUTRAL EXTRACTABLES 1

MDL 4 ML 5 Analyte CAS registry (ug/L) (ug/L)

Acenaphthene ...... 83–32–9 1.9 5.7 Acenaphthylene ...... 208–96–8 3.5 10.5 Anthracene ...... 120–12–7 1.9 5.7 Benzidine 2 ...... 92–87–5 44 132 Benzo(a)anthracene ...... 56–55–3 7.8 23.4 Benzo(a)pyrene ...... 50–32–8 2.5 7.5 Benzo(b)fluoranthene ...... 205–99–2 4.8 14.4 Benzo(k)fluoranthene ...... 207–08–9 2.5 7.5 Benzo(ghi)perylene ...... 191–24–2 4.1 12.3 Benzyl butyl phthalate ...... 85–68–7 2.5 7.5 bis(2-Chloroethoxy)methane ...... 111–91–1 5.3 15.9 bis(2-Ethylhexyl)phthalate ...... 117–81–7 2.5 7.5 bis(2-Chloroisopropyl) ether (2,2’-Oxybis[1-chloropropane]) ...... 108–60–1 5.7 17.1 4-Bromophenyl phenyl ether ...... 101–55–3 1.9 5.7 2-Chloronaphthalene ...... 91–58–7 1.9 5.7 4-Chlorophenyl phenyl ether ...... 7005–72–3 4.2 12.6 Chrysene ...... 218–01–9 2.5 7.5 Dibenz(a,h)anthracene ...... 53–70–3 2.5 7.5 Di-n-butylphthalate ...... 84–74–2 2.5 7.5 3,3’-Dichlorobenzidine ...... 91–94–1 16.5 49.5 Diethyl phthalate ...... 84–66–2 1.9 5.7 Dimethyl phthalate ...... 131–11–3 1.6 4.8 2,4-Dinitrotoluene ...... 121–14–2 5.7 17.1 2,6-Dinitrotoluene ...... 606–20–2 1.9 5.7 Di-n-octylphthalate ...... 117–84–0 2.5 7.5 Fluoranthene ...... 206–44–0 2.2 6.6 Fluorene ...... 86–73–7 1.9 5.7 Hexachlorobenzene ...... 118–74–1 1.9 5.7 Hexachlorobutadiene ...... 87–68–3 0.9 2.7 Hexachloroethane ...... 67–72–1 1.6 4.8 Indeno(1,2,3-cd)pyrene ...... 193–39–5 3.7 11.1

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TABLE 1—NON PESTICIDE/PCB BASE/NEUTRAL EXTRACTABLES 1—Continued

MDL 4 ML 5 Analyte CAS registry (ug/L) (ug/L)

Isophorone ...... 78–59–1 2.2 6.6 Naphthalene ...... 91–20–3 1.6 4.8 Nitrobenzene ...... 98–95–3 1.9 5.7 N-Nitrosodi-n-propylamine 3 ...... 621–64–7 — — Phenanthrene ...... 85–01–8 5.4 16.2 Pyrene ...... 129–00–0 1.9 5.7 1,2,4-Trichlorobenzene ...... 120–82–1 1.9 5.7 1 All analytes in this table are Priority Pollutants (40 CFR part 423, appendix A). 2 Included for tailing factor testing. 3 See section 1.2. 4 MDL values from the 1984 promulgated version of Method 625. 5 ML = Minimum Level—see Glossary for definition and derivation.

TABLE 2—ACID EXTRACTABLES 1

MDL 3 ML 4 Analyte CAS registry (ug/L) (ug/L)

4-Chloro-3-methylphenol ...... 59–50–7 3.0 9.0 2-Chlorophenol ...... 95–57–8 3.3 9.9 2,4-Dichlorophenol ...... 120–83–2 2.7 8.1 2,4-Dimethylphenol ...... 105–67–9 2.7 8.1 2,4-Dinitrophenol ...... 51–28–5 42 126 2-Methyl-4,6-dinitrophenol ...... 534–52–1 24 72 2-Nitrophenol ...... 88–75–5 3.6 10.8 4-Nitrophenol ...... 100–02–7 2.4 7.2 Pentachlorophenol 2 ...... 87–86–5 3.6 10.8 Phenol ...... 108–95–2 1.5 4.5 2,4,6-Trichlorophenol ...... 88–06–2 2.7 8.1 1 All analytes in this table are Priority Pollutants (40 CFR part 423, appendix A). 2 See section 1.2; included for tailing factor testing. 3 MDL values from the 1984 promulgated version of Method 625. 4 ML = Minimum Level—see Glossary for definition and derivation.

TABLE 3—ADDITIONAL EXTRACTABLE ANALYTES 12

MDL 7 ML 8 Analyte CAS registry (ug/L) (ug/L)

Acetophenone ...... 98–86–2 2-Acetylaminofluorene ...... 53–96–3 1-Acetyl-2-thiourea ...... 591–08–2 Alachlor ...... 15972–60–8 Aldrin 3 ...... 309–00–2 1.9 5.7 Ametryn ...... 834–12–8 2-Aminoanthraquinone ...... 117–79–3 Aminoazobenzene ...... 60–09–3 4-Aminobiphenyl ...... 92–67–1 3-Amino-9-ethylcarbazole ...... 132–32–1 Anilazine ...... 101–05–3 ...... 62–53–3 o-Anisidine ...... 90–04–0 Aramite ...... 140–57–8 Atraton ...... 1610–17–9 Atrazine ...... 1912–24–9 Azinphos-methyl ...... 86–50–0 Barban ...... 101–27–9 Benzanthrone ...... 82–05–3 Benzenethiol ...... 108–98–5 ...... 65–85–0 2,3-Benzofluorene ...... 243–17–4 p-Benzoquinone ...... 106–51–4 Benzyl alcohol ...... 100–51–6 alpha-BHC 34 ...... 319–84–6 beta-BHC 3 ...... 319–85–7 3.1 9.3 gamma-BHC (Lindane) 34 ...... 58–89–8 4.2 12.6 delta-BHC 3 ...... 319–86–8 Biphenyl ...... 92–52–4 Bromacil ...... 314–40–9 2-Bromochlorobenzene ...... 694–80–4 3-Bromochlorobenzene ...... 108–39–2

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TABLE 3—ADDITIONAL EXTRACTABLE ANALYTES 12—Continued

MDL 7 ML 8 Analyte CAS registry (ug/L) (ug/L)

Bromoxynil ...... 1689–84–5 Butachlor ...... 2318–4669 Butylate ...... 2008–41–5 n-C10 (n-decane) ...... 124–18–5 n-C12 (n-undecane) ...... 112–40–2 n-C14 (n-tetradecane) ...... 629–59–4 n-C16 (n-hexadecane) ...... 544–76–3 n-C18 (n-octadecane) ...... 593–45–3 n-C20 (n-eicosane) ...... 112–95–8 n-C22 (n-docosane) ...... 629–97–0 n-C24 (n-tetracosane) ...... 646–31–1 n-C26 (n-hexacosane) ...... 630–01–3 n-C28 (n-octacosane) ...... 630–02–4 n-C30 (n-triacontane) ...... 638–68–6 Captafol ...... 2425–06–1 Captan ...... 133–06–2 Carbaryl ...... 63–25–2 Carbazole ...... 86–74–8 Carbofuran ...... 1563–66–2 Carboxin ...... 5234–68 –4 Carbophenothion ...... 786–19–6 Chlordane 35 ...... 57–74–9 bis(2-Chloroethyl) ether 34 ...... 111–44–4 5.7 17.1 Chloroneb ...... 2675–77–6 4-Chloroaniline ...... 106–47–8 Chlorobenzilate ...... 510–15–6 ...... 470–90–6 4-Chloro-2-methylaniline ...... 95–69–2 3-(Chloromethyl)pyridine hydrochloride ...... 6959–48–4 4-Chloro-2-nitroaniline ...... 89–63–4 Chlorpropham ...... 101–21–3 Chlorothalonil ...... 1897–45–6 1-Chloronaphthalene ...... 90–13–1 3-Chloronitrobenzene ...... 121–73–3 4-Chloro-1,2-phenylenediamine ...... 95–83–0 4-Chloro-1,3-phenylenediamine ...... 5131–60–2 2-Chlorobiphenyl ...... 2051–60–7 Chlorpyrifos ...... 2921–88–2 ...... 56–72–4 m + p-Cresol ...... 65794–96–9 o-Cresol ...... 95–48–7 p-Cresidine ...... 120–71–8 Crotoxyphos ...... 7700–17–6 2-Cyclohexyl-4,6-dinitro-phenol ...... 131–89–5 Cyanazine ...... 21725–46–2 Cycloate ...... 1134–23–2 p-Cymene ...... 99–87–6 Dacthal (DCPA) ...... 1861–32–1 4,4’-DDD 3 ...... 72–54–8 2.8 8.4 4,4’-DDE 3 ...... 72–55–9 5.6 16.8 4,4’-DDT 3 ...... 50–29–3 4.7 14.1 Demeton-O ...... 298–03–3 Demeton-S ...... 126–75–0 Diallate (cis or trans) ...... 2303–16–4 2,4-Diaminotoluene ...... 95–80–7 Diazinon ...... 333–41–5 Dibenz(a,j)acridine ...... 224–42–0 Dibenzofuran ...... 132–64–9 Dibenzo(a,e)pyrene ...... 192–65–4 Dibenzothiophene ...... 132–65–0 1,2-Dibromo-3-chloropropane ...... 96–12–8 3,5-Dibromo-4-hydroxybenzonitrile ...... 1689–84–5 2,6-Di-tert-butyl-p-benzoquinone ...... 719–22–2 Dichlone ...... 117–80–6 2,3-Dichloroaniline ...... 608–27–5 2,3-Dichlorobiphenyl ...... 16605–91–7 2,6-Dichloro-4-nitroaniline ...... 99–30–9 2,3-Dichloronitrobenzene ...... 3209–22–1 1,3-Dichloro-2-propanol ...... 96–23–1 2,6-Dichlorophenol ...... 120–83–2 Dichlorvos ...... 62–73–7

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TABLE 3—ADDITIONAL EXTRACTABLE ANALYTES 12—Continued

MDL 7 ML 8 Analyte CAS registry (ug/L) (ug/L)

Dicrotophos ...... 141–66–2 Dieldrin 3 ...... 60–57–1 2.5 7.5 1,2:3,4-Diepoxybutane ...... 1464–53–5 Di(2-ethylhexyl) adipate ...... 103–23–1 ...... 56–53–1 Diethyl sulfate ...... 64–67–5 Dilantin (5,5-Diphenylhydantoin) ...... 57–41–0 ...... 60–51–5 3,3′-Dimethoxybenzidine ...... 119–90–4 Dimethylaminoazobenzene ...... 60–11–7 7,12-Dimethylbenz(a)anthracene ...... 57–97–6 3,3′-Dimethylbenzidine ...... 119–93–7 N,N-Dimethylformamide ...... 68–12–2 3,6-Dimethylphenathrene ...... 1576–67–6 alpha, alpha-Dimethylphenethylamine ...... 122–09–8 Dimethyl sulfone ...... 67–71–0 1,2-Dinitrobenzene ...... 528–29–0 1,3-Dinitrobenzene ...... 99–65–0 1,4-Dinitrobenzene ...... 100–25–4 Dinocap ...... 39300–45–3 Dinoseb ...... 88–85–7 Diphenylamine ...... 122–39–4 Diphenyl ether ...... 101–84–8 1,2-Diphenylhydrazine ...... 122–66–7 Diphenamid ...... 957–51–7 Diphenyldisulfide ...... 882–33–7 Disulfoton ...... 298–04–4 Disulfoton sulfoxide ...... 2497–07–6 Disulfoton sulfone ...... 2497–06–5 Endosulfan I 4 ...... 959–98–8 Endosulfan II 34 ...... 33213–65–9 Endosulfan sulfate 3 ...... 1031–07–8 5.6 16.8 Endrin 34 ...... 72–20–8 Endrin aldehyde 34 ...... 7421–93–4 Endrin ketone 34 ...... 53494–70–5 EPN ...... 2104–64–5 EPTC ...... 759–94–4 Ethion ...... 563–12–2 Ethoprop ...... 13194–48–4 Ethyl carbamate ...... 51–79–6 Ethyl methanesulfonate ...... 65–50–0 Ethylenethiourea ...... 96–45–7 Etridiazole ...... 2593–15–9 Ethynylestradiol-3-methyl ether ...... 72–33–3 Famphur ...... 52–85–7 ...... 22224–92–6 Fenarimol ...... 60168–88–9 Fensulfothion ...... 115–90–2 Fenthion ...... 55–38–9 Fluchloralin ...... 33245–39–5 Fluridone ...... 59756–60–4 Heptachlor 3 ...... 76–44–8 1.9 5.7 Heptachlor epoxide 3 ...... 1024–57–3 2.2 6.6 2,2′,3,3′,4,4′,6-Heptachlorobiphenyl ...... 52663–71–5 2,2′,4,4′,5′,6-Hexachlorobiphenyl ...... 60145–22–4 Hexachlorocyclopentadiene 34 ...... 77–47–4 Hexachlorophene ...... 70–30–4 Hexachloropropene ...... 1888–71–7 Hexamethylphosphoramide ...... 680–31–9 Hexanoic acid ...... 142–62–1 ...... 51235–04–2 Hydroquinone ...... 123–31–9 Isodrin ...... 465–73–6 2-Isopropylnaphthalene ...... 2027–17–0 Isosafrole ...... 120–58–1 Kepone ...... 143–50–0 ...... 21609–90–5 Longifolene ...... 475–20–7 Malachite green ...... 569–64–2 Malathion ...... 121–75–5 Maleic anhydride ...... 108–31–6

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TABLE 3—ADDITIONAL EXTRACTABLE ANALYTES 12—Continued

MDL 7 ML 8 Analyte CAS registry (ug/L) (ug/L)

Merphos ...... 150–50–5 Mestranol ...... 72–33–3 Methapyrilene ...... 91–80–5 Methoxychlor ...... 72–43–5 2-Methylbenzothioazole ...... 120–75–2 3-Methylcholanthrene ...... 56–49–5 4,4′-Methylenebis(2-chloroaniline) ...... 101–14–4 4,4′-Methylenebis(N,N-dimethylaniline) ...... 101–61–1 4,5-Methylenephenanthrene ...... 203–64–5 1-Methylfluorene ...... 1730–37–6 Methyl methanesulfonate ...... 66–27–3 2-Methylnaphthalene ...... 91–57–6 Methylparaoxon ...... 950–35–6 Methyl parathion ...... 298–00–0 1-Methylphenanthrene ...... 832–69–9 2-(Methylthio)benzothiazole ...... 615–22–5 Metolachlor ...... 5218–45–2 Metribuzin ...... 21087–64–9 Mevinphos ...... 7786–34–7 Mexacarbate ...... 315–18–4 MGK 264 ...... 113–48–4 Mirex ...... 2385–85–5 Molinate ...... 2212–67–1 ...... 6923–22–4 Naled ...... 300–76–5 Napropamide ...... 15299–99–7 1,4-Naphthoquinone ...... 130–15–4 1-Naphthylamine ...... 134–32–7 2-Naphthylamine ...... 91–59–8 1,5-Naphthalenediamine ...... 2243–62–1 Nicotine ...... 54–11–5 5-Nitroacenaphthene ...... 602–87–9 2-Nitroaniline ...... 88–74–4 3-Nitroaniline ...... 99–09–2 4-Nitroaniline ...... 100–01–6 5-Nitro-o-anisidine ...... 99–59–2 4-Nitrobiphenyl ...... 92–93–3 Nitrofen ...... 1836–75–5 5-Nitro-o-toluidine ...... 99–55–8 Nitroquinoline-1-oxide ...... 56–57–5 N-Nitrosodi-n-butylamine 4 ...... 924–16–3 N-Nitrosodiethylamine 4 ...... 55–18–5 N-Nitrosodimethylamine 34 ...... 62–75–9 N-Nitrosodiphenylamine 34 ...... 86–30–6 N-Nitrosomethylethylamine 4 ...... 10595–95–6 N-Nitrosomethylphenylamine 4 ...... 614–00–6 N-Nitrosomorpholine 4 ...... 59–89–2 N-Nitrosopiperidine 4 ...... 100–75–5 N-Nitrosopyrrolidine 4 ...... 930–55–2 trans-Nonachlor ...... 39765–80–5 Norflurazon ...... 27314–13–2 2,2′,3,3′,4,5′,6,6′-Octachlorobiphenyl ...... 40186–71–8 Octamethyl pyrophosphoramide ...... 152–16–9 4,4’-Oxydianiline ...... 101–80–4 Parathion ...... 56–38–2 PCB–1016 35 ...... 12674–11–2 PCB–1221 35 ...... 11104–28–2 30 90 PCB–1232 35 ...... 11141–16–5 PCB–1242 35 ...... 53469–21–9 PCB–1248 35 ...... 12672–29–6 PCB–1254 35 ...... 11097–69–1 36 108 PCB–1260 35 ...... 11098–82–5 PCB–1268 35 ...... 11100–14–4 Pebulate ...... 1114–71–2 Pentachlorobenzene ...... 608–93–5 Pentachloronitrobenzene ...... 82–68–8 2,2′,3,4′,6-Pentachlorobiphenyl ...... 68194–05–8 Pentachloroethane ...... 76–01–7 Pentamethylbenzene ...... 700–12–9 Perylene ...... 198–55–0 Phenacetin ...... 62–44–2

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TABLE 3—ADDITIONAL EXTRACTABLE ANALYTES 12—Continued

MDL 7 ML 8 Analyte CAS registry (ug/L) (ug/L)

cis-Permethrin ...... 61949–76–6 trans-Permethrin ...... 61949–77–7 Phenobarbital ...... 50–06–6 Phenothiazene ...... 92–84–2 1,4-Phenylenediamine ...... 624–18–0 1-Phenylnaphthalene ...... 605–02–7 2-Phenylnaphthalene ...... 612–94–2 Phorate ...... 298–02–2 ...... 2310–18–0 Phosmet ...... 732–11–6 ...... 13171–21–6 Phthalic anhydride ...... 85–44–9 alpha-Picoline (2-Methylpyridine) ...... 109–06–8 Piperonyl sulfoxide ...... 120–62–7 Prometon ...... 1610–18–0 Prometryn ...... 7287–19–6 Pronamide ...... 23950–58–5 Propachlor ...... 1918–16–7 Propazine ...... 139–40–2 Propylthiouracil ...... 51–52–5 Pyridine ...... 110–86–1 Resorcinol (1,3-Benzenediol) ...... 108–46–3 Safrole ...... 94–59–7 Simazine ...... 122–34–9 Simetryn ...... 1014–70–6 Squalene ...... 7683–64–9 Stirofos ...... 22248–79–9 ...... 57–24–9 Styrene 9 ...... 100–42–5 Sulfallate ...... 95–06–7 Tebuthiuron ...... 34014–18–1 Terbacil ...... 5902–51–2 Terbufos ...... 13071–79–9 Terbutryn ...... 886–50–0 alpha-Terpineol ...... 98–55–5 1,2,4,5-Tetrachlorobenzene ...... 95–94–3 2,2′,4,4′-Tetrachlorobiphenyl ...... 2437–79–8 2,3,7,8-Tetrachlorodibenzo-p-dioxin ...... 1746–01–6 2,3,4,6-Tetrachlorophenol ...... 58–90–2 ...... 22248–79–9 Tetraethyl dithiopyrophosphate ...... 3689–24–5 Tetraethyl pyrophosphate ...... 107–49–3 Thianaphthene (2,3-Benzothiophene) ...... 95–15–8 Thioacetamide ...... 62–55–5 Thionazin ...... 297–97–2 Thiophenol (Benzenethiol) ...... 108–98–5 Thioxanthone ...... 492–22–8 Toluene-1,3-diisocyanate ...... 26471–62–5 Toluene-2,4-diisocyanate ...... 584–84–9 o-Toluidine ...... 95–53–4 Toxaphene 35 ...... 8001–35–2 Triadimefon ...... 43121–43–3 1,2,3-Trichlorobenzene ...... 87–61–6 2,4,5-Trichlorobiphenyl ...... 15862–07–4 2,3,6-Trichlorophenol ...... 933–75–5 2,4,5-Trichlorophenol ...... 95–95–4 Tricyclazole ...... 41814–78–2 Trifluralin ...... 1582–09–8 1,2,3-Trimethoxybenzene ...... 634–36–6 2,4,5-Trimethylaniline ...... 137–17–7 Trimethyl phosphate ...... 512–56–1 Triphenylene ...... 217–59–4 Tripropyleneglycolmethyl ether ...... 20324–33–8 1,3,5-Trinitrobenzene ...... 99–35–4 Tris(2,3-dibromopropyl) phosphate ...... 126–72–7 Tri-p-tolyl phosphate ...... 78–32–0 O,O,O-Triethyl phosphorothioate ...... 126–68–1 Trithiane ...... 291–29–4 Vernolate ...... 1929–77–7 1 Compounds that have been demonstrated amenable to extraction and gas chromatography. 2 Determine each analyte in the fraction that gives the most accurate result.

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3 Priority Pollutant (40 CFR part 423, appendix A). 4 See section 1.2. 5 These compounds are mixtures of various isomers. 6 Detected as azobenzene. 7 MDL values from the 1984 promulgated version of Method 625. 8 ML = Minimum Level—see Glossary for definition and derivation. 9 Styrene may be susceptible to losses during sampling, preservation, and/or extraction of full-volume (1 L) water samples. However, styrene is not regulated at 40 CFR part 136, and it is also listed as an analyte in EPA Method 624.1 and EPA Method 1625C, where such losses may be less than using Method 625.1.

TABLE 4—CHROMATOGRAPHIC CONDITIONS AND CHARACTERISTIC m/z’s FOR BASE/NEUTRAL EXTRACTABLES

Characteristic m/z’s Retention Analyte time Electron impact ionization Chemical ionization (sec) 1 Primary Second Second Methane Methane Methane

N-Nitrosodimethylamine 385 42 74 44 ...... bis(2-Chloroethyl) ether 704 93 63 95 63 107 109 bis(2-Chloroisopropyl) ether ...... 799 45 77 79 77 135 137 Hexachloroethane ...... 823 117 201 199 199 201 203 N-Nitrosodi-n-propyl- amine ...... 830 130 42 101 ...... Nitrobenzene ...... 849 77 123 65 124 152 164 Isophorone ...... 889 82 95 138 139 167 178 bis(2-Chloroethoxy) methane ...... 939 93 95 123 65 107 137 1,2,4-Trichlorobenzene 958 180 182 145 181 183 209 Naphthalene ...... 967 128 129 127 129 157 169 Hexachlorobutadiene .... 1006 225 223 227 223 225 227 Hexachlorocyclopentadi- ene ...... 1142 237 235 272 235 237 239 2-Chloronaphthalene ..... 1200 162 164 127 163 191 203 Acenaphthylene ...... 1247 152 151 153 152 153 181 Dimethyl phthalate...... 1273 163 194 164 151 163 164 2,6-Dinitrotoluene ...... 1300 165 89 121 183 211 223 Acenaphthene ...... 1304 154 153 152 154 155 183 2,4-Dinitrotoluene ...... 1364 165 63 182 183 211 223 Fluorene ...... 1401 166 165 167 166 167 195 4-Chlorophenyl phenyl ether ...... 1409 204 206 141 ...... Diethyl phthalate...... 1414 149 177 150 177 223 251 N-Nitrosodiphenylamine 1464 169 168 167 169 170 198 4-Bromophenyl phenyl ether ...... 1498 248 250 141 249 251 277 alpha-BHC ...... 1514 183 181 109 ...... Hexachlorobenzene ...... 1522 284 142 249 284 286 288 beta-BHC ...... 1544 183 181 109 ...... gamma-BHC ...... 1557 181 183 109 ...... Phenanthrene ...... 1583 178 179 176 178 179 207 Anthracene ...... 1592 178 179 176 178 179 207 delta-BHC ...... 1599 183 109 181 ...... Heptachlor ...... 1683 100 272 274 ...... Di-n-butyl phthalate...... 1723 149 150 104 149 205 279 Aldrin ...... 1753 66 263 220 ...... Fluoranthene ...... 1817 202 101 100 203 231 243 Heptachlor epoxide ...... 1820 353 355 351 ...... gamma-Chlordane ...... 1834 373 375 377 ...... Pyrene ...... 1852 202 101 100 203 231 243 Benzidine2 ...... 1853 184 92 185 185 213 225 alpha-Chlordane ...... 1854 373 375 377 ...... Endosulfan I ...... 1855 237 339 341 ...... 4,4′-DDE ...... 1892 246 248 176 ...... Dieldrin ...... 1907 79 263 279 ...... Endrin ...... 1935 81 263 82 ...... Endosulfan II ...... 2014 237 339 341 ...... 4,4′-DDD ...... 2019 235 237 165 ...... Endrin aldehyde ...... 2031 67 345 250 ...... Butyl benzyl phthalate ... 2060 149 91 206 149 299 327 Endosulfan sulfate ...... 2068 272 387 422 ...... 4,4′-DDT ...... 2073 235 237 165 ...... Chrysene ...... 2083 228 226 229 228 229 257 3,3′-Dichlorobenzidine ... 2086 252 254 126 ...... Benzo(a)anthracene ...... 2090 228 229 226 228 229 257

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TABLE 4—CHROMATOGRAPHIC CONDITIONS AND CHARACTERISTIC m/z’s FOR BASE/NEUTRAL EXTRACTABLES—Continued

Characteristic m/z’s Retention Analyte time Electron impact ionization Chemical ionization (sec) 1 Primary Second Second Methane Methane Methane

bis(2-Ethylhexyl) phthal- ate ...... 2124 149 167 279 149 Di-n-octyl phthalate ...... 2240 149 43 57 ...... Benzo(b)fluoranthene .... 2286 252 253 125 252 253 281 Benzo(k)fluoranthene .... 2293 252 253 125 252 253 281 Benzo(a)pyrene ...... 2350 252 253 125 252 253 281 Indeno(1,2,3-cd) pyrene 2650 276 138 277 276 277 305 Dibenz(a,h)anthracene .. 2660 278 139 279 278 279 307 Benzo(ghi)perylene ...... 2750 276 138 277 276 277 305 Toxaphene ...... 159 231 233 ...... PCB 1016 ...... 224 260 294 ...... PCB 1221 ...... 190 224 260 ...... PCB 1232 ...... 190 224 260 ...... PCB 1242 ...... 224 260 294 ...... PCB 1248 ...... 294 330 262 ...... PCB 1254 ...... 294 330 362 ...... PCB 1260 ...... 330 362 394 1 Column: 30 m x 0.25 mm ID; 94% methyl, 5% phenyl, 1% vinyl bonded phase fused silica capillary. Conditions: 5 min at 30 °C; 30–280 at 8 °C per min; isothermal at 280 °C until benzo(ghi)perylene elutes. Gas velocity: 30 cm/sec at 30 °C (at constant pressure). 2 See section 1.2; included for tailing factor testing.

TABLE 5—CHROMATOGRAPHIC CONDITIONS AND CHARACTERISTIC m/z’s FOR ACID EXTRACTABLES

Characteristic m/z’s Retention Analyte Time Electron impact ionization Chemical ionization (sec) 1 Prime Second Second Methane Methane Methane

2-Chlorophenol ...... 705 128 64 130 129 131 157 Phenol ...... 700 94 65 66 95 123 135 2-Nitrophenol ...... 900 139 65 109 140 168 122 2,4-Dimethylphenol ...... 924 122 107 121 123 151 163 2,4-Dichlorophenol ...... 947 162 164 98 163 165 167 4-Chloro-3-methylphenol 1091 142 107 144 143 171 183 2,4,6-Trichlorophenol .... 1165 196 198 200 197 199 201 2,4-Dinitrophenol ...... 1325 184 63 154 185 213 225 4-Nitrophenol ...... 1354 65 139 109 140 168 122 2-Methyl-4,6- dinitrophenol ...... 1435 198 182 77 199 227 239 Pentachlorophenol ...... 1561 266 264 268 267 265 269 Column: 30 m x 0.25 mm ID; 94% methyl, 5% phenyl, 1% vinyl bonded phase fused silica capillary. Conditions: 5 min at 30 °C; 30–250 at 8 °C per min; isothermal at 280 °C until pentachlorophenol elutes. Gas velocity: 30 cm/sec at 30 °C (at constant pressure).

TABLE 6—QC ACCEPTANCE CRITERIA—METHOD 625 1

Range for Q Limit for s Range for Range for Limit for Analyte 2 3 3 3 (%) (%) X (%) P1, P2(%) RPD (%)

Acenaphthene ...... 70–130 29 60–132 47–145 48 Acenaphthylene ...... 60–130 45 54–126 33–145 74 Aldrin ...... 7–152 39 7–152 D–166 81 Anthracene ...... 58–130 40 43–120 27–133 66 Benzo(a)anthracene ...... 42–133 32 42–133 33–143 53 Benzo(b)fluoranthene ...... 42–140 43 42–140 24–159 71 Benzo(k)fluoranthene ...... 25–146 38 25–146 11–162 63 Benzo(a)pyrene ...... 32–148 43 32–148 17–163 72 Benzo(ghi)perylene ...... 13–195 61 D–195 D–219 97 Benzyl butyl phthalate ...... 43–140 36 D–140 D–152 60 beta-BHC ...... 42–131 37 42–131 24–149 61 delta-BHC ...... D–130 77 D–120 D–120 129 bis(2-Chloroethyl)ether ...... 52–130 65 43–126 12–158 108 bis(2-Chloroethoxy)methane ...... 52–164 32 49–165 33–184 54 bis(2-Chloroisopropyl) ether ...... 63–139 46 63–139 36–166 76 bis(2-Ethylhexyl) phthalate ...... 43–137 50 29–137 8–158 82

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TABLE 6—QC ACCEPTANCE CRITERIA—METHOD 625 1—Continued

Range for Q Limit for s Range for Range for Limit for Analyte 2 3 3 3 (%) (%) X (%) P1, P2(%) RPD (%)

4-Bromophenyl phenyl ether ...... 70–130 26 65–120 53–127 43 2-Chloronaphthalene ...... 70–130 15 65–120 60–120 24 4-Chlorophenyl phenyl ether ...... 57–145 36 38–145 25–158 61 Chrysene ...... 44–140 53 44–140 17–168 87 4,4′-DDD ...... D–135 56 D–135 D–145 93 4,4′-DDE ...... 19–130 46 19–120 4–136 77 4,4′-DDT ...... D–171 81 D–171 D–203 135 Dibenz(a,h)anthracene ...... 13–200 75 D–200 D–227 126 Di-n-butyl phthalate ...... 52–130 28 8–120 1–120 47 3,3′-Dichlorobenzidine ...... 18–213 65 8–213 D–262 108 Dieldrin ...... 70–130 38 44–119 29–136 62 Diethyl phthalate ...... 47–130 60 D–120 D–120 100 Dimethyl phthalate...... 50–130 110 D–120 D–120 183 2,4-Dinitrotoluene ...... 53–130 25 48–127 39–139 42 2,6-Dinitrotoluene ...... 68–137 29 68–137 50–158 48 Di-n-octyl phthalate ...... 21–132 42 19–132 4–146 69 Endosulfan sulfate ...... D–130 42 D–120 D–120 70 Endrin aldehyde ...... D–189 45 D–189 D–209 75 Fluoranthene ...... 47–130 40 43–121 26–137 66 Fluorene ...... 70–130 23 70–120 59–121 38 Heptachlor ...... D–172 44 D–172 D–192 74 Heptachlor epoxide ...... 70–130 61 71–120 26–155 101 Hexachlorobenzene ...... 38–142 33 8–142 D–152 55 Hexachlorobutadiene ...... 68–130 38 38–120 24–120 62 Hexachloroethane ...... 55–130 32 55–120 40–120 52 Indeno(1,2,3-cd)pyrene ...... 13–151 60 D–151 D–171 99 Isophorone ...... 52–180 56 47–180 21–196 93 Naphthalene ...... 70–130 39 36–120 21–133 65 Nitrobenzene ...... 54–158 37 54–158 35–180 62 N-Nitrosodi-n-propylamine ...... 59–170 52 14–198 D–230 87 PCB–1260 ...... 19–130 77 19–130 D–164 128 Phenanthrene ...... 67–130 24 65–120 54–120 39 Pyrene ...... 70–130 30 70–120 52–120 49 1,2,4-Trichlorobenzene ...... 61–130 30 57–130 44–142 50 4-Chloro-3-methylphenol ...... 68–130 44 41–128 22–147 73 2-Chlorophenol ...... 55–130 37 36–120 23–134 61 2,4-Dichlorophenol ...... 64–130 30 53–122 39–135 50 2,4-Dimethylphenol ...... 58–130 35 42–120 32–120 58 2,4-Dinitrophenol ...... 39–173 79 D–173 D–191 132 2-Methyl-4,6-dinitrophenol ...... 56–130 122 53–130 D–181 203 2-Nitrophenol ...... 61–163 33 45–167 29–182 55 4-Nitrophenol ...... 35–130 79 13–129 D–132 131 Pentachlorophenol ...... 42–152 52 38–152 14–176 86 Phenol ...... 48–130 39 17–120 5–120 64 2,4,6-Trichlorophenol ...... 69–130 35 52–129 37–144 58 1 Acceptance criteria are based upon method performance data in Table 7 and from EPA Method 1625. Where necessary, lim- its for recovery have been broadened to assure applicability to concentrations below those used to develop Table 7. 2 Test concentration = 100 μg/mL. 3 Test concentration = 100 μg/L. Q = Calibration verification (sections 7.3.1 and 13.4). s = Standard deviation for four recovery measurements in the DOC test (section 8.2.4). X = Average recovery for four recovery measurements in the DOC test (section 8.2.4). P1, P2 = MS/MSD recovery (section 8.3.2, section 8.4.2). RPD = MS/MSD relative percent difference (RPD; section 8.3.3). D = Detected; result must be greater than zero.

TABLE 7—PRECISION AND RECOVERY AS FUNCTIONS OF CONCENTRATION—METHOD 625 1

Single analyst Overall Recovery, X′ ′ ′ Analyte μ precision, sr precision, S ( g/L) (μg/L) (μg/L)

Acenaphthene ...... 0.96C + 0.19 0.15 X¥0.12 0.21 X¥0.67 Acenaphthylene ...... 0.89C + 0.74 0.24 X¥1.06 0.26 X¥0.54 Aldrin ...... 0.78C + 1.66 0.27 X¥1.28 0.43 X + 1.13 Anthracene ...... 0.80C + 0.68 0.21 X¥0.32 0.27 X¥0.64 Benzo(a)anthracene ...... 0.88C¥0.60 0.15 X + 0.93 0.26 X¥0.28 Benzo(b)fluoranthene ...... 0.93C¥1.80 0.22 X + 0.43 0.29 X + 0.96 Benzo(k)fluoranthene ...... 0.87C¥1.56 0.19 X + 1.03 0.35 X + 0.40 Benzo(a)pyrene ...... 0.90C¥0.13 0.22 X + 0.48 0.32 X + 1.35 Benzo(ghi)perylene ...... 0.98C¥0.86 0.29 X + 2.40 0.51 X¥0.44 Benzyl butyl phthalate ...... 0.66C¥1.68 0.18 X + 0.94 0.53 X + 0.92

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TABLE 7—PRECISION AND RECOVERY AS FUNCTIONS OF CONCENTRATION—METHOD 625 1— Continued

Single analyst Overall Recovery, X′ ′ ′ Analyte μ precision, sr precision, S ( g/L) (μg/L) (μg/L)

beta-BHC ...... 0.87C¥0.94 0.20 X¥0.58 0.30 X¥1.94 delta-BHC ...... 0.29C¥1.09 0.34 X + 0.86 0.93 X¥0.17 bis(2-Chloroethyl) ether ...... 0.86C¥1.54 0.35 X¥0.99 0.35 X + 0.10 bis(2-Chloroethoxy) methane ...... 1.12C¥5.04 0.16 X + 1.34 0.26 X + 2.01 bis(2-Chloroisopropyl) ether ...... 1.03C¥2.31 0.24 X + 0.28 0.25 X + 1.04 bis(2-Ethylhexyl) phthalate ...... 0.84C¥1.18 0.26 X + 0.73 0.36 X + 0.67 4-Bromophenyl phenyl ether ...... 0.91C¥1.34 0.13 X + 0.66 0.16 X + 0.66 2-Chloronaphthalene ...... 0.89C + 0.01 0.07 X + 0.52 0.13 X + 0.34 4-Chlorophenyl phenyl ether ...... 0.91C + 0.53 0.20 X¥0.94 0.30 X¥0.46 Chrysene ...... 0.93C¥1.00 0.28 X + 0.13 0.33 X¥0.09 4,4′-DDD ...... 0.56C¥0.40 0.29 X¥0.32 0.66 X¥0.96 4,4′-DDE ...... 0.70C¥0.54 0.26 X¥1.17 0.39 X¥1.04 4,4′-DDT ...... 0.79C¥3.28 0.42 X + 0.19 0.65 X¥0.58 Dibenz(a,h)anthracene ...... 0.88C + 4.72 0.30 X + 8.51 0.59 X + 0.25 Di-n-butyl phthalate ...... 0.59C + 0.71 0.13 X + 1.16 0.39 X + 0.60 3,3’-Dichlorobenzidine ...... 1.23C¥12.65 0.28 X + 7.33 0.47 X + 3.45 Dieldrin ...... 0.82C¥0.16 0.20 X¥0.16 0.26 X¥0.07 Diethyl phthalate ...... 0.43C + 1.00 0.28 X + 1.44 0.52 X + 0.22 Dimethyl phthalate ...... 0.20C + 1.03 0.54 X + 0.19 1.05 X¥0.92 2,4-Dinitrotoluene ...... 0.92C¥4.81 0.12 X + 1.06 0.21 X + 1.50 2,6-Dinitrotoluene ...... 1.06C¥3.60 0.14 X + 1.26 0.19 X + 0.35 Di-n-octyl phthalate ...... 0.76C¥0.79 0.21 X + 1.19 0.37 X + 1.19 Endosulfan sulfate ...... 0.39C + 0.41 0.12 X + 2.47 0.63 X¥1.03 Endrin aldehyde ...... 0.76C¥3.86 0.18 X + 3.91 0.73 X¥0.62 Fluoranthene ...... 0.81C + 1.10 0.22 X + 0.73 0.28 X¥0.60 Fluorene ...... 0.90C¥0.00 0.12 X + 0.26 0.13 X + 0.61 Heptachlor ...... 0.87C¥2.97 0.24 X¥0.56 0.50 X¥0.23 Heptachlor epoxide ...... 0.92C¥1.87 0.33 X¥0.46 0.28 X + 0.64 Hexachlorobenzene ...... 0.74C + 0.66 0.18 X¥0.10 0.43 X¥0.52 Hexachlorobutadiene ...... 0.71C¥1.01 0.19 X + 0.92 0.26 X + 0.49 Hexachloroethane ...... 0.73C¥0.83 0.17 X + 0.67 0.17 X + 0.80 Indeno(1,2,3-cd)pyrene ...... 0.78C¥3.10 0.29 X + 1.46 0.50 X + 0.44 Isophorone ...... 1.12C + 1.41 0.27 X + 0.77 0.33 X + 0.26 Naphthalene ...... 0.76C + 1.58 0.21 X¥0.41 0.30 X¥0.68 Nitrobenzene ...... 1.09C¥3.05 0.19 X + 0.92 0.27 X + 0.21 N-Nitrosodi-n-propylamine ...... 1.12C¥6.22 0.27 X + 0.68 0.44 X + 0.47 PCB–1260 ...... 0.81C¥10.86 0.35 X + 3.61 0.43 X + 1.82 Phenanthrene ...... 0.87C¥0.06 0.12 X + 0.57 0.15 X + 0.25 Pyrene ...... 0.84C¥0.16 0.16 X + 0.06 0.15 X + 0.31 1,2,4-Trichlorobenzene ...... 0.94C¥0.79 0.15 X + 0.85 0.21 X + 0.39 4-Chloro-3-methylphenol ...... 0.84C + 0.35 0.23 X + 0.75 0.29 X + 1.31 2-Chlorophenol ...... 0.78C + 0.29 0.18 X + 1.46 0.28 X + 0.97 2,4-Dichlorophenol ...... 0.87C + 0.13 0.15 X + 1.25 0.21 X + 1.28 2,4-Dimethylphenol ...... 0.71C + 4.41 0.16 X + 1.21 0.22 X + 1.31 2,4-Dinitrophenol ...... 0.81C¥18.04 0.38 X + 2.36 0.42 X + 26.29 2-Methyl-4,6-Dinitrophenol ...... 1.04C¥28.04 0.05 X + 42.29 0.26 X + 23.10 2-Nitrophenol ...... 1.07C¥1.15 0.16 X + 1.94 0.27 X + 2.60 4-Nitrophenol ...... 0.61C¥1.22 0.38 X + 2.57 0.44 X + 3.24 Pentachlorophenol ...... 0.93C + 1.99 0.24 X + 3.03 0.30 X + 4.33 Phenol ...... 0.43C + 1.26 0.26 X + 0.73 0.35 X + 0.58 2,4,6-Trichlorophenol ...... 0.91C¥0.18 0.16 X + 2.22 0.22 X + 1.81 1 Regressions based on data from Reference 2. X′ = Expected recovery for one or more measurements of a sample containing a concentration of C, in μg/L. sr′ = Expected single analyst standard deviation of measurements at an average concentration found of X, in μg/L. S′ = Expected interlaboratory standard deviation of measurements at an average concentration found of X, in μg/L. C = True value for the concentration, in μg/L. X = Average recovery found for measurements of samples containing a concentration of C, in μg/L.

TABLE 8—SUGGESTED INTERNAL AND SURROGATE STANDARDS

Range for surrogate recovery (%) 1 Base/neutral fraction Calibration Recovery from verification samples

Acenaphthalene-d8 ...... 66–152 33–168 Acenaphthene-d10 ...... 71–141 30–180 Aniline-d5. Anthracene-d10 ...... 58–171 23–142

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TABLE 8—SUGGESTED INTERNAL AND SURROGATE STANDARDS—Continued

Range for surrogate recovery (%) 1 Base/neutral fraction Calibration Recovery from verification samples

Benzo(a)anthracene-d12 ...... 28–357 22–329 Benzo(a)pyrene-d12 ...... 32–194 32–194 4-Chloroaniline-d4 ...... 1–145 1–145 bis(2-Chloroethyl) ether-d8 ...... 52–194 25–222 Chrysene-d12 ...... 23–290 23–290 Decafluorobiphenyl. 4,4′-Dibromobiphenyl. 4,4′-Dibromooctafluorobiphenyl. 1,4-Dichlorobenzene-d4 ...... 65–153 11–245 2,2′-Difluorobiphenyl. Dimethyl phthalate-d6 ...... 47–211 1–500 Fluoranthene-d10 ...... 47–215 30–187 Fluorene-d10 ...... 61–164 38–172 4-Fluoroaniline. 1-Fluoronaphthalene. 2-Fluoronaphthalene. 2-Methylnaphthalene-d10 ...... 50–150 50–150 Naphthalene-d8 ...... 71–141 22–192 Nitrobenzene-d5 ...... 46–219 15–314 2,3,4,5,6-Pentafluorobiphenyl. Perylene-d12. Phenanthrene-d10 ...... 67–149 34–168 Pyrene-d10 ...... 48–210 28–196 Pyridine-d5. Acid fraction. 2-Chlorophenol-d4 ...... 55–180 33–180 2,4-Dichlorophenol-d3 ...... 64–157 34–182 4,6-Dinitro-2-methylphenol-d2 ...... 56–177 22–307 2-Fluorophenol. 4-Methylphenol-d8 ...... 25–111 25–111 2-Nitrophenol-d4 ...... 61–163 37–163 4-Nitrophenol-d4 ...... 35–287 6–500 Pentafluorophenol. 2-Perfluoromethylphenol. Phenol-d5 ...... 48–208 8–424 1 Recovery from samples is the wider of the criteria in the CLP SOW for organics or in Method 1625.

TABLE 9A—DFTPP KEY m/z’s AND ABUNDANCE CRITERIA FOR QUADRUPOLE INSTRUMENTS 1

m/z Abundance criteria

51 ...... 30–60 percent of m/z 198. 68 ...... Less than 2 percent of m/z 69. 70 ...... Less than 2 percent of m/z 69. 127 ...... 40–60 percent of base peak m/z 198. 197 ...... Less than 1 percent of m/z 198. 198 ...... Base peak, 100 percent relative abundance. 199 ...... 5–9 percent of m/z 198. 275 ...... 10–30 percent of m/z 198. 365 ...... Greater than 1 percent of m/z 198. 441 ...... Present but less than m/z 443. 442 ...... 40–100 percent of m/z 198. 443 ...... 17–23 percent of m/z 442. 1 Criteria in these tables are for quadrupole and time-of-flight instruments. Alternative tuning criteria from other published EPA reference methods may be used provided method performance is not adversely affected. Alternative tuning criteria specified by an instrument manufacturer may also be used for another type of mass spectrometer, provided method performance is not ad- versely affected.

TABLE 9B—DFTPP KEY m/z’s AND ABUNDANCE CRITERIA FOR TIME-OF-FLIGHT INSTRUMENTS 1

m/z Abundance criteria

51 ...... 10–85 percent of the base peak. 68 ...... Less than 2 percent of m/z 69. 70 ...... Less than 2 percent of m/z 69. 127 ...... 10–80 percent of the base peak. 197 ...... Less than 2 percent of Mass 198.

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TABLE 9B—DFTPP KEY m/z’s AND ABUNDANCE CRITERIA FOR TIME-OF-FLIGHT INSTRUMENTS 1— Continued

m/z Abundance criteria

198 ...... Base peak, or greater than 50% of m/z 442. 199 ...... 5–9 percent of m/z 198. 275 ...... 10–60 percent of the base peak. 365 ...... Greater than 0.5 percent of m/z 198. 441 ...... Less than 150 percent of m/z 443. 442 ...... Base peak or greater than 30 percent of m/z 198. 443 ...... 15–24 percent of m/z 442. 1 Criteria in these tables are for quadrupole and time-of-flight instruments. Alternative tuning criteria from other published EPA reference methods may be used provided method performance is not adversely affected. Alternative tuning criteria specified by an instrument manufacturer may also be used for another type of mass spectrometer, or for an alternative carrier gas, provided method performance is not adversely affected.

21. FIGURES

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22. Glossary ppb part-per-billion These definitions and purposes are specific ppm part-per-million to this method but have been conformed to ppt part-per-trillion common usage to the extent possible. psig pounds-per-square inch gauge 22.1 Units of weight and measure and 22.2 Definitions and acronyms (in alpha- their abbreviations. betical order). 22.1.1 Symbols. Analyte—A compound or mixture of com- °C degrees Celsius pounds (e.g., PCBs) tested for by this meth- μg microgram od. The analytes are listed in Tables 1–3. μL microliter Batch—See Extraction. < less than Blank—An aliquot of reagent water that is > greater than treated exactly as a sample including expo- ≤ less than or equal to sure to all glassware, equipment, solvents, % percent reagents, internal standards, and surrogates 22.1.2 Abbreviations (in alphabetical that are used with samples. The blank is order). used to determine if analytes or inter- cm centimeter ferences are present in the laboratory envi- g gram h hour ronment, the reagents, or the apparatus. ID inside diameter Calibration—The process of determining in. inch the relationship between the output or re- L liter sponse of a measuring instrument and the m mass or meter value of an input standard. Historically, mg milligram EPA has referred to a multi-point calibra- min minute tion as the ‘‘initial calibration,’’ to differen- mL milliliter tiate it from a single-point calibration mm millimeter verification. ms millisecond Calibration standard—A solution prepared m/z mass-to-charge ratio from stock solutions and/or a secondary N normal; gram molecular weight of solute standards and containing the analytes of in- divided by hydrogen equivalent of solute, terest, surrogates, and internal standards. per liter of solution The calibration standard is used to calibrate ng nanogram the response of the GC/MS instrument pg picogram against analyte concentration.

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Calibration verification standard—The Laboratory Control Sample (LCS; labora- mid-point calibration standard used to verify tory fortified blank; section 8.4)—An aliquot calibration. See sections 7.3 and 13.4. of reagent water spiked with known quan- Descriptor—In SIM, the beginning and end- tities of the analytes of interest and surro- ing retention times for the RT window, the gates. The LCS is analyzed exactly like a m/z’s sampled in the RT window, and the sample. Its purpose is to assure that the re- dwell time at each m/z. sults produced by the laboratory remain Extracted ion current profile (EICP)—The within the limits specified in this method for line described by the signal at a given m/z. precision and recovery. Extraction Batch—A set of up to 20 field Laboratory fortified sample matrix—See samples (not including QC samples) started Matrix spike. through the extraction process on a given 24- Laboratory reagent blank—A blank run on hour shift (section 3.1). Each extraction laboratory reagents; e.g., methylene chloride batch must be accompanied by a blank (sec- (section 11.1.5). tion 8.5), a laboratory control sample (LCS, Matrix spike (MS) and matrix spike dupli- section 8.4), and a matrix spike and duplicate cate (MSD) (laboratory fortified sample ma- (MS/MSD; Section 8.3), resulting in a min- trix and duplicate)—Two aliquots of an envi- imum of five analyses (1 sample, 1 blank, 1 ronmental sample to which known quan- LCS, 1 MS, and 1 MSD) and a maximum of 24 tities of the analytes of interest and surro- analyses (20 field samples, 1 blank, 1 LCS, 1 gates are added in the laboratory. The MS/ MS, and 1 MSD) for the batch. If greater MSD are prepared and analyzed exactly like a field sample. Their purpose is to quantify than 20 samples are to be extracted in a 24- any additional bias and imprecision caused hour shift, the samples must be separated by the sample matrix. The background con- into extraction batches of 20 or fewer sam- centrations of the analytes in the sample ples. matrix must be determined in a separate ali- Field Duplicates—Two samples collected quot and the measured values in the MS/ at the same time and placed under identical MSD corrected for background concentra- conditions, and treated identically through- tions. out field and laboratory procedures. Results May—This action, activity, or procedural of analyses of the field duplicates provide an step is neither required nor prohibited. estimate of the precision associated with May not—This action, activity, or proce- sample collection, preservation, and storage, dural step is prohibited. as well as with laboratory procedures. Method blank—See blank. Field blank—An aliquot of reagent water Method detection limit (MDL)—A detec- or other reference matrix that is placed in a tion limit determined by the procedure at 40 sample container in the field, and treated as CFR part 136, appendix B. The MDLs deter- a sample in all respects, including exposure mined by EPA in the original version of the to sampling site conditions, storage, preser- method are listed in Tables 1, 2 and 3. As vation, and all analytical procedures. The noted in section 1.5, use the MDLs in Tables purpose of the field blank is to determine if 1, 2, and 3 in conjunction with current MDL the field or sample transporting procedures data from the laboratory actually analyzing and environments have contaminated the samples to assess the sensitivity of this pro- sample. cedure relative to project objectives and reg- GC—Gas chromatograph or gas chroma- ulatory requirements (where applicable). tography. Minimum level (ML)—The term ‘‘minimum Internal standard—A compound added to level’’ refers to either the sample concentra- an extract or standard solution in a known tion equivalent to the lowest calibration amount and used as a reference for quantita- point in a method or a multiple of the meth- tion of the analytes of interest and surro- od detection limit (MDL), whichever is high- gates. In this method the internal standards er. Minimum levels may be obtained in sev- are stable isotopically labeled analogs of se- eral ways: They may be published in a meth- lected method analytes (Table 8). Also see od; they may be based on the lowest accept- Internal standard quantitation. able calibration point used by a laboratory; Internal standard quantitation—A means or they may be calculated by multiplying of determining the concentration of an the MDL in a method, or the MDL deter- analyte of interest (Tables 1–3) by reference mined by a laboratory, by a factor of 3. For to a compound not expected to be found in a the purposes of NPDES compliance moni- sample. toring, EPA considers the following terms to DOC—Initial demonstration of capability be synonymous: ‘‘quantitation limit,’’ ‘‘re- (section 8.2); four aliquots of reagent water porting limit,’’ and ‘‘minimum level.’’ spiked with the analytes of interest and ana- MS—Mass spectrometer or mass spectrom- lyzed to establish the ability of the labora- etry, or matrix spike (a QC sample type). tory to generate acceptable precision and re- MSD—Matrix spike duplicate (a QC sample covery. A DOC is performed prior to the first type). time this method is used and any time the Must—This action, activity, or procedural method or instrumentation is modified. step is required.

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m/z—The ratio of the mass of an ion (m) a source that will attest to the purity, au- detected in the mass spectrometer to the thenticity, and concentration of the stand- charge (z) of that ion. ard. Preparation blank—See blank. Surrogate—A compound unlikely to be Quality control check sample (QCS)—See found in a sample, and which is spiked into Laboratory Control Sample. sample in a known amount before extraction Reagent water—Water demonstrated to be or other processing, and is quantitated with free from the analytes of interest and poten- the same procedures used to quantify other tially interfering substances at the MDLs for sample components. The purpose of the sur- the analytes in this method. rogate is to monitor method performance Regulatory compliance limit (or regu- with each sample. latory concentration limit)—A limit on the concentration or amount of a pollutant or METHOD 1613, REVISION B contaminant specified in a nationwide stand- ard, in a permit, or otherwise established by Tetra- Through Octa-Chlorinated Dioxins and a regulatory/control authority. Furans by Isotope Dilution HRGC/HRMS Relative retention time (RRT)—The ratio of the retention time of an analyte to the re- 1.0 Scope and Application tention time of its associated internal stand- ard. RRT compensates for small changes in 1.1 This method is for determination of the GC temperature program that can affect tetra- through octa-chlorinated dibenzo-p- the absolute retention times of the analyte dioxins (CDDs) and dibenzofurans (CDFs) in and internal standard. RRT is a unitless water, soil, sediment, sludge, tissue, and quantity. other sample matrices by high resolution gas Relative standard deviation (RSD)—The chromatography/high resolution mass spec- standard deviation times 100 divided by the trometry (HRGC/HRMS). The method is for mean. Also termed ‘‘coefficient of vari- use in EPA’s data gathering and monitoring ation.’’ programs associated with the Clean Water RF—Response factor. See section 7.2.2. Act, the Resource Conservation and Recov- RSD—See relative standard deviation. ery Act, the Comprehensive Environmental Safety Data Sheet (SDS)—Written infor- Response, Compensation and Liability Act, mation on a chemical’s toxicity, health haz- and the . The meth- ards, physical properties, fire, and reac- od is based on a compilation of EPA, indus- tivity, including storage, spill, and handling try, commercial laboratory, and academic precautions that meet the requirements of methods (References 1–6). OSHA, 29 CFR 1910.1200(g) and appendix D to 1.2 The seventeen 2,3,7,8-substituted § 1910.1200. United Nations Globally Har- CDDs/CDFs listed in Table 1 may be deter- monized System of Classification and Label- mined by this method. Specifications are ling of Chemicals (GHS), third revised edi- also provided for separate determination of tion, United Nations, 2009. 2,3,7,8-tetrachloro-dibenzo-p-dioxin (2,3,7,8- Selected Ion Monitoring (SIM)—An MS TCDD) and 2,3,7,8-tetrachloro-dibenzofuran technique in which a few m/z’s are mon- (2,3,7,8-TCDF). itored. When used with gas chromatography, 1.3 The detection limits and quantitation the m/z’s monitored are usually changed pe- levels in this method are usually dependent riodically throughout the chromatographic on the level of interferences rather than in- run, to correlate with the characteristic m/ strumental limitations. The minimum levels z’s of the analytes, surrogates, and internal (MLs) in Table 2 are the levels at which the standards as they elute from the CDDs/CDFs can be determined with no inter- chromatographic column. The technique is ferences present. The Method Detection often used to increase sensitivity and mini- Limit (MDL) for 2,3,7,8-TCDD has been deter- mize interferences. mined as 4.4 pg/L (parts-per-quadrillion) Signal-to-noise ratio (S/N)—The height of the signal as measured from the mean (aver- using this method. age) of the noise to the peak maximum di- 1.4 The GC/MS portions of this method vided by the width of the noise. are for use only by analysts experienced with Should—This action, activity, or proce- HRGC/HRMS or under the close supervision dural step is suggested but not required. of such qualified persons. Each laboratory SPE—Solid-phase extraction; an extrac- that uses this method must demonstrate the tion technique in which an analyte is ex- ability to generate acceptable results using tracted from an aqueous solution by passage the procedure in Section 9.2. over or through a material capable of revers- 1.5 This method is ‘‘performance-based’’. ibly adsorbing the analyte. Also termed liq- The analyst is permitted to modify the uid-solid extraction. method to overcome interferences or lower Stock solution—A solution containing an the cost of measurements, provided that all analyte that is prepared using a reference performance criteria in this method are met. material traceable to EPA, the National In- The requirements for establishing method stitute of Science and Technology (NIST), or equivalency are given in Section 9.1.2.

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1.6 Any modification of this method, be- agitated for 12–24 hours. The extract is evap- yond those expressly permitted, shall be con- orated to dryness, and the lipid content is sidered a major modification subject to ap- determined. plication and approval of alternate test pro- 37 2.2 After extraction, Cl4-labeled 2,3,7,8- cedures under 40 CFR 136.4 and 136.5. TCDD is added to each extract to measure 2.0 Summary of Method the efficiency of the cleanup process. Sample cleanups may include back-extraction with Flow charts that summarize procedures for acid and/or base, and gel permeation, alu- sample preparation, extraction, and analysis mina, silica gel, Florisil and activated car- are given in Figure 1 for aqueous and solid bon chromatography. High-performance liq- samples, Figure 2 for multi-phase samples, uid chromatography (HPLC) can be used for and Figure 3 for tissue samples. further isolation of the 2,3,7,8-isomers or 2.1 Extraction. other specific isomers or congeners. Prior to 2.1.1 Aqueous samples (samples con- the cleanup procedures cited above, tissue taining less than 1% solids)—Stable extracts are cleaned up using an anthropo- isotopically labeled analogs of 15 of the 2,3,7,8-substituted CDDs/CDFs are spiked genic isolation column, a batch silica gel ad- into a 1 L sample, and the sample is ex- sorption, or sulfuric acid and base back-ex- tracted by one of three procedures: traction, depending on the tissue extraction 2.1.1.1 Samples containing no visible par- procedure used. ticles are extracted with methylene chloride 2.3 After cleanup, the extract is con- in a separatory funnel or by the solid-phase centrated to near dryness. Immediately prior extraction technique summarized in Section to injection, internal standards are added to 2.1.1.3. The extract is concentrated for clean- each extract, and an aliquot of the extract is up. injected into the gas chromatograph. The 2.1.1.2 Samples containing visible par- analytes are separated by the GC and de- ticles are vacuum filtered through a glass- tected by a high-resolution (≥10,000) mass fiber filter. The filter is extracted in a Soxh- spectrometer. Two exact m/z’s are monitored let/Dean-Stark (SDS) extractor (Reference for each analyte. 7), and the filtrate is extracted with meth- 2.4 An individual CDD/CDF is identified ylene chloride in a separatory funnel. The by comparing the GC retention time and ion- methylene chloride extract is concentrated abundance ratio of two exact m/z’s with the and combined with the SDS extract prior to corresponding retention time of an authentic cleanup. standard and the theoretical or acquired ion- 2.1.1.3 The sample is vacuum filtered abundance ratio of the two exact m/z’s. The through a glass-fiber filter on top of a solid- non-2,3,7,8 substituted isomers and congeners phase extraction (SPE) disk. The filter and disk are extracted in an SDS extractor, and are identified when retention times and ion- the extract is concentrated for cleanup. abundance ratios agree within predefined 2.1.2 Solid, semi-solid, and multi-phase limits. Isomer specificity for 2,3,7,8-TCDD samples (but not tissue)—The labeled com- and 2,3,7,8-TCDF is achieved using GC col- pounds are spiked into a sample containing umns that resolve these isomers from the 10 g (dry weight) of solids. Samples con- other tetra-isomers. taining multiple phases are pressure filtered 2.5 Quantitative analysis is performed and any aqueous liquid is discarded. Coarse using selected ion current profile (SICP) solids are ground or homogenized. Any non- areas, in one of three ways: aqueous liquid from multi-phase samples is 2.5.1 For the 15 2,3,7,8-substituted CDDs/ combined with the solids and extracted in an CDFs with labeled analogs (see Table 1), the SDS extractor. The extract is concentrated GC/MS system is calibrated, and the con- for cleanup. centration of each compound is determined 2.1.3 Fish and other tissue—The sample is using the isotope dilution technique. extracted by one of two procedures: 2.5.2 For 1,2,3,7,8,9-HxCDD, OCDF, and the 2.1.3.1 Soxhlet or SDS extraction—A 20 g labeled compounds, the GC/MS system is aliquot of sample is homogenized, and a 10 g calibrated and the concentration of each aliquot is spiked with the labeled com- compound is determined using the internal pounds. The sample is mixed with sodium standard technique. sulfate, allowed to dry for 12–24 hours, and extracted for 18–24 hours using methylene 2.5.3 For non-2,3,7,8-substituted isomers chloride:hexane (1:1) in a Soxhlet extractor. and for all isomers at a given level of The extract is evaporated to dryness, and the chlorination (i.e., total TCDD), concentra- lipid content is determined. tions are determined using response factors 2.1.3.2 HCl digestion—A 20 g aliquot is ho- from calibration of the CDDs/CDFs at the mogenized, and a 10 g aliquot is placed in a same level of chlorination. bottle and spiked with the labeled com- 2.6 The quality of the analysis is assured pounds. After equilibration, 200 mL of hydro- through reproducible calibration and testing chloric acid and 200 mL of methylene chlo- of the extraction, cleanup, and GC/MS sys- ride:hexane (1:1) are added, and the bottle is tems.

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3.0 Definitions cise the cleanup systems when samples con- taining pentachloronaphthalene are ex- Definitions are given in the glossary at the pected. end of this method. 4.3.2 When a reference matrix that simu- 4.0 Contamination and Interferences lates the sample matrix under test is not available, reagent water (Section 7.6.1) can 4.1 Solvents, reagents, glassware, and be used to simulate water samples; play- other sample processing hardware may yield ground sand (Section 7.6.2) or white quartz artifacts and/or elevated baselines causing sand (Section 7.3.2) can be used to simulate misinterpretation of chromatograms (Ref- soils; filter paper (Section 7.6.3) can be used erences 8–9). Specific selection of reagents to simulate papers and similar materials; and purification of solvents by distillation in and corn oil (Section 7.6.4) can be used to all-glass systems may be required. Where simulate tissues. possible, reagents are cleaned by extraction 4.4 Interferences coextracted from sam- or solvent rinse. ples will vary considerably from source to 4.2 Proper cleaning of glassware is ex- source, depending on the diversity of the site tremely important, because glassware may being sampled. Interfering compounds may not only contaminate the samples but may be present at concentrations several orders also remove the analytes of interest by ad- of magnitude higher than the CDDs/CDFs. sorption on the glass surface. The most frequently encountered inter- 4.2.1 Glassware should be rinsed with sol- ferences are chlorinated biphenyls, methoxy vent and washed with a detergent solution as biphenyls, hydroxydiphenyl ethers, soon after use as is practical. Sonication of benzylphenyl ethers, polynuclear aromatics, glassware containing a detergent solution and pesticides. Because very low levels of for approximately 30 seconds may aid in CDDs/CDFs are measured by this method, cleaning. Glassware with removable parts, the elimination of interferences is essential. particularly separatory funnels with The cleanup steps given in Section 13 can be fluoropolymer stopcocks, must be disassem- used to reduce or eliminate these inter- bled prior to detergent washing. 4.2.2 After detergent washing, glassware ferences and thereby permit reliable deter- should be rinsed immediately, first with mination of the CDDs/CDFs at the levels methanol, then with hot tap water. The tap shown in Table 2. water rinse is followed by another methanol 4.5 Each piece of reusable glassware rinse, then acetone, and then methylene should be numbered to associate that glass- chloride. ware with the processing of a particular sam- 4.2.3 Do not bake reusable glassware in an ple. This will assist the laboratory in track- oven as a routine part of cleaning. Baking ing possible sources of contamination for in- may be warranted after particularly dirty dividual samples, identifying glassware asso- samples are encountered but should be mini- ciated with highly contaminated samples mized, as repeated baking of glassware may that may require extra cleaning, and deter- cause active sites on the glass surface that mining when glassware should be discarded. will irreversibly adsorb CDDs/CDFs. 4.6 Cleanup of tissue—The natural lipid 4.2.4 Immediately prior to use, the Soxh- content of tissue can interfere in the anal- let apparatus should be pre-extracted with ysis of tissue samples for the CDDs/CDFs. toluene for approximately three hours (see The lipid contents of different species and Sections 12.3.1 through 12.3.3). Separatory portions of tissue can vary widely. Lipids are funnels should be shaken with methylene soluble to varying degrees in various organic chloride/toluene (80/20 mixture) for two min- solvents and may be present in sufficient utes, drained, and then shaken with pure quantity to overwhelm the column methylene chloride for two minutes. chromatographic cleanup procedures used 4.3 All materials used in the analysis for cleanup of sample extracts. Lipids must shall be demonstrated to be free from inter- be removed by the lipid removal procedures ferences by running reference matrix method in Section 13.7, followed by alumina (Section blanks initially and with each sample batch 13.4) or Florisil (Section 13.8), and carbon (samples started through the extraction (Section 13.5) as minimum additional clean- process on a given 12-hour shift, to a max- up steps. If chlorodiphenyl ethers are de- imum of 20 samples). tected, as indicated by the presence of peaks 4.3.1 The reference matrix must simulate, at the exact m/z’s monitored for these as closely as possible, the sample matrix interferents, alumina and/or Florisil cleanup under test. Ideally, the reference matrix must be employed to eliminate these inter- should not contain the CDDs/CDFs in detect- ferences. able amounts, but should contain potential 5.0 Safety interferents in the concentrations expected to be found in the samples to be analyzed. 5.1 The toxicity or carcinogenicity of For example, a reference sample of human each compound or reagent used in this meth- adipose tissue containing od has not been precisely determined; how- pentachloronaphthalene can be used to exer- ever, each chemical compound should be

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treated as a potential health hazard. Expo- 5.3.2 Protective equipment—Disposable sure to these compounds should be reduced plastic gloves, apron or lab coat, safety to the lowest possible level. glasses or mask, and a glove box or fume 5.1.1 The 2,3,7,8-TCDD isomer has been hood adequate for radioactive work should found to be acnegenic, carcinogenic, and be used. During analytical operations that teratogenic in laboratory animal studies. It may give rise to aerosols or dusts, personnel is soluble in water to approximately 200 ppt should wear respirators equipped with acti- and in organic solvents to 0.14%. On the basis vated carbon filters. Eye protection equip- of the available toxicological and physical ment (preferably full face shields) must be properties of 2,3,7,8-TCDD, all of the CDDs/ worn while working with exposed samples or CDFs should be handled only by highly pure analytical standards. Latex gloves are trained personnel thoroughly familiar with commonly used to reduce exposure of the handling and cautionary procedures and the hands. When handling samples suspected or associated risks. known to contain high concentrations of the 5.1.2 It is recommended that the labora- CDDs/CDFs, an additional set of gloves can tory purchase dilute standard solutions of also be worn beneath the latex gloves. the analytes in this method. However, if pri- 5.3.3 Training—Workers must be trained mary solutions are prepared, they shall be in the proper method of removing contami- prepared in a hood, and a NIOSH/MESA ap- nated gloves and clothing without con- proved toxic gas respirator shall be worn tacting the exterior surfaces. when high concentrations are handled. 5.3.4 Personal hygiene—Hands and fore- 5.2 The laboratory is responsible for arms should be washed thoroughly after each maintaining a current awareness file of manipulation and before breaks (coffee, OSHA regulations regarding the safe han- lunch, and shift). dling of the chemicals specified in this meth- 5.3.5 Confinement—Isolated work areas od. A reference file of material safety data posted with signs, segregated glassware and sheets (MSDSs) should also be made avail- tools, and plastic absorbent paper on bench able to all personnel involved in these anal- tops will aid in confining contamination. yses. It is also suggested that the laboratory 5.3.6 Effluent vapors—The effluents of perform personal hygiene monitoring of each sample splitters from the gas chromatograph analyst who uses this method and that the (GC) and from roughing pumps on the mass results of this monitoring be made available spectrometer (MS) should pass through ei- to the analyst. Additional information on ther a column of activated charcoal or be laboratory safety can be found in References bubbled through a trap containing oil or 10–13. The references and bibliography at the high-boiling alcohols to condense CDD/CDF end of Reference 13 are particularly com- vapors. prehensive in dealing with the general sub- 5.3.7 Waste Handling—Good technique in- ject of laboratory safety. cludes minimizing contaminated waste. 5.3 The CDDs/CDFs and samples suspected Plastic bag liners should be used in waste to contain these compounds are handled cans. Janitors and other personnel must be using essentially the same techniques em- trained in the safe handling of waste. ployed in handling radioactive or infectious 5.3.8 Decontamination materials. Well-ventilated, controlled access 5.3.8.1 Decontamination of personnel—Use laboratories are required. Assistance in eval- any mild soap with plenty of scrubbing ac- uating the health hazards of particular lab- tion. oratory conditions may be obtained from 5.3.8.2 Glassware, tools, and surfaces— certain consulting laboratories and from Chlorothene NU Solvent is the least toxic State Departments of Health or Labor, many solvent shown to be effective. Satisfactory of which have an industrial health service. cleaning may be accomplished by rinsing The CDDs/CDFs are extremely toxic to lab- with Chlorothene, then washing with any de- oratory animals. Each laboratory must de- tergent and water. If glassware is first rinsed velop a strict safety program for handling with solvent, then the dish water may be dis- these compounds. The practices in Ref- posed of in the sewer. Given the cost of dis- erences 2 and 14 are highly recommended. posal, it is prudent to minimize solvent 5.3.1 Facility—When finely divided sam- wastes. ples (dusts, soils, dry chemicals) are handled, 5.3.9 Laundry—Clothing known to be con- all operations (including removal of samples taminated should be collected in plastic from sample containers, weighing, transfer- bags. Persons who convey the bags and laun- ring, and mixing) should be performed in a der the clothing should be advised of the haz- glove box demonstrated to be leak tight or in ard and trained in proper handling. The a fume hood demonstrated to have adequate clothing may be put into a washer without air flow. Gross losses to the laboratory ven- contact if the launderer knows of the poten- tilation system must not be allowed. Han- tial problem. The washer should be run dling of the dilute solutions normally used in through a cycle before being used again for analytical and animal work presents no in- other clothing. halation hazards except in the case of an ac- 5.3.10 Wipe tests—A useful method of de- cident. termining cleanliness of work surfaces and

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tools is to wipe the surface with a piece of in the pump only. Before use, the tubing filter paper. Extraction and analysis by GC shall be thoroughly rinsed with methanol, with an electron capture detector (ECD) can followed by repeated rinsing with reagent achieve a limit of detection of 0.1 μg per water to minimize sample contamination. wipe; analysis using this method can achieve An integrating flow meter is used to collect an even lower detection limit. Less than 0.1 proportional composite samples. μg per wipe indicates acceptable cleanliness; 6.2 Equipment for Glassware Cleaning— anything higher warrants further cleaning. Laboratory sink with overhead fume hood. More than 10 μg on a wipe constitutes an 6.3 Equipment for Sample Preparation acute hazard and requires prompt cleaning 6.3.1 Laboratory fume hood of sufficient before further use of the equipment or work size to contain the sample preparation equip- space, and indicates that unacceptable work ment listed below. practices have been employed. 6.3.2 Glove box (optional). 5.3.11 Table or wrist-action shaker—The 6.3.3 Tissue homogenizer—VirTis Model 45 use of a table or wrist-action shaker for ex- Macro homogenizer (American Scientific traction of tissues presents the possibility of Products H–3515, or equivalent) with stain- breakage of the extraction bottle and spill- less steel Macro-shaft and Turbo-shear blade. age of acid and flammable organic solvent. A 6.3.4 Meat grinder—Hobart, or equivalent, secondary containment system around the with 3–5 mm holes in inner plate. shaker is suggested to prevent the spread of 6.3.5 Equipment for determining percent acid and solvents in the event of such a moisture breakage. The speed and intensity of shaking 6.3.5.1 Oven—Capable of maintaining a action should also be adjusted to minimize temperature of 110 ±5 °C. the possibility of breakage. 6.3.5.2 Dessicator. 6.3.6 Balances 6.0 Apparatus and Materials 6.3.6.1 Analytical—Capable of weighing 0.1 NOTE: Brand names, suppliers, and part mg. numbers are for illustration purposes only 6.3.6.2 Top loading—Capable of weighing and no endorsement is implied. Equivalent 10 mg. performance may be achieved using appa- 6.4 Extraction Apparatus ratus and materials other than those speci- 6.4.1 Water samples fied here. Meeting the performance require- 6.4.1.1 pH meter, with combination glass ments of this method is the responsibility of electrode. the laboratory. 6.4.1.2 pH paper, wide range (Hydrion Pa- 6.1 Sampling Equipment for Discrete or pers, or equivalent). Composite Sampling 6.4.1.3 Graduated cylinder, 1 L capacity. 6.1.1 Sample bottles and caps 6.4.1.4 Liquid/liquid extraction—Sepa- 6.1.1.1 Liquid samples (waters, sludges and ratory funnels, 250 mL, 500 mL, and 2000 mL, similar materials containing 5% solids or with fluoropolymer stopcocks. less)—Sample bottle, amber glass, 1.1 L min- 6.4.1.5 Solid-phase extraction imum, with screw cap. 6.4.1.5.1 One liter filtration apparatus, in- 6.1.1.2 Solid samples (soils, sediments, cluding glass funnel, glass frit support, sludges, paper pulps, filter cake, compost, clamp, adapter, stopper, filtration flask, and and similar materials that contain more vacuum tubing (Figure 4). For wastewater than 5% solids)—Sample bottle, wide mouth, samples, the apparatus should accept 90 or amber glass, 500 mL minimum. 144 mm disks. For drinking water or other 6.1.1.3 If amber bottles are not available, samples containing low solids, smaller disks samples shall be protected from light. may be used. 6.1.1.4 Bottle caps—Threaded to fit sample 6.4.1.5.2 Vacuum source capable of main- bottles. Caps shall be lined with taining 25 in. Hg, equipped with shutoff valve fluoropolymer. and vacuum gauge. 6.1.1.5 Cleaning 6.4.1.5.3 Glass-fiber filter—Whatman GMF 6.1.1.5.1 Bottles are detergent water 150 (or equivalent), 1 micron pore size, to fit washed, then solvent rinsed before use. filtration apparatus in Section 6.4.1.5.1. 6.1.1.5.2 Liners are detergent water 6.4.1.5.4 Solid-phase extraction disk con- washed, rinsed with reagent water (Section taining octadecyl (C18) bonded silica uni- 7.6.1) followed by solvent, and baked at ap- formly enmeshed in an inert matrix—Fisher proximately 200 °C for a minimum of 1 hour Scientific 14–378F (or equivalent), to fit fil- prior to use. tration apparatus in Section 6.4.1.5.1. 6.1.2 Compositing equipment—Automatic 6.4.2 Soxhlet/Dean-Stark (SDS) extractor or manual compositing system incorporating (Figure 5)—For filters and solid/sludge sam- glass containers cleaned per bottle cleaning ples. procedure above. Only glass or fluoropolymer 6.4.2.1 Soxhlet—50 mm ID, 200 mL capac- tubing shall be used. If the sampler uses a ity with 500 mL flask (Cal-Glass LG–6900, or peristaltic pump, a minimum length of com- equivalent, except substitute 500 mL round- pressible silicone rubber tubing may be used bottom flask for 300 mL flat-bottom flask).

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6.4.2.2 Thimble—43 × 123 to fit Soxhlet 6.7.1.2 Syringe—10 mL, with Luer fitting. (Cal-Glass LG–6901–122, or equivalent). 6.7.1.3 Syringe filter holder—stainless 6.4.2.3 Moisture trap—Dean Stark or Bar- steel, and glass-fiber or fluoropolymer filters ret with fluoropolymer stopcock, to fit Soxh- (Gelman 4310, or equivalent). let. 6.7.1.4 UV detectors—254 nm, preparative 6.4.2.4 Heating mantle—Hemispherical, to or semi-preparative flow cell (Isco, Inc., fit 500 mL round-bottom flask (Cal-Glass LG– Type 6; Schmadzu, 5 mm path length; Beck- 8801–112, or equivalent). man-Altex 152W, 8 μL micro-prep flow cell, 2 6.4.2.5 Variable transformer—Powerstat mm path; Pharmacia UV–1, 3 mm flow cell; (or equivalent), 110 volt, 10 amp. LDC Milton-Roy UV–3, monitor #1203; or 6.4.3 Apparatus for extraction of tissue. equivalent). 6.4.3.1 Bottle for extraction (if digestion/ 6.7.2 Reverse-phase high-performance liq- extraction using HCl is used)’’ 500–600 mL uid chromatograph. wide-mouth clear glass, with fluoropolymer- 6.7.2.1 Column oven and detector—Perkin- lined cap. Elmer Model LC–65T (or equivalent) operated 6.4.3.2 Bottle for back-extraction—100–200 at 0.02 AUFS at 235 nm. mL narrow-mouth clear glass with 6.7.2.2 Injector—Rheodyne 7120 (or equiva- fluoropolymer-lined cap. lent) with 50 μL sample loop. 6.4.3.3 Mechanical shaker—Wrist-action 6.7.2.3 Column—Two 6.2 mm × 250 mm or platform-type rotary shaker that pro- Zorbax-ODS columns in series (DuPont In- duces vigorous agitation (Sybron struments Division, Wilmington, DE, or Thermolyne Model LE ‘‘Big Bill’’ rotator/ equivalent), operated at 50 °C with 2.0 mL/ shaker, or equivalent). min methanol isocratic effluent. 6.4.3.4 Rack attached to shaker table to 6.7.2.4 Pump—Altex 110A (or equivalent). permit agitation of four to nine samples si- 6.7.3 Pipets. multaneously. 6.7.3.1 Disposable, pasteur—150 mm long × 6.4.4 Beakers—400–500 mL. 5-mm ID (Fisher Scientific 13–678–6A, or 6.4.5 Spatulas—Stainless steel. equivalent). 6.5 Filtration Apparatus. 6.7.3.2 Disposable, serological—10 mL (6 6.5.1 Pyrex glass wool—Solvent-extracted mm ID). by SDS for three hours minimum. 6.7.4 Glass chromatographic columns. 6.7.4.1 150 mm long × 8-mm ID, (Kontes K– NOTE: Baking glass wool may cause active 420155, or equivalent) with coarse-glass frit sites that will irreversibly adsorb CDDs/ or glass-wool plug and 250 mL reservoir. CDFs. 6.7.4.2 200 mm long × 15 mm ID, with coarse-glass frit or glass-wool plug and 250 6.5.2 Glass funnel—125–250 mL. mL reservoir. 6.5.3 Glass-fiber filter paper—Whatman 6.7.4.3 300 mm long × 25 mm ID, with 300 GF/D (or equivalent), to fit glass funnel in mL reservoir and glass or fluoropolymer Section 6.5.2. stopcock. 6.5.4 Drying column—15–20 mm ID Pyrex 6.7.5 Stirring apparatus for batch silica chromatographic column equipped with cleanup of tissue extracts. coarse-glass frit or glass-wool plug. 6.7.5.1 Mechanical stirrer—Corning Model 6.5.5 Buchner funnel—15 cm. 320, or equivalent. 6.5.6 Glass-fiber filter paper—to fit 6.7.5.2 Bottle—500–600 mL wide-mouth Buchner funnel in Section 6.5.5. clear glass. 6.5.7 Filtration flasks—1.5–2.0 L, with side 6.7.6 Oven—For baking and storage of ad- arm. sorbents, capable of maintaining a constant 6.5.8 Pressure filtration apparatus— temperature (±5 °C) in the range of 105–250 °C. Millipore YT30 142 HW, or equivalent. 6.8 Concentration Apparatus. 6.6 Centrifuge Apparatus. 6.8.1 Rotary evaporator—Buchi/ 6.6.1 Centrifuge—Capable of rotating 500 Brinkman-American Scientific No. E5045–10 mL centrifuge bottles or 15 mL centrifuge or equivalent, equipped with a variable tem- tubes at 5,000 rpm minimum. perature water bath. 6.6.2 Centrifuge bottles—500 mL, with 6.8.1.1 Vacuum source for rotary evapo- screw-caps, to fit centrifuge. rator equipped with shutoff valve at the 6.6.3 Centrifuge tubes—12–15 mL, with evaporator and vacuum gauge. screw-caps, to fit centrifuge. 6.8.1.2 A recirculating water pump and 6.7 Cleanup Apparatus. chiller are recommended, as use of tap water 6.7.1 Automated gel permeation chro- for cooling the evaporator wastes large vol- matograph (Analytical Biochemical Labs, umes of water and can lead to inconsistent Inc, Columbia, MO, Model GPC Autoprep performance as water temperatures and pres- 1002, or equivalent). sures vary. 6.7.1.1 Column—600–700 mm long × 25 mm 6.8.1.3 Round-bottom flask—100 mL and ID, packed with 70 g of 500 mL or larger, with ground-glass fitting SX–3 Bio-beads (Bio-Rad Laboratories, Rich- compatible with the rotary evaporator. mond, CA, or equivalent). 6.8.2 Kuderna-Danish (K-D) Concentrator.

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6.8.2.1 Concentrator tube—10 mL, grad- 7.1.2 Sulfuric acid—Reagent grade (spe- uated (Kontes K–570050–1025, or equivalent) cific gravity 1.84). with calibration verified. Ground-glass stop- 7.1.3 Hydrochloric acid—Reagent grade, per (size 19/22 joint) is used to prevent evapo- 6N. ration of extracts. 7.1.4 Sodium chloride—Reagent grade, 6.8.2.2 Evaporation flask—500 mL (Kontes prepare at 5% (w/v) solution in reagent K–570001–0500, or equivalent), attached to water. concentrator tube with springs (Kontes K– 7.2 Solution Drying and Evaporation. 662750–0012 or equivalent). 7.2.1 Solution drying—Sodium sulfate, re- 6.8.2.3 Snyder column—Three-ball macro agent grade, granular, anhydrous (Baker (Kontes K–503000–0232, or equivalent). 3375, or equivalent), rinsed with methylene 6.8.2.4 Boiling chips. chloride (20 mL/g), baked at 400 °C for one 6.8.2.4.1 Glass or —Approxi- hour minimum, cooled in a dessicator, and mately 10/40 mesh, extracted with methylene stored in a pre-cleaned glass bottle with chloride and baked at 450 °C for one hour screw-cap that prevents moisture from en- minimum. tering. If, after heating, the sodium sulfate 6.8.2.4.2 Fluoropolymer (optional)—Ex- develops a noticeable grayish cast (due to tracted with methylene chloride. the presence of carbon in the crystal ma- 6.8.2.5 Water bath—Heated, with concen- trix), that batch of reagent is not suitable tric ring cover, capable of maintaining a for use and should be discarded. Extraction temperature within ±2 °C, installed in a fume with methylene chloride (as opposed to sim- hood. ple rinsing) and baking at a lower tempera- 6.8.3 Nitrogen blowdown apparatus— ture may produce sodium sulfate that is Equipped with water bath controlled in the suitable for use. range of 30–60 °C (N-Evap, Organomation As- 7.2.2 Tissue drying—Sodium sulfate, rea- sociates, Inc., South Berlin, MA, or equiva- gent grade, powdered, treated and stored as lent), installed in a fume hood. above. 6.8.4 Sample vials. 7.2.3 Prepurified nitrogen. 6.8.4.1 Amber glass—2–5 mL with 7.3 Extraction. fluoropolymer-lined screw-cap. 7.3.1 Solvents—Acetone, toluene, 6.8.4.2 Glass—0.3 mL, conical, with cyclohexane, hexane, methanol, methylene fluoropolymer-lined screw or crimp cap. chloride, and nonane; distilled in glass, pes- 6.9 Gas Chromatograph—Shall have ticide quality, lot-certified to be free of splitless or on-column injection port for cap- interferences. illary column, temperature program with 7.3.2 White quartz sand, 60/70 mesh—For isothermal hold, and shall meet all of the Soxhlet/Dean-Stark extraction (Aldrich performance specifications in Section 10. Chemical, Cat. No. 27–437–9, or equivalent). 6.9.1 GC column for CDDs/CDFs and for Bake at 450 °C for four hours minimum. isomer specificity for 2,3,7,8-TCDD—60 ±5 m 7.4 GPC Calibration Solution—Prepare a long × 0.32 ±0.02 mm ID; 0.25 μm 5% phenyl, solution containing 300 mg/mL corn oil, 15 94% methyl, 1% vinyl silicone bonded-phase mg/mL bis(2-ethylhexyl) phthalate, 1.4 mg/ fused-silica capillary column (J&W DB–5, or mL pentachlorophenol, 0.1 mg/mL perylene, equivalent). and 0.5 mg/mL sulfur. 6.9.2 GC column for isomer specificity for 7.5 Adsorbents for Sample Cleanup. 2,3,7,8-TCDF—30 ±5 m long × 0.32 ±0.02 mm ID; 7.5.1 Silica gel. 0.25 μm bonded-phase fused-silica capillary 7.5.1.1 Activated silica gel—100–200 mesh, column (J&W DB–225, or equivalent). Supelco 1–3651 (or equivalent), rinsed with 6.10 Mass Spectrometer—28–40 eV electron methylene chloride, baked at 180 °C for a impact ionization, shall be capable of repet- minimum of one hour, cooled in a dessicator, itively selectively monitoring 12 exact m/z’s and stored in a precleaned glass bottle with minimum at high resolution (≥10,000) during screw-cap that prevents moisture from en- a period of approximately one second, and tering. shall meet all of the performance specifica- 7.5.1.2 Acid silica gel (30% w/w)—Thor- tions in Section 10. oughly mix 44.0 g of concentrated sulfuric 6.11 GC/MS Interface—The mass spec- acid with 100.0 g of activated silica gel in a trometer (MS) shall be interfaced to the GC clean container. Break up aggregates with a such that the end of the capillary column stirring rod until a uniform mixture is ob- terminates within 1 cm of the ion source but tained. Store in a bottle with a does not intercept the electron or ion beams. fluoropolymer-lined screw-cap. 6.12 Data System—Capable of collecting, 7.5.1.3 Basic silica gel—Thoroughly mix 30 recording, and storing MS data. g of 1N sodium hydroxide with 100 g of acti- vated silica gel in a clean container. Break 7.0 Reagents and Standards up aggregates with a stirring rod until a uni- 7.1 pH Adjustment and Back-Extraction. form mixture is obtained. Store in a bottle 7.1.1 Potassium hydroxide—Dissolve 20 g with a fluoropolymer-lined screw-cap. reagent grade KOH in 100 mL reagent water. 7.5.1.4 Potassium silicate.

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7.5.1.4.1 Dissolve 56 g of high purity potas- 7.5.5.3 Activate in an oven at 130–150 °C for sium hydroxide (Aldrich, or equivalent) in a minimum of 24 hours and cool for 30 min- 300 mL of methanol in a 750–1000 mL flat-bot- utes. Use within 90 minutes of cooling. tom flask. 7.6 Reference Matrices—Matrices in 7.5.1.4.2 Add 100 g of silica gel and a stir- which the CDDs/CDFs and interfering com- ring bar, and stir on a hot plate at 60–70 °C pounds are not detected by this method. for one to two hours. 7.6.1 Reagent water—Bottled water pur- 7.5.1.4.3 Decant the liquid and rinse the chased locally, or prepared by passage potassium silicate twice with 100 mL por- through activated carbon. tions of methanol, followed by a single rinse 7.6.2 High-solids reference matrix—Play- with 100 mL of methylene chloride. ground sand or similar material. Prepared by 7.5.1.4.4 Spread the potassium silicate on extraction with methylene chloride and/or solvent-rinsed aluminum foil and dry for two baking at 450 °C for a minimum of four to four hours in a hood. hours. 7.5.1.4.5 Activate overnight at 200–250 °C. 7.6.3 Paper reference matrix—Glass-fiber 7.5.2 Alumina—Either one of two types of filter, Gelman Type A, or equivalent. Cut alumina, acid or basic, may be used in the paper to simulate the surface area of the cleanup of sample extracts, provided that the paper sample being tested. laboratory can meet the performance speci- 7.6.4 Tissue reference matrix—Corn or fications for the recovery of labeled com- other oil. May be prepared by ex- pounds described in Section 9.3. The same traction with methylene chloride. type of alumina must be used for all samples, 7.6.5 Other matrices—This method may be including those used to demonstrate initial verified on any reference matrix by per- precision and recovery (Section 9.2) and on- forming the tests given in Section 9.2. Ideal- going precision and recovery (Section 15.5). ly, the matrix should be free of the CDDs/ 7.5.2.1 Acid alumina—Supelco 19996–6C (or CDFs, but in no case shall the background equivalent). Activate by heating to 130 °C for level of the CDDs/CDFs in the reference ma- a minimum of 12 hours. trix exceed three times the minimum levels 7.5.2.2 Basic alumina—Supelco 19944–6C in Table 2. If low background levels of the (or equivalent). Activate by heating to 600 °C CDDs/CDFs are present in the reference ma- for a minimum of 24 hours. Alternatively, ac- trix, the spike level of the analytes used in tivate by heating in a tube furnace at 650–700 Section 9.2 should be increased to provide a °C under an air flow rate of approximately spike-to-background ratio in the range of 1:1 400 cc/minute. Do not heat over 700 °C, as this to 5:1 (Reference 15). can lead to reduced capacity for retaining 7.7 Standard Solutions—Purchased as so- the analytes. Store at 130 °C in a covered lutions or mixtures with certification to flask. Use within five days of baking. their purity, concentration, and authen- 7.5.3 Carbon. ticity, or prepared from materials of known 7.5.3.1 Carbopak C—(Supelco 1–0258, or purity and composition. If the chemical pu- equivalent). rity is 98% or greater, the weight may be 7.5.3.2 Celite 545—(Supelco 2–0199, or used without correction to compute the con- equivalent). centration of the standard. When not being 7.5.3.3 Thoroughly mix 9.0 g Carbopak C used, standards are stored in the dark at and 41.0 g Celite 545 to produce an 18% w/w room temperature in screw-capped vials with mixture. Activate the mixture at 130 °C for a fluoropolymer-lined caps. A mark is placed minimum of six hours. Store in a dessicator. on the vial at the level of the solution so 7.5.4 Anthropogenic isolation column— that solvent loss by evaporation can be de- Pack the column in Section 6.7.4.3 from bot- tected. If solvent loss has occurred, the solu- tom to top with the following: tion should be replaced. 7.5.4.1 2 g silica gel (Section 7.5.1.1). 7.8 Stock Solutions. 7.5.4.2 2 g potassium silicate (Section 7.8.1 Preparation—Prepare in nonane per 7.5.1.4). the steps below or purchase as dilute solu- 7.5.4.3 2 g granular anhydrous sodium sul- tions (Cambridge Isotope Laboratories (CIL), fate (Section 7.2.1). Woburn, MA, or equivalent). Observe the 7.5.4.4 10 g acid silica gel (Section 7.5.1.2). safety precautions in Section 5, and the rec- 7.5.4.5 2 g granular anhydrous sodium sul- ommendation in Section 5.1.2. fate. 7.8.2 Dissolve an appropriate amount of 7.5.5 Florisil column. assayed reference material in solvent. For 7.5.5.1 Florisil—60–100 mesh, Floridin Corp example, weigh 1–2 mg of 2,3,7,8-TCDD to (or equivalent). Soxhlet extract in 500 g por- three significant figures in a 10 mL ground- tions for 24 hours. glass-stoppered volumetric flask and fill to 7.5.5.2 Insert a glass wool plug into the ta- the mark with nonane. After the TCDD is pered end of a graduated serological pipet completely dissolved, transfer the solution (Section 6.7.3.2). Pack with 1.5 g (approx 2 to a clean 15 mL vial with fluoropolymer- mL) of Florisil topped with approx 1 mL of lined cap. sodium sulfate (Section 7.2.1) and a glass 7.8.3 Stock standard solutions should be wool plug. checked for signs of degradation prior to the

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preparation of calibration or performance trix for each sample batch. One mL each are test standards. Reference standards that can required for the blank and OPR with each be used to determine the accuracy of calibra- matrix in each batch. tion standards are available from CIL and 7.15 GC Retention Time Window Defining may be available from other vendors. Solution and Isomer Specificity Test Stand- 7.9 PAR Stock Solution ard—Used to define the beginning and ending 7.9.1 All CDDs/CDFs—Using the solutions retention times for the dioxin and furan iso- in Section 7.8, prepare the PAR stock solu- mers and to demonstrate isomer specificity tion to contain the CDDs/CDFs at the con- of the GC columns employed for determina- centrations shown in Table 3. When diluted, tion of 2,3,7,8-TCDD and 2,3,7,8-TCDF. The the solution will become the PAR (Section standard must contain the compounds listed 7.14). in Table 5 (CIL EDF—4006, or equivalent), at 7.9.2 If only 2,3,7,8-TCDD and 2,3,7,8-TCDF a minimum. It is not necessary to monitor are to be determined, prepare the PAR stock the window-defining compounds if only solution to contain these compounds only. 2,3,7,8-TCDD and 2,3,7,8-TCDF are to be de- 7.10 Labeled-Compound Spiking Solution. termined. In this case, an isomer-specificity 7.10.1 All CDDs/CDFs—From stock solu- test standard containing the most closely tions, or from purchased mixtures, prepare eluted isomers listed in Table 5 (CIL EDF- this solution to contain the labeled com- 4033, or equivalent) may be used. pounds in nonane at the concentrations 7.16 QC Check Sample—A QC Check Sam- shown in Table 3. This solution is diluted ple should be obtained from a source inde- with acetone prior to use (Section 7.10.3). pendent of the calibration standards. Ideally, 7.10.2 If only 2,3,7,8-TCDD and 2,3,7,8- this check sample would be a certified ref- TCDF are to be determined, prepare the la- erence material containing the CDDs/CDFs beled-compound solution to contain these in known concentrations in a sample matrix compounds only. This solution is diluted similar to the matrix under test. with acetone prior to use (Section 7.10.3). 7.17 Stability of Solutions—Standard so- 7.10.3 Dilute a sufficient volume of the la- lutions used for quantitative purposes (Sec- beled compound solution (Section 7.10.1 or tions 7.9 through 7.15) should be analyzed pe- 7.10.2) by a factor of 50 with acetone to pre- riodically, and should be assayed against ref- pare a diluted spiking solution. Each sample erence standards (Section 7.8.3) before fur- requires 1.0 mL of the diluted solution, but ther use. no more solution should be prepared than can be used in one day. 8.0 Sample Collection, Preservation, Storage, 7.11 Cleanup Standard—Prepare 37Cl4- and Holding Times 2,3,7,8-TCDD in nonane at the concentration 8.1 Collect samples in amber glass con- shown in Table 3. The cleanup standard is tainers following conventional sampling added to all extracts prior to cleanup to practices (Reference 16). Aqueous samples measure the efficiency of the cleanup proc- that flow freely are collected in refrigerated ess. bottles using automatic sampling equip- 7.12 Internal Standard(s). ment. Solid samples are collected as grab 7.12.1 All CDDs/CDFs—Prepare the inter- samples using wide-mouth jars. nal standard solution to contain 13C12-1,2,3,4- 8.2 Maintain aqueous samples in the dark TCDD and 13C2-1,2,3,7,8,9-HxCDD in nonane at at 0–4 °C from the time of collection until re- the concentration shown in Table 3. ceipt at the laboratory. If residual chlorine 7.12.2 If only 2,3,7,8-TCDD and 2,3,7,8- is present in aqueous samples, add 80 mg so- TCDF are to be determined, prepare the in- dium thiosulfate per liter of water. EPA ternal standard solution to contain 13C12- Methods 330.4 and 330.5 may be used to meas- 1,2,3,4-TCDD only. ure residual chlorine (Reference 17). If sam- 7.13 Calibration Standards (CS1 through ple pH is greater than 9, adjust to pH 7–9 CS5)—Combine the solutions in Sections 7.9 with sulfuric acid. through 7.12 to produce the five calibration Maintain solid, semi-solid, oily, and mixed- solutions shown in Table 4 in nonane. These phase samples in the dark at <4 °C from the solutions permit the relative response (la- time of collection until receipt at the labora- beled to native) and response factor to be tory. measured as a function of concentration. The Store aqueous samples in the dark at 0–4 CS3 standard is used for calibration °C. Store solid, semi-solid, oily, mixed-phase, verification (VER). If only 2,3,7,8-TCDD and and tissue samples in the dark at <¥10 °C. 2,3,7,8-TCDF are to be determined, combine 8.3 Fish and Tissue Samples. the solutions appropriate to these com- 8.3.1 Fish may be cleaned, filleted, or pounds. processed in other ways in the field, such 7.14 Precision and Recovery (PAR) Stand- that the laboratory may expect to receive ard—Used for determination of initial (Sec- whole fish, fish fillets, or other tissues for tion 9.2) and ongoing (Section 15.5) precision analysis. and recovery. Dilute 10 μL of the precision 8.3.2 Fish collected in the field should be and recovery standard (Section 7.9.1 or 7.9.2) wrapped in aluminum foil, and must be to 2.0 mL with acetone for each sample ma- maintained at a temperature less than 4 °C

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from the time of collection until receipt at 9.1.2.1 Each time a modification is made the laboratory. to this method, the analyst is required to re- 8.3.3 Samples must be frozen upon receipt peat the procedure in Section 9.2. If the de- at the laboratory and maintained in the dark tection limit of the method will be affected at <¥10 °C until prepared. Maintain unused by the change, the laboratory is required to sample in the dark at <¥10 °C. demonstrate that the MDL (40 CFR part 136, 8.4 Holding Times. appendix B) is lower than one-third the regu- 8.4.1 There are no demonstrated max- latory compliance level or one-third the ML imum holding times associated with CDDs/ in this method, whichever is higher. If cali- CDFs in aqueous, solid, semi-solid, tissues, bration will be affected by the change, the or other sample matrices. If stored in the analyst must recalibrate the instrument per dark at 0–4 °C and preserved as given above Section 10. (if required), aqueous samples may be stored 9.1.2.2 The laboratory is required to main- for up to one year. Similarly, if stored in the tain records of modifications made to this dark at <¥10 °C, solid, semi-solid, multi- method. These records include the following, phase, and tissue samples may be stored for at a minimum: up to one year. 9.1.2.2.1 The names, titles, addresses, and 8.4.2 Store sample extracts in the dark at telephone numbers of the analyst(s) who per- <¥10 °C until analyzed. If stored in the dark formed the analyses and modification, and of at <¥10 °C, sample extracts may be stored the quality control officer who witnessed and for up to one year. will verify the analyses and modifications. 9.1.2.2.2 A listing of pollutant(s) meas- 9.0 Quality Assurance/Quality Control ured, by name and CAS Registry number. 9.1 Each laboratory that uses this method 9.1.2.2.3 A narrative stating reason(s) for is required to operate a formal quality assur- the modifications. 9.1.2.2.4 Results from all quality control ance program (Reference 18). The minimum (QC) tests comparing the modified method to requirements of this program consist of an this method, including: initial demonstration of laboratory capa- (a) Calibration (Section 10.5 through 10.7). bility, analysis of samples spiked with la- (b) Calibration verification (Section 15.3). beled compounds to evaluate and document (c) Initial precision and recovery (Section data quality, and analysis of standards and 9.2). blanks as tests of continued performance. (d) Labeled compound recovery (Section Laboratory performance is compared to es- 9.3). tablished performance criteria to determine (e) Analysis of blanks (Section 9.5). if the results of analyses meet the perform- (f) Accuracy assessment (Section 9.4). ance characteristics of the method. 9.1.2.2.5 Data that will allow an inde- If the method is to be applied to sample pendent reviewer to validate each deter- matrix other than water (e.g., soils, filter mination by tracing the instrument output cake, compost, tissue) the most appropriate (peak height, area, or other signal) to the alternate matrix (Sections 7.6.2 through final result. These data are to include: 7.6.5) is substituted for the reagent water (a) Sample numbers and other identifiers. matrix (Section 7.6.1) in all performance (b) Extraction dates. tests. (c) Analysis dates and times. 9.1.1 The analyst shall make an initial (d) Analysis sequence/run chronology. demonstration of the ability to generate ac- (e) Sample weight or volume (Section 11). ceptable accuracy and precision with this (f) Extract volume prior to each cleanup method. This ability is established as de- step (Section 13). scribed in Section 9.2. (g) Extract volume after each cleanup step 9.1.2 In recognition of advances that are (Section 13). occurring in analytical technology, and to (h) Final extract volume prior to injection allow the analyst to overcome sample ma- (Section 14). trix interferences, the analyst is permitted (i) Injection volume (Section 14.3). certain options to improve separations or (j) Dilution data, differentiating between lower the costs of measurements. These op- dilution of a sample or extract (Section 17.5). tions include alternate extraction, con- (k) Instrument and operating conditions. centration, cleanup procedures, and changes (l) Column (dimensions, liquid phase, solid in columns and detectors. Alternate deter- support, film thickness, etc). minative techniques, such as the substi- (m) Operating conditions (temperatures, tution of spectroscopic or immuno-assay temperature program, flow rates). techniques, and changes that degrade meth- (n) Detector (type, operating conditions, od performance, are not allowed. If an ana- etc). lytical technique other than the techniques (o) Chromatograms, printer tapes, and specified in this method is used, that tech- other recordings of raw data. nique must have a specificity equal to or bet- (p) Quantitation reports, data system out- ter than the specificity of the techniques in puts, and other data to link the raw data to this method for the analytes of interest. the results reported.

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9.1.3 Analyses of method blanks are re- 9.3 The laboratory shall spike all samples quired to demonstrate freedom from con- with the diluted labeled compound spiking tamination (Section 4.3). The procedures and solution (Section 7.10.3) to assess method criteria for analysis of a method blank are performance on the sample matrix. described in Sections 9.5 and 15.6. 9.3.1 Analyze each sample according to 9.1.4 The laboratory shall spike all sam- the procedures in Sections 11 through 18. ples with labeled compounds to monitor 9.3.2 Compute the percent recovery of the method performance. This test is described labeled compounds and the cleanup standard in Section 9.3. When results of these spikes using the internal standard method (Section indicate atypical method performance for 17.2). samples, the samples are diluted to bring 9.3.3 The recovery of each labeled com- method performance within acceptable lim- pound must be within the limits in Table 7 its. Procedures for dilution are given in Sec- when all 2,3,7,8-substituted CDDs/CDFs are tion 17.5. determined, and within the limits in Table 9.1.5 The laboratory shall, on an ongoing 7a when only 2,3,7,8-TCDD and 2,3,7,8-TCDF basis, demonstrate through calibration are determined. If the recovery of any com- verification and the analysis of the ongoing pound falls outside of these limits, method precision and recovery aliquot that the ana- performance is unacceptable for that com- lytical system is in control. These proce- pound in that sample. To overcome such dif- dures are described in Sections 15.1 through ficulties, water samples are diluted and 15.5. smaller amounts of soils, sludges, sediments, 9.1.6 The laboratory shall maintain and other matrices are reanalyzed per Sec- records to define the quality of data that is tion 18.4. generated. Development of accuracy state- 9.4 Recovery of labeled compounds from ments is described in Section 9.4. samples should be assessed and records 9.2 Initial Precision and Recovery (IPR)— should be maintained. To establish the ability to generate accept- 9.4.1 After the analysis of five samples of able precision and recovery, the analyst a given matrix type (water, soil, sludge, shall perform the following operations. pulp, etc.) for which the labeled compounds 9.2.1 For low solids (aqueous) samples, ex- pass the tests in Section 9.3, compute the av- tract, concentrate, and analyze four 1 L erage percent recovery (R) and the standard aliquots of reagent water spiked with the di- deviation of the percent recovery (SR) for luted labeled compound spiking solution the labeled compounds only. Express the as- (Section 7.10.3) and the precision and recov- sessment as a percent recovery interval from ery standard (Section 7.14) according to the R¥2SR to R = 2SR for each matrix. For exam- procedures in Sections 11 through 18. For an ple, if R = 90% and SR = 10% for five analyses alternative sample matrix, four aliquots of of pulp, the recovery interval is expressed as the alternative reference matrix (Section 7.6) 70–110%. are used. All sample processing steps that 9.4.2 Update the accuracy assessment for are to be used for processing samples, includ- each labeled compound in each matrix on a ing preparation (Section 11), extraction (Sec- regular basis (e.g., after each 5–10 new meas- tion 12), and cleanup (Section 13), shall be in- urements). cluded in this test. 9.5 Method Blanks—Reference matrix 9.2.2 Using results of the set of four anal- method blanks are analyzed to demonstrate yses, compute the average concentration (X) freedom from contamination (Section 4.3). of the extracts in ng/mL and the standard de- 9.5.1 Prepare, extract, clean up, and con- viation of the concentration (s) in ng/mL for centrate a method blank with each sample each compound, by isotope dilution for batch (samples of the same matrix started CDDs/CDFs with a labeled analog, and by in- through the extraction process on the same ternal standard for 1,2,3,7,8,9-HxCDD, OCDF, 12-hour shift, to a maximum of 20 samples). and the labeled compounds. The matrix for the method blank shall be 9.2.3 For each CDD/CDF and labeled com- similar to sample matrix for the batch, e.g., pound, compare s and X with the cor- a 1 L reagent water blank (Section 7.6.1), responding limits for initial precision and re- high-solids reference matrix blank (Section covery in Table 6. If only 2,3,7,8-TCDD and 7.6.2), paper matrix blank (Section 7.6.3); tis- 2,3,7,8-TCDF are to be determined, compare s sue blank (Section 7.6.4) or alternative ref- and X with the corresponding limits for ini- erence matrix blank (Section 7.6.5). Analyze tial precision and recovery in Table 6a. If s the blank immediately after analysis of the and X for all compounds meet the acceptance OPR (Section 15.5) to demonstrate freedom criteria, system performance is acceptable from contamination. and analysis of blanks and samples may 9.5.2 If any 2,3,7,8-substituted CDD/CDF begin. If, however, any individual s exceeds (Table 1) is found in the blank at greater the precision limit or any individual X falls than the minimum level (Table 2) or one- outside the range for accuracy, system per- third the regulatory compliance level, formance is unacceptable for that compound. whichever is greater; or if any potentially Correct the problem and repeat the test (Sec- interfering compound is found in the blank tion 9.2). at the minimum level for each level of

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chlorination given in Table 2 (assuming a re- 10.1.2 Mass spectrometer (MS) resolu- 13 sponse factor of 1 relative to the C12-1,2,3,4- tion—Obtain a selected ion current profile TCDD internal standard for compounds not (SICP) of each analyte in Table 3 at the two listed in Table 1), analysis of samples is halt- exact m/z’s specified in Table 8 and at ≥10,000 ed until the blank associated with the sam- resolving power by injecting an authentic ple batch shows no evidence of contamina- standard of the CDDs/CDFs either singly or tion at this level. All samples must be asso- as part of a mixture in which there is no in- ciated with an uncontaminated method terference between closely eluted compo- blank before the results for those samples nents. may be reported for regulatory compliance 10.1.2.1 The analysis time for CDDs/CDFs purposes. may exceed the long-term mass stability of 9.6 QC Check Sample—Analyze the QC the mass spectrometer. Because the instru- Check Sample (Section 7.16) periodically to ment is operated in the high-resolution assure the accuracy of calibration standards mode, mass drifts of a few ppm (e.g., 5 ppm and the overall reliability of the analytical in mass) can have serious adverse effects on process. It is suggested that the QC Check instrument performance. Therefore, a mass- Sample be analyzed at least quarterly. drift correction is mandatory and a lock- 9.7 The specifications contained in this mass m/z from PFK is used for drift correc- method can be met if the apparatus used is tion. The lock-mass m/z is dependent on the calibrated properly and then maintained in a exact m/z’s monitored within each calibrated state. The standards used for cali- descriptor, as shown in Table 8. The level of bration (Section 10), calibration verification PFK metered into the HRMS during analyses (Section 15.3), and for initial (Section 9.2) should be adjusted so that the amplitude of and ongoing (Section 15.5) precision and re- the most intense selected lock-mass m/z sig- covery should be identical, so that the most nal (regardless of the descriptor number) precise results will be obtained. A GC/MS in- does not exceed 10% of the full-scale deflec- strument will provide the most reproducible tion for a given set of detector parameters. results if dedicated to the settings and condi- Under those conditions, sensitivity changes tions required for the analyses of CDDs/CDFs that might occur during the analysis can be by this method. more effectively monitored. 9.8 Depending on specific program re- quirements, field replicates may be collected NOTE: Excessive PFK (or any other ref- to determine the precision of the sampling erence substance) may cause noise problems technique, and spiked samples may be re- and contamination of the ion source necessi- quired to determine the accuracy of the tating increased frequency of source clean- analysis when the internal standard method ing. is used. 10.1.2.2 If the HRMS has the capability to 10.0 Calibration monitor resolution during the analysis, it is acceptable to terminate the analysis when 10.1 Establish the operating conditions the resolution falls below 10,000 to save rea- necessary to meet the minimum retention nalysis time. times for the internal standards in Section 10.1.2.3 Using a PFK molecular leak, tune 10.2.4 and the relative retention times for the the instrument to meet the minimum re- CDDs/CDFs in Table 2. quired resolving power of 10,000 (10% valley) 10.1.1 Suggested GC operating conditions: at m/z 304.9824 (PFK) or any other reference Injector temperature: 270 °C signal close to m/z 304 (from TCDF). For each Interface temperature: 290 °C descriptor (Table 8), monitor and record the Initial temperature: 200 °C resolution and exact m/z’s of three to five Initial time: Two minutes reference peaks covering the mass range of Temperature program: the descriptor. The resolution must be great- 200–220 °C, at 5 °C/minute er than or equal to 10,000, and the deviation 220 °C for 16 minutes between the exact m/z and the theoretical m/ 220–235 °C, at 5 °C/minute z (Table 8) for each exact m/z monitored 235 °C for seven minutes must be less than 5 ppm. 235–330 °C, at 5 °C/minute 10.2 Ion Abundance Ratios, Minimum Lev- els, Signal-to-Noise Ratios, and Absolute Re- NOTE: All portions of the column that con- tention Times—Choose an injection volume nect the GC to the ion source shall remain at of either 1 μL or 2 μL, consistent with the ca- or above the interface temperature specified pability of the HRGC/HRMS instrument. In- above during analysis to preclude condensa- ject a 1 μL or 2 μL aliquot of the CS1 calibra- tion of less volatile compounds. tion solution (Table 4) using the GC condi- Optimize GC conditions for compound sep- tions from Section 10.1.1. If only 2,3,7,8-TCDD aration and sensitivity. Once optimized, the and 2,3,7,8-TCDF are to be determined, the same GC conditions must be used for the operating conditions and specifications analysis of all standards, blanks, IPR and below apply to analysis of those compounds OPR aliquots, and samples. only.

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10.2.1 Measure the SICP areas for each using the optimized temperature program in analyte, and compute the ion abundance ra- Section 10.1. Table 5 gives the elution order tios at the exact m/z’s specified in Table 8. (first/last) of the window-defining com- Compare the computed ratio to the theo- pounds. If 2,3,7,8-TCDD and 2,3,7,8-TCDF only retical ratio given in Table 9. are to be analyzed, this test is not required. 10.2.1.1 The exact m/z’s to be monitored in 10.4 Isomer Specificity. each descriptor are shown in Table 8. Each 10.4.1 Analyze the isomer specificity test group or descriptor shall be monitored in standards (Section 7.15) using the procedure succession as a function of GC retention in Section 14 and the optimized conditions time to ensure that all CDDs/CDFs are de- for sample analysis (Section 10.1.1). tected. Additional m/z’s may be monitored in 10.4.2 Compute the percent valley between each descriptor, and the m/z’s may be divided the GC peaks that elute most closely to the among more than the five descriptors listed 2,3,7,8-TCDD and TCDF isomers, on their re- in Table 8, provided that the laboratory is spective columns, per Figures 6 and 7. able to monitor the m/z’s of all the CDDs/ 10.4.3 Verify that the height of the valley CDFs that may elute from the GC in a given between the most closely eluted isomers and retention-time window. If only 2,3,7,8-TCDD the 2,3,7,8-substituted isomers is less than and 2,3,7,8-TCDF are to be determined, the 25% (computed as 100 x/y in Figures 6 and 7). descriptors may be modified to include only If the valley exceeds 25%, adjust the analyt- the exact m/z’s for the tetra-and penta-iso- ical conditions and repeat the test or replace mers, the diphenyl ethers, and the lock m/ the GC column and recalibrate (Sections z’s. 10.1.2 through 10.7). 10.2.1.2 The mass spectrometer shall be 10.5 Calibration by Isotope Dilution—Iso- operated in a mass-drift correction mode, tope dilution calibration is used for the 15 using perfluorokerosene (PFK) to provide 2,3,7,8-substituted CDDs/CDFs for which la- lock m/z’s. The lock-mass for each group of beled compounds are added to samples prior m/z’s is shown in Table 8. Each lock mass to extraction. The reference compound for shall be monitored and shall not vary by each CDD/CDF compound is shown in Table ± more than 20% throughout its respective re- 2. tention time window. Variations of the lock 10.5.1 A calibration curve encompassing mass by more than 20% indicate the presence the concentration range is prepared for each of coeluting interferences that may signifi- compound to be determined. The relative re- cantly reduce the sensitivity of the mass sponse (RR) (labeled to native) vs. concentra- spectrometer. Reinjection of another aliquot tion in standard solutions is plotted or com- of the sample extract will not resolve the puted using a linear regression. Relative re- problem. Additional cleanup of the extract sponse is determined according to the proce- may be required to remove the interferences. dures described below. Five calibration 10.2.2 All CDDs/CDFs and labeled com- points are employed. pounds in the CS1 standard shall be within 10.5.2 The response of each CDD/CDF rel- the QC limits in Table 9 for their respective ative to its labeled analog is determined ion abundance ratios; otherwise, the mass using the area responses of both the primary spectrometer shall be adjusted and this test and secondary exact m/z’s specified in Table repeated until the m/z ratios fall within the 8, for each calibration standard, as follows: limits specified. If the adjustment alters the resolution of the mass spectrometer, resolu- tion shall be verified (Section 10.1.2) prior to ()AAC12+ RR = nnl repeat of the test. + 10.2.3 Verify that the HRGC/HRMS instru- ()AAC12lln ment meets the minimum levels in Table 2. where: The peaks representing the CDDs/CDFs and labeled compounds in the CS1 calibration A1n and A2n = The areas of the primary and standard must have signal-to-noise ratios (S/ secondary m/z’s for the CDD/CDF. N) greater than or equal to 10.0. Otherwise, A1l and A2l = The areas of the primary and the mass spectrometer shall be adjusted and secondary m/z’s for the labeled com- this test repeated until the minimum levels pound. in Table 2 are met. Cl = The concentration of the labeled com- 13 pound in the calibration standard (Table 10.2.4 The absolute retention time of C12- 1,2,3,4–TCDD (Section 7.12) shall exceed 25.0 4). minutes on the DB–5 column, and the reten- Cn = The concentration of the native com- 13 pound in the calibration standard (Table tion time of C12-1,2,3,4–TCDD shall exceed 15.0 minutes on the DB–225 column; other- 4). wise, the GC temperature program shall be 10.5.3 To calibrate the analytical system adjusted and this test repeated until the by isotope dilution, inject a volume of cali- above-stated minimum retention time cri- bration standards CS1 through CS5 (Section teria are met. 7.13 and Table 4) identical to the volume cho- 2010.3 Retention-Time Windows—Analyze sen in Section 10.2, using the procedure in the window defining mixtures (Section 7.15) Section 14 and the conditions in Section

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10.1.1 and Table 2. Compute the relative re- 10.8 Data Storage—MS data shall be col- sponse (RR) at each concentration. lected, recorded, and stored. 10.5.4 Linearity—If the relative response 10.8.1 Data acquisition—The signal at for any compound is constant (less than 20% each exact m/z shall be collected repetitively coefficient of variation) over the five-point throughout the monitoring period and stored calibration range, an averaged relative re- on a mass storage device. sponse may be used for that compound; oth- 10.8.2 Response factors and multipoint erwise, the complete calibration curve for calibrations—The data system shall be used that compound shall be used over the five- to record and maintain lists of response fac- point calibration range. tors (response ratios for isotope dilution) and 10.6 Calibration by Internal Standard— multipoint calibration curves. Computations The internal standard method is applied to of relative standard deviation (coefficient of determination of 1,2,3,7,8,9-HxCDD (Section variation) shall be used to test calibration 17.1.2), OCDF (Section 17.1.1), the non 2,3,7,8- linearity. Statistics on initial performance substituted compounds, and to the deter- (Section 9.2) and ongoing performance (Sec- mination of labeled compounds for tion 15.5) should be computed and main- intralaboratory statistics (Sections 9.4 and tained, either on the instrument data sys- 15.5.4). tem, or on a separate computer system. 10.6.1 Response factors—Calibration re- quires the determination of response factors 11.0 Sample Preparation (RF) defined by the following equation: 11.1 Sample preparation involves modi- fying the physical form of the sample so that ()AAC12+ RF = ssis the CDDs/CDFs can be extracted efficiently. + In general, the samples must be in a liquid ()AAC12is is s form or in the form of finely divided solids in where: order for efficient extraction to take place. Table 10 lists the phases and suggested quan- A1s and A2s = The areas of the primary and tities for extraction of various sample mat- secondary m/z’s for the CDD/CDF. rices. A1 and A2 = The areas of the primary and is is For samples known or expected to contain secondary m/z’s for the internal stand- high levels of the CDDs/CDFs, the smallest ard. sample size representative of the entire sam- C = The concentration of the internal stand- is ple should be used (see Section 17.5). ard (Table 4). For all samples, the blank and IPR/OPR C = The concentration of the compound in s aliquots must be processed through the same the calibration standard (Table 4). steps as the sample to check for contamina- 37 NOTE: There is only one m/z for Cl4-2,3,7,8- tion and losses in the preparation processes. TCDD. See Table 8. 11.1.1 For samples that contain particles, 10.6.2 To calibrate the analytical system percent solids and particle size are deter- by internal standard, inject 1.0 μL or 2.0 μL mined using the procedures in Sections 11.2 of calibration standards CS1 through CS5 and 11.3, respectively. (Section 7.13 and Table 4) using the proce- 11.1.2 Aqueous samples—Because CDDs/ dure in Section 14 and the conditions in Sec- CDFs may be bound to suspended particles, tion 10.1.1 and Table 2. Compute the response the preparation of aqueous samples is de- factor (RF) at each concentration. pendent on the solids content of the sample. 10.6.3 Linearity—If the response factor 11.1.2.1 Aqueous samples visibly absent (RF) for any compound is constant (less than particles are prepared per Section 11.4 and 35% coefficient of variation) over the five- extracted directly using the separatory fun- point calibration range, an averaged re- nel or SPE techniques in Sections 12.1 or sponse factor may be used for that com- 12.2, respectively. pound; otherwise, the complete calibration 11.1.2.2 Aqueous samples containing visi- curve for that compound shall be used over ble particles and containing one percent sus- the five-point range. pended solids or less are prepared using the 10.7 Combined Calibration—By using cali- procedure in Section 11.4. After preparation, bration solutions (Section 7.13 and Table 4) the sample is extracted directly using the containing the CDDs/CDFs and labeled com- SPE technique in 12.2 or filtered per Section pounds and the internal standards, a single 11.4.3. After filtration, the particles and fil- set of analyses can be used to produce cali- ter are extracted using the SDS procedure in bration curves for the isotope dilution and Section 12.3 and the filtrate is extracted internal standard methods. These curves are using the separatory funnel procedure in verified each shift (Section 15.3) by analyzing Section 12.1. the calibration verification standard (VER, 11.1.2.3 For aqueous samples containing Table 4). Recalibration is required if any of greater than one percent solids, a sample ali- the calibration verification criteria (Section quot sufficient to provide 10 g of dry solids is 15.3) cannot be met. used, as described in Section 11.5.

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11.1.3 Solid samples are prepared using 11.2 Determination of Percent Suspended the procedure described in Section 11.5 fol- Solids. lowed by extraction via the SDS procedure NOTE: This aliquot is used for determining in Section 12.3. the solids content of the sample, not for de- 11.1.4 Multiphase samples—The phase(s) termination of CDDs/CDFs. containing the CDDs/CDFs is separated from the non-CDD/CDF phase using pressure fil- 11.2.1 Aqueous liquids and multi-phase tration and centrifugation, as described in samples consisting of mainly an aqueous Section 11.6. The CDDs/CDFs will be in the phase. organic phase in a multiphase sample in 11.2.1.1 Dessicate and weigh a GF/D filter which an organic phase exists. (Section 6.5.3) to three significant figures. 11.1.5 Procedures for grinding, homogeni- 11.2.1.2 Filter 10.0 ±0.02 mL of well-mixed zation, and blending of various sample sample through the filter. phases are given in Section 11.7. 11.2.1.3 Dry the filter a minimum of 12 11.1.6 Tissue samples—Preparation proce- hours at 110 ±5 °C and cool in a dessicator. dures for fish and other tissues are given in 11.2.1.4 Calculate percent solids as fol- Section 11.8. lows:

weight of sample aliquot after drying (g)− weight of filter (g) % solids = ×100 10 g

11.2.2 Non-aqueous liquids, solids, semi- 11.2.2.2 Dry a minimum of 12 hours at 110 solid samples, and multi-phase samples in ±5 °C, and cool in a dessicator. which the main phase is not aqueous; but not 11.2.2.3 Calculate percent solids as fol- tissues. lows: 11.2.2.1 Weigh 5–10 g of sample to three significant figures in a tared beaker.

weight of sample aliquot after drying % solids =×100 weight of sample aliquot before drying

11.3 Determination of Particle Size. 11.4.2 Preparation of sample and QC 11.3.1 Spread the dried sample from Sec- aliquots. tion 11.2.2.2 on a piece of filter paper or alu- 11.4.2.1 Mark the original level of the minum foil in a fume hood or glove box. sample on the sample bottle for reference. 11.3.2 Estimate the size of the particles in Weigh the sample plus bottle to ±1. the sample. If the size of the largest particles 11.4.2.2 Spike 1.0 mL of the diluted la- is greater than 1 mm, the particle size must beled-compound spiking solution (Section be reduced to 1 mm or less prior to extrac- 7.10.3) into the sample bottle. Cap the bottle tion using the procedures in Section 11.7. and mix the sample by careful shaking. 11.4 Preparation of Aqueous Samples Con- Allow the sample to equilibrate for one to taining 1% Suspended Solids or Less. two hours, with occasional shaking. 11.4.1 Aqueous samples visibly absent par- 11.4.2.3 For each sample or sample batch ticles are prepared per the procedure below (to a maximum of 20 samples) to be extracted and extracted directly using the separatory during the same 12-hour shift, place two 1.0 L funnel or SPE techniques in Sections 12.1 or aliquots of reagent water in clean sample 12.2, respectively. Aqueous samples con- bottles or flasks. taining visible particles and one percent sus- 11.4.2.4 Spike 1.0 mL of the diluted la- pended solids or less are prepared using the beled-compound spiking solution (Section procedure below and extracted using either 7.10.3) into both reagent water aliquots. One the SPE technique in Section 12.2 or further of these aliquots will serve as the method prepared using the filtration procedure in blank. Section 11.4.3. The filtration procedure is fol- 11.4.2.5 Spike 1.0 mL of the PAR standard lowed by SDS extraction of the filter and (Section 7.14) into the remaining reagent particles (Section 12.3) and separatory funnel water aliquot. This aliquot will serve as the extraction of the filtrate (Section 12.1). The OPR (Section 15.5). SPE procedure is followed by SDS extraction 11.4.2.6 If SPE is to be used, add 5 mL of of the filter and disk. methanol to the sample, cap and shake the

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sample to mix thoroughly, and proceed to 11.6.1 Using the percent solids determined Section 12.2 for extraction. If SPE is not to in Section 11.2.1 or 11.2.2, determine the vol- be used, and the sample is visibly absent par- ume of sample that will provide 10 g of sol- ticles, proceed to Section 12.1 for extraction. ids, up to 1 L of sample. If SPE is not to be used and the sample con- 11.6.2 Pressure filter the amount of sam- tains visible particles, proceed to the fol- ple determined in Section 11.6.1 through lowing section for filtration of particles. Whatman GF/D glass-fiber filter paper (Sec- 11.4.3 Filtration of particles. tion 6.5.3). Pressure filter the blank and OPR 11.4.3.1 Assemble a Buchner funnel (Sec- aliquots through GF/D papers also. If nec- tion 6.5.5) on top of a clean filtration flask. essary to separate the phases and/or settle Apply vacuum to the flask, and pour the en- the solids, centrifuge these aliquots prior to tire contents of the sample bottle through a filtration. glass-fiber filter (Section 6.5.6) in the 11.6.3 Discard any aqueous phase (if Buchner funnel, swirling the sample remain- present). Remove any non-aqueous liquid ing in the bottle to suspend any particles. present and reserve the maximum amount 11.4.3.2 Rinse the sample bottle twice with filtered from the sample (Section 11.6.1) or 10 approximately 5 mL portions of reagent g, whichever is less, for combination with water to transfer any remaining particles the solid phase (Section 12.3.5). onto the filter. 11.6.4 If particles >1mm are present in the 11.4.3.3 Rinse any particles off the sides of sample (as determined in Section 11.3.2) and the Buchner funnel with small quantities of the sample is capable of being dried, spread reagent water. the sample and QC aliquots on clean alu- 11.4.3.4 Weigh the empty sample bottle to minum foil in a hood. After the aliquots are dry or if the sample cannot be dried, reduce ±1 g. Determine the weight of the sample by the particle size using the procedures in Sec- difference. Save the bottle for further use. tion 11.7 and extract the reduced particles 11.4.3.5 Extract the filtrate using the using the SDS procedure in Section 12.3. If separatory funnel procedure in Section 12.1. particles >1mm are not present, extract the 11.4.3.6 Extract the filter containing the particles and filter in the sample and QC particles using the SDS procedure in Section aliquots directly using the SDS procedure in 12.3. Section 12.3. 11.5 Preparation of Samples Containing 11.7 Sample grinding, homogenization, or Greater Than 1% Solids. blending—Samples with particle sizes great- 11.5.1 Weigh a well-mixed aliquot of each er than 1 mm (as determined in Section sample (of the same matrix type) sufficient 11.3.2) are subjected to grinding, homogeni- to provide 10 g of dry solids (based on the sol- zation, or blending. The method of reducing ids determination in Section 11.2) into a particle size to less than 1 mm is matrix-de- clean beaker or glass jar. pendent. In general, hard particles can be re- 11.5.2 Spike 1.0 mL of the diluted labeled duced by grinding with a mortar and pestle. compound spiking solution (Section 7.10.3) Softer particles can be reduced by grinding into the sample. in a Wiley mill or meat grinder, by homog- 11.5.3 For each sample or sample batch (to enization, or in a blender. a maximum of 20 samples) to be extracted 11.7.1 Each size-reducing preparation pro- during the same 12-hour shift, weigh two 10 g cedure on each matrix shall be verified by aliquots of the appropriate reference matrix running the tests in Section 9.2 before the (Section 7.6) into clean beakers or glass jars. procedure is employed routinely. 11.5.4 Spike 1.0 mL of the diluted labeled 11.7.2 The grinding, homogenization, or compound spiking solution (Section 7.10.3) blending procedures shall be carried out in a into each reference matrix aliquot. One ali- glove box or fume hood to prevent particles quot will serve as the method blank. Spike from contaminating the work environment. 1.0 mL of the PAR standard (Section 7.14) 11.7.3 Grinding—Certain papers and pulps, into the other reference matrix aliquot. This slurries, and amorphous solids can be ground aliquot will serve as the OPR (Section 15.5). in a Wiley mill or heavy duty meat grinder. 11.5.5 Stir or tumble and equilibrate the In some cases, reducing the temperature of aliquots for one to two hours. the sample to freezing or to dry ice or liquid 11.5.6 Decant excess water. If necessary to nitrogen temperatures can aid in the grind- remove water, filter the sample through a ing process. Grind the sample aliquots from glass-fiber filter and discard the aqueous liq- Section 11.5.7 or 11.6.4 in a clean grinder. Do uid. not allow the sample temperature to exceed 11.5.7 If particles >1mm are present in the 50 °C. Grind the blank and reference matrix sample (as determined in Section 11.3.2), aliquots using a clean grinder. spread the sample on clean aluminum foil in 11.7.4 Homogenization or blending—Par- a hood. After the sample is dry, grind to re- ticles that are not ground effectively, or par- duce the particle size (Section 11.7). ticles greater than 1 mm in size after grind- 11.5.8 Extract the sample and QC aliquots ing, can often be reduced in size by high using the SDS procedure in Section 12.3. speed homogenization or blending. Homog- 11.6 Multiphase Samples. enize and/or blend the particles or filter from

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Section 11.5.7 or 11.6.4 for the sample, blank, 11.8.3.1 Spike 1.0 mL of the labeled com- and OPR aliquots. pound spiking solution (Section 7.10.3) into 11.7.5 Extract the aliquots using the SDS the sample, blank, and OPR aliquot. procedure in Section 12.3. 11.8.3.2 Spike 1.0 mL of the PAR standard 11.8 Fish and Other Tissues—Prior to (Section 7.14) into the OPR aliquot. processing tissue samples, the laboratory 11.8.4 Extract the aliquots using the pro- must determine the exact tissue to be ana- cedures in Section 12.4. lyzed. Common requests for analysis of fish 12.0 Extraction and Concentration tissue include whole fish—skin on, whole fish—skin removed, edible fish fillets Extraction procedures include separatory (filleted in the field or by the laboratory), funnel (Section 12.1) and solid phase (Section specific organs, and other portions. Once the 12.2) for aqueous liquids; Soxhlet/Dean-Stark appropriate tissue has been determined, the (Section 12.3) for solids, filters, and SPE sample must be homogenized. disks; and Soxhlet extraction (Section 12.4.1) 11.8.1 Homogenization. and HCl digestion (Section 12.4.2) for tissues. 11.8.1.1 Samples are homogenized while Acid/base back-extraction (Section 12.5) is still frozen, where practical. If the labora- used for initial cleanup of extracts. tory must dissect the whole fish to obtain Macro-concentration procedures include the appropriate tissue for analysis, the un- rotary evaporation (Section 12.6.1), heating used tissues may be rapidly refrozen and mantle (Section 12.6.2), and Kuderna-Danish stored in a clean glass jar for subsequent use. (K-D) evaporation (Section 12.6.3). Micro- 11.8.1.2 Each analysis requires 10 g of tis- concentration uses nitrogen blowdown (Sec- sue (wet weight). Therefore, the laboratory tion 12.7). should homogenize at least 20 g of tissue to 12.1 Separatory funnel extraction of fil- allow for re-extraction of a second aliquot of trates and of aqueous samples visibly absent the same homogenized sample, if re-analysis particles. is required. When whole fish analysis is nec- 12.1.1 Pour the spiked sample (Section essary, the entire fish is homogenized. 11.4.2.2) or filtrate (Section 11.4.3.5) into a 2 L 11.8.1.3 Homogenize the sample in a tissue separatory funnel. Rinse the bottle or flask homogenizer (Section 6.3.3) or grind in a twice with 5 mL of reagent water and add meat grinder (Section 6.3.4). Cut tissue too these rinses to the separatory funnel. 12.1.2 Add 60 mL methylene chloride to large to feed into the grinder into smaller the empty sample bottle (Section 12.1.1), pieces. To assure homogeneity, grind three seal, and shake 60 seconds to rinse the inner times. surface. Transfer the solvent to the sepa- 11.8.1.4 Transfer approximately 10 g (wet ratory funnel, and extract the sample by weight) of homogenized tissue to a clean, shaking the funnel for two minutes with tared, 400–500 mL beaker. For the alternate periodic venting. Allow the organic layer to HCl digestion/extraction, transfer the tissue separate from the aqueous phase for a min- to a clean, tared 500–600 mL wide-mouth bot- imum of 10 minutes. If an emulsion forms tle. Record the weight to the nearest 10 mg. and is more than one-third the volume of the 11.8.1.5 Transfer the remaining homog- solvent layer, employ mechanical techniques enized tissue to a clean jar with a to complete the phase separation (see note fluoropolymer-lined lid. Seal the jar and below). Drain the methylene chloride extract ¥ ° store the tissue at < 10 C. Return any tis- through a solvent-rinsed glass funnel ap- sue that was not homogenized to its original proximately one-half full of granular anhy- ¥ ° container and store at < 10 C. drous sodium sulfate (Section 7.2.1) sup- 11.8.2 QC aliquots. ported on clean glass-fiber paper into a sol- 11.8.2.1 Prepare a method blank by adding vent-rinsed concentration device (Section approximately 10 g of the oily liquid ref- 12.6). erence matrix (Section 7.6.4) to a 400–500 mL beaker. For the alternate HCl digestion/ex- NOTE: If an emulsion forms, the analyst traction, add the reference matrix to a 500– must employ mechanical techniques to com- 600 mL wide-mouth bottle. Record the plete the phase separation. The optimum weight to the nearest 10 mg. technique depends upon the sample, but may 11.8.2.2 Prepare a precision and recovery include stirring, filtration through glass aliquot by adding approximately 10 g of the wool, use of phase separation paper, cen- oily liquid reference matrix (Section 7.6.4) to trifugation, use of an ultrasonic bath with a separate 400–500 mL beaker or wide-mouth ice, addition of NaCl, or other physical meth- bottle, depending on the extraction proce- ods. Alternatively, solid-phase or other ex- traction techniques may be used to prevent dure to be used. Record the weight to the emulsion formation. Any alternative tech- nearest 10 mg. If the initial precision and re- nique is acceptable so long as the require- covery test is to be performed, use four ments in Section 9 are met. aliquots; if the ongoing precision and recov- ery test is to be performed, use a single ali- Experience with aqueous samples high in quot. dissolved organic materials (e.g., paper mill 11.8.3 Spiking effluents) has shown that acidification of the

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sample prior to extraction may reduce the the filter. Do not allow the disk to go dry formation of emulsions. Paper industry from this point until the end of the extrac- methods suggest that the addition of up to tion. 400 mL of ethanol to a 1 L effluent sample 12.2.1.4 Rinse the filter/disk with two 50- may also reduce emulsion formation. How- mL portions of reagent water by adding the ever, studies by EPA suggest that the effect water to the reservoir and pulling most may be a result of sample dilution, and that through, leaving a layer of water on the sur- the addition of reagent water may serve the face of the filter. same function. Mechanical techniques may 12.2.2 Extraction. still be necessary to complete the phase sep- 12.2.2.1 Pour the spiked sample (Section aration. If either acidification or addition of 11.4.2.2), blank (Section 11.4.2.4), or IPR/OPR ethanol is utilized, the laboratory must per- aliquot (Section 11.4.2.5) into the reservoir form the startup tests described in Section and turn on the vacuum to begin the extrac- 9.2 using the same techniques. tion. Adjust the vacuum to complete the ex- 12.1.3 Extract the water sample two more traction in no less than 10 minutes. For sam- times with 60 mL portions of methylene ples containing a high concentration of par- chloride. Drain each portion through the so- ticles (suspended solids), filtration times dium sulfate into the concentrator. After the may be eight hours or longer. third extraction, rinse the separatory funnel with at least 20 mL of methylene chloride, 12.2.2.2 Before all of the sample has been and drain this rinse through the sodium sul- pulled through the filter/disk, rinse the sam- fate into the concentrator. Repeat this rinse ple bottle with approximately 50 mL of rea- at least twice. Set aside the funnel with so- gent water to remove any solids, and pour dium sulfate if the extract is to be combined into the reservoir. Pull through the filter/ with the extract from the particles. disk. Use additional reagent water rinses 12.1.4 Concentrate the extract using one until all visible solids are removed. of the macro-concentration procedures in 12.2.2.3 Before all of the sample and rinses Section 12.6. have been pulled through the filter/disk, 12.1.4.1 If the extract is from a sample rinse the sides of the reservoir with small visibly absent particles (Section 11.1.2.1), ad- portions of reagent water. just the final volume of the concentrated ex- 12.2.2.4 Allow the filter/disk to dry, then tract to approximately 10 mL with hexane, remove the filter and disk and place in a transfer to a 250 mL separatory funnel, and glass Petri dish. Extract the filter and disk back-extract using the procedure in Section per Section 12.3. 12.5. 12.3 SDS Extraction of Samples Con- 12.1.4.2 If the extract is from the aqueous taining Particles, and of Filters and/or filtrate (Section 11.4.3.5), set aside the con- Disks. centration apparatus for addition of the SDS 12.3.1 Charge a clean extraction thimble extract from the particles (Section 12.3.9.1.2). (Section 6.4.2.2) with 5.0 g of 100/200 mesh sili- 12.2 SPE of Samples Containing Less ca (Section 7.5.1.1) topped with 100 g of Than 1% Solids (References 19–20). quartz sand (Section 7.3.2). 12.2.1 Disk preparation. 12.2.1.1 Place an SPE disk on the base of NOTE: Do not disturb the silica layer the filter holder (Figure 4) and wet with tol- throughout the extraction process. uene. While holding a GMF 150 filter above the SPE disk with tweezers, wet the filter 12.3.2 Place the thimble in a clean extrac- with toluene and lay the filter on the SPE tor. Place 30–40 mL of toluene in the receiver disk, making sure that air is not trapped be- and 200–250 mL of toluene in the flask. tween the filter and disk. Clamp the filter 12.3.3 Pre-extract the glassware by heat- and SPE disk between the 1 L glass reservoir ing the flask until the toluene is boiling. and the vacuum filtration flask. When properly adjusted, one to two drops of 12.2.1.2 Rinse the sides of the filtration toluene will fall per second from the con- flask with approx 15 mL of toluene using a denser tip into the receiver. Extract the ap- squeeze bottle or syringe. Apply vacuum mo- paratus for a minimum of three hours. mentarily until a few drops appear at the 12.3.4 After pre-extraction, cool and dis- drip tip. Release the vacuum and allow the assemble the apparatus. Rinse the thimble filter/disk to soak for approx one minute. with toluene and allow to air dry. Apply vacuum and draw all of the toluene 12.3.5 Load the wet sample, filter, and/or through the filter/disk. Repeat the wash step disk from Section 11.4.3.6, 11.5.8, 11.6.4, 11.7.3, with approx 15 mL of acetone and allow the 11.7.4, or 12.2.2.4 and any nonaqueous liquid filter/disk to air dry. from Section 11.6.3 into the thimble and 12.2.1.3 Re-wet the filter/disk with ap- manually mix into the sand layer with a proximately 15 mL of methanol, allowing the clean metal spatula, carefully breaking up filter/disk to soak for approximately one any large lumps of sample. minute. Pull the methanol through the fil- 12.3.6 Reassemble the pre-extracted SDS ter/disk using the vacuum, but retain a layer apparatus, and add a fresh charge of toluene of methanol approximately 1 mm thick on to the receiver and reflux flask. Apply power

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to the heating mantle to begin refluxing. Ad- thimble, and install the thimble in the Soxh- just the reflux rate to match the rate of per- let apparatus. colation through the sand and silica beds 12.4.1.5 Rinse the beaker with several por- until water removal lessens the restriction tions of solvent mixture and add to the thim- to toluene flow. Frequently check the appa- ble. Fill the thimble/receiver with solvent. ratus for foaming during the first two hours Extract for 18–24 hours. of extraction. If foaming occurs, reduce the 12.4.1.6 After extraction, cool and dis- reflux rate until foaming subsides. assemble the apparatus. 12.3.7 Drain the water from the receiver 12.4.1.7 Quantitatively transfer the ex- at one to two hours and eight to nine hours, tract to a macro-concentration device (Sec- or sooner if the receiver fills with water. tion 12.6), and concentrate to near dryness. Reflux the sample for a total of 16–24 hours. Set aside the concentration apparatus for re- Cool and disassemble the apparatus. Record use. the total volume of water collected. 12.4.1.8 Complete the removal of the sol- 12.3.8 Remove the distilling flask. Drain vent using the nitrogen blowdown procedure the water from the Dean-Stark receiver and (Section 12.7) and a water bath temperature add any toluene in the receiver to the ex- of 60 °C. Weigh the receiver, record the tract in the flask. weight, and return the receiver to the blow- 12.3.9 Concentrate the extract using one down apparatus, concentrating the residue of the macro-concentration procedures in until a constant weight is obtained. Section 12.6 per the following: 12.4.1.9 Percent lipid determination—The 12.3.9.1 Extracts from the particles in an lipid content is determined by extraction of aqueous sample containing less than 1% sol- tissue with the same solvent system (meth- ids (Section 11.4.3.6). ylene chloride:hexane) that was used in 12.3.9.1.1 Concentrate the extract to ap- EPA’s National Dioxin Study (Reference 22) proximately 5 mL using the rotary evapo- so that lipid contents are consistent with rator or heating mantle procedures in Sec- that study. tion 12.6.1 or 12.6.2. 12.4.1.9.1 Redissolve the residue in the re- 12.3.9.1.2 Quantitatively transfer the ex- ceiver in hexane and spike 1.0 mL of the tract through the sodium sulfate (Section cleanup standard (Section 7.11) into the solu- 12.1.3) into the apparatus that was set aside tion. (Section 12.1.4.2) and reconcentrate to the 12.4.1.9.2 Transfer the residue/hexane to level of the toluene. the anthropogenic isolation column (Section 12.3.9.1.3 Adjust to approximately 10 mL 13.7.1) or bottle for the acidified silica gel with hexane, transfer to a 250 mL separatory batch cleanup (Section 13.7.2), retaining the funnel, and proceed with back-extraction boiling chips in the concentration apparatus. (Section 12.5). Use several rinses to assure that all material 12.3.9.2 Extracts from particles (Sections is transferred. If necessary, sonicate or heat 11.5 through 11.6) or from the SPE filter and the receiver slightly to assure that all mate- disk (Section 12.2.2.4)—Concentrate to ap- rial is re-dissolved. Allow the receiver to dry. proximately 10 mL using the rotary evapo- Weigh the receiver and boiling chips. rator or heating mantle (Section 12.6.1 or 12.4.1.9.3 Calculate the lipid content to 12.6.2), transfer to a 250 mL separatory fun- the nearest three significant figures as fol- nel, and proceed with back-extraction (Sec- lows: tion 12.5). 12.4 Extraction of Tissue—Two procedures Weight of residue(g) × are provided for tissue extraction. Percent lipid = 100 12.4.1 Soxhlet extraction (Reference 21). Weight of tissue (g) 12.4.1.1 Add 30–40 g of powdered anhydrous 12.4.1.9.4 It is not necessary to determine sodium sulfate to each of the beakers (Sec- the lipid content of the blank, IPR, or OPR tion 11.8.4) and mix thoroughly. Cover the aliquots. beakers with aluminum foil and allow to 12.4.2 HCl digestion/extraction and con- equilibrate for 12–24 hours. Remix prior to centration (References 23–26). extraction to prevent clumping. 12.4.2.1 Add 200 mL of 6 N HCl and 200 mL 12.4.1.2 Assemble and pre-extract the of methylene chloride:hexane (1:1) to the Soxhlet apparatus per Sections 12.3.1 sample and QC aliquots (Section 11.8.4). through 12.3.4, except use the methylene 12.4.2.2 Cap and shake each bottle one to chloride:hexane (1:1) mixture for the pre-ex- three times. Loosen the cap in a hood to vent traction and rinsing and omit the quartz excess pressure. Shake each bottle for 10–30 sand. The Dean-Stark moisture trap may seconds and vent. also be omitted, if desired. 12.4.2.3 Tightly cap and place on shaker. 12.4.1.3 Reassemble the pre-extracted Adjust the shaker action and speed so that Soxhlet apparatus and add a fresh charge of the acid, solvent, and tissue are in constant methylene chloride:hexane to the reflux motion. However, take care to avoid such flask. violent action that the bottle may be dis- 12.4.1.4 Transfer the sample/sodium sul- lodged from the shaker. Shake for 12–24 fate mixture (Section 12.4.1.1) to the Soxhlet hours.

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12.4.2.4 After digestion, remove the bot- 12.5.3 Partition the extract against 50 mL tles from the shaker. Allow the bottles to of sodium chloride solution (Section 7.1.4) in stand so that the solvent and acid layers sep- the same way as with base. Discard the aque- arate. ous layer. 12.4.2.5 Decant the solvent through a glass 12.5.4 Partition the extract against 50 mL funnel with glass-fiber filter (Sections 6.5.2 of sulfuric acid (Section 7.1.2) in the same through 6.5.3) containing approximately 10 g way as with base. Repeat the acid washing of granular anhydrous sodium sulfate (Sec- until no color is visible in the aqueous layer, tion 7.2.1) into a macro-concentration appa- to a maximum of four washings. ratus (Section 12.6). Rinse the contents of 12.5.5 Repeat the partitioning against so- the bottle with two 25 mL portions of hexane dium chloride solution and discard the aque- and pour through the sodium sulfate into the ous layer. apparatus. 12.5.6 Pour each extract through a drying 12.4.2.6 Concentrate the solvent to near column containing 7–10 cm of granular anhy- dryness using a macro-concentration proce- drous sodium sulfate (Section 7.2.1). Rinse dure (Section 12.6). the separatory funnel with 30–50 mL of sol- 12.4.2.7 Complete the removal of the sol- vent, and pour through the drying column. vent using the nitrogen blowdown apparatus Collect each extract in a round-bottom flask. (Section 12.7) and a water bath temperature Re-concentrate the sample and QC aliquots ° of 60 C. Weigh the receiver, record the per Sections 12.6 through 12.7, and clean up weight, and return the receiver to the blow- the samples and QC aliquots per Section 13. down apparatus, concentrating the residue 12.6 Macro-Concentration—Extracts in until a constant weight is obtained. toluene are concentrated using a rotary 12.4.2.8 Percent lipid determination—The evaporator or a heating mantle; extracts in lipid content is determined in the same sol- methylene chloride or hexane are con- vent system [methylene chloride:hexane centrated using a rotary evaporator, heating (1:1)] that was used in EPA’s National Dioxin mantle, or Kuderna-Danish apparatus. Study (Reference 22) so that lipid contents are consistent with that study. 12.6.1 Rotary evaporation—Concentrate 12.4.2.8.1 Redissolve the residue in the re- the extracts in separate round-bottom ceiver in hexane and spike 1.0 mL of the flasks. cleanup standard (Section 7.11) into the solu- 12.6.1.1 Assemble the rotary evaporator tion. according to manufacturer’s instructions, 12.4.2.8.2 Transfer the residue/hexane to and warm the water bath to 45 °C. On a daily the narrow-mouth 100–200 mL bottle retain- basis, preclean the rotary evaporator by con- ing the boiling chips in the receiver. Use sev- centrating 100 mL of clean extraction sol- eral rinses to assure that all material is vent through the system. Archive both the transferred, to a maximum hexane volume of concentrated solvent and the solvent in the approximately 70 mL. Allow the receiver to catch flask for a contamination check if nec- dry. Weigh the receiver and boiling chips. essary. Between samples, three 2–3 mL 12.4.2.8.3 Calculate the percent lipid per aliquots of solvent should be rinsed down the Section 12.4.1.9.3. It is not necessary to deter- feed tube into a waste beaker. mine the lipid content of the blank, IPR, or 12.6.1.2 Attach the round-bottom flask OPR aliquots. containing the sample extract to the rotary 12.4.2.9 Clean up the extract per Section evaporator. Slowly apply vacuum to the sys- 13.7.3. tem, and begin rotating the sample flask. 12.5 Back-Extraction with Base and Acid. 12.6.1.3 Lower the flask into the water 12.5.1 Spike 1.0 mL of the cleanup stand- bath, and adjust the speed of rotation and ard (Section 7.11) into the separatory funnels the temperature as required to complete con- containing the sample and QC extracts from centration in 15–20 minutes. At the proper Section 12.1.4.1, 12.3.9.1.3, or 12.3.9.2. rate of concentration, the flow of solvent 12.5.2 Partition the extract against 50 mL into the receiving flask will be steady, but of potassium hydroxide solution (Section no bumping or visible boiling of the extract 7.1.1). Shake for two minutes with periodic will occur. venting into a hood. Remove and discard the NOTE: If the rate of concentration is too aqueous layer. Repeat the base washing until fast, analyte loss may occur. no color is visible in the aqueous layer, to a maximum of four washings. Minimize con- 12.6.1.4 When the liquid in the concentra- tact time between the extract and the base tion flask has reached an apparent volume of to prevent degradation of the CDDs/CDFs. approximately 2 mL, remove the flask from Stronger potassium hydroxide solutions may the water bath and stop the rotation. Slowly be employed for back-extraction, provided and carefully admit air into the system. Be that the laboratory meets the specifications sure not to open the valve so quickly that for labeled compound recovery and dem- the sample is blown out of the flask. Rinse onstrates acceptable performance using the the feed tube with approximately 2 mL of procedure in Section 9.2. solvent.

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12.6.1.5 Proceed to Section 12.6.4 for prepa- the column will actively chatter but the ration for back-extraction or micro-con- chambers will not flood. centration and solvent exchange. 12.6.3.6 When the liquid reaches an appar- 12.6.2 Heating mantle—Concentrate the ent volume of 0.5 mL, remove the apparatus extracts in separate round-bottom flasks. from the water bath and allow to drain and 12.6.2.1 Add one or two clean boiling chips cool for at least 10 minutes. to the round-bottom flask, and attach a 12.6.3.7 Proceed to 12.6.4 for preparation three-ball macro Snyder column. Prewet the for back-extraction or micro-concentration column by adding approximately 1 mL of sol- and solvent exchange. vent through the top. Place the round-bot- 12.6.4 Preparation for back-extraction or tom flask in a heating mantle, and apply micro-concentration and solvent exchange. heat as required to complete the concentra- 12.6.4.1 For back-extraction (Section 12.5), tion in 15–20 minutes. At the proper rate of transfer the extract to a 250 mL separatory distillation, the balls of the column will ac- funnel. Rinse the concentration vessel with tively chatter, but the chambers will not small portions of hexane, adjust the hexane flood. volume in the separatory funnel to 10–20 mL, 12.6.2.2 When the liquid has reached an ap- and proceed to back-extraction (Section parent volume of approximately 10 mL, re- 12.5). move the round-bottom flask from the heat- 12.6.4.2 For determination of the weight of ing mantle and allow the solvent to drain residue in the extract, or for clean-up proce- and cool for at least 10 minutes. Remove the dures other than back-extraction, transfer Snyder column and rinse the glass joint into the extract to a blowdown vial using two to three rinses of solvent. Proceed with micro- the receiver with small portions of solvent. concentration and solvent exchange (Section 12.6.2.3 Proceed to Section 12.6.4 for prepa- 12.7). ration for back-extraction or micro-con- 12.7 Micro-Concentration and Solvent Ex- centration and solvent exchange. change. 12.6.3 Kuderna-Danish (K-D)—Concentrate 12.7.1 Extracts to be subjected to GPC or the extracts in separate 500 mL K-D flasks HPLC cleanup are exchanged into methylene equipped with 10 mL concentrator tubes. The chloride. Extracts to be cleaned up using sili- K-D technique is used for solvents such as ca gel, alumina, carbon, and/or Florisil are methylene chloride and hexane. Toluene is exchanged into hexane. difficult to concentrate using the K-D tech- 12.7.2 Transfer the vial containing the nique unless a water bath fed by a steam sample extract to a nitrogen blowdown de- generator is used. vice. Adjust the flow of nitrogen so that the 12.6.3.1 Add one to two clean boiling chips surface of the solvent is just visibly dis- to the receiver. Attach a three-ball macro turbed. Snyder column. Prewet the column by add- ing approximately 1 mL of solvent through NOTE: A large vortex in the solvent may the top. Place the K-D apparatus in a hot cause analyte loss. water bath so that the entire lower rounded 12.7.3 Lower the vial into a 45 °C water surface of the flask is bathed with steam. bath and continue concentrating. 12.6.3.2 Adjust the vertical position of the 12.7.3.1 If the extract is to be con- apparatus and the water temperature as re- centrated to dryness for weight determina- quired to complete the concentration in 15–20 tion (Sections 12.4.1.8, 12.4.2.7, and 13.7.1.4), minutes. At the proper rate of distillation, blow dry until a constant weight is obtained. the balls of the column will actively chatter 12.7.3.2 If the extract is to be con- but the chambers will not flood. centrated for injection into the GC/MS or the 12.6.3.3 When the liquid has reached an ap- solvent is to be exchanged for extract clean- parent volume of 1 mL, remove the K-D ap- up, proceed as follows: paratus from the bath and allow the solvent 12.7.4 When the volume of the liquid is ap- to drain and cool for at least 10 minutes. Re- proximately 100 L, add 2–3 mL of the desired move the Snyder column and rinse the flask solvent (methylene chloride for GPC and and its lower joint into the concentrator HPLC, or hexane for the other cleanups) and tube with 1–2 mL of solvent. A 5 mL syringe continue concentration to approximately 100 is recommended for this operation. μL. Repeat the addition of solvent and con- 12.6.3.4 Remove the three-ball Snyder col- centrate once more. umn, add a fresh boiling chip, and attach a 12.7.5 If the extract is to be cleaned up by two-ball micro Snyder column to the concen- GPC, adjust the volume of the extract to 5.0 trator tube. Prewet the column by adding mL with methylene chloride. If the extract approximately 0.5 mL of solvent through the is to be cleaned up by HPLC, further con- top. Place the apparatus in the hot water centrate the extract to 30 μL. Proceed with bath. GPC or HPLC cleanup (Section 13.2 or 13.6, 12.6.3.5 Adjust the vertical position and respectively). the water temperature as required to com- 12.7.6 If the extract is to be cleaned up by plete the concentration in 5–10 minutes. At column chromatography (alumina, silica gel, the proper rate of distillation, the balls of Carbopak/Celite, or Florisil), bring the final

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volume to 1.0 mL with hexane. Proceed with through the column, from bottom to top, at column cleanups (Sections 13.3 through 13.5 4.5–5.5 mL/minute prior to connecting the and 13.8). column to the detector. 12.7.7 If the extract is to be concentrated 13.2.1.4 After purging the column with sol- for injection into the GC/MS (Section 14), vent for one to two hours, adjust the column quantitatively transfer the extract to a 0.3 head pressure to 7–10 psig and purge for four mL conical vial for final concentration, rins- to five hours to remove air. Maintain a head ing the larger vial with hexane and adding pressure of 7–10 psig. Connect the column to the rinse to the conical vial. Reduce the vol- the detector (Section 6.7.1.4). ume to approximately 100 μL. Add 10 μL of 13.2.2 Column calibration. nonane to the vial, and evaporate the solvent 13.2.2.1 Load 5 mL of the calibration solu- to the level of the nonane. Seal the vial and tion (Section 7.4) into the sample loop. label with the sample number. Store in the 13.2.2.2 Inject the calibration solution and dark at room temperature until ready for record the signal from the detector. The GC/MS analysis. If GC/MS analysis will not elution pattern will be corn oil, bis(2-ethyl be performed on the same day, store the vial hexyl)phthalate, pentachlorophenol, pery- at <¥10 °C. lene, and sulfur. 13.2.2.3 Set the ‘‘dump time’’ to allow 13.0 Extract Cleanup >85% removal of the corn oil and >85% col- 13.1 Cleanup may not be necessary for rel- lection of the phthalate. atively clean samples (e.g., treated effluents, 13.2.2.4 Set the ‘‘collect time’’ to the peak groundwater, drinking water). If particular minimum between perylene and sulfur. circumstances require the use of a cleanup 13.2.2.5 Verify the calibration with the procedure, the analyst may use any or all of calibration solution after every 20 extracts. the procedures below or any other appro- Calibration is verified if the recovery of the priate procedure. Before using a cleanup pro- pentachlorophenol is greater than 85%. If cedure, the analyst must demonstrate that calibration is not verified, the system shall the requirements of Section 9.2 can be met be recalibrated using the calibration solu- using the cleanup procedure. If only 2,3,7,8- tion, and the previous 20 samples shall be re- TCDD and 2,3,7,8-TCDF are to be determined, extracted and cleaned up using the cali- the cleanup procedures may be optimized for brated GPC system. isolation of these two compounds. 13.2.3 Extract cleanup—GPC requires that 13.1.1 Gel permeation chromatography the column not be overloaded. The column (Section 13.2) removes high molecular weight specified in this method is designed to han- interferences that cause GC column perform- dle a maximum of 0.5 g of high molecular ance to degrade. It should be used for all soil weight material in a 5 mL extract. If the ex- and sediment extracts and may be used for tract is known or expected to contain more water extracts that are expected to contain than 0.5 g, the extract is split into aliquots high molecular weight organic compounds for GPC, and the aliquots are combined after (e.g., polymeric materials, humic acids). elution from the column. The residue con- 13.1.2 Acid, neutral, and basic silica gel tent of the extract may be obtained gravi- (Section 13.3), alumina (Section 13.4), and metrically by evaporating the solvent from a Florisil (Section 13.8) are used to remove 50 μL aliquot. nonpolar and polar interferences. Alumina 13.2.3.1 Filter the extract or load through and Florisil are used to remove the filter holder (Section 6.7.1.3) to remove chlorodiphenyl ethers. the particles. Load the 5.0 mL extract onto 13.1.3 Carbopak/Celite (Section 13.5) is the column. used to remove nonpolar interferences. 13.2.3.2 Elute the extract using the cali- 13.1.4 HPLC (Section 13.6) is used to pro- bration data determined in Section 13.2.2. vide specificity for the 2,3,7,8-substituted and Collect the eluate in a clean 400–500 mL other CDD and CDF isomers. beaker. 13.1.5 The anthropogenic isolation column 13.2.3.3 Rinse the sample loading tube (Section 13.7.1), acidified silica gel batch ad- thoroughly with methylene chloride between sorption procedure (Section 13.7.2), and sul- extracts to prepare for the next sample. furic acid and base back-extraction (Section 13.2.3.4 If a particularly dirty extract is 13.7.3) are used for removal of lipids from tis- encountered, a 5.0 mL methylene chloride sue samples. blank shall be run through the system to 13.2 Gel Permeation Chromatography check for carry-over. (GPC). 13.2.3.5 Concentrate the eluate per Sec- 13.2.1 Column packing. tions 12.6 and 12.7 for further cleanup or in- 13.2.1.1 Place 70–75 g of SX–3 Bio-beads jection into the GC/MS. (Section 6.7.1.1) in a 400–500 mL beaker. 13.3 Silica Gel Cleanup. 13.2.1.2 Cover the beads with methylene 13.3.1 Place a glass-wool plug in a 15 mm chloride and allow to swell overnight (a min- ID chromatography column (Section 6.7.4.2). imum of 12 hours). Pack the column bottom to top with: 1 g sili- 13.2.1.3 Transfer the swelled beads to the ca gel (Section 7.5.1.1), 4 g basic silica gel column (Section 6.7.1.1) and pump solvent (Section 7.5.1.3), 1 g silica gel, 8 g acid silica

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gel (Section 7.5.1.2), 2 g silica gel, and 4 g the column. Elute the interfering compounds granular anhydrous sodium sulfate (Section with 100 mL hexane and discard the eluate. 7.2.1). Tap the column to settle the adsorb- 13.4.7 The choice of eluting solvents will ents. depend on the choice of alumina (acid or 13.3.2 Pre-elute the column with 50–100 basic) made in Section 13.4.2. mL of hexane. Close the stopcock when the 13.4.7.1 If using acid alumina, elute the hexane is within 1 mm of the sodium sulfate. CDDs/CDFs from the column with 20 mL Discard the eluate. Check the column for methylene chloride:hexane (20:80 v/v). Collect channeling. If channeling is present, discard the eluate. the column and prepare another. 13.4.7.2 If using basic alumina, elute the 13.3.3 Apply the concentrated extract to CDDs/CDFs from the column with 20 mL the column. Open the stopcock until the ex- methylene chloride:hexane (50:50 v/v). Collect tract is within 1 mm of the sodium sulfate. the eluate. 13.3.4 Rinse the receiver twice with 1 mL 13.4.8 Concentrate the eluate per Sections portions of hexane, and apply separately to 12.6 and 12.7 for further cleanup or injection the column. Elute the CDDs/CDFs with 100 into the HPLC or GC/MS. mL hexane, and collect the eluate. 13.5 Carbon Column. 13.3.5 Concentrate the eluate per Sections 13.5.1 Cut both ends from a 10 mL dispos- able serological pipet (Section 6.7.3.2) to 12.6 and 12.7 for further cleanup or injection produce a 10 cm column. Fire-polish both into the HPLC or GC/MS. ends and flare both ends if desired. Insert a 13.3.6 For extracts of samples known to glass-wool plug at one end, and pack the col- contain large quantities of other organic umn with 0.55 g of Carbopak/Celite (Section compounds (such as paper mill effluents), it 7.5.3.3) to form an adsorbent bed approxi- may be advisable to increase the capacity of mately 2 cm long. Insert a glass-wool plug on the silica gel column. This may be accom- top of the bed to hold the adsorbent in place. plished by increasing the strengths of the 13.5.2 Pre-elute the column with 5 mL of acid and basic silica gels. The acid silica gel toluene followed by 2 mL of methylene chlo- (Section 7.5.1.2) may be increased in strength ride: methanol:toluene (15:4:1 v/v), 1 mL of to as much as 44% w/w (7.9 g sulfuric acid methylene chloride:cyclohexane (1:1 v/v), and added to 10 g silica gel). The basic silica gel 5 mL of hexane. If the flow rate of eluate ex- (Section 7.5.1.3) may be increased in strength ceeds 0.5 mL/minute, discard the column. to as much as 33% w/w (50 mL 1N NaOH 13.5.3 When the solvent is within 1 mm of added to 100 g silica gel), or the potassium the column packing, apply the sample ex- silicate (Section 7.5.1.4) may be used. tract to the column. Rinse the sample con- NOTE: The use of stronger acid silica gel tainer twice with 1 mL portions of hexane (44% w/w) may lead to charring of organic and apply separately to the column. Apply 2 compounds in some extracts. The charred mL of hexane to complete the transfer. material may retain some of the analytes 13.5.4 Elute the interfering compounds and lead to lower recoveries of CDDs/CDFs. with two 3 mL portions of hexane, 2 mL of Increasing the strengths of the acid and methylene chloride:cyclohexane (1:1 v/v), and basic silica gel may also require different 2 mL of methylene chlo- volumes of hexane than those specified above ride:methanol:toluene (15:4:1 v/v). Discard to elute the analytes off the column. There- the eluate. fore, the performance of the method after 13.5.5 Invert the column, and elute the such modifications must be verified by the CDDs/CDFs with 20 mL of toluene. If carbon procedure in Section 9.2. particles are present in the eluate, filter through glass-fiber filter paper. 13.4 Alumina Cleanup. 13.5.6 Concentrate the eluate per Sections 13.4.1 Place a glass-wool plug in a 15 mm 12.6 and 12.7 for further cleanup or injection ID chromatography column (Section 6.7.4.2). into the HPLC or GC/MS. 13.4.2 If using acid alumina, pack the col- 13.6 HPLC (Reference 6). umn by adding 6 g acid alumina (Section 13.6.1 Column calibration. 7.5.2.1). If using basic alumina, substitute 6 g 13.6.1.1 Prepare a calibration standard basic alumina (Section 7.5.2.2). Tap the col- containing the 2,3,7,8-substituted isomers umn to settle the adsorbents. and/or other isomers of interest at a con- 13.4.3 Pre-elute the column with 50–100 centration of approximately 500 pg/μL in mL of hexane. Close the stopcock when the methylene chloride. hexane is within 1 mm of the alumina. 13.6.1.2 Inject 30 μL of the calibration so- 13.4.4 Discard the eluate. Check the col- lution into the HPLC and record the signal umn for channeling. If channeling is present, from the detector. Collect the eluant for discard the column and prepare another. reuse. The elution order will be the tetra- 13.4.5 Apply the concentrated extract to through octa-isomers. the column. Open the stopcock until the ex- 13.6.1.3 Establish the collection time for tract is within 1 mm of the alumina. the tetra-isomers and for the other isomers 13.4.6 Rinse the receiver twice with 1 mL of interest. Following calibration, flush the portions of hexane and apply separately to injection system with copious quantities of

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methylene chloride, including a minimum of using a fresh anthropogenic isolation col- five 50 μL injections while the detector is umn. monitored, to ensure that residual CDDs/ 13.7.1.5 Redissolve the extract in a solvent CDFs are removed from the system. suitable for the additional cleanups to be 13.6.1.4 Verify the calibration with the used (Sections 13.2 through 13.6 and 13.8). calibration solution after every 20 extracts. 13.7.1.6 Spike 1.0 mL of the cleanup stand- Calibration is verified if the recovery of the ard (Section 7.11) into the residue/solvent. CDDs/CDFs from the calibration standard 13.7.1.7 Clean up the extract using the pro- (Section 13.6.1.1) is 75–125% compared to the cedures in Sections 13.2 through 13.6 and 13.8. calibration (Section 13.6.1.2). If calibration is Alumina (Section 13.4) or Florisil (Section not verified, the system shall be recalibrated 13.8) and carbon (Section 13.5) are rec- using the calibration solution, and the pre- ommended as minimum additional cleanup vious 20 samples shall be re-extracted and steps. cleaned up using the calibrated system. 13.7.1.8 Following cleanup, concentrate 13.6.2 Extract cleanup—HPLC requires the extract to 10 μL as described in Section that the column not be overloaded. The col- 12.7 and proceed with the analysis in Section umn specified in this method is designed to 14. handle a maximum of 30 μL of extract. If the 13.7.2 Acidified silica gel (Reference 28)— extract cannot be concentrated to less than Procedure alternate to the anthropogenic 30 μL, it is split into fractions and the frac- isolation column (Section 13.7.1) that is used tions are combined after elution from the for removal of lipids from the Soxhlet/SDS column. extraction (Section 12.4.1). 13.6.2.1 Rinse the sides of the vial twice 13.7.2.1 Adjust the volume of hexane in with 30 μL of methylene chloride and reduce the bottle (Section 12.4.1.9.2) to approxi- to 30 μL with the evaporation apparatus mately 200 mL. (Section 12.7). 13.7.2.2 Spike 1.0 mL of the cleanup stand- 13.6.2.2 Inject the 30 μL extract into the ard (Section 7.11) into the residue/solvent. HPLC. 13.7.2.3 Drop the stirring bar into the bot- 13.6.2.3 Elute the extract using the cali- tle, place the bottle on the stirring plate, bration data determined in Section 13.6.1. and begin stirring. Collect the fraction(s) in a clean 20 mL con- 13.7.2.4 Add 30–100 g of acid silica gel (Sec- centrator tube containing 5 mL of tion 7.5.1.2) to the bottle while stirring, hexane:acetone (1:1 v/v). keeping the silica gel in motion. Stir for two 13.6.2.4 If an extract containing greater to three hours. than 100 ng/mL of total CDD or CDF is en- NOTE: 30 grams of silica gel should be ade- countered, a 30 μL methylene chloride blank quate for most samples and will minimize shall be run through the system to check for contamination from this source. carry-over. 13.6.2.5 Concentrate the eluate per Sec- 13.7.2.5 After stirring, pour the extract tion 12.7 for injection into the GC/MS. through approximately 10 g of granular an- 13.7 Cleanup of Tissue Lipids—Lipids are hydrous sodium sulfate (Section 7.2.1) con- removed from the Soxhlet extract using ei- tained in a funnel with glass-fiber filter into ther the anthropogenic isolation column a macro contration device (Section 12.6). (Section 13.7.1) or acidified silica gel (Section Rinse the bottle and sodium sulfate with 13.7.2), or are removed from the HCl digested hexane to complete the transfer. extract using sulfuric acid and base back-ex- 13.7.2.6 Concentrate the extract per Sec- traction (Section 13.7.3). tions 12.6 through 12.7 and clean up the ex- 13.7.1 Anthropogenic isolation column tract using the procedures in Sections 13.2 (References 22 and 27)—Used for removal of through 13.6 and 13.8. Alumina (Section 13.4) lipids from the Soxhlet/SDS extraction (Sec- or Florisil (Section 13.8) and carbon (Section tion 12.4.1). 13.5) are recommended as minimum addi- 13.7.1.1 Prepare the column as given in tional cleanup steps. Section 7.5.4. 13.7.3 Sulfuric acid and base back-extrac- 13.7.1.2 Pre-elute the column with 100 mL tion. Used with HCl digested extracts (Sec- of hexane. Drain the hexane layer to the top tion 12.4.2). of the column, but do not expose the sodium 13.7.3.1 Spike 1.0 mL of the cleanup stand- sulfate. ard (Section 7.11) into the residue/solvent 13.7.1.3 Load the sample and rinses (Sec- (Section 12.4.2.8.2). tion 12.4.1.9.2) onto the column by draining 13.7.3.2 Add 10 mL of concentrated sul- each portion to the top of the bed. Elute the furic acid to the bottle. Immediately cap and CDDs/CDFs from the column into the appa- shake one to three times. Loosen cap in a ratus used for concentration (Section hood to vent excess pressure. Cap and shake 12.4.1.7) using 200 mL of hexane. the bottle so that the residue/solvent is ex- 13.7.1.4 Concentrate the cleaned up ex- posed to the acid for a total time of approxi- tract (Sections 12.6 through 12.7) to constant mately 45 seconds. weight per Section 12.7.3.1. If more than 500 13.7.3.3 Decant the hexane into a 250 mL mg of material remains, repeat the cleanup separatory funnel making sure that no acid

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is transferred. Complete the quantitative data collection after elution of these com- transfer with several hexane rinses. pounds. Return the column to the initial 13.7.3.4 Back extract the solvent/residue temperature for analysis of the next extract with 50 mL of potassium hydroxide solution or standard. per Section 12.5.2, followed by two reagent water rinses. 15.0 System and Laboratory Performance 13.7.3.5 Drain the extract through a filter 15.1 At the beginning of each 12-hour shift funnel containing approximately 10 g of during which analyses are performed, GC/MS granular anhydrous sodium sulfate in a system performance and calibration are glass-fiber filter into a macro concentration verified for all CDDs/CDFs and labeled com- device (Section 12.6). pounds. For these tests, analysis of the CS3 13.7.3.6 Concentrate the cleaned up ex- calibration verification (VER) standard (Sec- tract to a volume suitable for the additional tion 7.13 and Table 4) and the isomer speci- cleanups given in Sections 13.2 through 13.6 ficity test standards (Section 7.15 and Table and 13.8. Gel permeation chromatography 5) shall be used to verify all performance cri- (Section 13.2), alumina (Section 13.4) or teria. Adjustment and/or recalibration (Sec- Florisil (Section 13.8), and Carbopak/Celite tion 10) shall be performed until all perform- (Section 13.5) are recommended as minimum ance criteria are met. Only after all perform- additional cleanup steps. ance criteria are met may samples, blanks, 13.7.3.7 Following cleanup, concentrate IPRs, and OPRs be analyzed. the extract to 10 L as described in Section 15.2 MS Resolution—A static resolving 12.7 and proceed with analysis per Section 14. power of at least 10,000 (10% valley defini- 13.8 Florisil Cleanup (Reference 29). tion) must be demonstrated at the appro- 13.8.1 Pre-elute the activated Florisil col- priate m/z before any analysis is performed. umn (Section 7.5.3) with 10 mL of methylene Static resolving power checks must be per- chloride followed by 10 mL of formed at the beginning and at the end of hexane:methylene chloride (98:2 v/v) and dis- each 12-hour shift according to procedures in card the solvents. Section 10.1.2. Corrective actions must be 13.8.2 When the solvent is within 1 mm of implemented whenever the resolving power the packing, apply the sample extract (in does not meet the requirement. hexane) to the column. Rinse the sample 15.3 Calibration Verification. container twice with 1 mL portions of 15.3.1 Inject the VER standard using the hexane and apply to the column. procedure in Section 14. 13.8.3 Elute the interfering compounds 15.3.2 The m/z abundance ratios for all with 20 mL of hexane:methylene chloride CDDs/CDFs shall be within the limits in (98:2) and discard the eluate. Table 9; otherwise, the mass spectrometer 13.8.4 Elute the CDDs/CDFs with 35 mL of shall be adjusted until the m/z abundance ra- methylene chloride and collect the eluate. tios fall within the limits specified, and the Concentrate the eluate per Sections 12.6 verification test shall be repeated. If the ad- through 12.7 for further cleanup or for injec- justment alters the resolution of the mass tion into the HPLC or GC/MS. spectrometer, resolution shall be verified (Section 10.1.2) prior to repeat of the 14.0 HRGC/HRMS Analysis verification test. 14.1 Establish the operating conditions 15.3.3 The peaks representing each CDD/ given in Section 10.1. CDF and labeled compound in the VER 14.2 Add 10 uL of the appropriate internal standard must be present with S/N of at least standard solution (Section 7.12) to the sam- 10; otherwise, the mass spectrometer shall be ple extract immediately prior to injection to adjusted and the verification test repeated. minimize the possibility of loss by evapo- 15.3.4 Compute the concentration of each ration, adsorption, or reaction. If an extract CDD/CDF compound by isotope dilution is to be reanalyzed and evaporation has oc- (Section 10.5) for those compounds that have curred, do not add more instrument internal labeled analogs (Table 1). Compute the con- standard solution. Rather, bring the extract centration of the labeled compounds by the back to its previous volume (e.g., 19 L) with internal standard method (Section 10.6). pure nonane only (18 L if 2 L injections are These concentrations are computed based on used). the calibration data in Section 10. 14.3 Inject 1.0 μL or 2.0 μL of the con- 15.3.5 For each compound, compare the centrated extract containing the internal concentration with the calibration standard solution, using on-column or verification limit in Table 6. If only 2,3,7,8- splitless injection. The volume injected must TCDD and 2,3,7,8-TCDF are to be determined, be identical to the volume used for calibra- compare the concentration to the limit in tion (Section 10). Start the GC column ini- Table 6a. If all compounds meet the accept- tial isothermal hold upon injection. Start ance criteria, calibration has been verified MS data collection after the solvent peak and analysis of standards and sample ex- elutes. Stop data collection after the OCDD tracts may proceed. If, however, any com- and OCDF have eluted. If only 2,3,7,8-TCDD pound fails its respective limit, the measure- and 2,3,7,8-TCDF are to be determined, stop ment system is not performing properly for

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that compound. In this event, prepare a fresh ongoing data for each compound in each ma- calibration standard or correct the problem trix. Update QC charts to form a graphic rep- causing the failure and repeat the resolution resentation of continued laboratory perform- (Section 15.2) and verification (Section 15.3) ance. Develop a statement of laboratory ac- tests, or recalibrate (Section 10). curacy for each CDD/CDF in each matrix 15.4 Retention Times and GC Resolution. type by calculating the average percent re- 15.4.1 Retention times. covery (R) and the standard deviation of per- 15.4.1.1 Absolute—The absolute retention cent recovery (SR). Express the accuracy as a 13 13 times of the C12-1,2,3,4–TCDD and C12- recovery interval from R¥2SR to R = 2SR. 1,2,3,7,8,9-HxCDD GCMS internal standards For example, if R = 95% and SR = 5%, the ac- in the verification test (Section 15.3) shall be curacy is 85–105%. within ±15 seconds of the retention times ob- 15.6 Blank—Analyze the method blank ex- tained during calibration (Sections 10.2.1 and tracted with each sample batch immediately 10.2.4). following analysis of the OPR aliquot to 15.4.1.2 Relative—The relative retention demonstrate freedom from contamination times of CDDs/CDFs and labeled compounds and freedom from carryover from the OPR in the verification test (Section 15.3) shall be analysis. The results of the analysis of the within the limits given in Table 2. blank must meet the specifications in Sec- 15.4.2 GC resolution. tion 9.5.2 before sample analyses may pro- 15.4.2.1 Inject the isomer specificity ceed. standards (Section 7.15) on their respective columns. 16.0 Qualitative Determination 15.4.2.2 The valley height between 2,3,7,8- TCDD and the other tetra-dioxin isomers at A CDD, CDF, or labeled compound is iden- m/z 319.8965, and between 2,3,7,8-TCDF and tified in a standard, blank, or sample when the other tetra-furan isomers at m/z 303.9016 all of the criteria in Sections 16.1 through shall not exceed 25% on their respective col- 16.4 are met. umns (Figures 6 and 7). 16.1 The signals for the two exact m/z’s in 15.4.3 If the absolute retention time of Table 8 must be present and must maximize any compound is not within the limits speci- within the same two seconds. fied or if the 2,3,7,8-isomers are not resolved, 16.2 The signal-to-noise ratio (S/N) for the the GC is not performing properly. In this GC peak at each exact m/z must be greater event, adjust the GC and repeat the than or equal to 2.5 for each CDD or CDF de- verification test (Section 15.3) or recalibrate tected in a sample extract, and greater than (Section 10), or replace the GC column and or equal to 10 for all CDDs/CDFs in the cali- either verify calibration or recalibrate. bration standard (Sections 10.2.3 and 15.3.3). 15.5 Ongoing Precision and Recovery. 16.3 The ratio of the integrated areas of 15.5.1 Analyze the extract of the ongoing the two exact m/z’s specified in Table 8 must precision and recovery (OPR) aliquot (Sec- be within the limit in Table 9, or within ±10% tion 11.4.2.5, 11.5.4, 11.6.2, 11.7.4, or 11.8.3.2) of the ratio in the midpoint (CS3) calibration prior to analysis of samples from the same or calibration verification (VER), whichever batch. is most recent. 15.5.2 Compute the concentration of each 16.4 The relative retention time of the CDD/CDF by isotope dilution for those com- peak for a 2,3,7,8-substituted CDD or CDF pounds that have labeled analogs (Section must be within the limit in Table 2. The re- 10.5). Compute the concentration of tention time of peaks representing non- 1,2,3,7,8,9-HxCDD, OCDF, and each labeled 2,3,7,8-substituted CDDs/CDFs must be with- compound by the internal standard method in the retention time windows established in (Section 10.6). Section 10.3. 15.5.3 For each CDD/CDF and labeled com- 16.5 Confirmatory Analysis—Isomer speci- pound, compare the concentration to the ficity for 2,3,7,8-TCDF cannot be achieved on OPR limits given in Table 6. If only 2,3,7,8- the DB–5 column. Therefore, any sample in TCDD and 2,3,7,8-TCDF are to be determined, which 2,3,7,8-TCDF is identified by analysis compare the concentration to the limits in on a DB–5 column must have a confirmatory Table 6a. If all compounds meet the accept- analysis performed on a DB–225, SP–2330, or ance criteria, system performance is accept- equivalent GC column. The operating condi- able and analysis of blanks and samples may tions in Section 10.1.1 may be adjusted to op- proceed. If, however, any individual con- timize the analysis on the second GC col- centration falls outside of the range given, umn, but the GC/MS must meet the mass the extraction/concentration processes are resolution and calibration specifications in not being performed properly for that com- Section 10. pound. In this event, correct the problem, re- 16.6 If the criteria for identification in prepare, extract, and clean up the sample Sections 16.1 through 16.5 are not met, the batch and repeat the ongoing precision and CDD or CDF has not been identified and the recovery test (Section 15.5). results may not be reported for regulatory 15.5.4 Add results that pass the specifica- compliance purposes. If interferences pre- tions in Section 15.5.3 to initial and previous clude identification, a new aliquot of sample

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must be extracted, further cleaned up, and not added before extraction of the sample), it analyzed. cannot be used to quantitate the 1,2,3,7,8,9- 17.0 Quantitative Determination HxCDD by strict isotope dilution procedures. 17.1 Isotope Dilution Quantitation—By Therefore, 1,2,3,7,8,9-HxCDD is quantitated adding a known amount of a labeled com- using the averaged response of the labeled pound to every sample prior to extraction, analogs of the other two 2,3,7,8-substituted correction for recovery of the CDD/CDF can HxCDD’s: 1,2,3,4,7,8-HxCDD and 1,2,3,6,7,8- be made because the CDD/CDF and its la- HxCDD. As a result, the concentration of beled analog exhibit similar effects upon ex- 1,2,3,7,8,9-HxCDD is corrected for the average traction, concentration, and gas chroma- recovery of the other two HxCDD’s. tography. Relative response (RR) values are 17.1.3 Any peaks representing non-2,3,7,8- used in conjunction with the initial calibra- substituted CDDs/CDFs are quantitated tion data described in Section 10.5 to deter- using an average of the response factors from mine concentrations directly, so long as la- all of the labeled 2,3,7,8-isomers at the same beled compound spiking levels are constant, level of chlorination. using the following equation: 17.2 Internal Standard Quantitation and + Labeled Compound Recovery. ()AAC12nnl 17.2.1 Compute the concentrations of CngmL(/ )= 13 ex + 1,2,3,7,8,9-HxCDD, OCDF, the C-labeled ()AARR12ll analogs and the 37C-labeled cleanup standard where: in the extract using the response factors de- termined from the initial calibration data C = The concentration of the CDD/CDF in ex (Section 10.6) and the following equation: the extract, and the other terms are as defined in Section 10.5.2. ()AAC12+ 17.1.1 Because of a potential interference, CngmL(/ )= ssis the labeled analog of OCDF is not added to ex ()+ the sample. Therefore, OCDF is quantitated AARF12is is against labeled OCDD. As a result, the con- where: centration of OCDF is corrected for the re- covery of the labeled OCDD. In instances Cex = The concentration of the CDD/CDF in where OCDD and OCDF behave differently the extract, and the other terms are as during sample extraction, concentration, and defined in Section 10.6.1. cleanup procedures, this may decrease the NOTE: There is only one m/z for the 37Cl-la- accuracy of the OCDF results. However, beled standard. given the low toxicity of this compound rel- ative to the other dioxins and furans, the po- 17.2.2 Using the concentration in the ex- tential decrease in accuracy is not consid- tract determined above, compute the percent ered significant. recovery of the 13C-labeled compounds and 13 37 17.1.2 Because C12-1,2,3,7,8,9-HxCDD is the C-labeled cleanup standard using the used as an instrument internal standard (i.e., following equation:

Concentration found (μg /mL) Recovery (%) = ×100 Concentration spiked (μg /mL)

17.3 The concentration of a CDD/CDF in the extract and the weight of the solids (Sec- the solid phase of the sample is computed tion 11.5.1), as follows: using the concentration of the compound in

× ()Cex Vex Concentration in solid (ng/kg) = Ws

where: Ws = The sample weight (dry weight) in kg.

Cex = The concentration of the compound in 17.4 The concentration of a CDD/CDF in the extract. the aqueous phase of the sample is computed

Vex = The extract volume in mL. using the concentration of the compound in

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the extract and the volume of water ex- tracted (Section 11.4 or 11.5), as follows:

× ()Cex Vex Concentration inaqueous phase (pg/ L ) = Vs

where: quantitation m/z’s are within the calibration range (Section 17.5). Cex = The concentration of the compound in the extract. 17.6.3 For CDDs/CDFs having a labeled Vex = The extract volume in mL. analog, results are reported at the least di- Vs = The sample volume in liters. lute level at which the area at the quantita- tion m/z is within the calibration range (Sec- 17.5 If the SICP area at either quantita- tion m/z for any compound exceeds the cali- tion 17.5) and the labeled compound recovery bration range of the system, a smaller sam- is within the normal range for the method ple aliquot is extracted. (Section 9.3 and Tables 6, 6a, 7, and 7a). 17.5.1 For aqueous samples containing 1% 17.6.4 Additionally, if requested, the total solids or less, dilute 100 mL, 10 mL, etc., of concentration of all isomers in an individual sample to 1 L with reagent water and re-pre- level of chlorination (i.e., total TCDD, total pare, extract, clean up, and analyze per Sec- TCDF, total Paced, etc.) may be reported by tions 11 through 14. summing the concentrations of all isomers 17.5.2 For samples containing greater identified in that level of chlorination, in- than 1% solids, extract an amount of sample cluding both 2,3,7,8-substituted and non- equal to 1⁄10, 1⁄100, etc., of the amount used in 2,3,7,8-substituted isomers. Section 11.5.1. Re-prepare, extract, clean up, and analyze per Sections 11 through 14. 18.0 Analysis of Complex Samples 17.5.3 If a smaller sample size will not be 18.1 Some samples may contain high lev- representative of the entire sample, dilute els (>10 ng/L; >1000 ng/kg) of the compounds the sample extract by a factor of 10, adjust of interest, interfering compounds, and/or the concentration of the instrument internal polymeric materials. Some extracts will not μ standard to 100 pg/ L in the extract, and ana- concentrate to 10 μL (Section 12.7); others lyze an aliquot of this diluted extract by the may overload the GC column and/or mass internal standard method. spectrometer. 17.6 Results are reported to three signifi- 18.2 Analyze a smaller aliquot of the sam- cant figures for the CDDs/CDFs and labeled ple (Section 17.5) when the extract will not compounds found in all standards, blanks, concentrate to 10 μL after all cleanup proce- and samples. dures have been exhausted. 17.6.1 Reporting units and levels. 17.6.1.1 Aqueous samples—Report results 18.3 Chlorodiphenyl Ethers—If in pg/L (parts-per-quadrillion). chromatographic peaks are detected at the 17.6.1.2 Samples containing greater than retention time of any CDDs/CDFs in any of 1% solids (soils, sediments, filter cake, com- the m/z channels being monitored for the post)—Report results in ng/kg based on the chlorodiphenyl ethers (Table 8), cleanup pro- dry weight of the sample. Report the percent cedures must be employed until these inter- solids so that the result may be corrected. ferences are removed. Alumina (Section 13.4) 17.6.1.3 Tissues—Report results in ng/kg of and Florisil (Section 13.8) are recommended wet tissue, not on the basis of the lipid con- for removal of chlorodiphenyl ethers. tent of the sample. Report the percent lipid 18.4 Recovery of Labeled Compounds—In content, so that the data user can calculate most samples, recoveries of the labeled com- the concentration on a lipid basis if desired. pounds will be similar to those from reagent 17.6.1.4 Reporting level. water or from the alternate matrix (Section 17.6.1.4.1 Standards (VER, IPR, OPR) and 7.6). samples—Report results at or above the min- 18.4.1 If the recovery of any of the labeled imum level (Table 2). Report results below compounds is outside of the normal range the minimum level as not detected or as re- (Table 7), a diluted sample shall be analyzed quired by the regulatory authority. (Section 17.5). 17.6.1.4.2 Blanks—Report results above 18.4.2 If the recovery of any of the labeled one-third the ML. compounds in the diluted sample is outside 17.6.2 Results for CDDs/CDFs in samples of normal range, the calibration verification that have been diluted are reported at the standard (Section 7.13) shall be analyzed and least dilute level at which the areas at the calibration verified (Section 15.3).

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18.4.3 If the calibration cannot be verified, agement for Waste Reduction,’’ available a new calibration must be performed and the from the American Chemical Society’s De- original sample extract reanalyzed. partment of Government Relations and 18.4.4 If the calibration is verified and the Science Policy, 1155 16th Street N.W., Wash- diluted sample does not meet the limits for ington, D.C. 20036. labeled compound recovery, the method does not apply to the sample being analyzed and 21.0 Method Performance the result may not be reported for regu- Method performance was validated and latory compliance purposes. In this case, al- performance specifications were developed ternate extraction and cleanup procedures in using data from EPA’s international inter- this method must be employed to resolve the laboratory validation study (References 30– interference. If all cleanup procedures in this 31) and the EPA/paper industry Long-Term method have been employed and labeled Variability Study of discharges from the compound recovery remains outside of the pulp and paper industry (58 FR 66078). normal range, extraction and/or cleanup pro- cedures that are beyond this scope of this 22.0 References method will be required to analyze these samples. 1. Tondeur, Yves. ‘‘Method 8290: Analytical Procedures and Quality Assurance for Multi- 19.0 Pollution Prevention media Analysis of Polychlorinated Dibenzo- 19.1 The solvents used in this method pose p-dioxins and Dibenzofurans by High Resolu- little threat to the environment when man- tion Gas Chromatography/High Resolution aged properly. The solvent evaporation tech- Mass Spectrometry,’’ USEPA EMSL, Las niques used in this method are amenable to Vegas, Nevada, June 1987. solvent recovery, and it is recommended that 2. ‘‘Measurement of 2,3,7,8- the laboratory recover solvents wherever Tetrachlorinated Dibenzo-p-dioxin (TCDD) feasible. and 2,3,7,8-Tetrachlorinated Dibenzofuran 19.2 Standards should be prepared in vol- (TCDF) in Pulp, Sludges, Process Samples umes consistent with laboratory use to mini- and Wastewaters from Pulp and Paper mize disposal of standards. Mills,’’ Wright State University, Dayton, OH 45435, June 1988. 20.0 Waste Management 3. ‘‘NCASI Procedures for the Preparation 20.1 It is the laboratory’s responsibility to and Isomer Specific Analysis of Pulp and comply with all federal, state, and local reg- Paper Industry Samples for 2,3,7,8-TCDD and ulations governing waste management, par- 2,3,7,8-TCDF,’’ National Council of the Paper ticularly the hazardous waste identification Industry for Air and Stream Improvement rules and land disposal restrictions, and to Inc., 260 Madison Avenue, New York, NY protect the air, water, and land by mini- 10016, Technical Bulletin No. 551, Pre-Release mizing and controlling all releases from Copy, July 1988. fume hoods and bench operations. Compli- 4. ‘‘Analytical Procedures and Quality As- ance is also required with any sewage dis- surance Plan for the Determination of charge permits and regulations. PCDD/PCDF in Fish,’’ USEPA, Environ- 20.2 Samples containing HCl to pH <2 are mental Research Laboratory, 6201 Congdon hazardous and must be neutralized before Boulevard, Duluth, MN 55804, April 1988. being poured down a drain or must be han- 5. Tondeur, Yves. ‘‘Proposed GC/MS Meth- dled as hazardous waste. odology for the Analysis of PCDDs and 20.3 The CDDs/CDFs decompose above 800 PCDFs in Special Analytical Services Sam- °C. Low-level waste such as absorbent paper, ples,’’ Triangle Laboratories, Inc., 801–10 tissues, animal remains, and plastic gloves Capitola Dr, Research Triangle Park, NC may be burned in an appropriate incinerator. 27713, January 1988; updated by personal Gross quantities (milligrams) should be communication September 1988. packaged securely and disposed of through 6. Lamparski, L.L. and Nestrick, T.J. ‘‘De- commercial or governmental channels that termination of Tetra-, Hexa-, Hepta-, and are capable of handling extremely toxic Octachlorodibenzo-p-dioxin Isomers in Par- wastes. ticulate Samples at Parts per Trillion Lev- 20.4 Liquid or soluble waste should be dis- els,’’ Analytical Chemistry, 52: 2045–2054, solved in methanol or ethanol and irradiated 1980. with ultraviolet light with a wavelength 7. Lamparski, L.L. and Nestrick, T.J. shorter than 290 nm for several days. Use F40 ‘‘Novel Extraction Device for the Determina- BL or equivalent lamps. Analyze liquid tion of Chlorinated Dibenzo-p-dioxins wastes, and dispose of the solutions when the (PCDDs) and Dibenzofurans (PCDFs) in Mat- CDDs/CDFs can no longer be detected. rices Containing Water,’’ Chemosphere, 20.5 For further information on waste 19:27–31, 1989. management, consult ‘‘The Waste Manage- 8. Patterson, D.G., et. al. ‘‘Control of Inter- ment Manual for Laboratory Personnel’’ and ferences in the Analysis of Human Adipose ‘‘Less is Better—Laboratory Chemical Man- Tissue for 2,3,7,8-Tetrachlorodibenzo-p-

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dioxin,’’ Environmental Toxicological Chem- 22. ‘‘Analytical Procedures and Quality As- istry, 5:355–360, 1986. surance Plan for the Determination of 9. Stanley, John S. and Sack, Thomas M. PCDD/PCDF in Fish’’, U.S. Environmental ‘‘Protocol for the Analysis of 2,3,7,8- Protection Agency, Environmental Research Tetrachlorodibenzo-p-dioxin by High Resolu- Laboratory, Duluth, MN 55804, EPA/600/3–90/ tion Gas Chromatography/High Resolution 022, March 1990. Mass Spectrometry,’’ USEPA EMSL, Las 23. Afghan, B.K., Carron, J., Goulden, P.D., Vegas, Nevada 89114, EPA 600/4–86–004, Janu- Lawrence, J., Leger, D., Onuska, F., Sherry, ary 1986. J., and Wilkenson, R.J., ‘‘Recent Advances in 10. ‘‘Working with Carcinogens,’’ Depart- Ultratrace Analysis of Dioxins and Related ment of Health, Education, & Welfare, Public Halogenated Hydrocarbons’’, Can J. Chem., Health Service, Centers for Disease Control, 65: 1086–1097, 1987. NIOSH, Publication 77–206, August 1977, 24. Sherry, J.P. and Tse, H. ‘‘A Procedure NTIS PB–277256. for the Determination of Polychlorinated 11. ‘‘OSHA Safety and Health Standards, General Industry,’’ OSHA 2206, 29 CFR 1910. Dibenzo-p-dioxins in Fish’’, Chemosphere, 20: 12. ‘‘Safety in Academic Chemistry Labora- 865–872, 1990. tories,’’ ACS Committee on Chemical Safety, 25. ‘‘Preliminary Fish Tissue Study’’, Re- 1979. sults of Episode 4419, available from the EPA 13. ‘‘Standard Methods for the Examina- Sample Control Center operated by DynCorp tion of Water and Wastewater,’’ 18th edition Viar, Inc., 300 N Lee St, Alexandria, VA and later revisions, American Public Health 22314, 703–519–1140. Association, 1015 15th St, N.W., Washington, 26. Nestrick, Terry L. DOW Chemical Co., DC 20005, 1–35: Section 1090 (Safety), 1992. personal communication with D.R. 14. ‘‘Method 613—2,3,7,8- Rushneck, April 8, 1993. Details available Tetrachlorodibenzo-p-dioxin,’’ 40 CFR 136 (49 from the U.S. Environmental Protection FR 43234), October 26, 1984, Section 4.1. Agency Sample Control Center operated by 15. Provost, L.P. and Elder, R.S. ‘‘Inter- DynCorp Viar Inc, 300 N Lee St, Alexandria, pretation of Percent Recovery Data,’’ Amer- VA 22314, 703–519–1140. ican Laboratory, 15: 56–83, 1983. 27. Barnstadt, Michael. ‘‘Big Fish Col- 16. ‘‘Standard Practice for Sampling umn’’, Triangle Laboratories of RTP, Inc., Water,’’ ASTM Annual Book of Standards, SOP 129–90, 27 March 27, 1992. ASTM, 1916 Race Street, Philadelphia, PA 28. ‘‘Determination of Polychlorinated 19103–1187, 1980. Dibenzo-p-Dioxins (PCDD) and Dibenzofurans 17. ‘‘Methods 330.4 and 330.5 for Total Re- (PCDF) in Environmental Samples Using sidual Chlorine,’’ USEPA, EMSL, Cincinnati, EPA Method 1613’’, Chemical Sciences De- OH 45268, EPA 600/4–79–020, March 1979. partment, Midwest Research Institute, 425 18. ‘‘Handbook of Analytical Quality Con- trol in Water and Wastewater Laboratories,’’ Volker Boulevard, Kansas City, MO 44110– USEPA EMSL, Cincinnati, OH 45268, EPA– 2299, Standard Operating Procedure No. CS– 600/4–79–019, March 1979. 153, January 15, 1992. 19. Williams, Rick. Letter to Bill Telliard, 29. Ryan, John J. Raymonde Lizotte and June 4, 1993, available from the EPA Sample William H. Newsome, J. Chromatog. 303 Control Center operated by DynCorp Viar, (1984) 351-360. Inc., 300 N Lee St, Alexandria, VA 22314, 703– 30. Telliard, William A., McCarty, Harry 519–1140. B., and Riddick, Lynn S. ‘‘Results of the 20. Barkowski, Sarah. Fax to Sue Price, Interlaboratory Validation Study of USEPA August 6, 1992, available from the EPA Sam- Method 1613 for the Analysis of Tetra- ple Control Center operated by DynCorp through Octachlorinated Dioxins and Furans Viar, Inc., 300 N Lee St, Alexandria VA 22314, by Isotope Dilution GC/MS,’’ Chemosphere, 703–519–1140. 27, 41–46 (1993). 21. ‘‘Analysis of Multi-media, Multi-con- 31. ‘‘Results of the International Interlab- centration Samples for Dioxins and Furans, oratory Validation Study of USEPA Method PCDD/PCDF Analyses Data Package’’, Nar- 1613’’, October 1994, available from the EPA rative for Episode 4419, MRI Project No. 3091- Sample Control Center operated by DynCorp A, op.cit. February 12, 1993, Available from Viar, Inc., 300 N Lee St, Alexandria, VA the EPA Sample Control Center operated by 22314, 703–519–1140. DynCorp Viar Inc, 300 N Lee St, Alexandria, VA 22314 (703–519–1140). 23.0 Tables and Figures

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TABLE 1—CHLORINATED DIBENZO-P-DIOXINS AND FURANS DETERMINED BY ISOTOPE DILUTION AND INTERNAL STANDARD HIGH RESOLUTION GAS CHROMATOGRAPHY (HRGC)/HIGH RESOLUTION MASS SPECTROMETRY (HRMS)

CDDs/CDFs 1 CAS registry Labeled analog CAS registry

13 2,3,7,8-TCDD ...... 1746–01–6 C12-2,3,7,8-TCDD ...... 76523–40–5 37 Cl4-2,3,7,8-TCDD ...... 85508–50–5 Total TCDD ...... 41903–57–5 13 2,3,7,8-TCDF ...... 51207–31–9 C12-2,3,7,8-TCDF ...... 89059–46–1 Total-TCDF ...... 55722–27–5 13 1,2,3,7,8-PeCDD ...... 40321–76–4 C12-1,2,3,7,8-PeCDD ...... 109719–79–1 Total-PeCDD ...... 36088–22–9 13 1,2,3,7,8-PeCDF ...... 57117–41–6 C12-1,2,3,7,8-PeCDF ...... 109719–77–9 13 2,3,4,7,8-PeCDF ...... 57117–31–4 C12-2,3,4,7,8-PeCDF ...... 116843–02–8 Total-PeCDF ...... 30402–15–4 13 1,2,3,4,7,8-HxCDD ...... 39227–28–6 C12-1,2,3,4,7,8-HxCDD ...... 109719–80–4 13 1,2,3,6,7,8-HxCDD ...... 57653–85–7 C12-1,2,3,6,7,8-HxCDD ...... 109719–81–5 13 1,2,3,7,8,9-HxCDD ...... 19408–74–3 C12-1,2,3,7,8,9-HxCDD ...... 109719–82–6 Total-HxCDD ...... 34465–46–8 13 1,2,3,4,7,8-HxCDF ...... 70648–26–9 C12-1,2,3,4,7,8-HxCDF ...... 114423–98–2 13 1,2,3,6,7,8-HxCDF ...... 57117–44–9 C12-1,2,3,6,7,8-HxCDF ...... 116843–03–9 13 1,2,3,7,8,9-HxCDF ...... 72918–21–9 C12-1,2,3,7,8,9-HxCDF ...... 116843–04–0 13 2,3,4,6,7,8-HxCDF ...... 60851–34–5 C12-2,3,4,6,7,8-HxCDF ...... 116843–05–1 Total-HxCDF ...... 55684–94–1 13 1,2,3,4,6,7,8-HpCDD ...... 35822–46–9 C12-1,2,3,4,6,7,8-HpCDD ...... 109719–83–7 Total-HpCDD ...... 37871–00–4 13 1,2,3,4,6,7,8-HpCDF ...... 67562–39–4 C12-1,2,3,4,6,7,8-HpCDF ...... 109719–84–8 13 1,2,3,4,7,8,9-HpCDF ...... 55673–89–7 C12-1,2,3,4,7,8,9-HpCDF ...... 109719–94–0 Total-HpCDF ...... 38998–75–3 13 OCDD ...... 3268–87–9 C12-OCDD ...... 114423–97–1 OCDF ...... 39001–02–0 Not used. 1 Chlorinated dibenzo-p-dioxins and chlorinated dibenzofurans. TCDD = Tetrachlorodibenzo-p-dioxin. TCDF = Tetrachlorodibenzofuran. PeCDD = Pentachlorodibenzo-p-dioxin. PeCDF = Pentachlorodibenzofuran. HxCDD = Hexachlorodibenzo-p-dioxin. HxCDF = Hexachlorodibenzofuran. HpCDD = Heptachlorodibenzo-p-dioxin. HpCDF = Heptachlorodibenzofuran. OCDD = Octachlorodibenzo-p-dioxin. OCDF = Octachlorodibenzofuran.

TABLE 2—RETENTION TIME REFERENCES, QUANTITATION REFERENCES, RELATIVE RETENTION TIMES, AND MINIMUM LEVELS FOR CDDS AND DCFS

Minimum level 1 CDD/CDF Retention time and quantitation Relative reten- Water Extract reference tion time (pg/L; Solid (ng/ (pg/μL; ppq) kg; ppt) ppb)

Compounds using 13 C12–1,2,3,4-TCDD as the Injection Internal Standard

13 2,3,7,8-TCDF ...... C12-2,3,7,8-TCDF ...... 0.999–1.003 10 1 0.5 13 2,3,7,8-TCDD ...... C12-2,3,7,8-TCDD ...... 0.999–1.002 10 1 0.5 13 1,2,3,7,8-Pe ...... C12-1,2,3,7,8-PeCDF ...... 0.999–1.002 50 5 2.5 13 2,3,4,7,8-PeCDF ...... C12-2,3,4,7,8-PeCDF ...... 0.999–1.002 50 5 2.5 13 1,2,3,7,8-PeCDD ...... C12-1,2,3,7,8-PeCDD ...... 0.999–1.002 50 5 2.5 13 13 C12-2,3,7,8-TCDF ...... C12-1,2,3,4-TCDD ...... 0.923–1.103 13 13 C12-2,3,7,8-TCDD ...... C12-1,2,3,4-TCDD ...... 0.976–1.043 13 13 C12-2,3,7,8-TCDD ...... C12-1,2,3,4-TCDD ...... 0.989–1.052 13 13 C12-1,2,3,7,8-PeCDF ...... C12-1,2,3,4-TCDD ...... 1.000–1.425 13 13 C12-2,3,4,7,8-PeCDF ...... C12-1,2,3,4-TCDD ...... 1.001–1.526 13 13 C12-1,2,3,7,8-PeCDF ...... C12-1,2,3,4-TCDD ...... 1.000–1.567

Compounds using 13 C12–1,2,3,7,8,9-HxCDD as the Injection Internal Standard

13 1,2,3,4,7,8-HxCDF ...... C12-1,2,3,4,7,8-HxCDF ...... 0.999–1.001 50 5 2.5 13 1,2,3,6,7,8-HxCDF ...... C12-1,2,3,6,7,8-HxCDF ...... 0.997–1.005 50 5 2.5 13 1,2,3,7,8,9-HxCDF ...... C12-1,2,3,7,8,9-HxCDF ...... 0.999–1.001 50 5 2.5 13 2,3,4,6,7,8-HxCDF ...... C12-2,3,4,6,7,8-HxCDF ...... 0.999–1.001 50 5 2.5 13 1,2,3,4,7,8-HxCDD ...... C12-1,2,3,4,7,8-HxCDD ...... 0.999–1.001 50 5 2.5 13 1,2,3,6,7,8-HxCDD ...... C12-1,2,3,6,7,8-HxCDD ...... 0.998–1.004 50 5 2.5 1,2,3,7,8,9-HxCDD ...... (2) ...... 1.000–1.019 50 5 2.5 13 1,2,3,4,6,7,8-HpCDF ...... C12-1,2,3,4,6,7,8-HpCDF ...... 0.999–1.001 50 5 2.5

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TABLE 2—RETENTION TIME REFERENCES, QUANTITATION REFERENCES, RELATIVE RETENTION TIMES, AND MINIMUM LEVELS FOR CDDS AND DCFS—Continued

Minimum level 1 CDD/CDF Retention time and quantitation Relative reten- Water Extract reference tion time (pg/L; Solid (ng/ (pg/μL; ppq) kg; ppt) ppb)

13 1,2,3,4,7,8,9-HpCDF ...... C12-1,2,3,4,7,8,9-HpCDF ...... 0.999–1.001 50 5 2.5 13 1,2,3,4,6,7,8-HpCDD ...... C12-1,2,3,4,6,7,8-HpCDD ...... 0.999–1.001 50 5 2.5 13 OCDF ...... C12-OCDD ...... 0.999–1.001 100 10 5.0 13 OCDD ...... C12-OCDD ...... 0.999–1.001 100 10 5.0 13 1,2,3,4,6,7,8,-HxCDF ...... C12-1,2,3,7,8,9-HpCDD ...... 0.949–0.975 13 13 C121,2,3,7,8,9-HxCDF ...... C12-1,2,3,7,8,9-HpCDD ...... 0.977–1.047 13 13 C122,3,4,6,7,8,-HxCDF ...... C12-1,2,3,7,8,9-HpCDD ...... 0.959–1.021 13 13 C121,2,3,4,7,8,-HxCDF ...... C12-1,2,3,7,8,9-HpCDD ...... 0.977–1.000 13 13 C121,2,3,6,7,8,-HxCDF ...... C12-1,2,3,7,8,9-HpCDD ...... 0.981–1.003 13 13 C121,2,3,4,6,7,8-HxCDF ...... C12-1,2,3,7,8,9-HpCDD ...... 1.043–1.085 13 13 C121,2,3,4,7,8,9-HxCDF ...... C12-1,2,3,7,8,9-HpCDD ...... 1.057–1.151 13 13 C121,2,3,4,6,7,8-HxCDF ...... C12-1,2,3,7,8,9-HpCDD ...... 1.086–1.110 13 13 C12OCDD ...... C12-1,2,3,7,8,9-HpCDD ...... 1.032–1.311 1 The Minimum Level (ML) for each analyte is defined as the level at which the entire analytical system must give a recogniz- able signal and acceptable calibration point. It is equivalent to the concentration of the lowest calibration standard, assuming that all method-specified sample weights, volumes, and cleanup procedures have been employed. 2 13 The retention time reference for 1,2,3,7,8,9-HxCDD is C12-1,2,3,6,7,8-HxCDD, and 1,2,3,7,8,9-HxCDD is quantified using 13 13 the averaged responses for C12-1,2,3,4,7,8-HxCDD and C12-1,2,3,6,7,8-HxCDD.

TABLE 3—CONCENTRATION OF STOCK AND SPIKING SOLUTIONS CONTAINING CDDS/CDFS AND LABELED COMPOUNDS

Labeled com- Labeled pound stock compound PAR stock PAR spiking CDD/CDF spiking solu- solution 3 solution 4 solution 1 tion 2 (ng/mL) (ng/mL) (ng/mL) (ng/mL)

2,3,7,8-TCDD ...... 40 0.8 2,3,7,8-TCDF ...... 40 0 .8 1,2,3,7,8-PeCDD ...... 200 4 1,2,3,7,8-PeCDF ...... 200 4 2,3,4,7,8-PeCDF ...... 200 4 1,2,3,4,7,8-HxCDD ...... 200 4 1,2,3,6,7,8-HxCDD ...... 200 4 1,2,3,7,8,9-HxCDD ...... 200 4 1,2,3,4,7,8-HxCDF ...... 200 4 1,2,3,6,7,8-HxCDF ...... 200 4 1,2,3,7,8,9-HxCDF ...... 200 4 2,3,4,6,7,8-HxCDF ...... 200 4 1,2,3,4,6,7,8-HpCDD ...... 200 4 1,2,3,4,6,7,8-HpCDF ...... 200 4 1,2,3,4,7,8,9-HpCDF ...... 200 4 OCDD ...... 400 8 OCDF ...... 400 8 13 C12-2,3,7,8-TCDD ...... 100 2 13 C12-2,3,7,8-TCDF ...... 100 2 13 C12-1,2,3,7,8-PeCDD ...... 100 2 13 C12-1,2,3,7,8-PeCDF ...... 100 2 13 C12-2,3,4,7,8-PeCDF ...... 100 2 13 C12-1,2,3,4,7,8-HxCDD ...... 100 2 13 C12-1,2,3,6,7,8-HxCDD ...... 100 2 13 C12-1,2,3,4,7,8-HxCDF ...... 100 2 13 C12-1,2,3,6,7,8-HxCDF ...... 100 2 13 C12-1,2,3,7,8,9-HxCDF ...... 100 2 13 C12-2,3,4,6,7,8-HxCDF ...... 100 2 13 C12-1,2,3,4,6,7,8-HpCDD ...... 100 2 13 C12-1,2,3,4,6,7,8-HpCDF ...... 100 2 13 C12-1,2,3,4,7,8,9-HpCDF ...... 100 2 13 C12-OCDD ...... 200 4 Cleanup Standard 5 37 Cl4-2,3,7,8-TCDD ...... 0 .8 Internal Standards 6 13 C12-1,2,3,4-TCDD ...... 200 13 C12-1,2,3,7,8,9-HxCDD ...... 200 1 Section 7.10—prepared in nonane and diluted to prepare spiking solution. 2 Section 7.10.3—prepared in acetone from stock solution daily.

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3 Section 7.9—prepared in nonane and diluted to prepare spiking solution. 4 Section 7.14—prepared in acetone from stock solution daily. 5 Section 7.11—prepared in nonane and added to extract prior to cleanup. 6 Section 7.12—prepared in nonane and added to the concentrated extract immediately prior to injection into the GC (Section 14.2).

TABLE 4—CONCENTRATION OF CDDS/CDFS IN CALIBRATION AND CALIBRATION VERIFICATION SOLUTIONS 1 (SECTION 15.3)

CS2 CS3 CS4 CS5 CDD/CDF (ng/mL) (ng/mL) (ng/mL) (ng/mL)

2,3,7,8-TCDD ...... 0 .5 2 10 40 200 2,3,7,8-TCDF ...... 0.5 2 10 40 200 1,2,3,7,8-PeCDD ...... 2 .5 10 50 200 1000 1,2,3,7,8-PeCDF ...... 2 .5 10 50 200 1000 2,3,4,7,8-PeCDF ...... 2 .5 10 50 200 1000 1,2,3,4,7,8-HxCDD ...... 2.5 10 50 200 1000 1,2,3,6,7,8-HxCDD ...... 2.5 10 50 200 1000 1,2,3,7,8,9-HxCDD ...... 2.5 10 50 200 1000 1,2,3,4,7,8-HxCDF ...... 2 .5 10 50 200 1000 1,2,3,6,7,8-HxCDF ...... 2 .5 10 50 200 1000 1,2,3,7,8,9-HxCDF ...... 2 .5 10 50 200 1000 2,3,4,6,7,8-HxCDF ...... 2 .5 10 50 200 1000 1,2,3,4,6,7,8-HpCDD ...... 2.5 10 50 200 1000 1,2,3,4,6,7,8-HpCDF ...... 2 .5 10 50 200 1000 1,2,3,4,7,8,9-HpCDF ...... 2 .5 10 50 200 1000 OCDD ...... 5 .0 20 100 400 2000 OCDF ...... 5 .0 20 100 400 2000 13 C12-2,3,7,8-TCDD ...... 100 100 100 100 100 13 C12-2,3,7,8-TCDF ...... 100 100 100 100 100 13 C12-1,2,3,7,8-PeCDD ...... 100 100 100 100 100 13 C12-PeCDF ...... 100 100 100 100 100 13 C12-2,3,4,7,8-PeCDF ...... 100 100 100 100 100 13 C12-1,2,3,4,7,8-HxCDD ...... 100 100 100 100 100 13 C12-1,2,3,6,7,8-HxCDD ...... 100 100 100 100 100 13 C12-1,2,3,4,7,8-HxCDF ...... 100 100 100 100 100 13 C12-1,2,3,6,7,8-HxCDF ...... 100 100 100 100 100 13 C12-1,2,3,7,8,9-HxCDF ...... 100 100 100 100 100 13 C12-1,2,3,4,6,7,8-HpCDD ...... 100 100 100 100 100 13 C12-1,2,3,4,6,7,8-HpCDF ...... 100 100 100 100 100 13 C12-1,2,3,4,7,8,9-Hp CDF ...... 100 100 100 100 100 13 C12-OCDD ...... 200 200 200 200 200 Cleanup Standard: 37 C14-2,3,7,8-TCDD ...... 0.5 2 10 40 200 Internal Standards: 13 C12-1,2,3,4-TCDD ...... 100 100 100 100 100 13 C12-1,2,3,7,8,9-HxCDD ...... 100 100 100 100 100

TABLE 5—GC RETENTION TIME WINDOW DEFINING SOLUTION AND ISOMER SPECIFICITY TEST STANDARD (SECTION 7.15)

DB–5 column GC retention-time window defining solution CDD/CDF First eluted Last eluted

TCDF ...... 1,3,6,8- ...... 1,2,8,9- TCDD ...... 1,3,6,8- ...... 1,2,8,9- PeCDF ...... 1,3,4,6,8- ...... 1,2,3,8,9- PeCDD ...... 1,2,4,7,9- ...... 1,2,3,8,9- HxCDF ...... 1,2,3,4,6,8- ...... 1,2,3,4,8,9- HxCDD ...... 1,2,4,6,7,9- ...... 1,2,3,4,6,7- HpCDF ...... 1,2,3,4,6,7,8- ...... 1,2,3,4,7,8,9- HpCDD ...... 1,2,3,4,6,7,9- ...... 1,2,3,4,6,7,8-

DB–5 Column TCDD Specificity Test Standard 1,2,3,7 = 1,2,3,8-TCDD 2,3,7,8-TCDD 1,2,3,9-TCDD DB–225 Column TCDF Isomer Specificity Test Standard 2,3,4,7-TCDF 2,3,7,8-TCDF 1,2,3,9-TCDF

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TABLE 6—ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS WHEN ALL CDDS/CDFS ARE TESTED 1

IPR 23 Test conc. OPR VER CDD/CDF (ng/mL) s X (ng/mL) (ng/mL) (ng/mL) (ng/mL)

2,3,7,8-TCDD ...... 10 2.8 8.3–12.9 6.7 –15.8 7.8 –12.9 2,3,7,8-TCDF ...... 10 2 .0 8.7–13.7 7.5 –15.8 8.4 –12.0 1,2,3,7,8-PeCDD ...... 50 7 .5 38 –66 35 –71 39 –65 1,2,3,7,8-PeCDF ...... 50 7 .5 43 –62 40 –67 41 –60 2,3,4,7,8-PeCDF ...... 50 8 .6 36 –75 34 –80 41 –61 1,2,3,4,7,8-HxCDD ...... 50 9 .4 39 –76 35 –82 39 –64 1,2,3,6,7,8-HxCDD ...... 50 7 .7 42 –62 38 –67 39 –64 1,2,3,7,8,9-HxCDD ...... 50 11 .1 37 –71 32 –81 41 –61 1,2,3,4,7,8-HxCDF ...... 50 8 .7 41 –59 36 –67 45 –56 1,2,3,6,7,8-HxCDF ...... 50 6 .7 46 –60 42 –65 44 –57 1,2,3,7,8,9-HxCDF ...... 50 6 .4 42 –61 39 –65 45 –56 2,3,4,6,7,8-HxCDF ...... 50 7 .4 37 –74 35 –78 44 –57 1,2,3,4,6,7,8-HpCDD ...... 50 7 .7 38 –65 35 –70 43 –58 1,2,3,4,6,7,8-HpCDF ...... 50 6 .3 45 –56 41 –61 45 –55 1,2,3,4,7,8,9-HpCDF ...... 50 8 .1 43 –63 39 –69 43 –58 OCDD ...... 100 19 89–127 78–144 79 –126 OCDF ...... 100 27 74 –146 63 –170 63 –159 13 C12-2,3,7,8-TCDD ...... 100 37 28 –134 20–175 82–121 13 C12-2,3,7,8-TCDF ...... 100 35 31–113 22–152 71 –140 13 C12-1,2,3,7,8-PeCDD ...... 100 39 27–184 21–227 62 –160 13 C12-1,2,3,7,8-PeCDF ...... 100 34 27–156 21 –192 76 –130 13 C12-2,3,4,7,8-PeCDF ...... 100 38 16–279 13 –328 77 –130 13 C12-1,2,3,4,7,8-HxCDD ...... 100 41 29–147 21–193 85 –117 13 C12-1,2,3,6,7,8-HxCDD ...... 100 38 34–122 25–163 85 –118 13 C12-1,2,3,4,7,8-HxCDF ...... 100 43 27 –152 19 –202 76 –131 13 C12-1,2,3,6,7,8-HxCDF ...... 100 35 30 –122 21 –159 70 –143 13 C12-1,2,3,7,8,9-HxCDF ...... 100 40 24 –157 17 –205 74 –135 13 C12-2,3,4,6,7,8,-HxCDF ...... 100 37 29–136 22–176 73 –137 13 C12-1,2,3,4,6,7,8-HpCDD ...... 100 35 34–129 26–166 72 –138 13 C12-1,2,3,4,6,7,8-HpCDF ...... 100 41 32 –110 21 –158 78 –129 13 C12-1,2,3,4,7,8,9-HpCDF ...... 100 40 28 –141 20 –186 77 –129 13 C12-OCDD ...... 200 95 41–276 26–397 96 –415 37 Cl4-2,3,7,8-TCDD ...... 10 3.6 3.9–15.4 3.1 –19.1 7.9 –12.7 1 All specifications are given as concentration in the final extract, assuming a 20 μL volume. 2 s = standard deviation of the concentration. 3 X = average concentration.

TABLE 6A—ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS WHEN ONLY TETRA COMPOUNDS ARE TESTED 1

IPR 23 CDD/CDF Test Conc. OPR VER (ng/mL) s (ng/mL) X (ng/mL) (ng/mL) (ng/mL)

2,3,7,8-TCDD ...... 10 2.7 8.7–12.4 7.314.6 8.2–12.3 2,3,7,8-TCDF ...... 10 2.0 9.1–13.1 8.0 –14.7 8.6 –11.6 13 C12-2,3,7,8-TCDD ...... 100 35 32 –115 25–141 85–117 13 C12-2,3,7,8-TCDF ...... 100 34 35–99 26–126 76–131 37 Cl4-2,3,7,8-TCDD ...... 10 3.4 4.5–13.4 3.7 –15.8 8.3 –12.1 1 All specifications are given as concentration in the final extract, assuming a 20 μL volume. 2 s = standard deviation of the concentration. 3 X = average concentration.

TABLE 7—LABELED COMPOUNDS RECOVERY IN SAMPLES WHEN ALL CDDS/CDFS ARE TESTED

Labeled compound Test conc. recovery Compound (ng/mL) (ng/mL) 1 (%)

13 C12-2,3,7,8-TCDD ...... 100 25 –164 25–164 13 C12-2,3,7,8-TCDF ...... 100 24 –169 24–169 13 C12-1,2,3,7,8-PeCDD ...... 100 25 –181 25–181 13 C12-1,2,3,7,8-PeCDF ...... 100 24 –185 24–185 13 C12-2,3,4,7,8-PeCDF ...... 100 21 –178 21–178 13 C12-1,2,3,4,7,8-HxCDD ...... 100 32 –141 32–141 13 C12-1,2,3,6,7,8-HxCDD ...... 100 28 –130 28–130 13 C12-1,2,3,4,7,8-HxCDF ...... 100 26–152 26–152 13 C12-1,2,3,6,7,8-HxCDF ...... 100 26–123 26–123

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TABLE 7—LABELED COMPOUNDS RECOVERY IN SAMPLES WHEN ALL CDDS/CDFS ARE TESTED— Continued

Labeled compound Test conc. recovery Compound (ng/mL) (ng/mL) 1 (%)

13 C12-1,2,3,7,8,9-HxCDF ...... 100 29–147 29–147 13 C12-2,3,4,6,7,8-HxCDF ...... 100 28–136 28–136 13 C12-1,2,3,4,6,7,8-HpCDD ...... 100 23 –140 23–140 13 C12-1,2,3,4,6,7,8-HpCDF ...... 100 28–143 28–143 13 C12-1,2,3,4,7,8,9-HpCDF ...... 100 26–138 26–138 13 C12-OCDD ...... 200 34-313 17–157 37 Cl4-2,3,7,8-TCDD ...... 10 3.5–19.7 35–197 1 Specification given as concentration in the final extract, assuming a 20-μL volume.

TABLE 7A—LABELED COMPOUND RECOVERY IN SAMPLES WHEN ONLY TETRA COMPOUNDS ARE TESTED

Labeled compound Test conc. recovery Compound (ng/mL) (ng/mL) 1 (%)

13 C12-2,3,7,8-TCDD ...... 100 31 –137 31–137 13 C12-2,3,7,8-TCDF ...... 100 29 –140 29–140 37 Cl4-2,3,7,8-TCDD ...... 10 4.2–16.4 42–164 1 Specification given as concentration in the final extract, assuming a 20 μL volume.

TABLE 8—DESCRIPTORS, EXACT M/Z’S, M/Z TYPES, AND ELEMENTAL COMPOSITIONS OF THE CDDS AND CDFS

Descriptor Exact M/Z 1 M/Z type Elemental composition Substance 2

1 ...... 292.9825 Lock C7F11 ...... PFK 35 303.9016 M C12H4 Cl4O ...... TCDF 35 37 305.8987 M = 2 C12H4 Cl3 ClO ...... TCDF 13 35 3 315.9419 M C12H4 Cl4O ...... TCDF 13 35 37 3 317.9389 M = 2 C12H4 Cl3 ClO ...... TCDF 35 319.8965 M C12H4 Cl4O2 ...... TCDD 35 37 321.8936 M = 2 C12H4 Cl3 ClO2 ...... TCDD 37 4 327.8847 M C12H4 Cl4O2 ...... TCDD 330.9792 QC C7F13 ...... PFK 13 35 3 331.9368 M C12H4 Cl4O2 ...... TCDD 13 35 37 3 333.9339 M = 2 C12H4 Cl3 ClO2 ...... TCDD 35 37 375.8364 M = 2 C12H4 Cl5 ClO ...... HxCDPE 35 37 2 ...... 339.8597 M = 2 C12H3 Cl4 ClO ...... PeCDF 35 37 341.8567 M = 4 C12H3 Cl3 Cl2O ...... PeCDF 13 35 37 351.9000 M = 2 C12H3 Cl4 ClO ...... PeCDF 13 35 37 3 353.8970 M = 4 C12H3 Cl3 Cl2O ...... PeCDF 354.9792 Lock C9F13 ...... PFK 35 37 355.8546 M = 2 C12H3 Cl4 ClO2 ...... PeCDD 35 37 357.8516 M = 4 C12H3 Cl3 Cl2O2 ...... PeCDD 13 35 37 3 367.8949 M = 2 C12H3 Cl4 ClO2 ...... PeCDD 13 35 37 3 369.8919 M = 4 C12H3 Cl3 Cl2O2 ...... PeCDD 35 37 409.7974 M = 2 C12H3 Cl6 ClO ...... HpCDPE 35 37 3 ...... 373.8208 M = 2 C12H2 Cl5 ClO ...... HxCDF 35 37 375.8178 M = 4 C12H2 Cl4 Cl2O ...... HxCDF 13 35 3 383.8639 M C12H2 Cl6O ...... HxCDF 13 35 37 3 385.8610 M = 2 C12H2 Cl5 ClO ...... HxCDF 35 37 389.8157 M = 2 C12H2 Cl5 ClO2 ...... HxCDD 35 37 391.8127 M = 4 C12H2 Cl4 Cl2O2 ...... HxCDD 392.9760 Lock C9F15 ...... PFK 13 35 37 3 401.8559 M = 2 C12H2 Cl5 ClO2 ...... HxCDD 13 35 37 3 403.8529 M = 4 C12H2 Cl4 Cl2O2 ...... HxCDD 430.9729 QC C9F17 ...... PFK 35 37 445.7555 M = 4 C12H2 Cl6 Cl2O ...... OCDPE 35 37 4 ...... 407.7818 M = 2 C12H Cl6 ClO ...... HpCDF 35 37 409.7789 M = 4 C12H Cl5 Cl2O ...... HpCDF 13 35 3 417.8253 M C12H Cl7O ...... HpCDF 13 35 37 3 419.8220 M = 2 C12H Cl6 ClO ...... HpCDF 35 37 423.7766 M = 2 C12H Cl6 ClO2 ...... HpCDD 35 37 425.7737 M = 4 C12H Cl5 Cl2O2 ...... HpCDD

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TABLE 8—DESCRIPTORS, EXACT M/Z’S, M/Z TYPES, AND ELEMENTAL COMPOSITIONS OF THE CDDS AND CDFS—Continued

Descriptor Exact M/Z 1 M/Z type Elemental composition Substance 2

430.9729 Lock C9F17 ...... PFK 13 35 37 3 435.8169 M = 2 C12H Cl6 ClO2 ...... HpCDD 13 35 37 3 437.8140 M = 4 C12H Cl5 Cl2O2 ...... HpCDD 35 37 479.7165 M = 4 C12H Cl7 Cl2O ...... NCDPE 35 37 5 ...... 441.7428 M = 2 C12 Cl7 ClO ...... OCDF 442.9728 Lock C10F17 ...... PFK 35 37 443.7399 M = 4 C12 Cl6 Cl2O ...... OCDF 35 37 457.7377 M = 2 C12 Cl7 ClO2 ...... OCDD 35 37 459.7348 M = 4 C12 Cl6 Cl2O2 ...... OCDD 13 35 37 3 469.7779 M = 2 C12 Cl7 ClO2 ...... OCDD 13 35 37 3 471.7750 M = 4 C12 Cl6 Cl2O2 ...... OCDD 35 37 513.6775 M = 4 C12 Cl8 Cl2O ...... DCDPE 1 Nuclidic masses used: H = 1.007825. O = 15.994915. C = 12.00000. 35Cl = 34.968853. 13C = 13.003355. 37Cl = 36.965903. F = 18.9984. 2 TCDD = Tetrachlorodibenzo-p-dioxin. PeCDD = Pentachlorodibenzo-p-dioxin. HxCDD = Hexachlorodibenzo-p-dioxin. HpCDD = Heptachlorodibenzo-p-dioxin. OCDD = Octachlorodibenzo-p-dioxin. HxCDPE = Hexachlorodiphenyl ether. OCDPE = Octachlorodiphenyl ether. DCDPE = Decachlorodiphenyl ether. TCDF = Tetrachlorodibenzofuran. PeCDF = Pentachlorodibenzofuran. HxCDF = Hexachlorodibenzofuran. HpCDF = Heptachlorodibenzofuran. OCDF = Octachlorodibenzofuran. HpCDPE = Heptachlorodiphenyl ether. NCDPE = Nonachlorodiphenyl ether. PFK = Perfluorokerosene. 3 Labeled compound. 4 37 There is only one m/z for Cl4-2,3,7,8,-TCDD (cleanup standard).

TABLE 9—THEORETICAL ION ABUNDANCE RATIOS AND QC LIMITS

QC limit 1 Number of chlorine atoms M/Z’s forming ratio Theoretical ratio Lower Upper

4 2 ...... M/(M = 2) ...... 0.77 0.65 0.89 5 ...... (M = 2)/(M = 4) ...... 1.55 1.32 1.78 6 ...... (M = 2)/(M = 4) ...... 1.24 1.05 1.43 6 3 ...... M/(M = 2) ...... 0.51 0.43 0.59 7 ...... (M = 2)/(M = 4) ...... 1.05 0.88 1.20 7 4 ...... M/(M = 2) ...... 0.44 0.37 0.51 8 ...... (M = 2)/(M = 4) ...... 0.89 0.76 1.02 1 QC limits represent ±15% windows around the theoretical ion abundance ratios. 2 37 Does not apply to Cl4-2,3,7,8-TCDD (cleanup standard). 3 13 Used for C12-HxCDF only. 4 13 Used for C12-HpCDF only.

TABLE 10—SUGGESTED SAMPLE QUANTITIES TO BE EXTRACTED FOR VARIOUS MATRICES 1

2 Quantity ex- Sample Matrix Example Percent solids Phase tracted

Single-phase: Aqueous ...... Drinking water ...... <1 (3) ...... 1000 mL. Groundwater Treated wastewater Solid ...... Dry soil ...... >20 Solid ...... 10 g. Compost Ash Organic ...... Waste solvent ...... <1 Organic ...... 10 g. Waste oil Organic polymer Tissue ...... Fish ...... Organic ...... 10 g. Human adipose

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TABLE 10—SUGGESTED SAMPLE QUANTITIES TO BE EXTRACTED FOR VARIOUS MATRICES 1— Continued

2 Quantity ex- Sample Matrix Example Percent solids Phase tracted

Multi-phase: Liquid/Solid: Aqueous/Solid ...... Wet soil ...... 1–30 Solid ...... 10 g. Untreated effluent. Digested municipal sludge. Filter cake. Paper pulp. Organic/solid ...... Industrial sludge ...... 1–100 Both ...... 10 g. Oily waste Liquid/Liquid: Aqueous/organic ...... In-process effluent ...... <1 Organic ...... 10 g. Untreated effluent Drum waste Aqueous/organic/ Untreated effluent ...... >1 Organic and solid ...... 10 g. solid. Drum waste

1 The quantity of sample to be extracted is adjusted to provide 10 g of solids (dry weight). One liter of aqueous samples con- taining 1% solids will contain 10 g of solids. For aqueous samples containing greater than 1% solids, a lesser volume is used so that 10 g of solids (dry weight) will be extracted. 2 The sample matrix may be amorphous for some samples. In general, when the CDDs/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. 3 Aqueous samples are filtered after spiking with the labeled compounds. The filtrate and the materials trapped on the filter are extracted separately, and the extracts are combined for cleanup and analysis.

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24.0 Glossary of Definitions and Purposes amp—ampere cm—centimeter These definitions and purposes are specific to this method but have been conformed to g—gram common usage as much as possible. h—hour 24.1 Units of weight and Measure and D—inside diameter Their Abbreviations. in.—inch 24.1.1 Symbols: L—liter °C—degrees Celsius M—Molecular ion μL—microliter m—meter μm—micrometer mg—milligram <—less than min—minute >—greater than mL—milliliter %—percent mm—millimeter 24.1.2 Alphabetical abbreviations: m/z—mass-to-charge ratio

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N—normal; gram molecular weight of solute May—This action, activity, or procedural divided by hydrogen equivalent of solute, step is neither required nor prohibited. per liter of solution May Not—This action, activity, or proce- OD—outside diameter dural step is prohibited. pg—picogram Method Blank—An aliquot of reagent ppb—part-per-billion water that is treated exactly as a sample in- ppm—part-per-million cluding exposure to all glassware, equip- ppq—part-per-quadrillion ment, solvents, reagents, internal standards, ppt—part-per-trillion and surrogates that are used with samples. psig—pounds-per-square inch gauge The method blank is used to determine if v/v—volume per unit volume analytes or interferences are present in the w/v—weight per unit volume laboratory environment, the reagents, or the 24.2 Definitions and Acronyms (in Alpha- apparatus. betical Order). Minimum Level (ML)—The level at which Analyte—A CDD or CDF tested for by this the entire analytical system must give a rec- method. The analytes are listed in Table 1. ognizable signal and acceptable calibration Calibration Standard (CAL)—A solution point for the analyte. It is equivalent to the prepared from a secondary standard and/or concentration of the lowest calibration stock solutions and used to calibrate the re- standard, assuming that all method-specified sponse of the instrument with respect to sample weights, volumes, and cleanup proce- analyte concentration. dures have been employed. Calibration Verification Standard (VER)— MS—Mass spectrometer or mass spectrom- The mid-point calibration standard (CS3) etry. that is used in to verify calibration. See Must—This action, activity, or procedural Table 4. step is required. CDD—Chlorinated Dibenzo-p-ioxin—The OPR—Ongoing precision and recovery isomers and congeners of tetra-through octa- standard (OPR); a laboratory blank spiked chlorodibenzo-p-dioxin. with known quantities of analytes. The OPR CDF—Chlorinated Dibenzofuran—The iso- is analyzed exactly like a sample. Its purpose mers and congeners of tetra-through octa- is to assure that the results produced by the chlorodibenzofuran. laboratory remain within the limits speci- CS1, CS2, CS3, CS4, CS5—See Calibration fied in this method for precision and recov- standards and Table 4. ery. Field Blank—An aliquot of reagent water PAR—Precision and recovery standard; or other reference matrix that is placed in a secondary standard that is diluted and sample container in the laboratory or the spiked to form the IPR and OPR. field, and treated as a sample in all respects, including exposure to sampling site condi- PFK—Perfluorokerosene; the mixture of tions, storage, preservation, and all analyt- compounds used to calibrate the exact m/z ical procedures. The purpose of the field scale in the HRMS. blank is to determine if the field or sample Preparation Blank—See method blank. transporting procedures and environments Primary Dilution Standard—A solution have contaminated the sample. containing the specified analytes that is pur- GC—Gas chromatograph or gas chroma- chased or prepared from stock solutions and tography. diluted as needed to prepare calibration solu- GPC—Gel permeation chromatograph or tions and other solutions. gel permeation chromatography. Quality Control Check Sample (QCS)—A HPLC—High performance liquid chro- sample containing all or a subset of the matograph or high performance liquid chro- analytes at known concentrations. The QCS matography. is obtained from a source external to the lab- HRGC—High resolution GC. oratory or is prepared from a source of HRMS—High resolution MS. standards different from the source of cali- IPR—Initial precision and recovery; four bration standards. It is used to check labora- aliquots of the diluted PAR standard ana- tory performance with test materials pre- lyzed to establish the ability to generate ac- pared external to the normal preparation ceptable precision and accuracy. An IPR is process. performed prior to the first time this method Reagent Water—Water demonstrated to be is used and any time the method or instru- free from the analytes of interest and poten- mentation is modified. tially interfering substances at the method K-D—Kuderna-Danish concentrator; a de- detection limit for the analyte. vice used to concentrate the analytes in a Relative Standard Deviation (RSD)—The solvent. standard deviation times 100 divided by the Laboratory Blank—See method blank. mean. Also termed ‘‘coefficient of vari- Laboratory Control sample (LCS)—See on- ation.’’ going precision and recovery standard (OPR). RF—Response factor. See Section 10.6.1. Laboratory Reagent Blank—See method RR—Relative response. See Section 10.5.2. blank. RSD—See relative standard deviation.

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SDS—Soxhlet/Dean-Stark extractor; an ex- pounds are transferred from the aqueous traction device applied to the extraction of phase into the gaseous phase where they are solid and semi-solid materials (Reference 7). passed into a sorbent column and trapped. Should—This action, activity, or proce- After purging is completed, the trap is dural step is suggested but not required. backflushed and heated rapidly to desorb the SICP—Selected ion current profile; the compounds into a gas chromatograph (GC). line described by the signal at an exact m/z. The compounds are separated by the GC and SPE—Solid-phase extraction; an extrac- detected by a mass spectrometer (MS) (ref- tion technique in which an analyte is ex- erences 2 and 3). The labeled compounds tracted from an aqueous sample by passage serve to correct the variability of the analyt- over or through a material capable of revers- ical technique. ibly adsorbing the analyte. Also termed liq- 2.2 Identification of a compound (quali- uid-solid extraction. tative analysis) is performed by comparing Stock Solution—A solution containing an the GC retention time and the background analyte that is prepared using a reference corrected characteristic spectral masses material traceable to EPA, the National In- with those of authentic standards. stitute of Science and Technology (NIST), or 2.3 Quantitative analysis is performed by a source that will attest to the purity and GC/MS using extracted ion current profile authenticity of the reference material. (EICP) areas. Isotope dilution is used when TCDD—Tetrachlorodibenzo-p-dioxin. labeled compounds are available; otherwise, TCDF—Tetrachlorodibenzofuran. an internal standard method is used. VER—See calibration verification stand- 2.4 Quality is assured through reproduc- ard. ible calibration and testing of the purge and METHOD 1624 REVISION B—VOLATILE ORGANIC trap and GC/MS systems. COMPOUNDS BY ISOTOPE DILUTION GC/MS 3. Contamination and Interferences 1. Scope and Application 3.1 Impurities in the purge gas, organic 1.1 This method is designed to determine compounds out-gassing from the plumbing the volatile toxic organic pollutants associ- upstream of the trap, and solvent vapors in ated with the 1976 Consent Decree and addi- the laboratory account for the majority of tional compounds amenable to purge and contamination problems. The analytical sys- trap gas chromatography-mass spectrometry tem is demonstrated to be free from inter- (GC/MS). ferences under conditions of the analysis by 1.2 The chemical compounds listed in analyzing blanks initially and with each table 1 may be determined in municipal and sample lot (samples analyzed on the same 8 industrial discharges by this method. The hr shift), as described in Section 8.5. methmd is designed to meet the survey re- 3.2 Samples can be contaminated by diffu- quirements of Effluent Guidelines Division sion of volatile organic compounds (particu- (EGD) and the National Pollutants Dis- larly methylene chloride) through the bottle charge Elimination System (NPDES) under seal during shipment and storage. A field 40 CFR 136.1 and 136.5. Any modifications of blank prepared from reagent water and car- this method, beyond those expressly per- ried through the sampling and handling pro- mitted, shall be considered as major modi- tocol serves as a check on such contamina- fications subject to application and approval tion. of alternate test procedures under 40 CFR 3.3 Contamination by carry-over can 136.4 and 136.5. occur when high level and low level samples 1.3 The detection limit of this method is are analyzed sequentially. To reduce carry- usually dependent on the level of inter- over, the purging device and sample syringe ferences rather than instrumental limita- are rinsed between samples with reagent tions. The limits in table 2 represent the water. When an unusually concentrated sam- minimum quantity that can be detected with ple is encountered, it is followed by analysis no interferences present. of a reagent water blank to check for carry- 1.4 The GC/MS portions of this method over. For samples containing large amounts are for use only by analysts experienced with of water soluble materials, suspended solids, GC/MS or under the close supervision of such high boiling compounds, or high levels or qualified persons. Laboratories unfamiliar purgeable compounds, the purge device is with the analyses of environmental samples washed with soap solution, rinsed with tap by GC/MS should run the performance tests and distilled water, and dried in an oven at in reference 1 before beginning. 100–125 °C. The trap and other parts of the system are also subject to contamination; 2. Summary of Method therefore, frequent bakeout and purging of 2.1 Stable isotopically labeled analogs of the entire system may be required. the compounds of interest are added to a 5 3.4 Interferences resulting from samples mL water sample. The sample is purged at will vary considerably from source to source, 20–25 °C with an inert gas in a specially de- depending on the diversity of the industrial signed chamber. The volatile organic com- complex or municipality being sampled.

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4. Safety °C. The desorber shown in Figure 2 meets these specifications. 4.1 The toxicity or carcinogenicity of 5.2.4 The purge and trap device may be a each compound or reagent used in this meth- separate unit or coupled to a GC as shown in od has not been precisely determined; how- Figures 3 and 4. ever, each chemical compound should be 5.3 Gas chromatograph—shall be linearly treated as a potential health hazard. Expo- temperature programmable with initial and sure to these compounds should be reduced final holds, shall contain a glass jet sepa- to the lowest possible level. The laboratory rator as the MS interface, and shall produce is responsible for maintaining a current results which meet the calibration (Section awareness file of OSHA regulations regard- 7), quality assurance (Section 8), and per- ing the safe handling of the chemicals speci- formance tests (Section 11) of this method. fied in this method. A reference file of data ± × ± handling sheets should also be made avail- 5.3.1 Column—2.8 0.4 m 2 0.5 mm i. d. able to all personnel involved in these anal- glass, packekd with one percent SP–1000 on yses. Additional information on laboratory Carbopak B, 60/80 mesh, or equivalent. safety can be found in references 4–6. 5.4 Mass spectrometer—70 eV electron im- pact ionization; shall repetitively scan from 4.2 The following compounds covered by 20 to 250 amu every 2–3 seconds, and produce this method have been tentatively classified a unit resolution (valleys between m/z 174–176 as known or suspected human or mammalian less than 10 percent of the height of the m/z carcinogens: benzene, carbon tetrachloride, 175 peak), background corrected mass spec- chloroform, and vinyl chloride. Primary trum from 50 ng 4-bromo-fluorobenzene standards of these toxic compounds should (BFB) injected into the GC. The BFB spec- be prepared in a hood, and a NIOSH/MESA trum shall meet the mass-intensity criteria approved toxic gas respirator should be worn in Table 3. All portions of the GC column, when high concentrations are handled. transfer lines, and separator which connect 5. Apparatus and Materials the GC column to the ion source shall re- main at or above the column temperature 5.1 Sample bottles for discrete sampling. during analysis to preclude condensation of 5.1.1 Bottle—25 to 40 mL with screw cap less volatile compounds. (Pierce 13075, or equivalent). Detergent wash, 5.5 Data system—shall collect and record rinse with tap and distilled water, and dry at MS data, store mass intensity data in spec- >105 °C for one hr minimum before use. tral libraries, process GC/MS data and gen- 5.1.2 Septum—Teflon-faced silicone erate reports, and shall calculate and record (Pierce 12722, or equivalent), cleaned as response factors. above and baked at 100–200 °C, for one hour 5.5.1 Data acquisition—mass spectra shall minimum. be collected continuously throughout the 5.2 Purge and trap device—consists of analysis and stored on a mass storage device. purging device, trap, and desorber. Complete 5.5.2 Mass spectral libraries—user created devices are commercially available. libraries containing mass spectra obtained 5.2.1 Purging device—designed to accept 5 from analysis of authentic standards shall be mL samples with water column at least 3 cm employed to reverse search GC/MS runs for deep. The volume of the gaseous head space the compounds of interest (Section 7.2). between the water and trap shall be less than 5.5.3 Data processing—the data system 15 mL. The purge gas shall be introduced less shall be used to search, locate, identify, and than 5 mm from the base of the water col- quantify the compounds of interest in each umn and shall pass through the water as GC/MS analysis. Software routines shall be bubbles with a diameter less than 3 mm. The employed to compute retention times and purging device shown in Figure 1 meets these EICP areas. Displays of spectra, mass criteria. chromatograms, and library comparisons are 5.2.2 Trap—25 to 30 cm × 2.5 mm i.d. min- required to verify results. imum, containing the following: 5.5.4 Response factors and multipoint 5.2.2.1 Methyl silicone packing—one ±0.2 calibrations—the data system shall be used cm, 3 percent OV–1 on 60/80 mesh Chromosorb to record and maintain lists of response fac- W, or equivalent. tors (response ratios for isotope dilution) and 5.2.2.2 Porous polymer—15 ±1.0 cm, Tenax generate multi-point calibration curves (Sec- GC (2,6-diphenylene oxide polymer), 60/80 tion 7). Computations of relative standard mesh, chromatographic grade, or equivalent. deviation (coefficient of variation) are useful 5.2.2.3 Silica gel—8 ±1.0 cm, Davison for testing calibration linearity. Statistics Chemical, 35/60 mesh, grade 15, or equivalent. on initial and on-going performance shall be The trap shown in Figure 2 meets these spec- maintained (Sections 8 and 11). ifications. 5.6 Syringes—5 mL glass hypodermic, 5.2.3 Desorber—shall heat the trap to 175 with Luer-lok tips. ±5 °C in 45 seconds or less. The polymer sec- 5.7 Micro syringes—10, 25, and 100 uL. tion of the trap shall not exceed 180 °C, and 5.8 Syringe valves—2-way, with Luer ends the remaining sections shall not exceed 220 (Telfon or Kel-F).

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5.9 Syringe—5 mL, gas-tight, with shut- 6.5.3 Transfer the stock solution to a Tef- off valve. lon sealed screw-cap-bottle. Store, with 5.10 Bottles—15 mL., screw-cap with minimal headspace, in the dark at ¥10 to Telfon liner. ¥20 °C. 5.11 Balance—analytical, capable of 6.5.4 Prepare fresh standards weekly for weighing 0.1 mg. the gases and 2-chloroethylvinyl ether. All other standards are replaced after one 6. Reagents and Standards month, or sooner if comparison with check 6.1 Reagent water—water in which the standards indicate a change in concentra- compounds of interest and interfering com- tion. Quality control check standards that pounds are not detected by this method (Sec- can be used to determine the accuracy of tion 11.7). It may be generated by any of the calibration standards are available from the following methods: US Environmental Protection Agency, Envi- 6.1.1 Activated carbon—pass tap water ronmental Monitoring and Support Labora- through a carbon bed (Calgon Filtrasorb-300, tory, Cincinnati, Ohio. or equivalent). 6.6 Labeled compound spiking solution— 6.1.2 Water purifier—pass tap water from stock standard solutions prepared as through a purifier (Millipore Super Q, or above, or from mixtures, prepare the spiking equivalent). solution to contain a concentration such 6.1.3 Boil and purge—heat tap water to 90– that a 5–10 μL spike into each 5 mL sample, 100 °C and bubble contaminant free inert gas blank, or aqueous standard analyzed will re- through it for approx one hour. While still sult in a concentration of 20 μg/L of each la- hot, transfer the water to screw-cap bottles beled compound. For the gases and for the and seal with a Teflon-lined cap. water soluble compounds (acrolein, acrylo- 6.2 Sodium thiosulfate—ACS granular. nitrile, acetone, diethyl ether, and MEK), a 6.3 Methanol—pesticide quality or equiva- concentration of 100 μg/L may be used. In- lent. clude the internal standards (Section 7.5) in 6.4 Standard solutions—purchased as so- this solution so that a concentration of 20 μg/ lution or mixtures with certification to their L in each sample, blank, or aqueous standard purity, concentration, and authenticity, or will be produced. prepared from materials of known purity and 6.7 Secondary standards—using stock so- composition. If compound purity is 96 per- lutions, prepare a secondary standard in cent or greater, the weight may be used methanol to contain each pollutant at a con- without correction to calculate the con- centration of 500 μg/mL For the gases and centration of the standard. water soluble compounds (Section 6.6), a con- 6.5 Preparation of stock solutions—pre- centration of 2.5 mg/mL may be used. pare in methanol using liquid or gaseous 6.7.1 Aqueous calibration standards— standards per the steps below. Observe the using a 25 μL syringe, add 20 μL of the sec- safety precautions given in Section 4. ondary standard (Section 6.7) to 50, 100, 200, 6.5.1 Place approx 9.8 mL of methanol in a 500, and 1000 mL of reagent water to produce 10 mL ground glass stoppered volumetric concentrations of 200, 100, 50, 20, and 10 μg/L, flask. Allow the flask to stand unstoppered respectively. If the higher concentration for approximately 10 minutes or until all standard for the gases and water soluble methanol wetted surfaces have dried. In each compounds was chosen (Section 6.6), these case, weigh the flask, immediately add the compounds will be at concentrations of 1000, compound, then immediately reweigh to pre- 500, 250, 100, and 50 μg/L in the aqueous cali- vent evaporation losses from affecting the bration standards. measurement. 6.7.2 Aqueous performance standard—an 6.5.1.1 Liquids—using a 100 μL syringe, aqueous standard containing all pollutants, permit 2 drops of liquid to fall into the meth- internal standards, labeled compounds, and anol without contacting the leck of the BFB is prepared daily, and analyzed each flask. Alternatively, inject a known volume shift to demonstrate performance (Section of the compound into the methanol in the 11). This standard shall contain either 20 or flask using a micro-syringe. 100 μg/L of the labeled and pollutant gases 6.5.1.2 Gases (chloromethane, and water soluble compounds, 10 μg/L BFB, bromomethane, chloroethane, vinyl chlo- and 20 μg/L of all other pollutants, labeled ride)—fill a valved 5 mL gas-tight syringe compounds, and internal standards. It may with the compound. Lower the needle to ap- be the nominal 20 μg/L aqueous calibration proximately 5 mm above the methanol me- standard (Section 6.7.1). niscus. Slowly introduce the compound 6.7.3 A methanolic standard containing above the surface of the meniscus. The gas all pollutants and internal standards is pre- will dissolve rapidly in the methanol. pared to demonstrate recovery of these com- 6.5.2 Fill the flask to volume, stopper, pounds when syringe injection and purge and then mix by inverting several times. Cal- trap analyses are compared. This standard culate the concentration in mg/mL (μg/μL) shall contain either 100 μg/mL or 500 μg/mL from the weight gain (or density if a known of the gases and water soluble compounds, volume was injected). and 100 μg/mL of the remaining pollutants

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and internal standards (consistent with the other compounds. This recovery is dem- amounts in the aqueous performance stand- onstrated initially for each purge and trap ard in 6.7.2). GC/MS system. The test is repeated only if 6.7.4 Other standards which may be need- the purge and trap or GC/MS systems are ed are those for test of BFB performance modified in any way that might result in a (Section 7.1) and for collection of mass spec- change in recovery. tra for storage in spectral libraries (Section 7.3.2 Demonstrate that 100 ng toluene (or 7.2). toluene-d8) produces an area at m/z 91 (or 99) approx one-tenth that required to exceed the 7. Calibration linear range of the system. The exact value 7.1 Assemble the gas chromatographic ap- must be determined by experience for each paratus and establish operating conditions instrument. It is used to match the calibra- given in table 2. By injecting standards into tion range of the instrument to the analyt- the GC, demonstrate that the analytical sys- ical range and detection limits required. tem meets the detection limits in table 2 and 7.4 Calibration by isotope dilution—the the mass-intensity criteria in table 3 for 50 isotope dilution approach is used for the ng BFB. purgeable organic compounds when appro- 7.2 Mass spectral libraries—detection and priate labeled compounds are available and identification of the compound of interest when interferences do not preclude the anal- are dependent upon the spectra stored in ysis. If labeled compounds are not available, user created libraries. or interferences are present, internal stand- 7.2.1 Obtain a mass spectrum of each pol- ard methods (Section 7.5 or 7.6) are used. A lutant and labeled compound and each inter- calibration curve encompassing the con- nal standard by analyzing an authentic centration range of interest is prepared for standard either singly or as part of a mix- each compound determined. The relative re- ture in which there is no interference be- sponse (RR) vs concentration (μg/L) is plot- tween closely eluted components. That only ted or computed using a linear regression. a single compound is present is determined An example of a calibration curve for tol- by examination of the spectrum. Fragments uene using toluene-d8 is given in figure 5. not attributable to the compound under Also shown are the ±10 percent error limits study indicate the presence of an interfering (dotted lines). Relative response is deter- compound. Adjust the analytical conditions mined according to the procedures described and scan rate (for this test only) to produce below. A minimum of five data points are re- an undistorted spectrum at the GC peak quired for calibration (Section 7.4.4). maximum. An undistorted spectrum will 7.4.1 The relative response (RR) of pollut- usually be obtained if five complete spectra ant to labeled compound is determined from are collected across the upper half of the GC isotope ratio values calculated from acquired peak. Software algorithms designed to ‘‘en- data. Three isotope ratios are used in this hance’’ the spectrum may eliminate distor- process: tion, but may also eliminate authentic m/z’s R = the isotope ratio measured in the or introduce other distortion. X pure pollutant (figure 6A). 7.2.3 The authentic reference spectrum is R = the isotope ratio of pure labeled com- obtained under BFB tuning conditions (Sec- y pound (figure 6B). tion 7.1 and table 3) to normalize it to spec- R = the isotope ratio measured in the ana- tra from other instruments. m lytical mixture of the pollutant and la- 7.2.4 The spectrum is edited by saving the beled compounds (figure 6C). 5 most intense mass spectral peaks and all other mass spectral peaks greater than 10 The correct way to calculate RR is: RR = percent of the base peak. This spectrum is (Ry¥Rm) (RX + 1)/(Rm¥RX)(Ry + 1) If Rm is not stored for reverse search and for compound between 2Ry and 0.5RX, the method does not confirmation. apply and the sample is analyzed by internal 7.3 Assemble the purge and trap device. or external standard methods (Section 7.5 or Pack the trap as shown in Figure 2 and con- 7.6). dition overnight at 170–180 °C by 7.4.2 In most cases, the retention times of backflushing with an inert gas at a flow rate the pollutant and labeled compound are the of 20–30 mL/min. Condition traps daily for a same and isotope ratios (R’s) can be cal- minimum of 10 minutes prior to use. culated from the EICP areas, where: R = 7.3.1 Analyze the aqueous performance (area at m1/z)/(area at m2/z) If either of the standard (Section 6.7.2) according to the areas is zero, it is assigned a value of one in purge and trap procedure in Section 10. Com- the calculations; that is, if: area of m1/z = pute the area at the primary m/z (table 4) for 50721, and area of m2/z = 0, then R = 50721/1 = each compound. Compare these areas to 50720. The m/z’s are always selected such that those obtained by injecting one μL of the RX>Ry. When there is a difference in reten- methanolic standard (Section 6.7.3) to deter- tion times (RT) between the pollutant and mine compound recovery. The recovery shall labeled compounds, special precautions are be greater than 20 percent for the water solu- required to determine the isotope ratios. ble compounds, and 60–110 percent for all RX, Ry, and Rm are defined as follows: 351

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RX=[area m1/z (at RT1)]/1 point calibration range, an averaged re- Ry = 1/[area m2/z (at RT2)] sponse factor may be used for that com- Rm=[area m1/z (at RT1)]/[area m2/z (at RT2)] pound; otherwise, the complete calibration 7.4.3 An example of the above calculations curve for that compound shall be used over can be taken from the data plotted in figure the 5 point range. 6 for toluene and toluene-d8. For these data, 7.6 Combined calibration—by adding the RX = 168920/1 = 168900, Ry = 1/60960 = 0.00001640, isotopically labeled compounds and internal and Rm = 96868/82508 = 1.174. The RR for the standards (Section 6.6) to the aqueous cali- above data is then calculated using the equa- bration standards (Section 6.7.1), a single set tion given in Section 7.4.1. For the example, of analyses can be used to produce calibra- RR = 1.174. tion curves for the isotope dilution and in- NOTE: Not all labeled compounds elute be- ternal standard methods. These curves are fore their pollutant analogs. verified each shift (Section 11.5) by purging 7.4.4 To calibrate the analytical system the aqueous performance standard (Section by isotope dilution, analyze a 5 mL aliquot 6.7.2). Recalibration is required only if cali- of each of the aqueous calibration standards bration and on-going performance (Section (Section 6.7.1) spiked with an appropriate 11.5) criteria cannot be met. constant amount of the labeled compound spiking solution (Section 6.6), using the 8. Quality Assurance/Quality Control purge and trap procedure in section 10. Com- 8.1 Each laboratory that uses this method pute the RR at each concentration. is required to operate a formal quality assur- 7.4.5 Linearity—if the ratio of relative re- ance program. The minimum requirements sponse to concentration for any compound is of this program consist of an initial dem- constant (less than 20 percent coefficient of onstration of laboratory capability, analysis variation) over the 5 point calibration range, of samples spiked with labeled compounds to an averaged relative response/concentration evaluate and document data quality, and ratio may be used for that compound; other- analysis of standards and blanks as tests of wise, the complete calibration curve for that continued performance. Laboratory perform- compound shall be used over the 5 point cali- ance is compared to established performance bration range. criteria to determine if the results of anal- 7.5 Calibration by internal standard—used yses meet the performance characteristics of when criteria for isotope dilution (Section the method. 7.4) cannot be met. The method is applied to pollutants having no labeled analog and to 8.1.1 The analyst shall make an initial the labeled compounds. The internal stand- demonstration of the ability to generate ac- ards used for volatiles analyses are ceptable accuracy and precision with this bromochloromethane, 2-bromo-1- method. This ability is established as de- chloropropane, and 1,4-dichlorobutane. Con- scribed in Section 8.2. centrations of the labeled compounds and 8.1.2 The analyst is permitted to modify pollutants without labeled analogs are com- this method to improve separations or lower puted relative to the nearest eluted internal the costs of measurements, provided all per- standard, as shown in table 2. formance specifications are met. Each time a 7.5.1 Response factors—calibration re- modification is made to the method, the ana- quires the determination of response factors lyst is required to repeat the procedure in (RF) which are defined by the following Section 8.2 to demonstrate method perform- equation: ance. 8.1.3 Analyses of blanks are required to RF = (AsxCis)/(AisxCs), where As is the EICP area at the characteristic m/z for the com- demonstrate freedom from contamination pound in the daily standard. Ais is the EICP and that the compounds of interest and area at the characteristic m/z for the inter- interfering compounds have not been carried nal standard. over from a previous analysis (Section 3). Cis is the concentration (ug/L) of the inter- The procedures and criteria for analysis of a nal standard blank are described in Sections 8.5 and 11.7. Cs is the concentration of the pollutant in 8.1.4 The laboratory shall spike all sam- the daily standard. ples with labeled compounds to monitor 7.5.2 The response factor is determined at method performance. This test is described 10, 20, 50, 100, and 200 ug/L for the pollutants in Section 8.3. When results of these spikes (optionally at five times these concentra- indicate atypical method performance for tions for gases and water soluble pollut- samples, the samples are diluted to bring ants—see Section 6.7), in a way analogous to method performance within acceptable lim- that for calibration by isotope dilution (Sec- its (Section 14.2). tion 7.4.4). The RF is plotted against con- 8.1.5 The laboratory shall, on an on-going centration for each compound in the stand- basis, demonstrate through the analysis of ard (Cs) to produce a calibration curve. the aqueous performance standard (Section 7.5.3 Linearity—if the response factor 6.7.2) that the analysis system is in control. (RF) for any compound is constant (less than This procedure is described in Sections 11.1 35 percent coefficient of variation) over the 5 and 11.5.

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8.1.6 The laboratory shall maintain 8.4 As part of the QA program for the lab- records to define the quality of data that is oratory, method accuracy for wastewater generated. Development of accuracy state- samples shall be assessed and records shall ments is described in Sections 8.4 and 11.5.2. be maintained. After the analysis of five 8.2 Initial precision and accuracy—to es- wastewater samples for which the labeled tablish the ability to generate acceptable compounds pass the tests in Section 8.3.3, precision and accuracy, the analyst shall compute the average percent recovery (P) perform the following operations: and the standard deviation of the percent re- 8.2.1 Analyze two sets of four 5–mL covery (sp) for the labeled compounds only. aliquots (8 aliquots total) of the aqueous per- Express the accuracy assessment as a per- formance standard (Section 6.7.2) according cent recovery interval from P¥2sp to P + 2sp. to the method beginning in Section 10. For example, if P = 90% and sp = 10%, the ac- 8.2.2 Using results of the first set of four curacy interval is expressed as 70–110%. Up- analyses in Section 8.2.1, compute the aver- date the accuracy assessment for each com- ¯ μ age recovery (X) in g/L and the standard de- pound on a regular basis (e.g. after each 5–10 viation of the recovery (s) in μg/L for each new accuracy measurements). compound, by isotope dilution for 8.5 Blanks—reagent water blanks are ana- polluitants with a labeled analog, and by in- ternal standard for labeled compounds and lyzed to demonstrate freedom from carry- pollutants with no labeled analog. over (Section 3) and contamination. 8.2.3 For each compound, compare s and X¯ 8.5.1 The level at which the purge and with the corresponding limits for initial pre- trap system will carry greater than 5 μg/L of cision and accuracy found in table 5. If s and a pollutant of interest (table 1) into a suc- X¯ for all compounds meet the acceptance cri- ceeding blank shall be determined by ana- teria, system performance is acceptable and lyzing successively larger concentrations of analysis of blanks and samples may begin. If these compounds. When a sample contains individual X¯ falls outside the range for accu- this concentration or more, a blank shall be racy, system performance is unacceptable analyzed immediately following this sample for that compound. to demonstrate no carry-over at the 5 μg/L NOTE: The large number of compounds in level. table 5 present a substantial probability that 8.5.2 With each sample lot (samples ana- one or more will fail one of the acceptance lyzed on the same 8 hr shift), a blank shall be criteria when all compoulds are analyzed. To analyzed immediately after analysis of the determine if the analytical system is out of aqueous performance standard (Section 11.1) control, or if the failure can be attributed to to demonstrate freedom from contamina- probability, proceed as follows: tion. If any of the compounds of interest 8.2.4 Using the results of the second set of (table 1) or any potentially interfering com- ¯ four analyses, compute s and X for only pound is found in a blank at greater than 10 those compounds which failed the test of the μg/L (assuming a response factor of 1 relative first set of four analyses (Section 8.2.3). If to the nearest eluted internal standard for these compounds now pass, system perform- compounds not listed in table 1), analysis of ance is acceptable for all compounds and samples is halted until the source of con- analysis of blanks and samples may begin. If, tamination is eliminated and a blank shows however, any of the same compounds fail no evidence of contamination at this level. again, the analysis system is not performing 8.6 The specifications contained in this properly for the compound(s) in question. In method can be met if the apparatus used is this event, correct the problem and repeat calibrated properly, then maintained in a the entire test (Section 8.2.1). calibrated state. 8.3 The laboratory shall spike all samples with labeled compounds to assess method The standards used for calibration (Section performance on the sample matrix. 7), calibration verification (Section 11.5) and 8.3.1 Spike and analyze each sample ac- for initial (Section 8.2) and on-going (Section cording to the method beginning in Section 11.5) precision and accuracy should be iden- 10. tical, so that the most precise results will be 8.3.2 Compute the percent recovery (P) of obtained. The GC/MS instrument in par- the labeled compounds using the internal ticular will provide the most reproducible re- standard method (Section 7.5). sults if dedicated to the settings and condi- 8.3.3 Compare the percent recovery for tions required for the analyses of volatiles each compound with the corresponding la- by this method. beled compound recovery limit in table 5. If 8.7 Depending on specific program re- the recovery of any compound falls outside quirements, field replicates may be collected its warning limit, method performance is un- to determine the precision of the sampling acceptable for that compound in that sam- technique, and spiked samples may be re- ple. Therefore, the sample matrix is complex quired to determine the accuracy of the and the sample is to be diluted and reana- analysis when internal or external standard lyzed, per Section 14.2. methods are used.

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9. Sample Collection, Preservation, and tion 6.6) through the valve bore, then close Handling the valve. 10.4 Attach the syringe valve assembly to 9.1 Grab samples are collected in glass the syringe valve on the purging device. containers having a total volume greater Open both syringe valves and inject the sam- than 20 mL. Fill sample bottles so that no ple into the purging chamber. air bubbles pass through the sample as the 10.5 Close both valves and purge the sam- bottle is filled. Seal each bottle so that no ple for 11.0 ±0.1 minutes at 20–25 °C. air bubbles are entrapped. Maintain the her- 10.6 After the 11 minute purge time, at- metic seal on the sample bottle until time of tach the trap to the chromatograph and set analysis. the purge and trap apparatus to the desorb 9.2 Samples are maintained at 0–4 °C from mode (figure 4). Desorb the trapped com- the time of collection until analysis. If the pounds into the GC column by heating the sample contains residual chlorine, add so- trap to 170–180 °C while backflushing with dium thiosulfate preservative (10 mg/40 mL) carrier gas at 20–60 mL/min for four minutes. to the empty sample bottles just prior to Start MS data acquisition upon start of the shipment to the sample site. EPA Methods desorb cycle, and start the GC column tem- 330.4 and 330.5 may be used for measurement perature program 3 minutes later. Table 1 of residual chlorine (Reference 8). If preserv- summarizes the recommended operating con- ative has been added, shake bottle vigor- ditions for the gas chromatograph. Included ously for one minute immediately after fill- in this table are retention times and detec- ing. tion limits that were achieved under these 9.3 Experimental evidence indicates that conditions. Other columns may be used pro- some aromatic compounds, notably benzene, vided the requirements in Section 8 can be toluene, and ethyl benzene are susceptible to met. If the priority pollutant gases produce rapid biological degradation under certain GC peaks so broad that the precision and re- environmental conditions. Refrigeration covery specifications (Section 8.2) cannot be alone may not be adequate to preserve these met, the column may be cooled to ambient compounds in wastewaters for more than or sub-ambient temperatures to sharpen seven days. For this reason, a separate sam- these peaks. ple should be collected, acidified, and ana- 10.7 While analysis of the desorbed com- lyzed when these aromatics are to be deter- pounds proceeds, empty the purging chamber mined. Collect about 500 mL of sample in a using the sample introduction syringe. Wash clean container. the chamber with two 5-mL portions of rea- Adjust the pH of the sample to about 2 by gent water. After the purging device has adding HCl (1 + 1) while stirring. Check pH been emptied, allow the purge gas to vent with narrow range (1.4 to 2.8) pH paper. Fill through the chamber until the frit is dry, so a sample container as described in Section that it is ready for the next sample. 9.1. If residual chlorine is present, add so- 10.8 After desorbing the sample for four dium thiosulfate to a separate sample con- minutes, recondition the trap by returning tainer and fill as in Section 9.1. to the purge mode. Wait 15 seconds, then close the syringe valve on the purging device 9.4 All samples shall be analyzed within 14 to begin gas flow through the trap. Maintain days of collection. the trap temperature at 170–180 °C. After ap- 10. Purge, Trap, and GC/MS Analysis proximately seven minutes, turn off the trap heater and open the syringe valve to stop the 10.1 Remove standards and samples from gas flow through the trap. When cool, the cold storage and bring to 20–25 °. trap is ready for the next sample. 10.2 Adjust the purge gas flow rate to 40 ±4 mL/min. Attach the trap inlet to the purging 11. System Performance device and set the valve to the purge mode 11.1 At the beginning of each 8 hr shift (figure 3). Open the syringe valve located on during which analyses are performed, system the purging device sample introduction nee- calibration and performance shall be verified dle (figure 1). for all pollutants and labeled compounds. 10.3 Remove the plunger from a 5–mL sy- For these tests, analysis of the aqueous per- ringe and attach a closed syringe valve. Open formance standard (Section 6.7.2) shall be the sample bottle and carefully pour the used to verify all performance criteria. Ad- sample into the syringe barrel until it over- justment and/or recalibration (per Section 7) flows. Replace the plunger and compress the shall be performed until all performance cri- sample. Open the syringe valve and vent any teria are met. Only after all performance cri- residual air while adjusting the sample vol- teria are met may blanks and samples be ume to 5.0 mL. Because this process of tak- analyzed. ing an aliquot destroys the validity of the 11.2 BFB spectrum validity—the criteria sample for future analysis, fill a second sy- in table 3 shall be met. ringe at this time to protect against possible 11.3 Retention times—the absolute reten- loss of data. Add an appropriate amount of tion times of all compounds shall approxi- the labeled compound spiking solution (Sec- mate those given in Table 2.

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11.4 GC resolution—the valley height be- 12. Qualitative Determination—Accomplished tween toluene and toluene-d8 (at m/z 91 and by Comparison of Data from Analysis of a 99 plotted on the same graph) shall be less Sample or Blank with Data from Analysis of than 10 percent of the taller of the two the Shift Standard (Section 11.1). Identifica- peaks. tion is Confirmed When Spectra and Reten- 11.5 Calibration verification and on-going tion Times Agree Per the Criteria Below precision and accuracy—compute the con- 12.1 Labeled compounds and pollutants centration of each polutant (Table 1) by iso- having no labeled analog: tope dilution (Section 7.4) for those 12.1.1 The signals for all characteristic compmunds which have labeled analogs. masses stored in the spectral library (Sec- Compute the concentration of each pollutant tion 7.2.4) shall be present and shall maxi- (Table 1) which has no labeled analog by the mize within the same two consecutive scans. internal standard method (Section 7.5). Com- 12.1.2 Either (1) the background corrected pute the concentration of the labeled com- EICP areas, or (2) the corrected relative in- pounds by the internal standard method. tensities of the mass spectral peaks at the These concentrations are computed based on GC peak maximum shall agree within a fac- the calibration data determined in Section 7. tor of two (0.5 to 2 times) for all masses 11.5.1 For each pollutant and labeled com- stored in the library. pound, compare the concentration with the 12.1.3 The retention time relative to the corresponding limit for on-going accuracy in nearest eluted internal standard shall be Table 5. If all compmunds meet the accept- within ±7 scans or ±20 seconds, whichever is ance criteria, system performance is accept- greater. able and analysis of blanks and samples may 12.2 Pollutants having a labeled analog: continue. If any individual value falls out- 12.2.1 The signals for all characteristic side the range given, system performance is masses stored in the spectral library (Sec- unacceptable for that compound. tion 7.2.4) shall be present and shall maxi- NOTE: The large number of compounds in mize within the same two consecutive scans. Table 5 present a substantial probability 12.2.2 Either (1) the background corrected that one or more will fail the acceptance cri- EICP areas, or (2) the corrected relative in- teria when all compounds are analyzed. To tensities of the mass spectral peaks at the determine if the analytical system is out of GC peak maximum shall agree within a fac- control, or if the failure may be attributed tor of two for all masses stored in the spec- to probability, proceed as follows: tral library. 11.5.1.1 Analyze a second aliquot of the 12.2.3 The retention time difference be- aqueous performance standard (Section tween the pollutant and its labeled analog ± ± 6.7.2). shall agree within 2 scans or 6 seconds 11.5.1.2 Compute the concentration for (whichever is greater) of this difference in the shift standard (Section 11.1). only those compounds which failed the first test (Section 11.5.1). If these compounds now 12.3 Masses present in the experimental pass, system performance is acceptable for mass spectrum that are not present in the reference mass spectrum shall be accounted all compounds and analyses of blanks and for by contaminant or background ions. If samples may proceed. If, however, any of the the experimental mass spectrum is contami- compounds fail again, the measurement sys- nated, an experienced spectrometrist (Sec- tem is not performing properly for these tion 1.4) is to determine the presence or ab- compounds. In this event, locate and correct sence of the compound. the problem or recalibrate the system (Sec- tion 7), and repeat the entire test (Section 13. Quantitative Determination 11.1) for all compounds. 11.5.2 Add results which pass the speci- 13.1 Isotope dilution—by adding a known amount of a labeled compound to every sam- fication in 11.5.1.2 to initial (Section 8.2) and ple prior to purging, correction for recovery previous on-going data. Update QC charts to of the pollutant can be made because the pol- form a graphic representation of laboratory lutant and its labeled analog exhibit the performance (Figure 7). Develop a statement same effects upon purging, desorption, and of accuracy for each pollutant and labeled gas chromatography. Relative response (RR) compound by calculating the average per- values for sample mixtures are used in con- centage recovery (R) and the standard devi- junction with calibration curves described in ation of percent recovery (sr). Express the ac- Section 7.4 to determine concentrations di- ¥ curacy as a recovery interval from R 2sr to rectly, so long as labeled compound spiking R + 2sr. For example, if R = 95% and sr = 5%, levels are constant. For the toluene example the accuracy is 85–105 percent. given in Figure 6 (Section 7.4.3), RR would be equal to 1.174. For this RR value, the toluene calibration curve given in Figure 5 indicates a concentration of 31.8 μg/L.

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13.2 Internal standard—calculate the con- peaks greater than the height of the internal centration using the response factor deter- standard peaks. These peaks can obscure the mined from calibration data (Section 7.5) compounds of interest. and the following equation: Concentration = (As × Cis)/(Ais × RF) where 15. Method Performance the terms are as defined in Section 7.5.1. 15.1 The specifications for this method 13.3 If the EICP area at the quantitation were taken from the inter-laboratory valida- mass for any compound exceeds the calibra- tion of EPA Method 624 (reference 9). Method tion range of the system, the sample is di- 1624 has been shown to yield slightly better luted by successive factors of 10 and these di- performance on treated effluents than Meth- lutions are analyzed until the area is within od 624. Additional method performance data the calibration range. can be found in Reference 10. 13.4 Report results for all pollutants and labeled compounds (Table 1) found in all References standards, blanks, and samples, in μg/L to three significant figures. Results for samples 1. ‘‘Performance Tests for the Evaluation which have been diluted are reported at the of Computerized Gas Chromatography/Mass least dilute level at which the area at the Spectrometry Equipment and Laboratories,’’ quantitation mass is within the calibration USEPA, EMSL/Cincinnati, OH 45268, EPA– range (Section 13.3) and the labeled com- 600/4–80–025 (April 1980). pound recovery is within the normal range 2. Bellar, T.A. and Lichtenberg, J.J., for the Method (Section 14.2). ‘‘Journal American Water Works Associa- tion,’’ 66, 739 (1974). 14. Analysis of Complex Samples 3. Bellar, T.A. and Lichtenberg, J.J., 14.1 Untreated effluents and other sam- ‘‘Semi-automated Headspace Analysis of ples frequently contain high levels (>1000 μg/ Drinking Waters and Industrial Waters for L) of the compounds of interest and of inter- Purgeable Volatile Organic Compounds,’’ in fering compounds. Some samples will foam Measurement of Organic Pollutants Water and excessively when purged; others will over- Wastewater, C.E. VanHall, ed., American So- load the trap/or GC column. ciety for Testing Materials, Philadelphia, 14.2 Dilute 0.5 mL of sample with 4.5 mL PA, Special Technical Publication 686, (1978). of reagent water and analyze this diluted 4. ‘‘Working with Carcinogens,’’ DHEW, sample when labeled compound recovery is PHS, NIOSH, Publication 77–206 (1977). outside the range given in Table 5. If the re- 5. ‘‘OSHA Safety and Health Standards, covery remains outside of the range for this General Industry,’’ 29 CFR part 1910, OSHA diluted sample, the aqueous performance 2206, (1976). standard shall be analyzed (Section 11) and 6. ‘‘Safety in Academic Chemistry Labora- calibration verified (Section 11.5). If the re- tories,’’ American Chemical Society Publica- covery for the labeled compmund in the tion, Committee on Chemical Safety (1979). aqueous performance standard is outside the 7. ‘‘Handbook of Analytical Quality Con- range given in Table 5, the analytical system trol in Water and Wastewater Laboratories,’’ is out of control. In this case, the instrumelt USEPA, EMSL/Cincinnati, OH 45268, EPA–4– shall be repaired, the performance specifica- 79–019 (March 1979). tions in Section 11 shall be met, and the 8. ‘‘Methods 330.4 and 330.5 for Total Resid- analysis of the undiluted sample shall be re- ual Chlorine,’’ USEPA, EMSL/Cincinnati, OH peated. If the recovery for the aqueous per- 45268, EPA–4–79–020 (March 1979). formance standard is within the range given 9. ‘‘EPA Method Study 29 EPA Method in Table 5, the method does not work on the 624—Purgeables,’’ EPA 600/4–84–054, National sample being analyzed and the result may Technical Information Service, PB84–209915, not be reported for regulatory compliance Springfield, Virginia 22161, June 1984. purposes. 10. ‘‘Colby, B.N., Beimer, R.G., Rushneck, 14.3 Reverse search computer programs D.R., and Telliard, W.A., ‘‘Isotope Dilution can misinterpret the spectrum of Gas Chromatography-Mass Spectrometry for chromatographically unresolved pollutant the Determination of Priority Pollutants in and labeled compound pairs with overlapping Industrial Effluents,’’ USEPA, Effluent spectra when a high level of the pollutant is Guidelines Division, Washington, DC 20460 present. Examine each chromatogram for (1980).

TABLE 1—VOLATILE ORGANIC COMPOUNDS ANALYZED BY ISOTOPE DILUTION GC/MS

Compound Storet CAS reg- EPA- NPDES istry EGD

Acetone ...... 81552 67–64–1 516 V Acrolein ...... 34210 107–02–8 002 V 001 V Acrylonitrile ...... 34215 107–13–1 003 V 002 V Benzene ...... 34030 71–43–2 004 V 003 V Bromodichloromethane ...... 32101 75–27–4 048 V 012 V

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TABLE 1—VOLATILE ORGANIC COMPOUNDS ANALYZED BY ISOTOPE DILUTION GC/MS—Continued

CAS reg- EPA- Compound Storet istry EGD NPDES

Bromoform ...... 32104 75–25–2 047 V 005 V Bromomethane ...... 34413 74–83–9 046 V 020 V Carbon tetrachloride ...... 32102 56–23–5 006 V 006 V Chlorobenzene ...... 34301 108–90–7 007 V 007 V Chloroethane ...... 34311 75–00–3 016 V 009 V 2-chloroethylvinyl ether ...... 34576 110–75–8 019 V 010 V Chloroform ...... 32106 67–66–1 023 V 011 V Chloromethane ...... 34418 74–87–3 045 V 021 V Dibromochloromethane ...... 32105 124–48–1 051 V 008 V 1,1-dichloroethane ...... 34496 75–34–3 013 V 014 V 1,2-dichloroethane ...... 34536 107–06–2 010 V 015 V 1,1-dichloroethene ...... 34501 75–35–4 029 V 016 V Trans-1,2-dichloroethane ...... 34546 156–60–5 030 V 026 V 1,2-dichloropropane ...... 34541 78–87–5 032 V 017 V Cis-1,3-dichloropropene ...... 34704 10061–01–5 Trans-1,3-dichloropropene ...... 34699 10061–02–6 033 V Diethyl ether ...... 81576 60–29–7 515 V P-dioxane ...... 81582 123–91–1 527 V Ethylbenzene ...... 34371 100–41–4 038 V 019 V Methylene chloride ...... 34423 75–09–2 044 V 022 V Methyl ethyl ketone ...... 81595 78–93–3 514 V 1,1,2,2-tetrachloroethane ...... 34516 79–34–5 015 V 023 V Tetrachlorethene ...... 34475 127–18–4 085 V 024 V Toluene ...... 34010 108–88–3 086 V 025 V 1,1,1-trichloroethane ...... 34506 71–55–6 011 V 027 V 1,1,2-trichloroethane ...... 34511 79–00–5 014 V 028 V Trichloroethene ...... 39180 79–01–6 087 V 029 V Vinyl chloride ...... 39175 75–01–4 088 V 031 V

TABLE 2—GAS CHROMATOGRAPHY OF TABLE 2—GAS CHROMATOGRAPHY OF PURGEABLE ORGANIC COMPOUNDS BY ISO- PURGEABLE ORGANIC COMPOUNDS BY ISO- TOPE DILUTION GC/MS TOPE DILUTION GC/MS—Continued

Min- Min- Mean i Mean i EGD Ref re- mum EGD Ref re- mum No. Compound EGD ten- level No. Compound EGD ten- level (1) No. tion (2) (1) No. tion (2) time (μg/ time (μg/ (sec) L) (sec) L)

181 Bromochloromethane (I.S.)...... 181 730 10 211 1,1,1-trichloroethane-13C2 ...... 181 989 10 245 Chloromethane-d3 ...... 181 147 50 311 1,1,1-trichloroethane ...... 211 999 10 345 Chloromethane ...... 245 148 50 527 p-dioxane ...... 181 1001 10 246 Bromomethane-d3 ...... 181 243 50 206 Carbon tetrachloride-13C1...... 182 1018 10 346 Bromomethane ...... 246 246 50 306 Carbon tetrachloride...... 206 1018 10 288 Vinyl chloride-d3...... 181 301 50 248 Bromodichloromethane-13C1 ... 182 1045 10 388 Vinyl chloride...... 288 304 10 348 Bromodichloromethane ...... 248 1045 10 216 Chloroethane-d5 ...... 181 378 50 232 1,2-dichloropropane-d6 ...... 182 1123 10 316 Chloroethane ...... 216 386 50 332 1.2-dichloropropane ...... 232 1134 10 244 Methylene chloride-d2...... 181 512 10 233 Trans-1,3-dichloropropene-d4 .. 182 1138 10 344 Methylene chloride...... 244 517 10 333 Trans-1,3-dichloropropene ...... 233 1138 10 616 Acetone-d6 ...... 181 554 50 287 Trichloroethene-13C1 ...... 182 1172 10 716 Acetone ...... 616 565 50 387 Trichloroethene ...... 287 1187 10 002 Acrolein ...... 181 566 50 204 Benzene-d6 ...... 182 1200 10 203 Acrylonitrile-d3 ...... 181 606 50 304 Benzene ...... 204 1212 10 303 Acrylonitrile ...... 203 612 50 251 Chlorodibromemethane-13C1 .. 182 1222 10 229 1,1-dichloroethene-d2 ...... 181 696 10 351 Chlorodibromomethane ...... 251 1222 10 329 1,1-dichloroethene ...... 229 696 10 214 1,1,2-trichloroethane-13C2 ...... 182 1224 10 213 1,1-dichloroethane-d3 ...... 181 778 10 314 1,1,2-trichloroethane ...... 214 1224 10 313 1,1-dichloroethane ...... 213 786 10 019 2-chloroethylvinyl ether...... 182 1278 10 615 Diethyl ether-d10...... 181 804 50 182 2-bromo-1-chloropropane (I.S.) 182 1306 10 715 Diethyl ether...... 615 820 50 247 Bromoform-13C1 ...... 182 1386 10 230 Trans-1,2-dichloroethene-d2 .... 181 821 10 347 Bromoform ...... 247 1386 10 330 Trans-1,2-dichloroethene ...... 230 821 10 215 1,1,2,2-tetrachloroethane-d2 .... 183 1525 10 614 Methyl ethyl ketone-d3 ...... 181 840 50 315 1,1,2,2-tetrachloroethane ...... 215 1525 10 714 Methyl ethyl ketone ...... 614 848 50 285 Tetrachloroethene-13C2 ...... 183 1528 10 223 Chloroform-13C1 ...... 181 861 10 385 Tetrachloroethene ...... 285 1528 10 323 Chloroform ...... 223 861 10 183 1,4-dichlorobutale (int std) ...... 183 1555 10 210 1,2-dichloroethane-d4 ...... 181 901 10 286 Toluene-d8 ...... 183 1603 10 310 1,2-dichloroethane ...... 210 910 10 386 Toluene ...... 286 1619 10

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TABLE 2—GAS CHROMATOGRAPHY OF TABLE 4—VOLATILE PURGEABLE ORGANIC COMPOUNDS BY ISO- CHARACTERISTIC MASSES TOPE DILUTION GC/MS—Continued Labeled compound Analog Primary m/ Min- z’s Mean i re- Acetone ...... d6 58/64 EGD Ref ten- mum No. Compound EGD level Acrolein ...... d2 56/58 (1) No. tion (2) time (μg/ Acrylonitrile ...... d3 53/56 (sec) L) Benzene ...... d6 78/84 Bromodichloromethane ...... 13C 83/86 207 Chlorobenzene-d5 ...... 183 1679 10 Bromoform ...... 13C 173/176 307 Chlorobenzene ...... 207 1679 10 Bromomethale ...... d3 96/99 238 Ethylbenzene-d10 ...... 183 1802 10 Carbon tetrachloride ...... 13C 47/48 338 Ethylbenzene ...... 238 1820 10 185 Bromofluorobenzene ...... 183 1985 10 Chlorobenzene ...... d5 112/117 Chloroethane ...... d5 64/71 (1) Reference numbers beginning with 0, 1 or 5 indicate a 2-chloroethylvinyl ether ...... d7 106/113 pollutant quantified by the internal standard method; reference numbers beginning with 2 or 6 indicate a labeled compound Chloroform ...... 13C 85/86 quantified by the internal standard method; reference numbers Chloromethane ...... d3 50/53 beginning with 3 or 7 indicate a pollutant quantified by isotope Dibromochloromethane ...... 13C 129/130 dilution. 1,1-dichloroethane ...... d3 63/66 (2) This is a minimum level at which the analytical system shall give recognizable mass spectra (background corrected) 1,2-dichloroethane ...... d4 62/67 and acceptable calibration points. Column: 2.4m (8 ft) × 2 mm 1,1-dichloroethene ...... d2 61/65 i.d. glass, packed with one percent SP–1000 coated on 60/80 Trans-1,2-dichloroethene ...... d2 61/65 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 1,2-dichloropropane ...... d6 63/67 °C for 15 minutes. Cis-1,3-dichloropropene ...... d4 75/79 NOTE: The specifications in this table were developed from Trans-1,3-dichloropropene ...... d4 75/79 data collected from three wastewater laboratories. Diethyl ether ...... d10 74/84 p-dioxane ...... d8 88/96 TABLE 3—BFB MASS-INTENSITY SPECIFICATIONS Ethylbenzene ...... d10 106/116 Methylene chloride ...... d2 84/88 Mass Intensity required Methyl ethyl ketone ...... d3 72/75 1,1,2,2-tetrachloroethane ...... d2 83/84 50 15 to 40 percent of mass 95. 75 30 to 60 percent of mass 95. Tetrachloroethene ...... 13C2 166/172 95 base peak, 100 percent. Toluene ...... d8 92/99 96 5 to 9 percent of mass 95. 1,1,1-trichloroethane ...... d3 97/102 173 <2 percent of mass 174. 1,1,2-trichloroethane ...... 13C2 83/84 174 >50 percent of mass 95. Trichloroethene ...... 13C 95/133 175 5 to 9 percent of mass 174 Vinyl chloride ...... d3 62/65 176 95 to 101 percent of mass 174 177 5 to 9 percent of mass 176.

TABLE 5—ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS

Acceptance criteria at 20 μg/L Labeled compound On-going Compound Initial precision and accuracy recovery accuracy section 8.2.3 sec. 8.3 and sec. 11.5 14.2 s (μg/L) X¯ (μg/L) P (percent) R (μg/L)

Acetone ...... Note 1 Acrolein ...... Note 2 Acrylonitrile ...... Note 2 Benzene ...... 9.0 13.0–28.2 ns–196 4–33 Bromodichloromethane ...... 8.2 6.5–31.5 ns–199 4–34 Bromoform ...... 7.0 7.4–35.1 ns–214 6–36 Bromomethane ...... 25.0 d–54.3 ns–414 d–61 Carbon tetrachloride ...... 6.9 15.9–24.8 42–165 12–30 Chlorobenzene ...... 8.2 14.2–29.6 ns–205 4–35 Chloroethane ...... 14.8 2.1–46.7 ns–308 d–51 2–chloroethylvinyl ether ...... 36.0 d–69.8 ns–554 d–79 Chloroform ...... 7.9 11.6–26.3 18–172 8–30 Chloromethane ...... 26.0 d–55.5 ns–410 d–64 Dibromochloromethane ...... 7.9 11.2–29.1 16–185 8–32 1,1-dichloroethane ...... 6.7 11.4–31.4 23–191 9–33 1,2-dichloroethane ...... 7.7 11.6–30.1 12–192 8–33 1,1-dichloroethene ...... 11.7 d–49.8 ns–315 d–52 Trans-1,2–dichloroethene ...... 7.4 10.5–31.5 15–195 8–34

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TABLE 5—ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS—Continued

Acceptance criteria at 20 μg/L Labeled compound On-going Compound Initial precision and accuracy recovery accuracy section 8.2.3 sec. 8.3 and sec. 11.5 14.2 s (μg/L) X¯ (μg/L) P (percent) R (μg/L)

1,2-dichloropropane ...... 19.2 d–46.8 ns–343 d–51 Cis–1,3–dichloropropene ...... 22.1 d–51.0 ns–381 d–56 Trans–1,3–dichloropropene ...... 14.5 d–40.2 ns–284 d–44 Diethyl ether ...... Note 1 P-dioxane ...... Note 1 Ethyl benzene ...... 9.6 15.6–28.5 ns–203 5–35 Methylene chloride ...... 9.7 d–49.8 ns–316 d–50 Methyl ethyl ketone ...... Note 1 1,1,2,2-tetrachloroethane ...... 9.6 10.7–30.0 5–199 7–34 Tetrachloroethene ...... 6.6 15.1–28.5 31–181 11–32 Toluene ...... 6.3 14.5–28.7 4–193 6–33 1,1,1-trichloroethane ...... 5.9 10.5–33.4 12–200 8–35 1,1,2-trichloroethane ...... 7.1 11.8–29.7 21–184 9–32 Trichloroethene ...... 8.9 16.6–29.5 35–196 12–34 Vinyl chloride ...... 27.9 d–58.5 ns–452 d–65 d = detected; result must be greater than zero. ns = no specification; limit would be below detection limit. Note 1: Specifications not available for these compounds at time of release of this method. Note 2: Specifications not developed for these compounds; use method 603.

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METHOD 1625 REVISION B—SEMIVOLATILE OR- additional compounds amenable to extrac- GANIC COMPOUNDS BY ISOTOPE DILUTION GC/ tion and analysis by capillary column gas MS chromatography-mass spectrometry (GC/ MS). 1. Scope and Application 1.2 The chemical compounds listed in Ta- 1.1 This method is designed to determine bles 1 and 2 may be determined in municipal the semivolatile toxic organic pollutants as- and industrial discharges by this method. sociated with the 1976 Consent Decree and The method is designed to meet the survey

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requirements of Effluent Guidelines Division sible, reagents are cleaned by solvent rinse (EGD) and the National Pollutants Dis- and baking at 450 °C for one hour minimum. charge Elimination System (NPDES) under 3.2 Interferences coextracted from sam- 40 CFR 136.1. Any modifications of this meth- ples will vary considerably from source to od, beyond those expressly permitted, shall source, depending on the diversity of the in- be considered as major modifications subject dustrial complex or municipality being sam- to application and approval of alternate test ples. procedures under 40 CFR 136.4 and 136.5. 1.3 The detection limit of this method is 4. Safety usually dependent on the level of inter- 4.1 The toxicity or carcinogenicity of ferences rather than instrumental limita- each compound or reagent used in this meth- tions. The limits listed in Tables 3 and 4 rep- od has not been precisely determined; how- resent the minimum quantity that can be de- ever, each chemical compound should be tected with no interferences present. treated as a potential health hazard. Expo- 1.4 The GC/MS portions of this method sure to these compounds should be reduced are for use only by analysts experienced with to the lowest possible level. The laboratory GC/MS or under the close supervision of such is responsible for maintaining a current qualified persons. Laboratories unfamiliar awareness file of OSHA regulations regard- with analyses of environmental samples by ing the safe handling of the chemicals speci- GC/MS should run the performance tests in fied in this method. A reference file of data reference 1 before beginning. handling sheets should also be made avail- able to all personnel involved in these anal- 2. Summary of Method yses. Additional information on laboratory safety can be found in references 2–4. 2.1 Stable isotopically labeled analogs of 4.2 The following compounds covered by the compounds of interest are added to a one this method have been tentatively classified liter wastewater sample. The sample is ex- as known or suspected human or mammalian tracted at pH 12–13, then at pH <2 with meth- carcinogens: benzidine benzo(a)anthracene, ylene chloride using continuous extraction 3,3′-dichlorobenzidine, benzo(a)pyrene, techniques. The extract is dried over sodium dibenzo(a,h)anthracene, N- sulfate and concentrated to a volume of one nitrosodimethylamine, and -naphtylamine. mL. An internal standard is added to the ex- b Primary standards of these compounds shall tract, and the extract is injected into the gas be prepared in a hood, and a NIOSH/MESA chromatograph (GC). The compounds are approved toxic gas respirator should be worn separated by GC and detected by a mass when high concentrations are handled. spectrometer (MS). The labeled compounds serve to correct the variability of the analyt- 5. Apparatus and Materials ical technique. 2.2 Identification of a compound (quali- 5.1 Sampling equipment for discrete or tative analysis) is performed by comparing composite sampling. the GC retention time and background cor- 5.1.1 Sample bottle, amber glass, 1.1 liters rected characteristic spectral masses with minimum. If amber bottles are not available, those of authentic standards. samples shall be protected from light. Bot- tles are detergent water washed, then sol- 2.3 Quantitative analysis is performed by vent rinsed or baked at 450 °C for one hour GC/MS using extracted ion current profile minimum before use. (EICP) areas. Isotope dilution is used when 5.1.2 Bottle caps—threaded to fit sample labeled compounds are available; otherwise, bottles. Caps are lined with Teflon. Alu- an internal standard method is used. minum foil may be substituted if the sample 2.4 Quality is assured through reproduc- is not corrosive. Liners are detergent water ible calibration and testing of the extraction washed, then reagent water (Section 6.5) and and GC/MS systems. solvent rinsed, and baked at approximately ° 3. Contamination and Interferences 200 C for one hour minimum before use. 5.1.3 Compositing equipment—automatic 3.1 Solvents, reagents, glassware, and or manual compositing system incorporating other sample processing hardware may yield glass containers for collection of a minimum artifacts and/or elevated baselines causing 1.1 liters. Sample containers are kept at 0 to misinterpretation of chromatograms and 4 °C during sampling. Glass or Teflon tubing spectra. All materials shall be demonstrated only shall be used. If the sampler uses a peri- to be free from interferences under the con- staltic pump, a minimum length of com- ditions of analysis by running method blanks pressible silicone rubber tubing may be used initially and with each sample lot (samples in the pump only. Before use, the tubing is started through the extraction process on a thoroughly rinsed with methanol, followed given 8 hr shift, to a maximum of 20). Spe- by repeated rinsings with reagent water cific selection of reagents and purification of (Section 6.5) to minimize sample contamina- solvents by distillation in all-glass systems tion. An integrating flow meter is used to may be required. Glassware and, where pos- collect proportional composite samples.

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5.2 Continuous liquid-liquid extractor— 5.10.1 Data acquisition—mass spectra Teflon or glass conncecting joints and stop- shall be collected continuously throughout cocks without lubrication (Hershberg-Wolf the analysis and stored on a mass storage de- Extractor) one liter capacity, Ace Glass 6841– vice. 10, or equivalent. 5.10.2 Mass spectral libraries—user cre- 5.3 Drying column—15 to 20 mm i.d. Pyrex ated libraries containing mass spectra ob- chromatographic column equipped with tained from analysis of authentic standards coarse glass frit or glass wool plug. shall be employed to reverse search GC/MS 5.4 Kuderna-Danish (K-D) apparatus runs for the compounds of interest (Section 5.4.1 Concentrator tube—10mL, graduated 7.2). (Kontes K–570050–1025, or equivalent) with 5.10.3 Data processing—the data system calibration verified. Ground glass stopper shall be used to search, locate, identify, and (size 19/22 joint) is used to prevent evapo- quantify the compounds of interest in each ration of extracts. GC/MS analysis. Software routines shall be 5.4.2 Evaporation flask—500 mL (Kontes employed to compute retention times and K–570001–0500, or equivalent), attached to peak areas. Displays of spectra, mass concentrator tube with springs (Kontes K– chromatograms, and library comparisons are 662750–0012). required to verify results. 5.4.3 Snyder column—three ball macro 5.10.4 Response factors and multipoint (Kontes K–503000–0232, or equivalent). calibrations—the data system shall be used to record and maintain lists of response fac- 5.4.4 Snyder column—two ball micro tors (response ratios for isotope dilution) and (Kontes K–469002–0219, or equivalent). multipoint calibration curves (Section 7). 5.4.5 Boiling chips—approx 10/40 mesh, ex- Computations of relative standard deviation tracted with methylene chloride and baked (coefficient of variation) are useful for test- at 450 °C for one hr minimum. ing calibration linearity. Statistics on ini- 5.5 Water bath—heated, with concentric tial (Section 8.2) and on-going (Section 12.7) ± ring cover, capable of temperature control 2 performance shall be computed and main- ° C, installed in a fume hood. tained. 5.6 Sample vials—amber glass, 2–5 mL with Teflon-lined screw cap. 6. Reagents and Standards 5.7 Analytical balance—capable of weigh- 6.1 Sodium hydroxide—reagent grade, 6N ing 0.1 mg. in reagent water. 5.8 Gas chromatograph—shall have 6.2 Sulfuric acid—reagent grade, 6N in re- splitless or on-column injection port for cap- agent water. illary column, temperature program with 30 ° 6.3 Sodium sulfate—reagent grade, granu- C hold, and shall meet all of the perform- lar anhydrous, rinsed with methylene chlo- ance specifications in Section 12. ride (20 mL/g) and conditioned at 450 °C for ± × ± 5.8.1 Column—30 5 m 0.25 0.02 mm i.d. one hour minimum. 5% phenyl, 94% methyl, 1% vinyl silicone 6.4 Methylene chloride—distilled in glass bonded phase fused silica capillary column (J (Burdick and Jackson, or equivalent). & W DB–5, or equivalent). 6.5 Reagent water—water in which the 5.9 Mass spectrometer—70 eV electron im- compounds of interest and interfering com- pact ionization, shall repetitively scan from pounds are not detected by this method. 35 to 450 amu in 0.95 to 1.00 second, and shall 6.6 Standard solutions—purchased as so- produce a unit resolution (valleys between lutions or mixtures with certification to m/z 441–442 less than 10 percent of the height their purity, concentration, and authen- of the 441 peak), backgound corrected mass ticity, or prepared from materials of known spectrum from 50 ng purity and composition. If compound purity decafluorotriphenylphosphine (DFTPP) in- is 96 percent or greater, the weight may be troduced through the GC inlet. The spectrum used without correction to compute the con- shall meet the mass-intensity criteria in centration of the standard. When not being Table 5 (reference 5). The mass spectrometer used, standards are stored in the dark at ¥20 shall be interfaced to the GC such that the to ¥10 °C in screw-capped vials with Teflon- end of the capillary column terminates with- lined lids. A mark is placed on the vial at the in one centimeter of the ion source but does level of the solution so that solvent evapo- not intercept the electron or ion beams. All ration loss can be detected. The vials are portions of the column which connect the GC brought to room temperature prior to use. to the ion source shall remain at or above Any precipitate is redissolved and solvent is the column temperature during analysis to added if solvent loss has occurred. preclude condensation of less volatile com- 6.7 Preparation of stock solutions—pre- pounds. pare in methylene chloride, benzene, p- 5.10 Data system—shall collect and record dioxane, or a mixture of these solvents per MS data, store mass-intensity data in spec- the steps below. Observe the safety pre- tral libraries, process GC/MS data, generate cautions in Section 4. The large number of reports, and shall compute and record re- labeled and unlabeled acid, base/neutral, and sponse factors. Appendix C compounds used for combined

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calibration (Section 7) and calibration 6.14 Precision and recovery standard— verification (12.5) require high used for determination of initial (Section 8.2) concentratimns (approx 40 mg/mL) when in- and on-going (Section 12.7) precision and re- dividual stock solutions are prepared, so covery. This solution shall contain the pol- that dilutions of mixtures will permit cali- lutants and labeled compounds at a nominal bration with all compounds in a single set of concentration of 100 μg/mL. solutions. The working range for most com- 6.15 Stability of solutions—all standard pounds is 10–200 μg/mL. Compounds with a re- solutions (Sections 6.8–6.14) shall be analyzed duced MS response may be prepared at high- within 48 hours of preparation and on a er concentrations. monthly basis thereafter for signs of deg- 6.7.1 Dissolve an appropriate amount of radation. Standards will remain acceptable assayed reference material in a suitable sol- if the peak area at the quantitation mass vent. For example, weigh 400 mg naphthalene relative to the DFB internal standard re- in a 10 mL ground glass stoppered volumetric mains within ±15 percent of the area ob- flask and fill to the mark with benzene. tained in the initial analysis of the standard. After the naphthalene is completely dis- 7. Calibration solved, transfer the solution to a 15 mL vial with Teflon-lined cap. 7.1 Assemble the GC/MS and establish the 6.7.2 Stock standard solutions should be operating conditions in Table 3. Analyze checked for signs of degradation prior to the standards per the procedure in Section 11 to preparation of calibration or performance demonstrate that the analytical system test standards. Quality control check sam- meets the detection limits in Tables 3 and 4, ples that can be used to determine the accu- and the mass-intensity criteria in Table 5 for racy of calibration standards are available 50 ng DFTPP. from the US Environmental Protection 7.2 Mass spectral libraries—detection and Agency, Environmental Monitoring and Sup- identification of compounds of interest are port Laboratory, Cincinnati, Ohio 45268. dependent upon spectra stored in user cre- 6.7.3 Stock standard solutions shall be re- ated libraries. placed after six months, or sooner if com- 7.2.1 Obtain a mass spectrum of each pol- parison with quality control check samples lutant, labeled compound, and the internal indicates a change in concentration. standard by analyzing an authentic standard 6.8 Labeled compound spiking solution— either singly or as part of a mixture in which from stock standard solutions prepared as there is no interference between closely above, or from mixtures, prepare the spiking eluted components. That only a single com- solution at a concentration of 200 μg/mL, or pound is present is determined by examina- at a concentration appropriate to the MS re- tion of the spectrum. Fragments not attrib- sponse of each compound. utable to the compound under study indicate the presence of an interfering compound. 6.9 Secondary standard—using stock solu- 7.2.2 Adjust the analytical conditions and tions (Section 6.7), prepare a secondary scan rate (for this test only) to produce an standard containing all of the compounds in undistorted spectrum at the GC peak max- Tables 1 and 2 at a concentration of 400 μg/ imum. An undistorted spectrum will usually mL, or higher concentration appropriate to be obtained if five complete spectra are col- the MS response of the compound. lected across the upper half of the GC peak. 6.10 Internal standard solution—prepare Software algorithms designed to ‘‘enhance’’ ′ 2,2 -difluorobiphenyl (DFB) at a concentra- the spectrum may eliminate distortion, but tion of 10 mg/mL in benzene. may also eliminate authentic masses or in- 6.11 DFTPP solution—prepare at 50 μg/mL troduce other distortion. in acetone. 7.2.3 The authentic reference spectrum is 6.12 Solutions for obtaining authentic obtained under DFTPP tuning conditions mass spectra (Section 7.2)—prepare mixtures (Section 7.1 and Table 5) to normalize it to of compounds at concentrations which will spectra from other instruments. assure authentic spectra are obtained for 7.2.4 The spectrum is edited by saving the storage in libraries. 5 most intense mass spectral peaks and all 6.13 Calibration solutions—combine 0.5 other mass spectral peaks greater than 10 mL of the solution in Section 6.8 with 25, 50, percent of the base peak. This edited spec- 125, 250, and 500 uL of the solution in section trum is stored for reverse search and for 6.9 and bring to 1.00 mL total volume each. compound confirmation. This will produce calibration solutions of 7.3 Analytical range—demonstrate that 20 nominal 10, 20, 50, 100, and 200 μg/mL of the ng anthracene or phenanthrene produces an pollutants and a constant nominal 100 μg/mL area at m/z 178 approx one-tenth that re- of the labeled compounds. Spike each solu- quired to exceed the linear range of the sys- tion with 10 μL of the internal standard solu- tem. The exact value must be determined by tion (Section 6.10). These solutions permit experience for each instrument. It is used to the relative response (labeled to unlabeled) match the calibration range of the instru- to be measured as a function of concentra- ment to the analytical range and detection tion (Section 7.4). limits required, and to diagnose instrument

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sensitivity problems (Section 15.4). The 20 or operator to remove the contributions of ug/mL calibration standard (Section 6.13) can the compounds to each other, the equations be used to demonstrate this performance. in Section 7.4.3 apply. This usually occurs 7.3.1 Polar compound detection—dem- when the height of the valley between the onstrate that unlabeled pentachlorophenol two GC peaks at the same m/z is less than 10 and benzidine are detectable at the 50 μg/mL percent of the height of the shorter of the level (per all criteria in Section 13). The 50 two peaks. If significant GC and spectral μg/mL calibration standard (Section 6.13) can overlap occur, RR is computed using the fol- be used to demonstrate this performance. lowing equation: 7.4 Calibration with isotope dilution—iso- RR = (Ry ¥ Rm) (RX + 1)/(Rm ¥ RX) (Ry + 1), tope dilution is used when (1) labeled com- where RX is measured as shown in Figure 3A, pounds are available, (2) interferences do not Ry is measured as shown in Figure 3B, and preclude its use, and (3) the quantitation Rm is measured as shown in Figure 3C. For mass extracted ion current profile (EICP) example, RX = 46100/4780 = 9.644, Ry = 2650/ area for the compound is in the calibration 43600 = 0.0608, Rm = 49200/48300 = 1.019. amd RR range. If any of these conditions preclude = 1.114. isotope dilution, internal standard methods (Section 7.5 or 7.6) are used. 7.4.5 To calibrate the analytical system μ 7.4.1 A calibration curve encompassing by isotope dilution, analyze a 1.0 L aliquot the concentration range is prepared for each of each of the calibration standards (Section compound to be determined. The relative re- 6.13) using the procedure in Section 11. Com- sponse (pollutant to labeled) vs concentra- pute the RR at each concentration. tion in standard solutions is plotted or com- 7.4.6 Linearity—if the ratio of relative re- puted using a linear regression. The example sponse to concentration for any compound is in Figure 1 shows a calibration curve for phe- constant (less than 20 percent coefficient of nol using phenol-d5 as the isotopic diluent. variation) over the 5 point calibration range, Also shown are the ±10 percent error limits and averaged relative response/concentration (dotted lines). Relative Reponse (RR) is de- ratio may be used for that compound; other- termined according to the procedures de- wise, the complete calibration curve for that scribed below. A minimum of five data compound shall be used over the 5 point cali- points are employed for calibration. bration range. 7.4.2 The relative response of a pollutant 7.5 Calibration by internal standard—used to its labeled analog is determined from iso- when criteria for istope dilution (Section 7.4) tope ratio values computed from acquired cannot be met. The internal standard to be data. Three isotope ratios are used in this used for both acid and base/neutral analyses process: is 2,2′–difluorobiphenyl. The internal stand- ard method is also applied to determination R = the isotope ratio measured for the X of compounds having no labeled analog, and pure pollutant. to measurement of labeled compounds for R = the isotope ratio measured for the la- y intra-laboratory statistics (Sections 8.4 and beled compound. 12.7.4). R = the isotope ratio of an analytical m 7.5.1 Response factors—calibration re- mixture of pollutant and labeled compounds. quires the determination of response factors The m/z’s are selected such that RX>Ry. If (RF) which are defined by the following Rm is not between 2Ry and 0.5RX, the method equation: does not apply and the sample is analyzed by × × internal or external standard methods. RF = (As Cis)/(Ais Cs), where 7.4.3 Capillary columns usually separate As is the area of the characteristic mass for the pollutant-labeled pair, with the labeled the compmund in the daily standard compound eluted first (Figure 2). For this Ais is the area of the characteristic mass for the internal standard case, RX = [area m1/z]/1, at the retention time of the pollutant (RT ). R = 1/[area m /z, at Cis is the concentration of the internal 2 y 2 μ the retention time of the labeled compound standard ( g/mL) Cs is the concentration of the compound in RT1). Rm = [area at m1/z (at RT2)]/[area at the daily standard (μg/mL) RT1)], as measured in the mixture of the pol- lutant and labeled compounds (Figure 2), and 7.5.1.1 The response factor is determined RR = Rm. for at least five concentrations appropriate 7.4.4 Special precautions are taken when to the response of each compound (Section the pollutant-labeled pair is not separated, 6.13); nominally, 10, 20, 50, 100, and 200 μg/mL. or when another labeled compound with The amount of internal standard added to interfering spectral masses overlaps the pol- each extract is the same (100 μg/mL) so that lutant (a case which can occur with isomeric Cis remains constant. The RF is plotted vs compounds). In this case, it is necessary to concentration for each compound in the determine the respective contributions of standard (Cs) to produce a calibration curve. the pollutant and labeled compounds to the 7.5.1.2 Linearity—if the response factor respective EICP areas. If the peaks are sepa- (RF) for any compound is constant (less than rated well enough to permit the data system 35 percent coefficient of variation) over the 5

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point calibration range, an averaged re- generated. Development of accuracy state- sponse factor may be used for that com- ments is described in Section 8.4. pound; otherwise, the complete calibration 8.2 Initial precision and accuracy—to es- curve for that compound shall be used over tablish the ability to generate acceptable the 5 point range. precision and accuracy, the analyst shall 7.6 Combined calibration—by using cali- perform the following operations: bration solutions (Section 6.13) containing 8.2.1 Extract, concentrate, and analyze the pollutants, labeled compounds, and the two sets of four one-liter aliquots (8 aliquots internal standard, a single set of analyses total) of the precision and recovery standard can be used to produce calibration curves for (Section 6.14) according to the procedure in the isotope dilution and internal standard Section 10. methods. These curves are verified each shift 8.2.2 Using results of the first set of four (Section 12.5) by analyzing the 100 μg/mL analyses, compute the average recovery (X¯ ) calibration standard (Section 6.13). Re- in μg/mL and the standard deviation of the calibration is required only if calibration recovery (s) in qg/μL for each compound, by verification (Section 12.5) criteria cannot be isotope dilution for pollutants with a labeled met. analog, and by internal standard for labeled compounds and pollutants with no labeled 8. Quality Assurance/Quality Control analog. 8.2.3 For each compound, compare s and X¯ 8.1 Each laboratory that uses this method with the corresponding limits for initial pre- is required to operate a formal quality assur- cision and accuracy in Table 8. If s and X¯ for ance program. The minimum requirements all compounds meet the acceptance criteria, of this program consist of an initial dem- system performance is acceptable and anal- onstration of laboratory capability, analysis ysis of blanks and samples may begin. If, of samples spiked with labeled compounds to however, any individual s exceeds the preci- evaluate and document data quality, and sion limit or any individual X¯ falls outside analysis of standards and blanks as tests of the range for accuracy, system performance continued performance. Laboratory perform- is unacceptable for that compound. ance is compared to established performance NOTE: The large number of compounds in criteria to determine if the results of anal- Table 8 present a substantial probability yses meet the performance characteristics of that one or more will fail the acceptance cri- the method. teria when all compounds are analyzed. To 8.1.1 The analyst shall make an initial determine if the analytical system is out of demonstration of the ability to generate ac- control, or if the failure can be attributed to ceptable accuracy and precision with this probability, proceed as follows: method. This ability is established as de- 8.2.4 Using the results of the second set of scribed in Section 8.2. four analyses, compute s and X¯ for only 8.1.2 The analyst is permitted to modify those compounds which failed the test of the this method to improve separations or lower first set of four analyses (Section 8.2.3). If the costs of measurements, provided all per- these compounds now pass, system perform- formance specifications are met. Each time a ance is acceptable for all compounds and modification is made to the method, the ana- analysis of blanks and samples may begin. If, lyst is required to repeat the procedure in however, any of the same compoulds fail Section 8.2 to demonstrate method perform- again, the analysis system is not performing ance. properly for these compounds. In this event, 8.1.3 Analyses of blanks are required to correct the problem and repeat the entire demonstrate freedom from contamination. test (Section 8.2.1). The procedures and criteria for analysis of a 8.3 The laboratory shall spike all samples blank are described in Section 8.5. with labeled compounds to assess method 8.1.4 The laboratory shall spike all sam- performance on the sample matrix. ples with labeled compounds to monitor 8.3.1 Analyze each sample according to method performance. This test is described the method in Section 10. in Section 8.3. When results of these spikes 8.3.2 Compute the percent recovery (P) of indicate atypical method performance for the labeled compounds using the internal samples, the samples are diluted to bring standard methmd (Section 7.5). method performance within acceptable lim- 8.3.3 Compare the labeled compound re- its (Section 15). covery for each compound with the cor- 8.1.5 The laboratory shall, on an on-going responding limits in Table 8. If the recovery basis, demonstrate through calibration of any compounds falls outside its warning verification and the analysis of the precision limit, method performance is unacceptable and recovery standard (Section 6.14) that the for that compound in that sample, Therefore, analysis system is in control. These proce- the sample is complex and is to be diluted dures are described in Sections 12.1, 12.5, and and reanalyzed per Section 15.4. 12.7. 8.4 As part of the QA program for the lab- 8.1.6 The laboratory shall maintain oratory, method accuracy for wastewater records to define the quality of data that is samples shall be assessed and records shall

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be maintained. After the analysis of five 330.4 and 330.5 may be used to measure resid- wastewater samples for which the labeled ual chlorine (Reference 8). compounds pass the tests in Section 8.3, 9.3 Begin sample extraction within seven compute the average percent recovery (P) days of collection, and analyze all extracts and the standard deviation of the percent re- within 40 days of extraction. covery (sp) for the labeled compounds only. Express the accuracy assessment as a per- 10. Sample Extraction and Concentration (See Figure 4) cent recovery interval from P—2 sp to P + 2sp. For example, if P = 90% and sp = 10%, the ac- 10.1 Labeled compound spiking—measure curacy interval is expressed as 70–100%. Up- 1.00 ±0.01 liter of sample into a glass con- date the accuracy assessment for each com- tainer. For untreated effluents, and samples pound on a regular basis (e.g. after each 5–10 which are expected to be difficult to extract new accuracy measurements). and/or concentrate, measure an additional 8.5 Blanks—reagent water blanks are ana- 10.0 ±0.1 mL and dilute to a final volume of lyzed to demonstrate freedom from contami- 1.00 ±0.01 liter with reagent water in a glass nation. container. 8.5.1 Extract and concentrate a blank 10.1.1 For each sample or sample lot (to a with each sample lot (samples started maximum of 20) to be extracted at the same through the extraction process on the same time, place three 1.00 ±0.10 liter aliquots of 8 hr shift, to a maximum of 20 samples). Ana- reagent water in glass containers. lyze the blank immediately after analysis of 10.1.2 Spike 0.5 mL of the labeled com- the precision and recovery standard (Section pound spiking solution (Section 6.8) into all 6.14) to demonstrate freedom from contami- samples and one reagant water aliquot. nation. 10.1.3 Spike 1.0 mL of the precision and 8.5.2 If any of the compounds of interest recovery standard (Section 6.14) into the two (Tables 1 and 2) or any potentially inter- remaining reagent water aliquots. fering compound is found in a blank at great- 10.1.4 Stir and equilibrate all solutions for er than 10 μg/L (assuming a response factor 1–2 hr. of 1 relative to the internal standard for 10.2 Base/neutral extraction—place 100–150 compounds not listed in Tables 1 and 2), mL methylene chloride in each continuous analysis of samples is halted until the source extractor and 200–300 in each distilling flask. of contamination is eliminated and a blank 10.2.1 Pour the sample(s), blank, and shows no evidence of contamination at this standard aliquots into the extractors. Rinse level. the glass containers with 50–100 mL meth- 8.6 The specifications contained in this ylene chloride and add to the respective ex- method can be met if the apparatus used is tractor. calibrated properly, then maintained in a 10.2.2 Adjust the pH of the waters in the calibrated state. The standards used for cali- extractors to 12–13 with 6N NaOH while mon- bration (Section 7), calibration verification itoring with a pH meter. Begin the extrac- (Section 12.5), and for initial (Section 8.2) tion by heating the flask until the meth- and on-going (Section 12.7) precision and re- ylene chloride is boiling. When properly ad- covery should be identical, so that the most justed, 1–2 drops of methylene chloride per precise results will be obtained. The GC/MS second will fall from the condensor tip into instrument in particular will provide the the water. After 1–2 hours of extraction, test most reproducible results if dedicated to the the pH and readjust to 12–13 if required. Ex- settings and conditions required for the anal- tract for 18–24 hours. ysis of semi-volatiles by this method. 10.2.3 Remove the distilling flask, esti- 8.7 Depending on specific program re- mate and record the volume of extract (to quirements, field replicates may be collected the nearest 100 mL), and pour the contents to determine the precision of the sampling through a drying column containing 7 to 10 technique, and spiked samples may be re- cm anhydrous sodium sulfate. Rinse the dis- quired to determine the accuracy of the tilling flask with 30–50 mL of methylene analysis when internal or external standard chloride and pour through the drying col- methods are used. umn. Collect the solution in a 500 mL K-D evaporator flask equipped with a 10 mL con- 9. Sample Collection, Preservation, and centrator tube. Seal, label as the base/neu- Handling tral fraction, and concentrate per Sections 9.1 Collect samples in glass containers 10.4 to 10.5. following conventional sampling practices 10.3 Acid extraction—adjust the pH of the (Reference 7). Composite samples are col- waters in the extractors to 2 or less using 6N lected in refrigerated glass containers (Sec- sulfuric acid. Charge clean distilling flasks tion 5.1.3) in accordance with the require- with 300–400 mL of methylene chloride. Test ments of the sampling program. and adjust the pH of the waters after the 9.2 Maintain samples at 0–4 °C from the first 1–2 hr of extraction. Extract for 18–24 time collectimn until extraction. If residual hours. chlorine is present, add 80 mg sodium 10.3.1 Repeat Section 10.2.3, except label thiosulfate per liter of water. EPA Methods as the acid fraction.

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10.4 Concentration—concentrate the ex- and bring to the mark with solvent if re- tracts in separate 500 mL K-D flasks quired. equipped with 10 mL concentrator tubes. 11.3 Add the internal standard solution 10.4.1 Add 1 to 2 clean boiling chips to the (Section 6.10) to the extract (use 1.0 uL of so- flask and attach a three-ball macro Snyder lution per 0.1 mL of extract) immediately column. Prewet the column by adding ap- prior to injection to minimize the possibility proximately one mL of methylene chloride of loss by evaporation, adsorption, or reac- through the top. Place the K-D apparatus in tion. Mix thoroughly. a hot water bath so that the entire lower 11.4 Inject a volume of the standard solu- rounded surface of the flask is bathed with tion or extract such that 100 ng of the inter- steam. Adjust the vertical position of the ap- nal standard will be injected, using on-col- paratus and the water temperature as re- umn or splitless injection. For 1 mL ex- quired to complete the concentration in 15 to tracts, this volume will be 1.0 uL. Start the 20 minutes. At the proper rate of distillation, GC column initial isothermal hold upon in- the balls of the column will actively chatter jection. Start MS data collection after the but the chambers will not flood. When the solvent peak elutes. Stop data collection liquid has reached an apparent volume of 1 after the benzo (ghi) perylene or mL, remove the K-D apparatus from the bath pentachlorophenol peak elutes for the base/ and allow the solvent to drain and cool for at neutral or acid fraction, respectively. Return least 10 minutes. Remove the Snyder column the column to the initial temperature for and rinse the flask and its lower joint into analysis of the next sample. the concentrator tube with 1–2 mL of meth- ylene chloride. A 5-mL syringe is rec- 12. System and Laboratory Performance ommended for this operation. 10.4.2 For performance standards (Sec- 12.1 At the beginning of each 8 hr shift tions 8.2 and 12.7) and for blanks (Section during which analyses are performed, GC/MS 8.5), combine the acid and base/neutral ex- system performance and calibration are tracts for each at this point. Do not combine verified for all pollutants and labeled com- the acid and base/neutral extracts for sam- pounds. For these tests, analysis of the 100 ples. μg/mL calibration standard (Section 6.13) 10.5 Add a clean boiling chip and attach a shall be used to verify all performance cri- two ball micro Snyder column to the concen- teria. Adjustment and/or recalibration (per trator tube. Prewet the column by adding Section 7) shall be performed until all per- approx 0.5 mL methylene chloride through formance criteria are met. Only after all per- the top. Place the apparatus in the hot water formance criteria are met may samples, bath. Adjust the vertical position and the blanks, and precision and recovery standards water temperature as required to complete be analyzed. the concentration in 5–10 minutes. At the 12.2 DFTPP spectrum validity—inject 1 proper rate of distillation, the balls of the μL of the DFTPP solution (Section 6.11) ei- column will actively chatter but the cham- ther separately or within a few seconds of in- bers will not flood. When the liquid reaches jection of the standard (Section 12.1) ana- an apparent volume of approx 0.5 mL, re- lyzed at the beginning of each shift. The cri- move the apparatus from the water bath and teria in Table 5 shall be met. allow to drain and cool for at least 10 min- 12.3 Retention times—the absolute reten- utes. Remove the micro Snyder column and tion time of 2,2′-difluorobiphenyl shall be rinse its lower joint into the concentrator within the range of 1078 to 1248 seconds and tube with approx 0.2 mL of methylene chlo- the relative retention times of all pollutants ride. Adjust the final volume to 1.0 mL. and labeled compounds shall fall within the 10.6 Transfer the concentrated extract to limits given in Tables 3 and 4. a clean screw-cap vial. Seal the vial with a 12.4 GC resolution—the valley height be- Teflon-lined lid, and mark the level on the tween anthracene and phenanthrene at m/z vial. Label with the sample number and frac- 178 (or the analogs at m/z 188) shall not ex- tion, and store in the dark at ¥20 to ¥10 °C ceed 10 percent of the taller of the two peaks. until ready for analysis. 12.5 Calibration verification—compute the concentration of each pollutant (Tables 1 11. GC/MS Analysis and 2) by isotope dilution (Section 7.4) for 11.1 Establish the operating conditions those compounds which have labeled given in Table 3 or 4 for analysis of the base/ analogs. Compute the concentration of each neutral or acid extracts, respectively. For pollutant which has no labeled analog by the analysis of combined extracts (Section internal standard method (Section 7.5). Com- 10.4.2), use the operating conditions in Table pute the concentration of the labeled com- 3. pounds by the internal standard method. 11.2 Bring the concentrated extract (Sec- These concentrations are computed based on tion 10.6) or standard (Sections 6.13 through the calibration data determined in Section 7. 6.14) to room temperature and verify that 12.5.1 For each pollutant and labeled com- any precipitate has redissolved. Verify the pound being tested, compare the concentra- level on the extract (Sections 6.6 and 10.6) tion with the calibration verification limit

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in Table 8. If all compounds meet the accept- ery (sr). Express the accuracy as a recovery ance criteria, calibration has been verified interval from R¥2sr to R + 2sr. For example, and analysis of blanks, samples, and preci- if R = 95% and sr = 5%, the accuracy is sion and recovery standards may proceed. If, 85¥105%. however, any compound fails, the measure- ment system is not performing properly for 13. Qualitative Determination that compound. In this event, prepare a fresh 13.1 Qualititative determination is ac- calibration standard or correct the problem complished by comparison of data from anal- causing the failure and repeat the test (Sec- ysis of a sample or blank with data from tion 12.1), or recalibrate (Section 7). analysis of the shift standard (Section 12.1) 12.6 Multiple peaks—each compound in- and with data stored in the spectral libraries jected shall give a single, distinct GC peak. (Section 7.2.4). Identification is confirmed 12.7 On-going precision and accuracy. when spectra and retention times agree per 12.7.1 Analyze the extract of one of the the criteria below. pair of precision and recovery standards 13.2 Labeled compounds and pollutants (Section 10.1.3) prior to analysis of samples having no labeled analog: from the same lot. 13.2.1 The signals for all characteristic 12.7.2 Compute the concentration of each masses stored in the spectral library (Sec- pollutant (Tables 1 and 2) by isotope dilution tion 7.2.4) shall be present and shall maxi- (Section 7.4) for those compounds which have mize within the same two consecutive scans. labeled analogs. Compute the concentration 13.2.2 Either (1) the background corrected of each pollutant which has no labeled ana- EICP areas, or (2) the corrected relative in- log by the internal standard method (Section tensities of the mass spectral peaks at the 7.5). Compute the concentration of the la- GC peak maximum shall agree within a fac- beled compounds by the internal standard tor of two (0.5 to 2 times) for all masses method. 12.7.3 For each pollutant and labeled com- stored in the library. pound, compare the concentration with the 13.2.3 The retention time relative to the limits for on-going accuracy in Table 8. If all nearest eluted internal standard shall be ± ± compounds meet the acceptance criteria, within 15 scans or 15 seconds, whichever is system performance is acceptable and anal- greater of this difference in the shift stand- ysis of blanks and samples may proceed. If, ard (Section 12.1). however, any individual concentration falls 13.3 Pollutants having a labled analog: outside of the range given, system perform- 13.3.1 The signals for all characteristic ance is unacceptable for that compound. masses stored in the spectral library (Sec- NOTE: The large number of compounds in tion 7.2.4) shall be present and shall maxi- Table 8 present a substantial probability mize within the same two consecutive scans. that one or more will fail when all com- 13.3.2. Either (1) the background corrected pounds are analyzed. To determine if the ex- EICP areas, or (2) the corrected relative in- traction/concentration system is out of con- tensities of the mass spectral peaks at the trol or if the failure is caused by probability, GC peak maximum shall agree within a fac- proceed as follows: tor of two for all masses stored in the spec- 12.7.3.1 Analyze the second aliquot of the tral library. pair of precision and recovery standard (Sec- 13.3.3. The retention time difference be- tion 10.1.3). tween the pollutant and its labeled analog 12.7.3.2 Compute the concentration of shall agree within ±6 scans or ±6 seconds only those pollutants or labeled compounds (whichever is greater) of this difference in that failed the previous test (Section 12.7.3). the shift standard (Section 12.1). If these compounds now pass, the extraction/ 13.4 Masses present in the experimental concentration processes are in control and mass spectrum that are not present in the analysis of blanks and samples may proceed. reference mass spectrum shall be accounted If, however, any of the same compounds fail for by contaminant or background ions. If again, the extraction/concentration proc- the experimental mass spectrum is contami- esses are not being performed properly for nated, an experienced spectrometrist (Sec- these compounds. In this event, correct the tion 1.4) is to determine the presence or ab- problem, re-extract the sample lot (Section sence of the cmmpound. 10) and repeat the on-going precision and re- 14. Quantitative Determination covery test (Section 12.7). 12.7.4 Add results which pass the speci- 14.1 Isotope dilution—by adding a known fications in Section 12.7.2 to initial and pre- amount of a labeled compound to every sam- vious on-going data. Update QC charts to ple prior to extraction, correction for recov- perform a graphic representation of contin- ery of the pollutant can be made because the ued laboratory performance (Figure 5). De- pollutant and its labeled analog exhibit the velop a statement of laboratory accuracy for same effects upon extraction, concentration, each pollutant and labeled compound by cal- and gas chromatography. Relative response culating the average percent recovery (R) (RR) values for mixtures are used in conjunc- and the standard deviation of percent recov- tion with calibration curves described in

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Section 7.4 to determine concentrations di- of the labeled compounds are within a factor rectly, so long as labeled compound spiking of two of the respective areas in the shift levels are constant. For the phenml example standard, and the internal standard area is given in Figure 1 (Section 7.4.1), RR would be less than one-half of its respective area, then equal to 1.114. For this RR value, the phenol internal standard loss in the extract has oc- calibration curve given in Figure 1 indicates curred. In this case, use one of the labeled a concentration of 27 μg/mL in the sample ex- compounds (perferably a polynuclear aro- tract (Cex). matic hydrocarbon) to compute the con- 14.2 Internal standard—compute the con- centration of a pollutant with no labeled centration in the extract using the response analog. factor determined from calibration data 15.4 Recovery of labeled compounds—in (Section 7.5) and the following equation: most samples, labeled compound recoveries Cex(μg/mL) = (As × Cis/(Ais × RF) where Cex is will be similar to those from reagent water the concentration of the compound in the ex- (Section 12.7). If the labeled compound recov- tract, and the other terms are as defined in ery is outside the limits given in Table 8, the Section 7.5.1. dilute extract (Section 10.1) is analyzed as in 14.3 The concentration of the pollutant in Section 14.4. If the recoveries of all labeled water is computed using the volumes of the compounds and the internal staldard are low original water sample (Section 10.1) and the (per the criteria above), then a loss in instru- final extract volume (Section 10.5), as fol- ment sensitivity is the most likely cause. In μ × lows: Concentration in water ( g/L) = (Cex this case, the 100 μg/mL calibration standard Vex)/Vs where Vex is the extract volume in (Section 12.1) shall be analyzed and calibra- mL, and Vs is the sample volume in liters. tion verified (Section 12.5). If a loss in sensi- 14.4 If the EICP area at the quantitiation tivity has occurred, the instrument shall be mass for any compound exceeds the calibra- repaired, the performance specifications in tion range of the system, the extract of the Section 12 shall be met, and the extract re- dilute aliquot (Section 10.1) is analyzed by analyzed. If a loss in instrument sensitivity isotope dilution; otherwise, the extract is di- has not occurred, the method does not work μ luted by a factor of 10, 9 L of internal stand- on the sample being analyzed and the result ard solution (Section 6.10) are added to a 1.0 may not be reported for regulatory compli- mL aliquot, and this diluted extract is ana- ance purposes. lyzed by the internal standard method (Sec- tion 14.2). Quantify each compound at the 16. Method Performance highest concentration level within the cali- bration range. 16.1 Interlaboratory performance for this 14.5 Report results for all pollutants and method is detailed in references 9 and 10. labeled compounds (Tables 1 and 2) found in 16.2 A chromatogram of the 100 μg/mL all standards, blanks, and samples in μg/L, to acid/base/neutral calibration standard (Sec- three significant figures. Results for samples tion 6.13) is shown in Figure 6. which have been diluted are reported at the REFERENCES least dilute level at which the area at the quantitation mass is within the calibration 1. ‘‘Performance Tests for the Evaluation range (Section 14.4) and the labeled com- of Computerized Gas Chromatography/Mass pound recovery is within the normal range Spectrometry Equipment and Laboratories’’ for the method (Section 15.4). USEPA, EMSL/Cincinnati, OH 45268, EPA– 600/4–80–025 (April 1980). 15. Analysis of Complex Samples 2. ‘‘Working with Carcinogens,’’ DHEW, 15.1 Untreated effluents and other sam- PHS, CDC, NIOSH, Publication 77–206, (Au- ples frequently contain high levels (>1000 μg/ gust 1977). L) of the compounds of interest, interfering 3. ‘‘OSHA Safety and Health Standards, compounds, and/or polymeric materials. General Industry’’ OSHA 2206, 29 CFR part Some samples will not concentrate to one 1910 (January 1976). mL (Section 10.5); others will overload the 4. ‘‘Safety in Academic Chemistry Labora- GC column and/or mass spectrometer. tories, ’’ ACS Committee on Chemical Safety 15.2 Analyze the dilute aliquot (Section (1979). 10.1) when the sample will not concentrate to 5. ‘‘Reference Compound to Calibrate Ion 1.0 mL. If a dilute aliquot was not extracted, Abundance Measurement in Gas Chroma- and the sample holding time (Section 9.3) has tography-Mass Spectrometry Systems,’’ J.W. not been exceeded, dilute an aliquot of the Eichelberger, L.E. Harris, and W.L. Budde, sample with reagent water and re-extract Anal. Chem., 47, 955 (1975). (Section 10.1); otherwise, dilute the extract 6. ‘‘Handbook of Analytical Quality Con- (Section 14.4) and analyze by the internal trol in Water and Wastewater Laboratories,’’ standard method (Section 14.2). USEPA, EMSL/Cincinnati, OH 45268, EPA– 15.3 Recovery of internal standard—the 600/4–79–019 (March 1979). EICP area of the internal standard should be 7. ‘‘Standard Practice for Sampling within a factor of two of the area in the shift Water,’’ ASTM Annual Book of Standards, standard (Section 12.1). If the absolute areas ASTM, Philadelphia, PA, 76 (1980).

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8. ‘‘Methods 330.4 and 330.5 for Total Resid- Industrial Effluents.’’ USEPA, Effluent ual Chlorine,’’ USEPA, EMSL/ Cincinnati, Guidelines Division, Washington, DC 20460 OH 45268, EPA 600/4–70–020 (March 1979). (1980). 9. Colby, B.N., Beimer, R.G., Rushneck, 10. ‘‘Inter-laboratory Validation of US En- D.R., and Telliard, W.A., ‘‘Isotope Dilution vironmental Protection Agency Method Gas Chromatography-Mass Spectrometry for 1625,’’ USEPA, Effluent Guidelines Division, the determination of Priority Pollutants in Washington, DC 20460 (June 15, 1984).

TABLE 1—BASE/NEUTRAL EXTRACTABLE COMPOUNDS

CAS reg- EPA- Compound STORET istry EGD NPDES

Acenaphthene ...... 34205 83–32–9 001 B 001 B Acenaphthylene ...... 34200 208–96–8 077 B 002 B Anthracene ...... 34220 120–12–7 078 B 003 B Benzidine ...... 39120 92–87–5 005 B 004 B Benzo(a)anthracene ...... 34526 56–55–3 072 B 005 B Benzo(b)fluoranthene ...... 34230 205–99–2 074 B 007 B Benzo(k)fluoranthene ...... 34242 207–08–9 075 B 009 B Benzo(a)pyrene ...... 34247 50–32–8 073 B 006 B Benzo(ghi)perylene ...... 34521 191–24–2 079 B 008 B Biphenyl (Appendix C) ...... 81513 92–52–4 512 B Bis(2-chloroethyl) ether ...... 34273 111–44–4 018 B 011 B Bis(2-chloroethyoxy)methane ...... 34278 111–91–1 043 B 010 B Bis(2-chloroisopropyl) ether ...... 34283 108–60–1 042 B 012 B Bis(2-ethylhexyl) phthalate ...... 39100 117–81–7 066 B 013 B 4-bromophenyl phenyl ether ...... 34636 101–55–3 041 B 014 B Butyl benzyl phthalate ...... 34292 85–68–7 067 B 015 B n-C10 (Appendix C) ...... 77427 124–18–5 517 B n-C12 (Appendix C) ...... 77588 112–40–2 506 B n-C14 (Appendix C) ...... 77691 629–59–4 518 B n-C16 (Appendix C) ...... 77757 544–76–3 519 B n-C18 (Appendix C) ...... 77804 593–45–3 520 B n-C20 (Appendix C) ...... 77830 112–95–8 521 B n-C22 (Appendix C) ...... 77859 629–97–0 522 B n-C24 (Appendix C) ...... 77886 646–31–1 523 B n-C26 (Appendix C) ...... 77901 630–01–3 524 B n-C28 (Appendix C) ...... 78116 630–02–4 525 B n-C30 (Appendix C) ...... 78117 638–68–6 526 B Carbazole (4c) ...... 77571 86–74–8 528 B 2-chloronaphthalene ...... 34581 91–58–7 020 B 016 B 4-chlorophenyl phenyl ether ...... 34641 7005–72–3 040 B 017 B Chrysene ...... 34320 218–01–9 076 B 018 B P-cymene (Appendix C) ...... 77356 99–87–6 513 B Dibenzo(a,h)anthracene ...... 34556 53–70–3 082 B 019 B Dibenzofuran (Appendix C and 4c) ...... 81302 132–64–9 505 B Dibenzothiophene (Synfuel) ...... 77639 132–65–0 504 B Di-n-butyl phthalate ...... 39110 84–74–2 068 B 026 B 1,2-dichlorobenzene ...... 34536 95–50–1 025 B 020 B 1,3-dichlorobenzene ...... 34566 541–73–1 026 B 021 B 1,4-dichlorobenzene ...... 34571 106–46–7 027 B 022 B 3,3′-dichlorobenzidine ...... 34631 91–94–1 028 B 023 B Diethyl phthalate ...... 34336 84–66–2 070 B 024 B 2,4-dimethylphenol ...... 34606 105–67–9 034 A 003 A Dimethyl phthalate ...... 34341 131–11–3 071 B 025 B 2,4-dinitrotoluene ...... 34611 121–14–2 035 B 027 B 2,6-dinitrotoluene ...... 34626 606–20–2 036 B 028 B Di-n-octyl phthalate ...... 34596 117–84–0 069 B 029 B Diphenylamine (Appendix C) ...... 77579 122–39–4 507 B Diphenyl ether (Appendix C) ...... 77587 101–84–8 508 B 1,2-diphenylhydrazine ...... 34346 122–66–7 037 B 030 B Fluoranthene ...... 34376 206–44–0 039 B 031 B Fluorene ...... 34381 86–73–7 080 B 032 B Hexachlorobenzene ...... 39700 118–74–1 009 B 033 B Hexachlorobutadiene ...... 34391 87–68–3 052 B 034 B Hexachloroethane ...... 34396 67–72–1 012 B 036 B Hexachlorocyclopentadiene ...... 34386 77–47–4 053 B 035 B Indeno(1,2,3-cd)pyrene ...... 34403 193–39–5 083 B 037 B Isophorone ...... 34408 78–59–1 054 B 038 B Naphthalene ...... 34696 91–20–3 055 B 039 B B-naphthylamine (Appendix C) ...... 82553 91–59–8 502 B Nitrobenzene ...... 34447 98–95–3 056 B 040 B N-nitrosodimethylamine ...... 34438 62–75–9 061 B 041 B N-nitrosodi-n-propylamine ...... 34428 621–64–7 063 B 042 B N-nitrosodiphenylamine ...... 34433 86–30–3 062 B 043 B

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TABLE 1—BASE/NEUTRAL EXTRACTABLE COMPOUNDS—Continued

CAS reg- EPA- Compound STORET istry EGD NPDES

Phenanthrene ...... 34461 85–01–8 081 B 044 B Phenol ...... 34694 108–95–2 065 A 010 A a-Picoline (Synfuel) ...... 77088 109–06–89 503 B Pyrene ...... 34469 129–00–0 084 B 045 B styrene (Appendix C) ...... 77128 100–42–5 510 B a-terpineol (Appendix C) ...... 77493 98–55–5 509 B 1,2,3-trichlorobenzene (4c) ...... 77613 87–61–6 529 B 1,2,4-trichlorobenzene ...... 34551 120–82–1 008 B 046 B

TABLE 2—ACID EXTRACTABLE COMPOUNDS

CAS reg- EPA- Compound STORET istry EGD NPDES

4-chloro-3-methylphenol ...... 34452 59–50–7 022 A 008 A 2-chlorophenol ...... 34586 95–57–8 024 A 001 A 2,4-dichlorophenol ...... 34601 120–83–2 031 A 002 A 2,4-dinitrophenol ...... 34616 51–28–5 059 A 005 A 2-methyl-4,6-dinitrophenol ...... 34657 534–52–1 060 A 004 A 2-nitrophenol ...... 34591 88–75–5 057 A 006 A 4-nitrophenol ...... 34646 100–02–7 058 A 007 A Pentachlorophenol ...... 39032 87–86–5 064 A 009 A 2,3,6-trichlorophenol (4c) ...... 77688 93–37–55 530 A 2,4,5-trichlorophenol (4c) ...... 95–95–4 531 A 2,4,6-trichlorophenol ...... 34621 88–06–2 021 A 011 A

TABLE 3—GAS CHROMATOGRAPHY OF BASE/NEUTRAL EXTRACTABLE COMPOUNDS

Retention time Detec- EGD Compound tion No. 1 Mean EGD limit 2 (sec) Ref Relative (μg/L)

164 2,2′-difluorobiphenyl (int std) ...... 1163 164 1.000–1.000 10 061 N-nitrosodimethylamine ...... 385 164 ns 50 603 alpha picoline-d7...... 417 164 0.326–0.393 50 703 alpha picoline...... 426 603 1.006–1.028 50 610 styrene-d5 ...... 546 164 0.450–0.488 10 710 styrene ...... 549 610 1.002–1.009 10 613 p-cymene-d14 ...... 742 164 0.624–0.652 10 713 p-cymene ...... 755 613 1.008–1.023 10 265 phenol-d5 ...... 696 164 0.584–0.613 10 365 phenol ...... 700 265 0.995–1.010 10 218 bis(2-chloroethyl) ether-d8...... 696 164 0.584–0.607 10 318 bis(2-chloroethyl) ether...... 704 218 1.007–1.016 10 617 n-decane-d22 ...... 698 164 0.585–0.615 10 717 n-decane ...... 720 617 1.022–1.038 10 226 1,3-dichlorobenzene-d4 ...... 722 164 0.605–0.636 10 326 1,3-dichlorobenzene ...... 724 226 0.998–1.008 10 227 1,4-dichlorobenzene-d4 ...... 737 164 0.601–0.666 10 327 1,4-dichlorobenzene ...... 740 227 0.997–1.009 10 225 1,2-dichlorobenzene-d4 ...... 758 164 0.632–0.667 10 325 1,2-dichlorobenzene ...... 760 225 0.995–1.008 10 242 bis(2-chloroisopropyl) ether-d12...... 788 164 0.664–0.691 10 342 bis(2-chloroisopropyl) ether...... 799 242 1.010–1.016 10 212 hexachloroethane-13C ...... 819 164 0.690–0.717 10 312 hexachloroethane ...... 823 212 0.999–1.001 10 063 N-nitrosodi-n-propylamine ...... 830 164 ns 20 256 nitrobenzene-d5 ...... 845 164 0.706–0.727 10 356 nitrobenzene ...... 849 256 1.002–1.007 10 254 isophorone-d8 ...... 881 164 0.747–0.767 10 354 isophorone ...... 889 254 0.999–1.017 10 234 2,4-dimethyl phenol-d3...... 921 164 0.781–0.803 10 334 2,4-dimethylphenol ...... 924 234 0.999–1.003 10 043 bis(2-chloroethoxy) methane...... 939 164 ns 10 208 1,2,4-trichlorobenzene-d3 ...... 955 164 0.813–0.830 10 308 1,2,4-trichlorobenzene ...... 958 208 1.000–1.005 10 255 naphthalene-d8 ...... 963 164 0.819–0.836 10 355 naphthalene ...... 967 255 1.001–1.006 10 609 alpha-terpineol-d3 ...... 973 164 0.829–0.844 10

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TABLE 3—GAS CHROMATOGRAPHY OF BASE/NEUTRAL EXTRACTABLE COMPOUNDS—Continued

Retention time Detec- EGD Compound tion No. 1 Mean EGD limit 2 (sec) Ref Relative (μg/L)

709 alpha-terpineol ...... 975 609 0.998–1.008 10 606 n-dodecane-d26 ...... 953 164 0.730–0.908 10 706 n-dodecane ...... 981 606 0.986–1.051 10 529 1,2,3-trichlorobenzene ...... 1003 164 ns 10 252 hexachlorobutadiene-13C4 ...... 1005 164 0.856–0.871 10 352 hexachlorobutadiene ...... 1006 252 0.999–1.002 10 253 hexachlorocyclopentadiene-13C4 ...... 1147 164 0.976–0.986 10 353 hexachlorocyclopentadiene ...... 1142 253 0.999–1.001 10 220 2-chloronaphthalene-d7 ...... 1185 164 1.014–1.024 10 320 2-chloronaphthalene ...... 1200 220 0.997–1.007 10 518 n-tetradecane ...... 1203 164 ns 10 612 Biphenyl-d10 ...... 1205 164 1.016–1.027 10 712 Biphenyl ...... 1195 612 1.001–1.006 10 608 Diphenyl ether-d10...... 1211 164 1.036–1.047 10 708 Diphenyl ether...... 1216 608 0.997–1.009 10 277 Acenaphthylene-d8 ...... 1265 164 1.080–1.095 10 377 Acenaphthylene ...... 1247 277 1.000–1.004 10 271 Dimethyl phthalate-d4...... 1269 164 1.083–1.102 10 371 Dimethyl phthalate...... 1273 271 0.998–1.005 10 236 2,6-dinitrotoluene-d3 ...... 1283 164 1.090–1.112 10 336 2,6-dinitrotoluene ...... 1300 236 1.001–1.005 10 201 Acenaphthene-d10 ...... 1298 164 1.107–1.125 10 301 Acenaphthene ...... 1304 201 0.999–1.009 10 605 Dibenzofuran-d8 ...... 1331 164 1.134–1.155 10 705 Dibenzofuran ...... 1335 605 0.998–1.007 10 602 Beta-naphthylamine-d7 ...... 1368 164 1.163–1.189 50 702 Beta-naphthylamine ...... 1371 602 0.996–1.007 50 280 Fluorene-d10 ...... 1395 164 1.185–1.214 10 380 Fluorene ...... 1401 281 0.999–1.008 10 240 4-chlorophenyl phenyl ether-d5 ...... 1406 164 1.194–1.223 10 340 4-chlorophenyl phenyl ether ...... 1409 240 0.990–1.015 10 270 Diethyl phthalate-d4...... 1409 164 1.197–1.229 10 370 Diethyl phthalate...... 1414 270 0.996–1.006 10 619 n-hexadecane-d34 ...... 1447 164 1.010–1.478 10 719 n-hexadecane ...... 1469 619 1.013–1.020 10 235 2,4-dinitrotoluene-d3 ...... 1359 164 1.152–1.181 10 335 2,4-dinitrotoluene ...... 1344 235 1.000–1.002 10 237 1,2-diphenylhydrazine-d8 ...... 1433 164 1.216–1.248 20 337 1,2-diphenylhydrazine (3) ...... 1439 237 0.999–1.009 20 607 Diphenylamine-d10 ...... 1437 164 1.213–1.249 20 707 Diphenylamine ...... 1439 607 1.000–1.007 20 262 N-nitrosodiphenylamine-d6 ...... 1447 164 1.225–1.252 20 362 N-nitrosodiphenylamine (4) ...... 1464 262 1.000–1.002 20 041 4-bromophenyl phenyl ether ...... 1498 164 1.271–1.307 10 209 Hexachlorobenzene-13C6 ...... 1521 164 1.288–1.327 10 309 Hexachlorobenzene ...... 1522 209 0.999–1.001 10 281 Phenanthrene-d10 ...... 1578 164 1.334–1.380 10 520 n-octadecane ...... 1580 164 ns 10 381 Phenanthrene ...... 1583 281 1.000–1.005 10 278 Anthracene-d10 ...... 1588 164 1.342–1.388 10 378 Anthracene ...... 1592 278 0.998–1.006 10 604 Dibenzothiophene-d8 ...... 1559 164 1.314–1.361 10 704 Dibenzothiophene ...... 1564 604 1.000–1.006 10 528 Carbazole ...... 1650 164 ns 20 621 n-eicosane-d42 ...... 1655 164 1.184–1.662 10 721 n-eicosane ...... 1677 621 1.010–1.021 10 268 Di-n-butyl phthalate-d4...... 1719 164 1.446–1.510 10 368 Di-n-butyl phthalate...... 1723 268 1.000–1.003 10 239 Fluoranthene-d10 ...... 1813 164 1.522–1.596 10 339 Fluoranthene ...... 1817 239 1.000–1.004 10 284 Pyrene-d10 ...... 1844 164 1.523–1.644 10 384 Pyrene ...... 1852 284 1.001–1.003 10 205 Benzidine-d8 ...... 1854 164 1.549–1.632 50 305 Benzidine ...... 1853 205 1.000–1.002 50 522 n-docosane ...... 1889 164 ns 10 623 n-tetracosane-d50 ...... 1997 164 1.671–1.764 10 723 n-tetracosane ...... 2025 612 1.012–1.015 10 067 Butylbenzyl phthalate...... 2060 164 ns 10 276 Chrysene-d12 ...... 2081 164 1.743–1.837 10 376 Chrysene ...... 2083 276 1.000–1.004 10

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TABLE 3—GAS CHROMATOGRAPHY OF BASE/NEUTRAL EXTRACTABLE COMPOUNDS—Continued

Retention time Detec- EGD Compound tion No. 1 Mean EGD limit 2 (sec) Ref Relative (μg/L)

272 Benzo(a)anthracene-d12 ...... 2082 164 1.735–1.846 10 372 Benzo(a)anthracene ...... 2090 272 0.999–1.007 10 228 3,3′-dichlorobenzidine-d6 ...... 2088 164 1.744–1.848 50 328 3,3′-dichlorobenzidine ...... 2086 228 1.000–1.001 50 266 Bis(2-ethylhexyl) phthalate-d4...... 2123 164 1.771–1.880 10 366 Bis(2-ethylhexyl) phthalate...... 2124 266 1.000–1.002 10 524 n-hexacosane ...... 2147 164 ns 10 269 di-n-octyl phthalate-d4...... 2239 164 1.867–1.982 10 369 di-n-octyl phthalate...... 2240 269 1.000–1.002 10 525 n-octacosane ...... 2272 164 ns 10 274 Benzo(b)fluoranthene-d12 ...... 2281 164 1.902–2.025 10 354 Benzo(b)fluoranthene ...... 2293 274 1.000–1.005 10 275 Benzo(k)fluoranthene-d12 ...... 2287 164 1.906–2.033 10 375 Benzo(k)fluoranthene ...... 2293 275 1.000–1.005 10 273 Benzo(a)pyrene-d12 ...... 2351 164 1.954–2.088 10 373 Benzo(a)pyrene ...... 2350 273 1.000–1.004 10 626 N-triacontane-d62 ...... 2384 164 1.972–2.127 10 726 N-triacontane ...... 2429 626 1.011–1.028 10 083 Indeno(1,2,3-cd)pyrene ...... 2650 164 ns 20 082 Dibenzo(a,h)anthracene ...... 2660 164 ns 20 279 Benzo(ghi)perylene-d12 ...... 2741 164 2.187–2.524 20 379 Benzo(ghi)perylene ...... 2750 279 1.001–1.006 20 1 Reference numbers beginning with 0, 1 or 5 indicate a pollutant quantified by the internal standard method; reference num- bers 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 This is a minimum level at which the entire GC/MS system must give recognizable mass spectra (background corrected) and acceptable calibration points. 3 Detected as azobenzene. 4 Detected as diphenylamine. ns = specification not available at time of release of method. Column: 30 ±2 m × 0.25 ±0.02 mm i.d. 94% methyl, 4% phenyl, 1% vinyl bonded phase fused silica capillary. Temperature program: 5 min at 30 °C; 30 – 280 °C at 8 °C per min; isothermal at 280 °C until benzo(ghi)perylene elutes. Gas velocity: 30 ±5 cm/sec.

TABLE 4—GAS CHROMATOGRAPHY OF ACID EXTRACTABLE COMPOUNDS

Retention time Detec- EGD Compound tion No. 1 Mean EGD limit 2 (sec) Ref Relative (μg/L)

164 2,2′-difluorobiphenyl (int std) ...... 1163 164 1.000–1.000 10 224 2-chlorophenol-d4 ...... 701 164 0.587–0.618 10 324 2-chlorophenol ...... 705 224 0.997–1.010 10 257 2-nitrophenol-d4 ...... 898 164 0.761–0.783 20 357 2-nitrophenol ...... 900 257 0.994–1.009 20 231 2,4-dichlorophenol-d3 ...... 944 164 0.802–0.822 10 331 2,4-dichlorophenol ...... 947 231 0.997–1.006 10 222 4-chloro-3-methylphenol-d2 ...... 1086 164 0.930–0.943 10 322 4-chloro-3-methylphenol ...... 1091 222 0.998–1.003 10 221 2,4,6-trichlorophenol-d2 ...... 1162 164 0.994–1.005 10 321 2,4,6-trichlorophenol ...... 1165 221 0.998–1.004 10 531 2,4,5-trichlorophenol ...... 1170 164 ns 10 530 2,3,6-trichlorophenol ...... 1195 164 ns 10 259 2,4-dinitrophenol-d3 ...... 1323 164 1.127–1.149 50 359 2,4-dinitrophenol ...... 1325 259 1.000–1.005 50 258 4-nitrophenol-d4 ...... 1349 164 1.147–1.175 50 358 4-nitrophenol ...... 1354 258 0.997–1.006 50 260 2-methyl-4,6-dinitrophenol-d2 ...... 1433 164 1.216–1.249 20 360 2-methyl-4,6-dinitrophenol ...... 1435 260 1.000–1.002 20 264 Pentachlorophenol-13C6 ...... 1559 164 1.320–1.363 50 364 Pentachlorophenol ...... 1561 264 0.998–1.002 50 1 Reference numbers beginning with 0, 1 or 5 indicate a pollutant quantified by the internal standard method; reference num- bers 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 This is a minimum level at which the entire GC/MS system must give recognizable mass spectra (background corrected) and acceptable calibration points. ns = specification not available at time of release of method. Column: 30 ±2m × 0.25 ±0.02mm i.d. 94% methyl, 4% phenyl, 1% vinyl bonded phase fused silica capillary. Temperature program: 5 min at 30 °C; 8 °C/min. to 250 °C or until pentachlorophenol elutes. Gas velocity: 30 ±5 cm/sec.

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TABLE 5—DFTPP MASS INTENSITY TABLE 6—BASE/NEUTRAL EXTRACTABLE COM- SPECIFICATIONS POUND CHARACTERISTIC MASSES—Continued

Mass Intensity required Labeled Primary m/ Compound analog z 51 30–60 percent of mass 198. 68 Less than 2 percent of mass 69. 1,4-dichlorobenzene ...... d4 146 /152 70 Less than 2 percent of mass 69. 3,3′-dichlorobenzidine ...... d6 252 /258 127 40–60 percent of mass 198. Diethyl phthalate ...... d4 149/153 197 Less than 1 percent of mass 198. 2,4-dimethylphenol ...... d3 122/125 199 5–9 percent of mass 198. Dimethyl phthalate ...... d4 163 /167 275 10–30 percent of mass 198. 2,4-dinitrotoluene ...... d3 164/168 365 greater than 1 percent of mass 198 2,6-dinitrotoluene ...... d3 165/167 441 present and less than mass 443 Di-n-octyl phthalate ...... d4 149 /153 442 40–100 percent of mass 198. Diphenylamine ...... d10 169/179 443 17–23 percent of mass 442. Diphenyl ether ...... d10 170/180 1,2-diphenylhydrazine 1 ...... d10 77/82 TABLE 6—BASE/NEUTRAL EXTRACTABLE Fluoranthene ...... d10 202/212 Fluorene ...... d10 166/176 COMPOUND CHARACTERISTIC MASSES Hexachlorobenzene ...... 13C6 284 /292 Hexachlorobutadiene ...... 13C4 225 /231 Labeled Primary m/ Hexachloroethane ...... 13C 201 /204 Compound analog z Hexachlorocyclopentadiene ...... 13C4 237 /241 Acenaphthene ...... d10 154/164 Ideno(1,2,3-cd)pyrene ...... 276 Acenaphthylene ...... d8 152 /160 Isophorone ...... d8 82 /88 Anthracene ...... d10 178 /188 Naphthalene ...... d8 128 /136 Benzidine ...... d8 184 /192 B-naphthylamine ...... d7 143/150 Benzo(a)anthracene ...... d12 228/240 Nitrobenzene ...... d5 123 /128 Benzo(b)fluoranthene ...... d12 252/264 N-nitrosodimethylamine ...... 74 Benzo(k)fluoranthene ...... d12 252/264 N-nitrosodi-n-propylamine ...... 70 Benzo(a)pyrene ...... d12 252 /264 N-nitrosodiphenylamile 2 ...... d6 169/175 Benzo(ghi)perylene ...... d12 276/288 Phenanthrene ...... d10 178/188 Biphenyl ...... d10 154 /164 Phenol ...... d5 94 /71 Bis(2-chloroethyl) ether ...... d8 93 /101 a-picoline ...... d7 93/100 Bis(2-chloroethoxy)methane ...... 93 Pyrene ...... d10 202 /212 Bis(2-chloroisopropyl) ether ...... d12 121 /131 Styrene ...... d5 104/109 Bis(2-ethylhexyl) phthalate ...... d4 149/153 a-terpineol ...... d3 59 /62 4-bromophenyl phenyl ether ...... 248 1,2,3-trichlorobenzene ...... d3 180/183 Butyl benzyl phthalate ...... 149 1,2,4-trichlorobenzene ...... d3 180/183 n-C10 ...... d22 55/66 1 n-C12 ...... d26 55/66 Detected as azobenzene. 2 n-C14 ...... 55 Detected as diphenylamine. n-C16 ...... d34 55/66 n-C18 ...... 55 TABLE 7—ACID EXTRACTABLE COMPOUND n-C20 ...... d42 55/66 CHARACTERISTIC MASSES n-C22 ...... 55 n-C24 ...... d50 55/66 Labeled Primary m/ n-C26 ...... 55 Compound analog z n-C28 ...... 55 n-C30 ...... d62 55/66 4-chloro-3-methylphenol ...... d2 107 /109 Carbazole ...... d8 167 /175 2-chlorophenol ...... d4 128 /132 2-chloronaphthalene ...... d7 162/169 2,4-dichlorophenol ...... d3 162/167 4-chlorophenyl phenyl ether ...... d5 204/209 2,4-dinitrophenol ...... d3 184/187 Chrysene ...... d12 228/240 2-methyl-4,6-dinitrophenol ...... d2 198/200 p-cymene ...... d14 114/130 2-nitrophenol ...... d4 139/143 Dibenzo(a,h)anthracene ...... 278 4-nitrophenol ...... d4 139/143 Dibenzofuran ...... d8 168 /176 Pentachlorophenol ...... 13C6 266/272 Dibenzothiophene ...... d8 184 /192 2,3,6-trichlorophenol ...... d2 196/200 Di-n-butyl phthalate ...... d4 149 /153 2,4,5-trichlorophenol ...... d2 196/200 1,2-dichlorobenzene ...... d4 146 /152 2,4,6-trichlorophenol ...... d2 196/200 1,3-dichlorobenzene ...... d4 146 /152

TABLE 8—ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS

Acceptance criteria Initial precision and ac- Labeled com- EGD Compound curacy section 8.2.3 pound recov- Calibration On-going No. 1 μ verification accuracy ( g/L) ery sec. 8.3 sec. 12.5 sec. 11.6 R and 14.2 P μ μ s X (percent) ( g/mL) ( g/L)

301 Acenaphthene ...... 21 79–134 ...... 80–125 72–144 201 Acenaphthene-d10...... 38 38–147 20–270 71–141 30–180 377 Acenaphtylene ...... 38 69–186 ...... 60–166 61–207 277 Acenaphthylene-d8...... 31 38–146 23–239 66–152 33–168

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TABLE 8—ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS—Continued

Acceptance criteria Initial precision and ac- Labeled com- EGD Compound curacy section 8.2.3 pound recov- Calibration On-going No. 1 μ verification accuracy ( g/L) ery sec. 8.3 sec. 12.5 sec. 11.6 R and 14.2 P μ μ s X (percent) ( g/mL) ( g/L)

378 Anthracene ...... 41 58–174 ...... 60–168 50–199 278 Anthracene-d10 ...... 49 31–194 14–419 58–171 23–242 305 Benzidine ...... 119 16–518 ...... 34–296 11–672 205 Benzidine-d8 ...... 269 ns-ns ns-ns ns-ns ns-ns 372 Benzo(a)anthracene ...... 20 65–168 ...... 70–142 62–176 272 Benzo(a)anthracene-d12 ...... 41 25–298 12–605 28–357 22–329 374 Benzo(b)fluoranthene ...... 183 32–545 ...... 61–164 20–ns 274 Benzo(b)fluoranthene-d12 ...... 168 11–577 ns-ns 14–ns ns-ns 375 Benzo(k)fluoranthene ...... 26 59–143 ...... 13–ns 53–155 275 Benzo(k)fluoranthene-d12 ...... 114 15–514 ns-ns 13–ns ns–685 373 Benzo(a)pyrene ...... 26 62–195 ...... 78–129 59–206 273 Benzo(a)pyrene-d12 ...... 24 35–181 21–290 12–ns 32–194 379 Benzo(ghi)perylene ...... 21 72–160 ...... 69–145 58–168 279 Benzo(ghi)perylene-d12 ...... 45 29–268 14–529 13–ns 25–303 712 Biphenyl (Appendix C) ...... 41 75–148 ...... 58–171 62–176 612 Biphenyl-d12 ...... 43 28–165 ns-ns 52–192 17–267 318 Bis(2-chloroethyl) ether...... 34 55–196 ...... 61–164 50–213 218 Bis(2-chloroethyl) ether-d8...... 33 29–196 15–372 52–194 25–222 043 Bis(2-chloroethoxy)methane* ...... 27 43–153 ...... 44–228 39–166 342 Bis(2-chloroisopropyl) ether...... 17 81–138 ...... 67–148 77–145 242 Bis(2-chloroisopropyl)ether-d12 ...... 27 35–149 20–260 44–229 30–169 366 Bis(2-ethylhexyl) phthalate...... 31 69–220 ...... 76–131 64–232 266 Bis(2-ethylhexyl) phthalate-d4...... 29 32–205 18–364 43–232 28–224 041 4-bromophenyl phenyl ether* ...... 44 44–140 ...... 52–193 35–172 067 Butyl benzyl phthalate* ...... 31 19–233 ...... 22–450 35–170 717 n-C10 (Appendix C) ...... 51 24–195 ...... 42–235 19–237 617 n-C10-d22 ...... 70 ns–298 ns-ns 44–227 ns–504 706 n-C12 (Appendix C) ...... 74 35–369 ...... 60–166 29–424 606 n-C12-d26 ...... 53 ns–331 ns-ns 41–242 ns–408 518 n-C14 (Appendix C)* ...... 109 ns–985 ...... 37–268 ns-ns 719 n-C16 (Appendix C) ...... 33 80–162 ...... 72–138 71–181 619 n-C16-d34 ...... 46 37–162 18–308 54–186 28–202 520 n-C18 (Appendix C)* ...... 39 42–131 ...... 40–249 35–167 721 n-C20 (Appendix C) ...... 59 53–263 ...... 54–184 46–301 621 n-C20-d42 ...... 34 34–172 19–306 62–162 29–198 522 n-C22 (Appendix C)* ...... 31 45–152 ...... 40–249 39–195 723 n-C24 (Appendix C) ...... 11 80–139 ...... 65–154 78–142 623 n-C24-d50 ...... 28 27–211 15–376 50–199 25–229 524 n-C26 (Appendix C)* ...... 35 35–193 ...... 26–392 31–212 525 n-C28 (Appendix C)* ...... 35 35–193 ...... 26–392 31–212 726 n-C30 (Appendix C) ...... 32 61–200 ...... 66–152 56–215 626 n-C30-d62 ...... 41 27–242 13–479 24–423 23–274 528 Carbazole (4c)*...... 38 36–165 ...... 44–227 31–188 320 2-chloronaphthalene ...... 100 46–357 ...... 58–171 35–442 220 2-chloronaphthalene-d7 ...... 41 30–168 15–324 72–139 24–204 322 4-chloro-3-methylphenol ...... 37 76–131 ...... 85–115 62–159 222 4-chloro-3-methylphenol-d2 ...... 111 30–174 ns–613 68–147 14–314 324 2-chlorophenol ...... 13 79–135 ...... 78–129 76–138 224 2-chlorophenol-d4 ...... 24 36–162 23–255 55–180 33–176 340 4-chlorophenyl phenyl ether ...... 42 75–166 ...... 71–142 63–194 240 4-chlorophenyl phenyl ether-d5 ...... 52 40–161 19–325 57–175 29–212 376 Chrysene ...... 51 59–186 ...... 70–142 48–221 276 Chrysene-d12...... 69 33–219 13–512 24–411 23–290 713 p-cymene (Appendix C) ...... 18 76–140 ...... 79–127 72–147 613 p-cymene-d14 ...... 67 ns–359 ns-ns 66–152 ns–468 082 Dibenzo(a,h)anthracene* ...... 55 23–299 ...... 13–761 19–340 705 Dibenzofuran (Appendix C) ...... 20 85–136 ...... 73–136 79–146 605 Dibenzofuran-d8 ...... 31 47–136 28–220 66–150 39–160 704 Dibenzothiophene (Synfuel)...... 31 79–150 ...... 72–140 70–168 604 Dibenzothiophene-d8 ...... 31 48–130 29–215 69–145 40–156 368 Di-n-butyl phthalate...... 15 76–165 ...... 71–142 74–169 268 Di-n-butyl phthalate-d4...... 23 23–195 13–346 52–192 22–209 325 1,2-dichlorobenzene ...... 17 73–146 ...... 74–135 70–152 225 1,2-dichlorobenzene-d4 ...... 35 14–212 ns–494 61–164 11–247 326 1,3-dichlorobenzene ...... 43 63–201 ...... 65–154 55–225 226 1,3-dichlorobenzene-d4 ...... 48 13–203 ns–550 52–192 ns–260 327 1,4-dichlorobenzene ...... 42 61–194 ...... 62–161 53–219

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TABLE 8—ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS—Continued

Acceptance criteria Initial precision and ac- Labeled com- EGD Compound curacy section 8.2.3 pound recov- Calibration On-going No. 1 μ verification accuracy ( g/L) ery sec. 8.3 sec. 12.5 sec. 11.6 R and 14.2 P μ μ s X (percent) ( g/mL) ( g/L)

227 1,4-dichlorobenzene-d4 ...... 48 15–193 ns–474 65–153 11–245 328 3,3′-dichlorobenzidine ...... 26 68–174 ...... 77–130 64–185 228 3,3′-dichlorobenzidine-d6 ...... 80 ns–562 ns-ns 18–558 ns-ns 331 2,4-dichlorophenol ...... 12 85–131 ...... 67–149 83–135 231 2,4-dichlorophenol-d3 ...... 28 38–164 24–260 64–157 34–182 370 Diethyl phthalate...... 44 75–196 ...... 74–135 65–222 270 Diethyl phthalate-d4...... 78 ns–260 ns-ns 47–211 ns-ns 334 2,4-dimethylphenol ...... 13 62–153 ...... 67–150 60–156 234 2,4-dimethylphenol-d3 ...... 22 15–228 ns–449 58–172 14–242 371 Dimethyl phthalate...... 36 74–188 ...... 73–137 67–207 271 Dimethyl phthalate-d4...... 108 ns–640 ns-ns 50–201 ns-ns 359 2,4-dinitrophenol ...... 18 72–134 ...... 75–133 68–141 259 2,4-dinitrophenol-d3 ...... 66 22–308 ns-ns 39–256 17–378 335 2,4-dinitrotoluene ...... 18 75–158 ...... 79–127 72–164 235 2,4-dinitrotoluene-d3 ...... 37 22–245 10–514 53–187 19–275 336 2,6-dinitrotoluene ...... 30 80–141 ...... 55–183 70–159 236 2,6-dinitrotoluene-d3 ...... 59 44–184 17–442 36–278 31–250 369 Di-n-octyl phthalate...... 16 77–161 ...... 71–140 74–166 269 Di-n-octyl phthalate-d4...... 46 12–383 ns-ns 21–467 10–433 707 Diphenylamine (Appendix C) ...... 45 58–205 ...... 57–176 51–231 607 Diphenylamine-d10 ...... 42 27–206 11–488 59–169 21–249 708 Diphenyl ether (Appendix C) ...... 19 82–136 ...... 83–120 77–144 608 Diphenyl ether-d10...... 37 36–155 19–281 77–129 29–186 337 1,2-diphenylhydrazine ...... 73 49–308 ...... 75–134 40–360 237 1,2-diphenylhydrazine-d10...... 35 31–173 17–316 58–174 26–200 339 Fluoranthene ...... 33 71–177 ...... 67–149 64–194 239 Fluoranthene-d10...... 35 36–161 20–278 47–215 30–187 380 Fluorene ...... 29 81–132 ...... 74–135 70–151 280 Fluorene-d10...... 43 51–131 27–238 61–164 38–172 309 Hexachlorobenzene ...... 16 90–124 ...... 78–128 85–132 209 Hexachlorobenzene-13C6...... 81 36–228 13–595 38–265 23–321 352 hexachlorobutadiene ...... 56 51–251 ...... 74–135 43–287 252 hexachlorobutadiene-13C4 ...... 63 ns–316 ns-ns 68–148 ns–413 312 hexachloroethane ...... 227 21–ns ...... 71–141 13–ns 212 hexachloroethane-13C1 ...... 77 ns–400 ns-ns 47–212 ns–563 353 hexachlorocyclopentadiene ...... 15 69–144 ...... 77–129 67–148 253 hexachlorocyclopentadiene-13C4 ...... 60 ns-ns ns-ns 47–211 ns-ns 083 ideno(1,2,3-cd)pyrene* ...... 55 23–299 ...... 13–761 19–340 354 isophorone ...... 25 76–156 ...... 70–142 70–168 254 isophorone-d8 ...... 23 49–133 33–193 52–194 44–147 360 2-methyl-4,6-dinitrophenol ...... 19 77–133 ...... 69–145 72–142 260 2-methyl-4,6-dinitrophenol-d2...... 64 36–247 16–527 56–177 28–307 355 naphthalene ...... 20 80–139 ...... 73–137 75–149 255 naphthalene-d8 ...... 39 28–157 14–305 71–141 22–192 702 B-naphthylamine (Appendix C) ...... 49 10–ns ...... 39–256 ns-ns 602 B-naphthylamine-d7 ...... 33 ns-ns ns-ns 44–230 ns-ns 356 nitrobenzene ...... 25 69–161 ...... 85–115 65–169 256 nitrobenzene-d5 ...... 28 18–265 ns-ns 46–219 15–314 357 2-nitrophenol ...... 15 78–140 ...... 77–129 75–145 257 2-nitrophenol-d4 ...... 23 41–145 27–217 61–163 37–158 358 4-nitrophenol ...... 42 62–146 ...... 55–183 51–175 258 4-nitrophenol-d4 ...... 188 14–398 ns-ns 35–287 ns-ns 061 N-nitrosodimethylamile* ...... 198 21–472 ...... 40–249 12–807 063 N-nitrosodi-n-proplyamine* ...... 198 21–472 ...... 40–249 12–807 362 N-nitrosodiphenylamine ...... 45 65–142 ...... 68–148 53–173 262 N-nitrosodiphenylamine-d6 ...... 37 54–126 26–256 59–170 40–166 364 pentachlorophenol ...... 21 76–140 ...... 77–130 71–150 264 pentachlorophenol-13C6...... 49 37–212 18–412 42–237 29–254 381 phenanthrene ...... 13 93–119 ...... 75–133 87–126 281 phenanthrene-d10 ...... 40 45–130 24–241 67–149 34–168 365 phenol ...... 36 77–127 ...... 65–155 62–154 265 phenol-d5 ...... 161 21–210 ns-ns 48–208 ns-ns 703 a-picoline (Synfuel)...... 38 59–149 ...... 60–165 50–174 603 a-picoline-d7 ...... 138 11–380 ns-ns 31–324 ns–608 384 pyrene ...... 19 76–152 ...... 76–132 72–159 284 pyrene-d10 ...... 29 32–176 18–303 48–210 28–196 710 styrene (Appendix C) ...... 42 53–221 ...... 65–153 48–244

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TABLE 8—ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS—Continued

Acceptance criteria Initial precision and ac- Labeled com- EGD Compound curacy section 8.2.3 pound recov- Calibration On-going No. 1 μ verification accuracy ( g/L) ery sec. 8.3 sec. 12.5 sec. 11.6 R and 14.2 P μ μ s X (percent) ( g/mL) ( g/L)

610 styrene-d5 ...... 49 ns–281 ns-ns 44–228 ns–348 709 a-terpineol (Appendix C) ...... 44 42–234 ...... 54–186 38–258 609 a-terpineol-d3 ...... 48 22–292 ns–672 20–502 18–339 529 1,2,3-trichlorobenzene (4c)*...... 69 15–229 ...... 60–167 11–297 308 1,2,4-trichlorobenzene ...... 19 82–136 ...... 78–128 77–144 208 1,2,4-trichlorobenzene-d3 ...... 57 15–212 ns–592 61–163 10–282 530 2,3,6-trichlorophenol (4c)*...... 30 58–137 ...... 56–180 51–153 531 2,4,5-trichlorophenol (4c)*...... 30 58–137 ...... 56–180 51–153 321 2,4,6-trichlorophenol ...... 57 59–205 ...... 81–123 48–244 221 2,4,6-trichlorophenol-d2...... 47 43–183 21–363 69–144 34–226

1 Reference numbers beginning with 0, 1 or 5 indicate a pollutant quantified by the internal standard method; reference num- bers 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. * Measured by internal standard; specification derived from related compound. ns = no specification; limit is outside the range that can be measured reliably.

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ATTACHMENT 1 TO METHOD 1625 Section 6.12 The solutions for obtaining au- thentic mass spectra are to include all INTRODUCTION additional analytes listed in Tables 1 and To support measurement of several 2 of this attachment. semivolatile pollutants, EPA has developed Section 6.13 The calibration solutions are this attachment to EPA Method 1625B. 1 The modified to include the analytes listed in modifications listed in this attachment are Tables 1 and 2 and the labeled compounds approved only for monitoring wastestreams listed in Tables 5 and 6 of this attach- from the Centralized Waste Treatment Point ment. Source Category (40 CFR part 437) and the Section 6.14 The precision and recovery Landfills Point Source Category (40 CFR standard is modified to include the part 445). EPA Method 1625B (the Method) analytes listed in Tables 1 and 2 and the employs sample extraction with methylene labeled compounds listed in Tables 5 and chloride followed by analysis of the extract 6 of this attachment. using capillary column gas chromatography- Section 6.15 The solutions containing the mass spectrometry (GC/MS). This attach- additional analytes listed in Tables 1 and ment addresses the addition of the 2 of this attachment are to be analyzed semivolatile pollutants listed in Tables 1 and for stability. 2 to all applicable standard, stock, and spik- Section 7.2.1 This section is modified to in- ing solutions utilized for the determination clude the analytes listed in Tables 1 and of semivolatile organic compounds by EPA 2 and the labeled compounds listed in Ta- Method 1625B. bles 5 and 6 of this attachment. 1.0 EPA METHOD 1625 REVISION B Section 7.4.5 This section is modified to in- MODIFICATION SUMMARY clude the analytes listed in Tables 1 and 2 and the labeled compounds listed in Ta- The additional semivolatile organic com- bles 5 and 6 in the calibration. pounds listed in Tables 1 and 2 are added to Section 8.2 The initial precision and recov- all applicable calibration, spiking, and other ery (IPR) requirements are modified to solutions utilized in the determination of include the analytes listed in Tables 1 semivolatile compounds by EPA Method and 2 and the labeled compounds listed in 1625. The instrument is to be calibrated with Tables 5 and 6 of this attachment. Addi- these compounds, and all procedures and tional IPR performance criteria are sup- quality control tests described in the Method plied in Table 7 of this attachment. must be performed. Section 8.3 The labeled compounds listed in 2.0 SECTION MODIFICATIONS Tables 3 and 4 of this attachment are to be included in the method performance NOTE: All section and figure numbers in tests. Additional method performance this Attachment reference section and figure criteria are supplied in Table 7 of this at- numbers in EPA Method 1625 Revision B un- tachment. less noted otherwise. Sections not listed here Section 8.5.2 The acceptance criteria for remain unchanged. blanks includes the analytes listed in Ta- Section 6.7 The stock standard solutions de- bles 1 and 2 of this attachment. scribed in this section are modified such that the analytes in Tables 1 and 2 of this Section 10.1.2 The labeled compound solu- attachment are required in addition to tion must include the labeled compounds those specified in the Method. listed in Tables 5 and 6 of this attach- Section 6.8 The labeled compound spiking ment. solution in this section is modified to in- Section 10.1.3 The precision and recovery clude the labeled compounds listed in Ta- standard must include the analytes list- bles 5 and 6 of this attachment. ed in Tables 1 and 2 and the labeled com- Section 6.9 The secondary standard is modi- pounds listed in Tables 5 and 6 of this at- fied to include the additional analytes tachment. listed in Tables 1 and 2 of this attach- Section 12.5 Additional QC requirements for ment. calibration verification are supplied in Table 7 of this attachment. 1 EPA Method 1625 Revision B, Section 12.7 Additional QC requirements for Semivolatile Organic Compounds by Isotope ongoing precision and recovery are sup- Dilution GC/MS, 40 CFR part 136, appendix A. plied in Table 7 of this attachment.

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TABLE 1—BASE/NEUTRAL EXTRACTABLE COMPOUNDS

Pollutant Compound CAS Registry EPA-EGD

acetophenone 1 ...... 98–86–2 758 aniline 2 ...... 62–53–3 757 -2,3-dichloroaniline 1 ...... 608–27–5 578 -o-cresol 1 ...... 95–48–7 771 pyridine 2 ...... 110–86–1 1330 CAS = Chemical Abstracts Registry. EGD = Effluent Guidelines Division. 1 Analysis of this pollutant is approved only for the Centralized Waste Treatment industry. 2 Analysis of this pollutant is approved only for the Centralized Waste Treatment and Landfills industries.

TABLE 2—ACID EXTRACTABLE COMPOUNDS

Pollutant Compound CAS Registry EPA-EGD

p-cresol 1 ...... 106–44–5 1744 CAS = Chemical Abstracts Registry. EGD = Effluent Guidelines Division. 1 Analysis of this pollutant is approved only for the Centralized Waste Treatment and Landfills industries.

TABLE 3—GAS CHROMATOGRAPHY 1 OF BASE/NEUTRAL EXTRACTABLE COMPOUNDS

2 Retention time Minimum EGD No. Compound level 3 Mean μ (sec) EGD Ref Relative ( g/L)

758 ...... acetophenone 4 ...... 818 658 1.003–1.005 10 757 ...... aniline 5 ...... 694 657 0.994–1.023 10 578 ...... 2,3-dichloroaniline 4 ...... 1160 164 1.003–1.007 10 771 ...... o-cresol 4 ...... 814 671 1.005–1.009 10 1330 ...... pyridine 5 ...... 378 1230 1.005–1.011 10 EGD = Effluent Guidelines Division. 1 The data presented in this table were obtained under the chromatographic conditions given in the footnote to Table 3 of EPA Method 1625B. 2 Retention times are approximate and are intended to be consistent with the retention times for the analytes in EPA Method 1625B. 3 See the definition in footnote 2 to Table 3 of EPA Method 1625B. 4 Analysis of this pollutant is approved only for the Centralized Waste Treatment industry. 5 Analysis of this pollutant is approved only for the Centralized Waste Treatment and Landfills industries.

TABLE 4—GAS CHROMATOGRAPHY 1 OF ACID EXTRACTABLE COMPOUNDS

2 Retention time Minimum EGD No. Compound level Mean μ 3 (sec) EGD Ref Relative ( /L)

1744 ...... p-cresol 4 ...... 834 1644 1.004–1.008 20 EGD = Effluent Guidelines Division. 1 The data presented in this table were obtained under the chromatographic conditions given in the footnote to Table 4 of EPA Method 1625B. 2 Retention times are approximate and are intended to be consistent with the retention times for the analytes in EPA Method 1625B. 3 See the definition in footnote 2 to Table 4 of EPA Method 1625B. 4 Analysis of this pollutant is approved only for the Centralized Waste Treatment and Landfills industries.

TABLE 5—BASE/NEUTRAL EXTRACTABLE COMPOUND CHARACTERISTIC M/Z’S

Compound Labeled Ana- Primary log m/z 1

2 acetophenone ...... d5 105/110 3 aniline ...... d7 93/100 2 o-cresol ...... d7 108/116 2,3-dichloroaniline 2 ...... n/a 161 3 pyridine ...... d5 79/84 m/z = mass to charge ratio.

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1 Native/labeled. 2 Analysis of this pollutant is approved only for the Centralized Waste Treatment industry. 3 Analysis of this pollutant is approved only for the Centralized Waste Treatment and Landfills industries.

TABLE 6—ACID EXTRACTABLE COMPOUND CHARACTERISTIC M/Z’S

Compound Labeled Ana- Primary log m/z 1

2 p-cresol ...... d7 108/116 m/z = mass to charge ratio. 1 Native/labeled. 2 Analysis of this pollutant is approved only for the Centralized Waste Treatment and Landfills industries.

TABLE 7—ACCEPTANCE CRITERIA FOR PERFORMANCE TESTS

Acceptance criteria Initial precision and accu- Labeled Calibration On-going racy section 8.2 compound verification accuracy EGD No. Compound (μg/L) recovery sec. 12.5 sec. 12.7 R sec. 8.3 and μg/mL) (μg/L) s 14.2 P (μg/L) X (percent)

758 ...... acetophenone 1 ...... 34 44–167 ...... 85–115 45–162 1 658 ...... acetophenone-d 5 ...... 51 23–254 45–162 85–115 22–264 757 ...... aniline 2 ...... 32 30–171 ...... 85–115 33–154 2 657 ...... aniline-d 7 ...... 71 15–278 33–154 85–115 12–344 771 ...... o-cresol 1 ...... 40 31–226 ...... 85–115 35–196 1 671 ...... o-cresol-d 7 ...... 23 30–146 35–196 85–115 31–142 1744 ...... p-cresol 2 ...... 59 54–140 ...... 85–115 37–203 2 1644 ...... p-cresol-d7 ...... 22 11–618 37–203 85–115 16–415 578 ...... 2,3-dichloroaniline 1 ...... 13 40–160 ...... 85–115 44–144 1330 ...... pyridine 2 ...... 28 10–421 ...... 83–117 18–238 2 1230 ...... pyridine-d 5 ...... ns 7–392 19–238 85–115 4–621 s = Standard deviation of four recovery measurements. X = Average recovery for four recovery measurements. EGD = Effluent Guidelines Division. ns = no specification; limit is outside the range that can be measured reliably. 1 Analysis of this pollutant is approved only for the Centralized Waste Treatment industry. 2 Analysis of this pollutant is approved only for the Centralized Waste Treatment and Landfills industries.

[49 FR 43261, Oct. 26, 1984; 50 FR 692, 695, Jan. 4, 1985, as amended at 51 FR 23702, June 30, 1986; 62 FR 48405, Sept. 15, 1997; 65 FR 3044, Jan. 19, 2000; 65 FR 81295, 81298, Dec. 22, 2000; 82 FR 40875, Aug. 28, 2017]

APPENDIX B TO PART 136—DEFINITION laboratory be included in the determination AND PROCEDURE FOR THE DETER- of the method detection limit. MINATION OF THE METHOD DETEC- (2) The MDL procedure is not applicable to methods that do not produce results with a TION LIMIT—REVISION 2 continuous distribution, such as, but not limited to, methods for whole effluent tox- Definition icity, presence/absence methods, and micro- The method detection limit (MDL) is de- biological methods that involve counting colonies. The MDL procedure also is not ap- fined as the minimum measured concentra- plicable to measurements such as, but not tion of a substance that can be reported with limited to, biochemical oxygen demand, 99% confidence that the measured concentra- color, pH, specific conductance, many titra- tion is distinguishable from method blank tion methods, and any method where low- results. level spiked samples cannot be prepared. Ex- cept as described in the addendum, for the I. Scope and Application purposes of this procedure, ‘‘spiked samples’’ (1) The MDL procedure is designed to be a are prepared from a clean reference matrix, straightforward technique for estimation of such as reagent water, spiked with a known the detection limit for a broad variety of and consistent quantity of the analyte. MDL physical and chemical methods. The proce- determinations using spiked samples may dure requires a complete, specific, and well- not be appropriate for all gravimetric meth- defined analytical method. It is essential ods (e.g., residue or total suspended solids), that all sample processing steps used by the but an MDL based on method blanks can be determined in such instances.

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II. Procedure available. (The rationale for removal of spe- cific outliers must be documented and main- (1) Estimate the initial MDL using one or tained on file with the results of the MDL more of the following: determination.) (a) The mean determined concentration (i) If there are multiple instruments that plus three times the standard deviation of a will be assigned the same MDL, then the set of method blanks. sample analyses must be distributed across (b) The concentration value that cor- all of the instruments. responds to an instrument signal-to-noise (ii) A minimum of two spiked samples and ratio in the range of 3 to 5. two method blank samples prepared and ana- (c) The concentration equivalent to three lyzed on different calendar dates is required times the standard deviation of replicate in- for each instrument. Each analytical batch strumental measurements of spiked blanks. may contain one spiked sample and one (d) That region of the calibration where method blank sample run together. A spiked there is a significant change in sensitivity, sample and a method blank sample may be i.e., a break in the slope of the calibration. analyzed in the same batch, but are not re- (e) Instrumental limitations. quired to be. (f) Previously determined MDL. (iii) The same prepared extract may be NOTE: It is recognized that the experience analyzed on multiple instruments so long as of the analyst is important to this process. the minimum requirement of seven prepara- However, the analyst should include some or tions in at least three separate batches is all of the above considerations in the initial maintained. estimate of the MDL. (c) Evaluate the spiking level: If any result (2) Determine the initial MDL. for any individual analyte from the spiked samples does not meet the method quali- NOTE: The Initial MDL is used when the tative identification criteria or does not pro- laboratory does not have adequate data to vide a numerical result greater than zero, perform the Ongoing Annual Verification then repeat the spiked samples at a higher specified in Section (4), typically when a new concentration. (Qualitative identification method is implemented or if a method was criteria are a set of rules or guidelines for es- rarely used in the last 24 months. tablishing the identification or presence of (a) Select a spiking level, typically 2—10 an analyte using a measurement system. times the estimated MDL in Section 1. Spik- Qualitative identification does not ensure ing levels in excess of 10 times the estimated that quantitative results for the analyte can detection limit may be required for analytes be obtained.) with very poor recovery (e.g., for an analyte (d) Make all computations as specified in with 10% recovery, spiked at 100 micrograms/ the analytical method and express the final L, with mean recovery of 10 micrograms/L; results in the method-specified reporting the calculated MDL may be around 3 units. micrograms/L. Therefore, in this example, (i) Calculate the sample standard deviation the spiking level would be 33 times the MDL, (S) of the replicate spiked sample measure- but spiking lower may result in no recovery ments and the sample standard deviation of at all). the replicate method blank measurements (b) Process a minimum of seven spiked from all instruments to which the MDL will samples and seven method blank samples be applied. through all steps of the method. The samples (ii) Compute the MDLs (the MDL based on used for the MDL must be prepared in at spiked samples) as follows: least three batches on three separate cal- MDL = t( ¥ ¥α= )S endar dates and analyzed on three separate S n 1, 1 0.99 s calendar dates. (Preparation and analysis Where: may be on the same day.) Existing data may MDLs = the method detection limit based on be used, if compliant with the requirements spiked samples for at least three batches, and generated t(n-1, 1¥α=0.99) = the Student’s t-value appro- within the last twenty four months. The priate for a single-tailed 99th percentile t most recent available data for method statistic and a standard deviation esti- blanks and spiked samples must be used. mate with n-1 degrees of freedom. See Statistical outlier removal procedures Addendum Table 1. should not be used to remove data for the Ss = sample standard deviation of the rep- initial MDL determination, since the total licate spiked sample analyses. number of observations is small and the pur- (iii) Compute the MDLb (the MDL based on pose of the MDL procedure is to capture rou- method blanks) as follows: tine method variability. However, docu- (A) If none of the method blanks give nu- mented instances of gross failures (e.g., in- merical results for an individual analyte, the strument malfunctions, mislabeled samples, MDLb does not apply. A numerical result in- cracked vials) may be excluded from the cal- cludes both positive and negative results, in- culations, provided that at least seven cluding results below the current MDL, but spiked samples and seven method blanks are not results of ‘‘ND’’ (not detected) commonly

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observed when a peak is not present in (b) Ensure that at least seven spiked sam- chromatographic analysis. ples and seven method blanks are completed (B) If some (but not all) of the method for the annual verification. If only one in- blanks for an individual analyte give numer- strument is in use, a minimum of seven ical results, set the MDLb equal to the high- spikes are still required, but they may be est method blank result. If more than 100 drawn from the last two years of data collec- method blanks are available, set MDLb to tion. the level that is no less than the 99th per- (c) At least once per year, re-evaluate the centile of the method blank results. For ‘‘n’’ spiking level. method blanks where n ≥ 100, sort the meth- (i) If more than 5% of the spiked samples od blanks in rank order. The (n * 0.99) ranked do not return positive numerical results that method blank result (round to the nearest meet all method qualitative identification whole number) is the MDLb. For example, to criteria, then the spiking level must be in- find MDLb from a set of 164 method blanks creased and the initial MDL re-determined where the highest ranked method blank re- following the procedure in section 2. sults are . . . 1.5, 1.7, 1.9, 5.0, and 10, then 164 (ii) [Reserved] × 0.99 = 162.36 which rounds to the 162nd (d) If the method is altered in a way that method blank result. Therefore, MDLb is 1.9 can be reasonably expected to change its sen- for n = 164 (10 is the 164th result, 5.0 is the sitivity, then re-determine the initial MDL 163rd result, and 1.9 is the 162nd result). Al- according to section 2, and the restart the ternatively, you may use spreadsheet algo- ongoing data collection. rithms to calculate the 99th percentile to in- (e) If a new instrument is added to a group terpolate between the ranks more precisely. of instruments whose data are being pooled (C) If all of the method blanks for an indi- to create a single MDL, analyze a minimum vidual analyte give numerical results, then of two spiked replicates and two method calculate the MDLb as: blank replicates on the new instrument. If both method blank results are below the ex- MDLb = X + tn¥1,1¥α=(0.99)Sb Where: isting MDL, then the existing MDLb is vali- dated. Combine the new spiked sample re- MDL = the MDL based on method blanks b sults to the existing spiked sample results X = mean of the method blank results (use and recalculate the MDL as in Section 4. If zero in place of the mean if the mean is s the recalculated MDL does not vary by more negative) s than the factor specified in section 4(f) of t( ¥ α= ) = the Student’s t-value appro- n 1, 1 0.99 this procedure, then the existing MDL is priate for the single-tailed 99th per- s validated. If either of these two conditions is centile t statistic and a standard devi- not met, then calculate a new MDL following ation estimate with n¥1 degrees of free- the instructions in section 2. dom. See Addendum Table 1. (4) Ongoing Annual Verification. S = sample standard deviation of the rep- b (a) At least once every thirteen months, licate method blank sample analyses. re-calculate MDLs and MDLb from the col- NOTE: If 100 or more method blanks are lected spiked samples and method blank re- available, as an option, MDLb may be set to sults using the equations in section 2. the concentration that is greater than or (b) Include data generated within the last equal to the 99th percentile of the method twenty four months, but only data with the blank results, as described in Section same spiking level. Only documented in- (2)(d)(iii)(B). stances of gross failures (e.g., instrument (e) Select the greater of MDLs or MDLb as malfunctions, mislabeled samples, cracked the initial MDL. vials) may be excluded from the calcula- (3) Ongoing Data Collection. tions. (The rationale for removal of specific (a) During any quarter in which samples outliers must be documented and maintained are being analyzed, prepare and analyze a on file with the results of the MDL deter- minimum of two spiked samples on each in- mination.) If the laboratory believes the sen- strument, in separate batches, using the sitivity of the method has changed signifi- same spiking concentration used in Section cantly, then the most recent data available 2. If any analytes are repeatedly not detected may be used, maintaining compliance with in the quarterly spiked sample analyses, or the requirement for at least seven replicates do not meet the qualitative identification in three separate batches on three separate criteria of the method (see section 2(c) of days (see section 2b). this procedure), then this is an indication (c) Include the initial MDL spiked samples, that the spiking level is not high enough and if the data were generated within twenty should be adjusted upward. Note that it is four months. not necessary to analyze additional method (d) Only use data associated with accept- blanks together with the spiked samples, the able calibrations and batch QC. Include all method blank population should include all routine data, with the exception of batches of the routine method blanks analyzed with that are rejected and the associated samples each batch during the course of sample anal- reanalyzed. If the method has been altered in ysis. a way that can be reasonably expected to

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change its sensitivity, then use only data ADDENDUM TO SECTION II: DETERMINATION OF collected after the change. THE MDL FOR A SPECIFIC MATRIX (e) Ideally, use all method blank results The MDL may be determined in a specific from the last 24 months for the MDLb cal- sample matrix as well as in reagent water. culation. The laboratory has the option to (1) Analyze the sample matrix to deter- use only the last six months of method blank mine the native (background) concentration data or the fifty most recent method blanks, of the analyte(s) of interest. whichever criteria yields the greater number (2) If the response for the native concentra- of method blanks. tion is at a signal-to-noise ratio of approxi- (f) The verified MDL is the greater of the mately 5–20, determine the matrix-specific

MDLs or MDLb. If the verified MDL is within MDL according to Section 2 but without 0.5 to 2.0 times the existing MDL, and fewer spiking additional analyte. than 3% of the method blank results (for the (3) Calculate MDLb using the method individual analyte) have numerical results blanks, not the sample matrix. above the existing MDL, then the existing (4) If the signal-to-noise ratio is less than MDL may optionally be left unchanged. Oth- 5, then the analyte(s) should be spiked into erwise, adjust the MDL to the new the sample matrix to obtain a concentration that will give results with a signal-to-noise verification MDL. (The range of 0.5 to 2.0 ap- ratio of approximately 10–20. proximates the 95th percentile confidence in- (5) If the analytes(s) of interest have sig- terval for the initial MDL determination nal-to-noise ratio(s) greater than approxi- with six degrees of freedom.) mately 20, then the resulting MDL is likely to be biased high.

TABLE 1—SINGLE-TAILED 99th PERCENTILE t STATISTIC

Degrees of Number of replicates freedom t (n¥1, 0.99) (n¥1)

7 ...... 6 3.143 8 ...... 7 2.998 9 ...... 8 2.896 10 ...... 9 2.821 11 ...... 10 2.764 16 ...... 15 2.602 21 ...... 20 2.528 26 ...... 25 2.485 31 ...... 30 2.457 32 ...... 31 2.453 48 ...... 47 2.408 50 ...... 49 2.405 61 ...... 60 2.390 64 ...... 63 2.387 80 ...... 79 2.374 96 ...... 95 2.366 100 ...... 99 2.365

III. Documentation APPENDIX C TO PART 136—DETERMINA- The analytical method used must be spe- TION OF METALS AND TRACE ELE- cifically identified by number or title and MENTS IN WATER AND WASTES BY IN- the MDL for each analyte expressed in the DUCTIVELY COUPLED PLASMA-ATOM- appropriate method reporting units. Data IC EMISSION SPECTROMETRY METHOD and calculations used to establish the MDL 200.7 must be able to be reconstructed upon re- quest. The sample matrix used to determine 1.0 Scope and Application the MDL must also be identified with MDL 1.1 Inductively coupled plasma-atomic value. Document the mean spiked and recov- emission spectrometry (ICP–AES) is used to ered analyte levels with the MDL. The ra- determine metals and some nonmetals in so- tionale for removal of outlier results, if any, lution. This method is a consolidation of ex- must be documented and maintained on file isting methods for water, wastewater, and with the results of the MDL determination. solid wastes.1–4 (For analysis of petroleum [82 FR 40939, Aug. 28, 2017] products see References 5 and 6, Section 16.0). This method is applicable to the fol- lowing analytes:

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Chemical abstract a digestion/extraction is required prior to Analyte services registry analysis when the elements are not in solu- number (CASRN) tion (e.g., soils, sludges, sediments and aque- Aluminum (Al) ...... 7429–90–5 ous samples that may contain particulate Antimony (Sb) ...... 7440–36–0 and suspended solids). Aqueous samples con- Arsenic (As) ...... 7440–38–2 taining suspended or particulate material 1% Barium (Ba) ...... 7440–39–3 (w/v) should be extracted as a solid type sam- Beryllium (Be) ...... 7440–41–7 ple. Boron (B) ...... 7440–42–8 Cadmium (Cd) ...... 7440–43–9 1.6 When determining boron and silica in Calcium (Ca) ...... 7440–70–2 aqueous samples, only plastic, PTFE or Cerium a (Cr) ...... 7440–45–1 quartz labware should be used from time of Chromium (Cr) ...... 7440–47–3 sample collection to completion of analysis. Cobalt (Co) ...... 7440–48–4 For accurate determination of boron in solid Copper (Cu) ...... 7440–50–8 samples only quartz or PTFE beakers should Iron (Fe) ...... 7439–89–6 Lead (Pb) ...... 7439–92–1 be used during acid extraction with imme- Lithium (Li) ...... 7439–93–2 diate transfer of an extract aliquot to a plas- Magnesium (Mg) ...... 7439–95–4 tic centrifuge tube following dilution of the Manganese (Mn) ...... 7439–96–5 extract to volume. When possible, Mercury (Hg) ...... 7439–97–6 borosilicate glass should be avoided to pre- Molybdenum (Mo) ...... 7439–98–7 Nickel (Ni) ...... 7440–02–0 vent contamination of these analytes. Phosphorus (P) ...... 7723–14–0 1.7 Silver is only slightly soluble in the Potassium (K) ...... 7440–09–7 presence of chloride unless there is a suffi- Selenium (Se) ...... 7782–49–2 cient chloride concentration to form the b Silica (Si02) ...... 7631–86–9 soluble chloride complex. Therefore, low re- Silver (Ag) ...... 7440–22–4 Sodium (Na) ...... 7440–23–5 coveries of silver may occur in samples, for- Strontium (Sr) ...... 7440–24–6 tified sample matrices and even fortified Thallium (Tl) ...... 7440–28–0 blanks if determined as a dissolved analyte Tin (Sn) ...... 7440–31–5 or by ‘‘direct analysis’’ where the sample has Titanium (Ti) ...... 7440–32–6 not been processed using the total recover- Vanadium (V) ...... 7440–62–2 able mixed acid digestion. For this reason it Zinc (Zn) ...... 7440–66–6 is recommended that samples be digested a Cerium has been included as method analyte for correc- prior to the determination of silver. The tion of potential interelement spectral interference. b This method is not suitable for the determination of silica total recoverable sample digestion procedure in solids. given in this method is suitable for the de- termination of silver in aqueous samples 1.2 For reference where this method is ap- containing concentrations up to 0.1 mg/L. proved for use in compliance monitoring pro- For the analysis of wastewater samples con- grams [e.g., Clean Water Act (NPDES) or taining higher concentrations of silver, suc- Safe Drinking Water Act (SDWA)] consult both the appropriate sections of the Code of ceeding smaller volume, well mixed aliquots Federal Regulation (40 CFR Part 136 Table should be prepared until the analysis solu- 1B for NPDES, and Part 141 § 141.23 for drink- tion contains <0.1 mg/L silver. The extrac- tion of solid samples containing concentra- ing water), and the latest FEDERAL REGISTER announcements. tions of silver >50 mg/kg should be treated in 1.3 ICP–AES can be used to determine dis- a similar manner. Also, the extraction of tin solved analytes in aqueous samples after from solid samples should be prepared again suitable filtration and acid preservation. To using aliquots <1 g when determined sample reduce potential interferences, dissolved sol- concentrations exceed 1%. ids should be <0.2% (w/v) (Section 4.2). 1.8 The total recoverable sample digestion 1.4 With the exception of silver, where procedure given in this method will solu- this method is approved for the determina- bilize and hold in solution only minimal con- tion of certain metal and metalloid contami- centrations of barium in the presence of free nants in drinking water, samples may be sulfate. For the analysis of barium in sam- analyzed directly by pneumatic nebulization ples having varying and unknown concentra- without acid digestion if the sample has been tions of sulfate, analysis should be com- properly preserved with acid and has tur- pleted as soon as possible after sample prepa- bidity of <1 NTU at the time of analysis. ration. This total recoverable determination proce- 1.9 The total recoverable sample digestion dure is referred to as ‘‘direct analysis’’. How- procedure given in this method is not suit- ever, in the determination of some primary able for the determination of volatile drinking water metal contaminants, organo-mercury compounds. However, if di- preconcentration of the sample may be re- gestion is not required (turbidity <1 NTU), quired prior to analysis in order to meet the combined concentrations of inorganic drinking water acceptance performance cri- and organo-mercury in solution can be deter- teria (Sections 11.2.2 through 11.2.7). mined by ‘‘direct analysis’’ pneumatic 1.5 For the determination of total recov- nebulization provided the sample solution is erable analytes in aqueous and solid samples adjusted to contain the same mixed acid

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(HNO3 + HCl) matrix as the total recoverable trix as in the calibration standards. The cali- calibration standards and blank solutions. bration blank is a zero standard and is used 1.10 Detection limits and linear ranges for to calibrate the ICP instrument (Section the elements will vary with the wavelength 7.10.1). selected, the spectrometer, and the matrices. 3.2 Calibration Standard (CAL)—A solu- Table 1 provides estimated instrument detec- tion prepared from the dilution of stock tion limits for the listed wavelengths.7 How- standard solutions. The CAL solutions are ever, actual method detection limits and lin- used to calibrate the instrument response ear working ranges will be dependent on the with respect to analyte concentration (Sec- sample matrix, instrumentation, and se- tion 7.9). lected operating conditions. 3.3 Dissolved Analyte—The concentration 1.11 Users of the method data should state of analyte in an aqueous sample that will the data-quality objectives prior to analysis. pass through a 0.45 μm membrane filter as- Users of the method must document and sembly prior to sample acidification (Section have on file the required initial demonstra- 11.1). tion performance data described in Section 3.4 Field Reagent Blank (FRB)—An ali- 9.2 prior to using the method for analysis. quot of reagent water or other blank matrix that is placed in a sample container in the 2.0 Summary of Method laboratory and treated as a sample in all re- 2.1 An aliquot of a well mixed, homo- spects, including shipment to the sampling site, exposure to the sampling site condi- geneous aqueous or solid sample is accu- tions, storage, preservation, and all analyt- rately weighed or measured for sample proc- ical procedures. The purpose of the FRB is to essing. For total recoverable analysis of a determine if method analytes or other inter- solid or an aqueous sample containing ferences are present in the field environment undissolved material, analytes are first solu- (Section 8.5). bilized by gentle refluxing with nitric and 3.5 Instrument Detection Limit (IDL)— hydrochloric acids. After cooling, the sample The concentration equivalent to the analyte is made up to volume, is mixed and signal which is equal to three times the centrifuged or allowed to settle overnight standard deviation of a series of 10 replicate prior to analysis. For the determination of measurements of the calibration blank sig- dissolved analytes in a filtered aqueous sam- nal at the same wavelength (Table 1.). ple aliquot, or for the ‘‘direct analysis’’ total 3.6 Instrument Performance Check (IPC) recoverable determination of analytes in Solution—A solution of method analytes, drinking water where sample turbidity is <1 used to evaluate the performance of the in- NTU, the sample is made ready for analysis strument system with respect to a defined by the appropriate addition of nitric acid, set of method criteria (Sections 7.11 and and then diluted to a predetermined volume 9.3.4). and mixed before analysis. 3.7 Internal Standard—Pure analyte(s) 2.2 The analysis described in this method added to a sample, extract, or standard solu- involves multielemental determinations by tion in known amount(s) and used to meas- ICP–AES using sequential or simultaneous ure the relative responses of other method instruments. The instruments measure char- analytes that are components of the same acteristic atomic-line emission spectra by sample or solution. The internal standard optical spectrometry. Samples are nebulized must be an analyte that is not a sample com- and the resulting aerosol is transported to ponent (Section 11.5). the plasma torch. Element specific emission 3.8 Laboratory Duplicates (LD1 and spectra are produced by a radio-frequency in- LD2)—Two aliquots of the same sample ductively coupled plasma. The spectra are taken in the laboratory and analyzed sepa- dispersed by a grating spectrometer, and the rately with identical procedures. Analyses of intensities of the line spectra are monitored LD1 and LD2 indicate precision associated at specific wavelengths by a photosensitive with laboratory procedures, but not with device. Photocurrents from the photosensi- sample collection, preservation, or storage tive device are processed and controlled by a procedures. computer system. A background correction 3.9 Laboratory Fortified Blank (LFB)—An technique is required to compensate for vari- aliquot of LRB to which known quantities of able background contribution to the deter- the method analytes are added in the labora- mination of the analytes. Background must tory. The LFB is analyzed exactly like a be measured adjacent to the analyte wave- sample, and its purpose is to determine length during analysis. Various interferences whether the methodology is in control and must be considered and addressed appro- whether the laboratory is capable of making priately as discussed in Sections 4.0, 7.0, 9.0, accurate and precise measurements (Sec- 10.0, and 11.0. tions 7.10.3 and 9.3.2). 3.10 Laboratory Fortified Sample Matrix 3.0 Definitions (LFM)—An aliquot of an environmental sam- 3.1 Calibration Blank—A volume of rea- ple to which known quantities of the method gent water acidified with the same acid ma- analytes are added in the laboratory. The

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LFM is analyzed exactly like a sample, and ‘‘direct analysis’’ of an unfiltered acid pre- its purpose is to determine whether the sam- served drinking water sample with turbidity ple matrix contributes bias to the analytical of <1 NTU (Section 11.2.1), or by analysis of results. The background concentrations of the solution extract of a solid sample or an the analytes in the sample matrix must be unfiltered aqueous sample following diges- determined in a separate aliquot and the tion by refluxing with hot dilute mineral measured values in the LFM corrected for acid(s) as specified in the method (Sections background concentrations (Section 9.4). 11.2 and 11.3). 3.11 Laboratory Reagent Blank (LRB)— 3.21 Water Sample—For the purpose of An aliquot of reagent water or other blank this method, a sample taken from one of the matrices that are treated exactly as a sam- following sources: drinking, surface, ground, ple including exposure to all glassware, storm runoff, industrial or domestic waste- equipment, solvents, reagents, and internal water. standards that are used with other samples. The LRB is used to determine if method 4.0 Interferences analytes or other interferences are present 4.1 Spectral interferences are caused by in the laboratory environment, reagents, or background emission from continuous or re- apparatus (Sections 7.10.2 and 9.3.1). combination phenomena, stray light from 3.12 Linear Dynamic Range (LDR)—The the line emission of high concentration ele- concentration range over which the instru- ments, overlap of a spectral line from an- ment response to an analyte is linear (Sec- other element, or unresolved overlap of mo- tion 9.2.2). lecular band spectra. 3.13 Method Detection Limit (MDL)—The 4.1.1 Background emission and stray light minimum concentration of an analyte that can usually be compensated for by sub- can be identified, measured, and reported tracting the background emission deter- with 99% confidence that the analyte con- mined by measurement(s) adjacent to the centration is greater than zero (Section 9.2.4 analyte wavelength peak. Spectral scans of and Table 4.). samples or single element solutions in the 3.14 Plasma Solution—A solution that is analyte regions may indicate not only when used to determine the optimum height above alternate wavelengths are desirable because the work coil for viewing the plasma (Sec- of severe spectral interference, but also will tions 7.15 and 10.2.3). show whether the most appropriate estimate 3.15 Quality Control Sample (QCS)—A so- of the background emission is provided by an lution of method analytes of known con- interpolation from measurements on both centrations which is used to fortify an ali- sides of the wavelength peak or by the meas- quot of LRB or sample matrix. The QCS is ured emission on one side or the other. The obtained from a source external to the lab- location(s) selected for the measurement of oratory and different from the source of cali- background intensity will be determined by bration standards. It is used to check either the complexity of the spectrum adjacent to laboratory or instrument performance (Sec- the wavelength peak. The location(s) used tions 7.12 and 9.2.3). for routine measurement must be free of off- 3.16 Solid Sample—For the purpose of this line spectral interference (interelement or method, a sample taken from material clas- molecular) or adequately corrected to reflect sified as soil, sediment or sludge. the same change in background intensity as 3.17 Spectral Interference Check (SIC) So- occurs at the wavelength peak. lution—A solution of selected method 4.1.2 Spectral overlaps may be avoided by analytes of higher concentrations which is using an alternate wavelength or can be used to evaluate the procedural routine for compensated for by equations that correct correcting known interelement spectral for interelement contributions, which in- interferences with respect to a defined set of volves measuring the interfering elements. method criteria (Sections 7.13, 7.14 and 9.3.5). Some potential on-line spectral interferences 3.18 Standard Addition—The addition of a observed for the recommended wavelengths known amount of analyte to the sample in are given in Table 2. When operative and un- order to determine the relative response of corrected, these interferences will produce the detector to an analyte within the sample false-positive determinations and be re- matrix. The relative response is then used to ported as analyte concentrations. The inter- assess either an operative matrix effect or ferences listed are only those that occur be- the sample analyte concentration (Sections tween method analytes. Only interferences 9.5.1 and 11.5). of a direct overlap nature that were observed 3.19 Stock Standard Solution—A con- with a single instrument having a working centrated solution containing one or more resolution of 0.035 nm are listed. More exten- method analytes prepared in the laboratory sive information on interferant effects at using assayed reference materials or pur- various wavelengths and resolutions is avail- chased from a reputable commercial source able in Boumans’ Tables.8 Users may apply (Section 7.8). interelement correction factors determined 3.20 Total Recoverable Analyte—The con- on their instruments within tested con- centration of analyte determined either by centration ranges to compensate (off-line or

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on-line) for the effects of interfering ele- or a computer software routine must be em- ments. ployed for comparing the determinative data 4.1.3 When interelement corrections are to limits files for notifying the analyst when applied, there is a need to verify their accu- an interfering element is detected in the racy by analyzing spectral interference sample at a concentration that will produce check solutions as described in Section 7.13. either an apparent false positive concentra- Interelement corrections will vary for the tion, greater than the analyte IDL, or false same emission line among instruments be- negative analyte concentration, less than cause of differences in resolution, as deter- the 99% lower control limit of the calibra- mined by the grating plus the entrance and tion blank. When the interference accounts exit slit widths, and by the order of disper- for 10% or more of the analyte concentra- sion. Interelement corrections will also vary tion, either an alternate wavelength free of depending upon the choice of background interference or another approved test proce- correction points. Selecting a background dure must be used to complete the analysis. correction point where an interfering emis- For example, the copper peak at 213.853 nm sion line may appear should be avoided when practical. Interelement corrections that con- could be mistaken for the zinc peak at 213.856 stitute a major portion of an emission signal nm in solutions with high copper and low may not yield accurate data. Users should zinc concentrations. For this example, a not forget that some samples may contain spectral scan in the 213.8 nm region would uncommon elements that could contribute not reveal the misidentification because a spectral interferences.78 single peak near the zinc location would be 4.1.4 The interference effects must be observed. The possibility of this evaluated for each individual instrument misidentification of copper for the zinc peak whether configured as a sequential or simul- at 213.856 nm can be identified by measuring taneous instrument. For each instrument, the copper at another emission line, e.g., intensities will vary not only with optical 324.754 nm. Users should be aware that, de- resolution but also with operating conditions pending upon the instrumental resolution, (such as power, viewing height and argon alternate wavelengths with adequate sensi- flow rate). When using the recommended tivity and freedom from interference may wavelengths given in Table 1, the analyst is not be available for all matrices. In these required to determine and document for each circumstances the analyte must be deter- wavelength the effect from the known inter- mined using another approved test proce- ferences given in Table 2, and to utilize a dure. computer routine for their automatic correc- 4.2 Physical interferences are effects asso- tion on all analyses. To determine the appro- ciated with the sample nebulization and priate location for off-line background cor- transport processes. Changes in viscosity and rection, the user must scan the area on ei- surface tension can cause significant inac- ther side adjacent to the wavelength and curacies, especially in samples containing record the apparent emission intensity from high dissolved solids or high acid concentra- all other method analytes. This spectral in- tions. If physical interferences are present, formation must be documented and kept on they must be reduced by such means as a file. The location selected for background high-solids nebulizer, diluting the sample, correction must be either free of off-line interelement spectral interference or a com- using a peristaltic pump, or using an appro- puter routine must be used for their auto- priate internal standard element. Another matic correction on all determinations. If a problem that can occur with high dissolved wavelength other than the recommended solids is buildup at the tip of the wavelength is used, the user must determine nebulizer, which affects aerosol flow rate and and document both the on-line and off-line causes instrumental drift. This problem can spectral interference effect from all method be controlled by a high-solids nebulizer, wet- analytes and provide for their automatic cor- ting the argon prior to nebulization, using a rection on all analyses. Tests to determine tip washer, or diluting the sample. Also, it the spectral interference must be done using has been reported that better control of the analyte concentrations that will adequately argon flow rates, especially for the nebulizer, describe the interference. Normally, 100 mg/ improves instrument stability and precision; L single element solutions are sufficient, this is accomplished with the use of mass however, for analytes such as iron that may flow controllers. be found at high concentration a more appro- 4.3 Chemical interferences include molec- priate test would be to use a concentration ular-compound formation, ionization effects, near the upper LDR limit. See Section 10.4 and solute-vaporization effects. Normally, for required spectral interference test cri- these effects are not significant with the teria. ICP–AES technique. If observed, they can be 4.1.5 When interelement corrections are minimized by careful selection of operating not used, either on-going SIC solutions (Sec- conditions (such as incident power and obser- tion 7.14) must be analyzed to verify the ab- vation height), by buffering of the sample, by sence of interelement spectral interference matrix matching, and by standard-addition

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procedures. Chemical interferences are high- contact with human waste should be immu- ly dependent on matrix type and the specific nized against known disease causative analyte element. agents. 4.4 Memory interferences result when 5.4 The inductively coupled plasma should analytes in a previous sample contribute to only be viewed with proper eye protection the signals measured in a new sample. Mem- from the ultraviolet emissions. ory effects can result from sample deposition 5.5 It is the responsibility of the user of on the uptake tubing to the nebulizer, and this method to comply with relevant dis- from the buildup of sample material in the posal and waste regulations. For guidance plasma torch and spray chamber. The site see Sections 14.0 and 15.0. where these effects occur is dependent on the element and can be minimized by flushing 6.0 Equipment and Supplies the system with a rinse blank between sam- ples (Section 7.10.4). The possibility of mem- 6.1 Inductively coupled plasma emission ory interferences should be recognized with- spectrometer: in an analytical run and suitable rinse times 6.1.1 Computer-controlled emission spec- should be used to reduce them. The rinse trometer with background-correction capa- times necessary for a particular element bility. must be estimated prior to analysis. This The spectrometer must be capable of meet- may be achieved by aspirating a standard ing and complying with the requirements de- containing elements corresponding to either scribed and referenced in Section 2.2. their LDR or a concentration ten times 6.1.2 Radio-frequency generator compliant those usually encountered. The aspiration with FCC regulations. time should be the same as a normal sample 6.1.3 Argon gas supply—High purity grade analysis period, followed by analysis of the (99.99%). When analyses are conducted fre- rinse blank at designated intervals. The quently, liquid argon is more economical and length of time required to reduce analyte requires less frequent replacement of tanks signals to within a factor of two of the meth- than compressed argon in conventional cyl- od detection limit, should be noted. Until the inders. required rinse time is established, this meth- 6.1.4 A variable speed peristaltic pump is od requires a rinse period of at least 60 sec- required to deliver both standard and sample onds between samples and standards. If a solutions to the nebulizer. memory interference is suspected, the sam- 6.1.5 (Optional) Mass flow controllers to ple must be re-analyzed after a long rinse pe- riod. regulate the argon flow rates, especially the aerosol transport gas, are highly rec- 5.0 Safety ommended. Their use will provide more ex- acting control of reproducible plasma condi- 5.1 The toxicity or carcinogenicity of tions. each reagent used in this method have not 6.2 Analytical balance, with capability to been fully established. Each chemical should measure to 0.1 mg, for use in weighing solids, be regarded as a potential health hazard and for preparing standards, and for determining exposure to these compounds should be as dissolved solids in digests or extracts. low as reasonably achievable. Each labora- 6.3 A temperature adjustable hot plate ca- tory is responsible for maintaining a current pable of maintaining a temperature of 95 °C. awareness file of OSHA regulations regard- ing the safe handling of the chemicals speci- 6.4 (Optional) A temperature adjustable block digester capable of maintaining a tem- fied in this method.9–12 A reference file of ° material data handling sheets should also be perature of 95 C and equipped with 250 mL made available to all personnel involved in constricted digestion tubes. the chemical analysis. Specifically, con- 6.5 (Optional) A steel cabinet centrifuge centrated nitric and hydrochloric acids with guard bowl, electric timer and brake. present various hazards and are moderately 6.6 A gravity convection drying oven with toxic and extremely irritating to skin and thermostatic control capable of maintaining mucus membranes. Use these reagents in a 180 °C ±5 °C. fume hood whenever possible and if eye or 6.7 (Optional) An air displacement skin contact occurs, flush with large vol- pipetter capable of delivering volumes rang- umes of water. Always wear safety glasses or ing from 0.1–2500 μL with an assortment of a shield for eye protection, protective cloth- high quality disposable pipet tips. ing and observe proper mixing when working 6.8 Mortar and pestle, ceramic or non- with these reagents. metallic material. 5.2 The acidification of samples con- 6.9 Polypropylene sieve, 5-mesh (4 mm taining reactive materials may result in the opening). release of toxic gases, such as cyanides or 6.10 Labware—For determination of trace sulfides. Acidification of samples should be levels of elements, contamination and loss done in a fume hood. are of prime consideration. Potential con- 5.3 All personnel handling environmental tamination sources include improperly samples known to contain or to have been in cleaned laboratory apparatus and general

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contamination within the laboratory envi- 7.2 Hydrochloric acid, concentrated (sp.gr. ronment from dust, etc. A clean laboratory 1.19)—HCl. work area designated for trace element sam- 7.2.1 Hydrochloric acid (1 + 1)—Add 500 ple handling must be used. Sample con- mL concentrated HCl to 400 mL reagent tainers can introduce positive and negative water and dilute to 1 L. errors in the determination of trace ele- 7.2.2 Hydrochloric acid (1 + 4)—Add 200 ments by contributing contaminants mL concentrated HCl to 400 mL reagent through surface desorption or leaching, or water and dilute to 1 L. depleting element concentrations through 7.2.3 Hydrochloric acid (1 + 20)—Add 10 adsorption processes. All reusable labware mL concentrated HCl to 200 mL reagent (glass, quartz, polyethylene, PTFE, FEP, water. etc.) should be sufficiently clean for the task 7.3 Nitric acid, concentrated (sp.gr. 1.41)— objectives. Several procedures found to pro- HNO3. vide clean labware include washing with a 7.3.1 Nitric acid (1 + 1)—Add 500 mL con- detergent solution, rinsing with tap water, centrated HNO3 to 400 mL reagent water and soaking for four hours or more in 20% (v/v) dilute to 1 L. nitric acid or a mixture of HNO3 and HCl (1 7.3.2 Nitric acid (1 + 2)—Add 100 mL con- + 2 + 9), rinsing with reagent water and stor- centrated HNO3 to 200 mL reagent water. ing clean. 23 Chromic acid cleaning solutions 7.3.3 Nitric acid (1 + 5)—Add 50 mL con- must be avoided because chromium is an centrated HNO3 to 250 mL reagent water. analyte. 7.3.4 Nitric acid (1 + 9)—Add 10 mL con- 6.10.1 Glassware—Volumetric flasks, grad- centrated HNO3 to 90 mL reagent water. uated cylinders, funnels and centrifuge tubes 7.4 Reagent water. All references to water (glass and/or metal-free plastic). in this method refer to ASTM Type I grade 6.10.2 Assorted calibrated pipettes. water.14 6.10.3 Conical Phillips beakers (Corning 7.5 Ammonium hydroxide, concentrated 1080–250 or equivalent), 250 mL with 50 mm (sp.gr. 0.902). watch glasses. 7.6 Tartaric acid, ACS reagent grade. 6.10.4 Griffin beakers, 250 mL with 75 mm 7.7 , 50%, stabilized cer- watch glasses and (optional) 75 mm ribbed tified reagent grade. watch glasses. 7.8 Standard Stock Solutions—Stock 6.10.5 (Optional) PTFE and/or quartz Grif- standards may be purchased or prepared fin beakers, 250 mL with PTFE covers. from ultra-high purity grade chemicals 6.10.6 Evaporating dishes or high-form (99.99–99.999% pure). All compounds must be crucibles, porcelain, 100 mL capacity. dried for one hour at 105 °C, unless otherwise 6.10.7 Narrow-mouth storage bottles, FEP specified. It is recommended that stock solu- (fluorinated ethylene propylene) with screw tions be stored in FEP bottles. Replace stock closure, 125 mL to 1 L capacities. standards when succeeding dilutions for 6.10.8 One-piece stem FEP wash bottle preparation of calibration standards cannot with screw closure, 125 mL capacity. be verified. CAUTION: Many of these chemicals are ex- 7.0 Reagents and Standards tremely toxic if inhaled or swallowed (Sec- 7.1 Reagents may contain elemental im- tion 5.1). Wash hands thoroughly after han- purities which might affect analytical data. dling. Only high-purity reagents that conform to Typical stock solution preparation proce- the American Chemical Society specifica- dures follow for 1 L quantities, but for the tions 13 should be used whenever possible. If purpose of pollution prevention, the analyst the purity of a reagent is in question, ana- is encouraged to prepare smaller quantities lyze for contamination. All acids used for when possible. Concentrations are calculated this method must be of ultra high-purity based upon the weight of the pure element or grade or equivalent. Suitable acids are avail- upon the weight of the compound multiplied able from a number of manufacturers. Redis- by the fraction of the analyte in the com- tilled acids prepared by sub-boiling distilla- pound tion are acceptable. From pure element,

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where: gravimetric factor = the weight frac- glass volumetric container. Use of a nonglass tion of the analyte in the compound volumetric flask is recommended to avoid 7.8.1 Aluminum solution, stock, 1 mL = boron contamination from glassware. 1000 μg Al: Dissolve 1.000 g of aluminum 7.8.7 Cadmium solution, stock, 1 mL = μ metal, weighed accurately to at least four 1000 g Cd: Dissolve 1.000 g Cd metal, acid significant figures, in an acid mixture of 4.0 cleaned with (1 + 9) HNO3, weighed accu- mL of (1 + 1) HCl and 1 mL of concentrated rately to at least four significant figures, in 50 mL (1 + 1) HNO3 with heating to effect dis- HNO3 in a beaker. Warm beaker slowly to ef- fect solution. When dissolution is complete, solution. Let solution cool and dilute with transfer solution quantitatively to a 1 L reagent water in a 1 L volumetric flask. flask, add an additional 10.0 mL of (1 + 1) HCl 7.8.8 Calcium solution, stock, 1 mL = 1000 μg Ca: Suspend 2.498 g CaCO (Ca fraction = and dilute to volume with reagent water. 3 0.4005), dried at 180 °C for one hour before 7.8.2 Antimony solution, stock, 1 mL = weighing, weighed accurately to at least four 1000 μg Sb: Dissolve 1.000 g of antimony pow- significant figures, in reagent water and dis- der, weighed accurately to at least four sig- solve cautiously with a minimum amount of nificant figures, in 20.0 mL (1 + 1) HNO and 3 (1 + 1) HNO . Add 10.0 mL concentrated HNO 10.0 mL concentrated HCl. Add 100 mL rea- 3 3 and dilute to volume in a 1 L volumetric gent water and 1.50 g tartaric acid. Warm so- flask with reagent water. lution slightly to effect complete dissolu- 7.8.9 Cerium solution, stock, 1 mL = 1000 tion. Cool solution and add reagent water to μg Ce: Slurry 1.228 g CeO2 (Ce fraction = volume in a 1 L volumetric flask. 0.8141), weighed accurately to at least four 7.8.3 Arsenic solution, stock, 1 mL = 1000 significant figures, in 100 mL concentrated μ g As: Dissolve 1.320 g of As2O3 (As fraction HNO and evaporate to dryness. Slurry the = 0.7574), weighed accurately to at least four 3 residue in 20 mL H2O, add 50 mL con- significant figures, in 100 mL of reagent centrated HNO , with heat and stirring add water containing 10.0 mL concentrated 3 60 mL 50% H2O2 dropwise in 1 mL increments NH4OH. Warm the solution gently to effect allowing periods of stirring between the 1 dissolution. Acidify the solution with 20.0 mL additions. Boil off excess H2O2 before di- mL concentrated HNO3 and dilute to volume luting to volume in a 1 L volumetric flask in a 1 L volumetric flask with reagent water. with reagent water. 7.8.4 Barium solution, stock, 1 mL = 1000 7.8.10 Chromium solution, stock, 1 mL = μg Ba: Dissolve 1.437 g BaCO (Ba fraction = 3 1000 μg Cr: Dissolve 1.923 g CrO3 (Cr fraction 0.6960), weighed accurately to at least four = 0.5200), weighed accurately to at least four significant figures, in 150 mL (1 + 2) HNO3 significant figures, in 120 mL (1 + 5) HNO3. with heating and stirring to degas and dis- When solution is complete, dilute to volume solve compound. Let solution cool and dilute in a 1 L volumetric flask with reagent water. with reagent water in 1 L volumetric flask. 7.8.11 Cobalt solution, stock, 1 mL = 1000 7.8.5 Beryllium solution, stock, 1 mL = μg Co: Dissolve 1.000 g Co metal, acid cleaned μ 1000 g Be: DO NOT DRY. Dissolve 19.66 g with (1 + 9) HNO3, weighed accurately to at BeSO4•4H2O (Be fraction = 0.0509), weighed least four significant figures, in 50.0 mL (1 + accurately to at least four significant fig- 1) HNO3. Let solution cool and dilute to vol- ures, in reagent water, add 10.0 mL con- ume in a 1 L volumetric flask with reagent centrated HNO3, and dilute to volume in a 1 water. L volumetric flask with reagent water. 7.8.12 Copper solution, stock, 1 mL = 1000 7.8.6 Boron solution, stock, 1 mL = 1000 μg μg Cu: Dissolve 1.000 g Cu metal, acid cleaned B: DO NOT DRY. Dissolve 5.716 g anhydrous with (1 + 9) HNO3, weighed accurately to at H3BO3 (B fraction = 0.1749), weighed accu- least four significant figures, in 50.0 mL (1 + rately to at least four significant figures, in 1) HNO3 with heating to effect dissolution. reagent water and dilute in a 1 L volumetric Let solution cool and dilute in a 1 L volu- flask with reagent water. Transfer imme- metric flask with reagent water. diately after mixing to a clean FEP bottle to 7.8.13 Iron solution, stock, 1 mL = 1000 μg minimize any leaching of boron from the Fe: Dissolve 1.000 g Fe metal, acid cleaned

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with (1 + 1) HCl, weighed accurately to four = 0.7116), weighed accurately to at least four significant figures, in 100 mL (1 + 1) HCl with significant figures, in 200 mL reagent water heating to effect dissolution. Let solution and dilute to volume in a 1 L volumetric cool and dilute with reagent water in a 1 L flask with reagent water. volumetric flask. 7.8.24 Silica solution, stock, 1 mL = 1000 7.8.14 Lead solution, stock, 1 mL = 1000 μg μg SiO2: DO NOT DRY. Dissolve 2.964 g Pb: Dissolve 1.599 g Pb(NO3)2 (Pb fraction = (NH4)2SiF6, weighed accurately to at least 0.6256), weighed accurately to at least four four significant figures, in 200 mL (1 + 20) significant figures, in a minimum amount of HCl with heating at 85 °C to effect dissolu- (1 + 1) HNO3. Add 20.0 mL (1 + 1) HNO3 and di- tion. Let solution cool and dilute to volume lute to volume in a 1 L volumetric flask with in a 1 L volumetric flask with reagent water. reagent water. 7.8.25 Silver solution, stock, 1 mL = 1000 7.8.15 Lithium solution, stock, 1 mL = 1000 μg Ag: Dissolve 1.000 g Ag metal, weighed ac- μg Li: Dissolve 5.324 g Li2CO3 (Li fraction = curately to at least four significant figures, 0.1878), weighed accurately to at least four in 80 mL (1 + 1) HNO3 with heating to effect significant figures, in a minimum amount of dissolution. Let solution cool and dilute with (1 + 1) HCl and dilute to volume in a 1 L vol- reagent water in a 1 L volumetric flask. umetric flask with reagent water. Store solution in amber bottle or wrap bottle 7.8.16 Magnesium solution, stock, 1 mL = completely with aluminum foil to protect so- 1000 μg Mg: Dissolve 1.000 g cleanly polished lution from light. Mg ribbon, accurately weighed to at least 7.8.26 Sodium solution, stock, 1 mL = 1000 four significant figures, in slowly added 5.0 μg Na: Dissolve 2.542 g NaCl (Na fraction = mL (1 + 1) HCl (CAUTION: reaction is vig- 0.3934), weighed accurately to at least four orous). Add 20.0 mL (1 + 1) HNO3 and dilute significant figures, in reagent water. Add 10.0 to volume in a 1 L volumetric flask with rea- mL concentrated HNO3 and dilute to volume gent water. in a 1 L volumetric flask with reagent water. 7.8.17 Manganese solution, stock, 1 mL = 7.8.27 Strontium solution, stock, 1 mL = 1000 μg Mn: Dissolve 1.000 g of manganese 1000 μg Sr: Dissolve 1.685 g SrCO3 (Sr fraction metal, weighed accurately to at least four = 0.5935), weighed accurately to at least four significant figures, in 50 mL (1 + 1) HNO3 and significant figures, in 200 mL reagent water dilute to volume in a 1 L volumetric flask with dropwise addition of 100 mL (1 + 1) HCl. with reagent water. Dilute to volume in a 1 L volumetric flask 7.8.18 Mercury solution, stock, 1 mL = 1000 with reagent water. μg Hg: DO NOT DRY. CAUTION: highly toxic 7.8.28 Thallium solution, stock, 1 mL = element. Dissolve 1.354 g HgCl2 (Hg fraction = 1000 μg Tl: Dissolve 1.303 g TlNO3 (Tl fraction 0.7388) in reagent water. Add 50.0 mL con- = 0.7672), weighed accurately to at least four centrated HNO3 and dilute to volume in 1 L significant figures, in reagent water. Add 10.0 volumetric flask with reagent water. mL concentrated HNO3 and dilute to volume 7.8.19 Molybdenum solution, stock, 1 mL in a 1 L volumetric flask with reagent water. = 1000 μg Mo: Dissolve 1.500 g MoO3 (Mo frac- 7.8.29 Tin solution, stock, 1 mL = 1000 μg tion = 0.6666), weighed accurately to at least Sn: Dissolve 1.000 g Sn shot, weighed accu- four significant figures, in a mixture of 100 rately to at least four significant figures, in mL reagent water and 10.0 mL concentrated an acid mixture of 10.0 mL concentrated HCl NH4OH, heating to effect dissolution. Let so- and 2.0 mL (1 + 1) HNO3 with heating to ef- lution cool and dilute with reagent water in fect dissolution. Let solution cool, add 200 a 1 L volumetric flask. mL concentrated HCl, and dilute to volume 7.8.20 Nickel solution, stock, 1 mL = 1000 in a 1 L volumetric flask with reagent water. μg Ni: Dissolve 1.000 g of nickel metal, 7.8.30 Titanium solution, stock, 1 mL = weighed accurately to at least four signifi- 1000 μg Ti: DO NOT DRY. Dissolve 6.138 g cant figures, in 20.0 mL hot concentrated (NH4)2TiO(C2O4)2•H2O (Ti fraction = 0.1629), HNO3, cool, and dilute to volume in a 1 L vol- weighed accurately to at least four signifi- umetric flask with reagent water. cant figures, in 100 mL reagent water. Dilute 7.8.21 Phosphorus solution, stock, 1 mL = to volume in a 1 L volumetric flask with rea- 1000 μg P: Dissolve 3.745 g NH4H2PO4 (P frac- gent water. tion = 0.2696), weighed accurately to at least 7.8.31 Vanadium solution, stock, 1 mL = four significant figures, in 200 mL reagent 1000 μg V: Dissolve 1.000 g V metal, acid water and dilute to volume in a 1 L volu- cleaned with (1 + 9) HNO3, weighed accu- metric flask with reagent water. rately to at least four significant figures, in 7.8.22 Potassium solution, stock, 1 mL = 50 mL (1 + 1) HNO3 with heating to effect dis- 1000 μg K: Dissolve 1.907 g KCl (K fraction = solution. Let solution cool and dilute with 0.5244) dried at 110 °C, weighed accurately to reagent water to volume in a 1 L volumetric at least four significant figures, in reagent flask. water, add 20 mL (1 + 1) HCl and dilute to 7.8.32 Yttrium solution, stock 1 mL = 1000 volume in a 1 L volumetric flask with rea- μg Y: Dissolve 1.270 g Y2O3 (Y fraction = gent water. 0.7875), weighed accurately to at least four 7.8.23 Selenium solution, stock, 1 mL = significant figures, in 50 mL (1 + 1) HNO3, 1000 μg Se: Dissolve 1.405 g SeO2 (Se fraction heating to effect dissolution. Cool and dilute 395

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to volume in a 1 L volumetric flask with rea- the same entire preparation scheme as the gent water. samples including sample digestion, when 7.8.33 Zinc solution, stock, 1 mL = 1000 μg applicable. Zn: Dissolve 1.000 g Zn metal, acid cleaned 7.10.3 The laboratory fortified blank with (1 + 9) HNO3, weighed accurately to at (LFB) is prepared by fortifying an aliquot of least four significant figures, in 50 mL (1 + 1) the laboratory reagent blank with all HNO3 with heating to effect dissolution. Let analytes to a suitable concentration using solution cool and dilute with reagent water the following recommended criteria: Ag 0.1 to volume in a 1 L volumetric flask. mg/L, K 5.0 mg/L and all other analytes 0.2 7.9 Mixed Calibration Standard Solu- mg/L or a concentration approximately 100 tions—For the analysis of total recoverable times their respective MDL, whichever is digested samples prepare mixed calibration greater. The LFB must be carried through standard solutions (see Table 3) by com- the same entire preparation scheme as the bining appropriate volumes of the stock so- samples including sample digestion, when lutions in 500 mL volumetric flasks con- applicable. taining 20 mL (1 + 1) HNO3 and 20 mL (1 + 1) 7.10.4 The rinse blank is prepared by HCl and dilute to volume with reagent acidifying reagent water to the same con- water. Prior to preparing the mixed stand- centrations of acids as used in the calibra- ards, each stock solution should be analyzed tion blank and stored in a convenient man- separately to determine possible spectral ner. interferences or the presence of impurities. 7.11 Instrument Performance Check (IPC) Care should be taken when preparing the Solution—The IPC solution is used to peri- mixed standards to ensure that the elements odically verify instrument performance dur- are compatible and stable together. To mini- ing analysis. It should be prepared in the mize the opportunity for contamination by same acid mixture as the calibration stand- the containers, it is recommended to trans- ards by combining method analytes at appro- fer the mixed-standard solutions to acid- priate concentrations. Silver must be lim- cleaned, never-used FEP (FEP) ited to <0.5 mg/L; while potassium and phos- bottles for storage. Fresh mixed standards phorus because of higher MDLs and silica be- should be prepared, as needed, with the real- cause of potential contamination should be ization that concentrations can change on at concentrations of 10 mg/L. For other aging. Calibration standards not prepared analytes a concentration of 2 mg/L is rec- from primary standards must be initially ommended. The IPC solution should be pre- verified using a certified reference solution. pared from the same standard stock solu- For the recommended wavelengths listed in tions used to prepare the calibration stand- Table 1 some typical calibration standard ards and stored in an FEP bottle. Agency combinations are given in Table 3. programs may specify or request that addi- tional instrument performance check solu- Note: If the addition of silver to the rec- tions be prepared at specified concentrations ommended mixed-acid calibration standard in order to meet particular program needs. results in an initial precipitation, add 15 mL 7.12 Quality Control Sample (QCS)—Anal- of reagent water and warm the flask until ysis of a QCS is required for initial and peri- the solution clears. For this acid combina- odic verification of calibration standards or tion, the silver concentration should be lim- stock standard solutions in order to verify ited to 0.5 mg/L. instrument performance. The QCS must be 7.10 Blanks—Four types of blanks are re- obtained from an outside source different quired for the analysis. The calibration from the standard stock solutions and pre- blank is used in establishing the analytical pared in the same acid mixture as the cali- curve, the laboratory reagent blank is used bration standards. The concentration of the to assess possible contamination from the analytes in the QCS solution should be 1 mg/ sample preparation procedure, the labora- L, except silver, which must be limited to a tory fortified blank is used to assess routine concentration of 0.5 mg/L for solution sta- laboratory performance and a rinse blank is bility. The QCS solution should be stored in used to flush the instrument uptake system a FEP bottle and analyzed as needed to meet and nebulizer between standards, check solu- data-quality needs. A fresh solution should tions, and samples to reduce memory inter- be prepared quarterly or more frequently as ferences. needed. 7.10.1 The calibration blank for aqueous 7.13 Spectral Interference Check (SIC) So- samples and extracts is prepared by lutions—When interelement corrections are acidifying reagent water to the same con- applied, SIC solutions are needed containing centrations of the acids as used for the concentrations of the interfering elements at standards. The calibration blank should be levels that will provide an adequate test of stored in a FEP bottle. the correction factors. 7.10.2 The laboratory reagent blank (LRB) 7.13.1 SIC solutions containing (a) 300 mg/ must contain all the reagents in the same L Fe; (b) 200 mg/L AL; (c) 50 mg/L Ba; (d) 50 volumes as used in the processing of the mg/L Be; (e) 50 mg/L Cd; (f) 50 mg/L Ce; (g) 50 samples. The LRB must be carried through mg/L Co; (h) 50 mg/L Cr; (i) 50 mg/L Cu; (j) 50

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mg/L Mn; (k) 50 mg/L Mo; (l) 50 mg/L Ni; (m) daily verification is not required; however, 50 mg/L Sn; (n) 50 mg/L SiO2; (o) 50 mg/L Ti; all interelement spectral correction factors (p) 50 mg/L Tl and (q) 50 mg/L V should be must be verified annually and updated, if prepared in the same acid mixture as the necessary. calibration standards and stored in FEP bot- 7.13.6 If the instrument does not display tles. These solutions can be used to periodi- negative concentration values, fortify the cally verify a partial list of the on-line (and SIC solutions with the elements of interest possible off-line) interelement spectral cor- at 1 mg/L and test for analyte recoveries rection factors for the recommended wave- that are below 95%. In the absence of meas- lengths given in Table 1. Other solutions urable analyte, over-correction could go un- could achieve the same objective as well. detected because a negative value could be (Multielement SIC solutions3 may be pre- reported as zero. pared and substituted for the single element 7.14 For instruments without interele- solutions provided an analyte is not subject ment correction capability or when interele- to interference from more than one ment corrections are not used, SIC solutions interferant in the solution.) (containing similar concentrations of the Note: If wavelengths other than those rec- major components in the samples, e.g., 10 ommended in Table 1 are used, other solu- mg/L) can serve to verify the absence of ef- tions different from those above (a through fects at the wavelengths selected. These data q) may be required. must be kept on file with the sample anal- 7.13.2 For interferences from iron and alu- ysis data. If the SIC solution confirms an op- minum, only those correction factors (posi- erative interference that is 10% of the tive or negative) when multiplied by 100 to analyte concentration, the analyte must be calculate apparent analyte concentrations determined using a wavelength and back- that exceed the determined analyte IDL or ground correction location free of the inter- fall below the lower 3-sigma control limit of ference or by another approved test proce- the calibration blank need be tested on a dure. Users are advised that high salt con- daily basis. centrations can cause analyte signal sup- 7.13.3 For the other interfering elements, pressions and confuse interference tests. only those correction factors (positive or 7.15 Plasma Solution—The plasma solu- negative) when multiplied by 10 to calculate tion is used for determining the optimum apparent analyte concentrations that exceed viewing height of the plasma above the work the determined analyte IDL or fall below the coil prior to using the method (Section 10.2). lower 3-sigma control limit of the calibra- The solution is prepared by adding a 5 mL al- tion blank need be tested on a daily basis. iquot from each of the stock standard solu- 7.13.4 If the correction routine is oper- tions of arsenic, lead, selenium, and thallium ating properly, the determined apparent to a mixture of 20 mL (1 + 1) nitric acid and analyte(s) concentration from analysis of 20 mL (1 + 1) hydrochloric acid and diluting each interference solution (a through q) to 500 mL with reagent water. Store in a should fall within a specific concentration FEP bottle. range bracketing the calibration blank. This concentration range is calculated by multi- 8.0 Sample Collection, Preservation, and plying the concentration of the interfering Storage element by the value of the correction factor 8.1 Prior to the collection of an aqueous being tested and dividing by 10. If after sub- sample, consideration should be given to the traction of the calibration blank the appar- type of data required, (i.e., dissolved or total ent analyte concentration is outside (above recoverable), so that appropriate preserva- or below) this range, a change in the correc- tion and pretreatment steps can be taken. tion factor of more than 10% should be sus- The pH of all aqueous samples must be test- pected. The cause of the change should be de- ed immediately prior to aliquoting for proc- termined and corrected and the correction essing or ‘‘direct analysis’’ to ensure the factor should be updated. sample has been properly preserved. If prop- Note: The SIC solution should be analyzed erly acid preserved, the sample can be held more than once to confirm a change has oc- up to six months before analysis. curred with adequate rinse time between so- 8.2 For the determination of the dissolved lutions and before subsequent analysis of the elements, the sample must be filtered calibration blank. through a 0.45 μm pore diameter membrane 7.13.5 If the correction factors tested on a filter at the time of collection or as soon daily basis are found to be within the 10% thereafter as practically possible. (Glass or criteria for five consecutive days, the re- plastic filtering apparatus are recommended quired verification frequency of those factors to avoid possible contamination. Only plas- in compliance may be extended to a weekly tic apparatus should be used when the deter- basis. Also, if the nature of the samples ana- minations of boron and silica are critical.) lyzed is such (e.g., finished drinking water) Use a portion of the filtered sample to rinse that they do not contain concentrations of the filter flask, discard this portion and col- the interfering elements at the 10 mg/L level, lect the required volume of filtrate. Acidify

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the filtrate with (1 + 1) nitric acid imme- analyte concentration is no more than 10% diately following filtration to pH <2. below the stated concentration of the stand- 8.3 For the determination of total recov- ard. Determined LDRs must be documented erable elements in aqueous samples, samples and kept on file. The LDR which may be are not filtered, but acidified with (1 + 1) ni- used for the analysis of samples should be tric acid to pH <2 (normally, 3 mL of (1 + 1) judged by the analyst from the resulting acid per liter of sample is sufficient for most data. Determined sample analyte concentra- ambient and drinking water samples). Pres- tions that are greater than 90% of the deter- ervation may be done at the time of collec- mined upper LDR limit must be diluted and tion, however, to avoid the hazards of strong reanalyzed. The LDRs should be verified an- acids in the field, transport restrictions, and nually or whenever, in the judgment of the possible contamination it is recommended analyst, a change in analytical performance that the samples be returned to the labora- caused by either a change in instrument tory within two weeks of collection and acid hardware or operating conditions would dic- preserved upon receipt in the laboratory. tate they be redetermined. Following acidification, the sample should 9.2.3 Quality control sample (QCS)—When be mixed, held for 16 hours, and then verified beginning the use of this method, on a quar- to be pH <2 just prior withdrawing an aliquot terly basis, after the preparation of stock or for processing or ‘‘direct analysis’’. If for calibration standard solutions or as required some reason such as high alkalinity the sam- to meet data-quality needs, verify the cali- ple pH is verified to be >2, more acid must be bration standards and acceptable instrument added and the sample held for 16 hours until performance with the preparation and anal- verified to be pH <2. See Section 8.1. yses of a QCS (Section 7.12). To verify the Note: When the nature of the sample is ei- calibration standards the determined mean ther unknown or is known to be hazardous, concentrations from three analyses of the acidification should be done in a fume hood. QCS must be within 5% of the stated values. See Section 5.2. If the calibration standard cannot be verified, performance of the determinative 8.4 Solid samples require no preservation step of the method is unacceptable. The ° prior to analysis other than storage at 4 C. source of the problem must be identified and There is no established holding time limita- corrected before either proceeding on with tion for solid samples. the initial determination of method detec- 8.5 For aqueous samples, a field blank tion limits or continuing with on-going anal- should be prepared and analyzed as required yses. by the data user. Use the same container and 9.2.4 Method detection limit (MDL)— acid as used in sample collection. MDLs must be established for all wave- lengths utilized, using reagent water (blank) 9.0 Quality Control fortified at a concentration of two to three 9.1 Each laboratory using this method is times the estimated instrument detection required to operate a formal quality control limit.15 To determine MDL values, take (QC) program. The minimum requirements of seven replicate aliquots of the fortified rea- this program consist of an initial demonstra- gent water and process through the entire tion of laboratory capability, and the peri- analytical method. Perform all calculations odic analysis of laboratory reagent blanks, defined in the method and report the con- fortified blanks and other laboratory solu- centration values in the appropriate units. tions as a continuing check on performance. Calculate the MDL as follows: The laboratory is required to maintain per- MDL = (t) × (S) formance records that define the quality of where: the data thus generated. 9.2 Initial Demonstration of Performance t = students’ t value for a 99% confidence (mandatory). level and a standard deviation estimate 9.2.1 The initial demonstration of per- with n-1 degrees of freedom [t = 3.14 for formance is used to characterize instrument seven replicates] performance (determination of linear dy- S = standard deviation of the replicate anal- namic ranges and analysis of quality control yses samples) and laboratory performance (deter- Note: If additional confirmation is desired, mination of method detection limits) prior reanalyze the seven replicate aliquots on two to analyses conducted by this method. more nonconsecutive days and again cal- 9.2.2 Linear dynamic range (LDR)—The culate the MDL values for each day. An aver- upper limit of the LDR must be established age of the three MDL values for each analyte for each wavelength utilized. It must be de- may provide for a more appropriate MDL es- termined from a linear calibration prepared timate. If the relative standard deviation in the normal manner using the established (RSD) from the analyses of the seven analytical operating procedure for the in- aliquots is <10%, the concentration used to strument. The LDR should be determined by determine the analyte MDL may have been analyzing succeedingly higher standard con- inappropriately high for the determination. centrations of the analyte until the observed If so, this could result in the calculation of

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an unrealistically low MDL. Concurrently, 9.3.1 Laboratory reagent blank (LRB)— determination of MDL in reagent water rep- The laboratory must analyze at least one resents a best case situation and does not re- LRB (Section 7.10.2) with each batch of 20 or flect possible matrix effects of real world fewer samples of the same matrix. LRB data samples. However, successful analyses of are used to assess contamination from the LFMs (Section 9.4) and the analyte addition laboratory environment. LRB values that ex- test described in Section 9.5.1 can give con- ceed the MDL indicate laboratory or reagent fidence to the MDL value determined in rea- contamination should be suspected. When gent water. Typical single laboratory MDL LRB values constitute 10% or more of the values using this method are given in Table 4. analyte level determined for a sample or is The MDLs must be sufficient to detect 2.2 times the analyte MDL whichever is analytes at the required levels according to greater, fresh aliquots of the samples must compliance monitoring regulation (Section be prepared and analyzed again for the af- 1.2). MDLs should be determined annually, fected analytes after the source of contami- when a new operator begins work or when- nation has been corrected and acceptable ever, in the judgment of the analyst, a LRB values have been obtained. change in analytical performance caused by 9.3.2 Laboratory fortified blank (LFB)— either a change in instrument hardware or The laboratory must analyze at least one operating conditions would dictate they be LFB (Section 7.10.3) with each batch of sam- redetermined. ples. Calculate accuracy as percent recovery 9.3 Assessing Laboratory Performance using the following equation: (mandatory)

where: must be kept on file and be available for re- R = percent recovery view. LFB = laboratory fortified blank 9.3.4 Instrument performance check (IPC) LRB = laboratory reagent blank solution—For all determinations the labora- s = concentration equivalent of analyte tory must analyze the IPC solution (Section added to fortify the LBR solution 7.11) and a calibration blank immediately following daily calibration, after every 10th If the recovery of any analyte falls outside sample (or more frequently, if required) and the required control limits of 85–115%, that at the end of the sample run. Analysis of the analyte is judged out of control, and the calibration blank should always be

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9.3.5 Spectral interference check (SIC) so- one gram aliquot by multiplying the added lution—For all determinations the labora- analyte concentration (mg/L) in solution by tory must periodically verify the interele- the conversion factor 100 (mg/L × 0.1L/0.001kg ment spectral interference correction rou- = 100, Section 12.5). (For notes on Ag, Ba, and tine by analyzing SIC solutions. The prepa- Sn see Sections 1.7 and 1.8.) Over time, sam- ration and required periodic analysis of SIC ples from all routine sample sources should solutions and test criteria for verifying the be fortified. interelement interference correction routine Note: The concentration of calcium, mag- are given in Section 7.13. Special cases where nesium, sodium and strontium in environ- on-going verification is required are de- mental waters, along with iron and alu- scribed in Section 7.14. minum in solids can vary greatly and are not 9.4 Assessing Analyte Recovery and Data Quality. necessarily predictable. Fortifying these 9.4.1 Sample homogeneity and the chem- analytes in routine samples at the same con- ical nature of the sample matrix can affect centration used for the LFB may prove to be analyte recovery and the quality of the data. of little use in assessing data quality for Taking separate aliquots from the sample for these analytes. For these analytes sample di- replicate and fortified analyses can in some lution and reanalysis using the criteria given cases assess the effect. Unless otherwise in Section 9.5.2 is recommended. Also, if specified by the data user, laboratory or pro- specified by the data user, laboratory or pro- gram, the following laboratory fortified ma- gram, samples can be fortified at higher con- trix (LFM) procedure (Section 9.4.2) is re- centrations, but even major constituents quired. Also, other tests such as the analyte should be limited to <25 mg/L so as not to addition test (Section 9.5.1) and sample dilu- alter the sample matrix and affect the anal- tion test (Section 9.5.2) can indicate if ma- ysis. trix effects are operative. 9.4.3 Calculate the percent recovery for 9.4.2 The laboratory must add a known each analyte, corrected for background con- amount of each analyte to a minimum of centrations measured in the unfortified sam- 10% of the routine samples. In each case the ple, and compare these values to the des- LFM aliquot must be a duplicate of the ali- ignated LFM recovery range of 70–130% or a quot used for sample analysis and for total 3-sigma recovery range calculated from the recoverable determinations added prior to regression equations given in Table 9.16 Re- sample preparation. For water samples, the covery calculations are not required if the added analyte concentration must be the concentration added is less than 30% of the same as that used in the laboratory fortified sample background concentration. Percent blank (Section 7.10.3). For solid samples, recovery may be calculated in units appro- however, the concentration added should be priate to the matrix, using the following expressed as mg/kg and is calculated for a equation:

where: 9.4.5 Where reference materials are avail- R = percent recovery able, they should be analyzed to provide ad- Cs = fortified sample concentration ditional performance data. The analysis of C = sample background concentration reference samples is a valuable tool for dem- s = concentration equivalent of analyte onstrating the ability to perform the method added to fortify the sample acceptably. Reference materials containing high concentrations of analytes can provide 9.4.4 If the recovery of any analyte falls additional information on the performance outside the designated LFM recovery range, and the laboratory performance for that of the spectral interference correction rou- analyte is shown to be in control (Section tine. 9.3), the recovery problem encountered with 9.5 Assess the possible need for the meth- the fortified sample is judged to be matrix od of standard additions (MSA) or internal related, not system related. The data user standard elements by the following tests. Di- should be informed that the result for that rections for using MSA or internal stand- analyte in the unfortified sample is suspect ard(s) are given in Section 11.5. due to either the heterogeneous nature of 9.5.1 Analyte addition test: An analyte(s) the sample or matrix effects and analysis by standard added to a portion of a prepared method of standard addition or the use of an sample, or its dilution, should be recovered internal standard(s) (Section 11.5) should be to within 85% to 115% of the known value. considered. The analyte(s) addition should produce a

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minimum level of 20 times and a maximum 10.2.1 Ignite the plasma and select an ap- of 100 times the method detection limit. If propriate incident rf power with minimum the analyte addition is <20% of the sample reflected power. Allow the instrument to be- analyte concentration, the following dilu- come thermally stable before beginning. tion test should be used. If recovery of the This usually requires at least 30 to 60 min- analyte(s) is not within the specified limits, utes of operation. While aspirating the 1000 a matrix effect should be suspected, and the μg/mL solution of yttrium (Section 7.8.32), associated data flagged accordingly. The follow the instrument manufacturer’s in- method of additions or the use of an appro- structions and adjust the aerosol carrier gas priate internal standard element may pro- flow rate through the nebulizer so a defini- vide more accurate data. tive blue emission region of the plasma ex- 9.5.2 Dilution test: If the analyte con- tends approximately from 5–20 mm above the centration is sufficiently high (minimally, a top of the work coil.18 Record the nebulizer factor of 50 above the instrument detection gas flow rate or pressure setting for future limit in the original solution but <90% of the reference. linear limit), an analysis of a 1 + 4 dilution 10.2.2 After establishing the nebulizer gas should agree (after correction for the fivefold flow rate, determine the solution uptake dilution) within 10% of the original deter- rate of the nebulizer in mL/min. by aspi- mination. If not, a chemical or physical in- rating a known volume calibration blank for terference effect should be suspected and the a period of at least three minutes. Divide the associated data flagged accordingly. The spent volume by the aspiration time (in min- method of standard additions or the use of utes) and record the uptake rate. Set the an internal-standard element may provide peristaltic pump to deliver the uptake rate more accurate data for samples failing this in a steady even flow. test. 10.2.3 After horizontally aligning the plas- ma and/or optically profiling the spectrom- 10.0 Calibration and Standardization eter, use the selected instrument conditions from Sections 10.2.1 and 10.2.2, and aspirate 10.1 Specific wavelengths are listed in the plasma solution (Section 7.15), con- Table 1. Other wavelengths may be sub- taining 10 μg/mL each of As, Pb, Se and Tl. stituted if they can provide the needed sensi- Collect intensity data at the wavelength tivity and are corrected for spectral inter- peak for each analyte at 1 mm intervals from ference. However, because of the difference 14–18 mm above the top of the work coil. among various makes and models of spec- (This region of the plasma is commonly re- trometers, specific instrument operating ferred to as the analytical zone.)19 Repeat conditions cannot be given. The instrument the process using the calibration blank. De- and operating conditions utilized for deter- termine the net signal to blank intensity mination must be capable of providing data ratio for each analyte for each viewing of acceptable quality to the program and height setting. Choose the height for viewing data user. The analyst should follow the in- the plasma that provides the largest inten- structions provided by the instrument manu- sity ratio for the least sensitive element of facturer unless other conditions provide the four analytes. If more than one position similar or better performance for a task. Op- provides the same ratio, select the position erating conditions for aqueous solutions usu- that provides the highest net intensity ally vary from 1100–1200 watts forward power, counts for the least sensitive element or ac- 15–16 mm viewing height, 15–19 L/min. argon cept a compromise position of the intensity coolant flow, 0.6–1 L/min. argon aerosol flow, ratios of all four analytes. 1–1.8 mL/min. sample pumping rate with a 10.2.4 The instrument operating condition one minute preflush time and measurement finally selected as being optimum should time near 1 s per wavelength peak (for se- provide the lowest reliable instrument detec- quential instruments) and near 10 s per sam- tion limits and method detection limits. ple (for simultaneous instruments). Use of Refer to Tables 1 and 4 for comparison of the Cu/Mn intensity ratio at 324.754 nm and IDLs and MDLs, respectively. 257.610 nm (by adjusting the argon aerosol 10.2.5 If either the instrument operating flow) has been recommended as a way to conditions, such as incident power and/or achieve repeatable interference correction nebulizer gas flow rate are changed, or a new factors.17 torch injector tube having a different orifice 10.2 Prior to using this method optimize i.d. is installed, the plasma and plasma view- the plasma operating conditions. The fol- ing height should be reoptimized. lowing procedure is recommended for 10.2.6 Before daily calibration and after vertically configured plasmas. The purpose the instrument warmup period, the nebulizer of plasma optimization is to provide a max- gas flow must be reset to the determined op- imum signal-to-background ratio for the timized flow. If a mass flow controller is least sensitive element in the analytical being used, it should be reset to the recorded array. The use of a mass flow controller to optimized flow rate. In order to maintain regulate the nebulizer gas flow rate greatly valid spectral interelement correction rou- facilitates the procedure. tines the nebulizer gas flow rate should be

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the same from day-to-day (<2% change). The calculations. (If mercury is to be deter- change in signal intensity with a change in mined, a separate aliquot must be addition- nebulizer gas flow rate for both ‘‘hard’’ (Pb ally acidified to contain 1% (v/v) HCl to 220.353 nm) and ‘‘soft’’ (Cu 324.754) lines is il- match the signal response of mercury in the lustrated in Figure 1. calibration standard and reduce memory in- 10.3 Before using the procedure (Section terference effects. Section 1.9). 11.0) to analyze samples, there must be data NOTE: If a precipitate is formed during available documenting initial demonstration acidification, transport, or storage, the sam- of performance. The required data and proce- ple aliquot must be treated using the proce- dure is described in Section 9.2. This data must be generated using the same instru- dure described in Sections 11.2.2 through ment operating conditions and calibration 11.2.7 prior to analysis. routine (Section 11.4) to be used for sample analysis. These documented data must be 11.2 Aqueous Sample Preparation—Total kept on file and be available for review by Recoverable Analytes the data user. 11.2.1 For the ‘‘direct analysis’’ of total 10.4 After completing the initial dem- recoverable analytes in drinking water sam- onstration of performance, but before ana- ples containing turbidity <1 NTU, treat an lyzing samples, the laboratory must estab- unfiltered acid preserved sample aliquot lish and initially verify an interelement using the sample preparation procedure de- spectral interference correction routine to scribed in Section 11.1.1 while making allow- be used during sample analysis. A general de- ance for sample dilution in the data calcula- scription concerning spectral interference tion (Section 1.2). For the determination of and the analytical requirements for back- total recoverable analytes in all other aque- ground correction and for correction of ous samples or for preconcentrating drinking interelement spectral interference in par- ticular are given in Section 4.1. To determine water samples prior to analysis follow the the appropriate location for background cor- procedure given in Sections 11.2.2 through rection and to establish the interelement in- 11.2.7. terference correction routine, repeated spec- 11.2.2 For the determination of total re- tral scan about the analyte wavelength and coverable analytes in aqueous samples (other repeated analyses of the single element solu- than drinking water with <1 NTU turbidity), tions may be required. Criteria for deter- transfer a 100 mL (1 mL) aliquot from a well mining an interelement spectral interference mixed, acid preserved sample to a 250 mL is an apparent positive or negative con- Griffin beaker (Sections 1.2, 1.3, 1.6, 1.7, 1.8, centration on the analyte that is outside the and 1.9). (When necessary, smaller sample al- 3-sigma control limits of the calibration iquot volumes may be used.) blank for the analyte. (The upper-control NOTE: If the sample contains undissolved limit is the analyte IDL.) Once established, solids >1%, a well mixed, acid preserved ali- the entire routine must be initially and peri- quot containing no more than 1 g particulate odically verified annually, or whenever there material should be cautiously evaporated to is a change in instrument operating condi- near 10 mL and extracted using the acid-mix- tions (Section 10.2.5). Only a portion of the ture procedure described in Sections 11.3.3 correction routine must be verified more fre- through 11.3.6. quently or on a daily basis. Test criteria and required solutions are described in Section 11.2.3 Add 2 mL (1 + 1) nitric acid and 1.0 7.13. Initial and periodic verification data of mL of (1 + 1) hydrochloric acid to the beaker the routine should be kept on file. Special containing the measured volume of sample. cases where on-going verification are re- Place the beaker on the hot plate for solu- quired is described in Section 7.14. tion evaporation. The hot plate should be lo- cated in a fume hood and previously adjusted 11.0 Procedure to provide evaporation at a temperature of approximately but no higher than 85 °C. (See 11.1 Aqueous Sample Preparation— the following note.) The beaker should be Dissolved Analytes covered with an elevated watch glass or 11.1.1 For the determination of dissolved other necessary steps should be taken to pre- analytes in ground and surface waters, pipet vent sample contamination from the fume an aliquot (20 mL) of the filtered, acid pre- hood environment. served sample into a 50 mL polypropylene NOTE: For proper heating adjust the tem- centrifuge tube. Add an appropriate volume perature control of the hot plate such that of (1 + 1) nitric acid to adjust the acid con- an uncovered Griffin beaker containing 50 centration of the aliquot to approximate a mL of water placed in the center of the hot 1% (v/v) nitric acid solution (e.g., add 0.4 mL plate can be maintained at a temperature ap- (1 + 1) HNO3 to a 20 mL aliquot of sample). proximately but no higher than 85 °C. (Once Cap the tube and mix. The sample is now the beaker is covered with a watch glass the ready for analysis (Section 1.3). Allowance temperature of the water will rise to ap- for sample dilution should be made in the proximately 95 °C.)

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11.2.4 Reduce the volume of the sample al- provide a reflux temperature of approxi- iquot to about 20 mL by gentle heating at 85 mately 95 °C. (See the following note.) °C. DO NOT BOIL. This step takes about two NOTE: For proper heating adjust the tem- hours for a 100 mL aliquot with the rate of perature control of the hot plate such that evaporation rapidly increasing as the sample an uncovered Griffin beaker containing 50 volume approaches 20 mL. (A spare beaker mL of water placed in the center of the hot containing 20 mL of water can be used as a gauge.) plate can be maintained at a temperature ap- ° 11.2.5 Cover the lip of the beaker with a proximately but no higher than 85 C. (Once watch glass to reduce additional evaporation the beaker is covered with a watch glass the and gently reflux the sample for 30 minutes. temperature of the water will rise to ap- (Slight boiling may occur, but vigorous boil- proximately 95 °C.) Also, a block digester ca- ing must be avoided to prevent loss of the pable of maintaining a temperature of 95 °C HCl-H2O azeotrope.) and equipped with 250 mL constricted volu- 11.2.6 Allow the beaker to cool. Quan- metric digestion tubes may be substituted titatively transfer the sample solution to a for the hot plate and conical beakers in the 50 mL volumetric flask, make to volume extraction step. with reagent water, stopper and mix. 11.3.4 Heat the sample and gently reflux 11.2.7 Allow any undissolved material to for 30 minutes. Very slight boiling may settle overnight, or centrifuge a portion of occur, however vigorous boiling must be the prepared sample until clear. (If after avoided to prevent loss of the HCl-H O centrifuging or standing overnight the sam- 2 azeotrope. Some solution evaporation will ple contains suspended solids that would clog the nebulizer, a portion of the sample may be occur (3–4 mL). filtered for their removal prior to analysis. 11.3.5 Allow the sample to cool and quan- However, care should be exercised to avoid titatively transfer the extract to a 100 mL potential contamination from filtration.) volumetric flask. Dilute to volume with rea- The sample is now ready for analysis. Be- gent water, stopper and mix. cause the effects of various matrices on the 11.3.6 Allow the sample extract solution stability of diluted samples cannot be char- to stand overnight to separate insoluble ma- acterized, all analyses should be performed terial or centrifuge a portion of the sample as soon as possible after the completed prep- solution until clear. (If after centrifuging or aration. standing overnight the extract solution con- tains suspended solids that would clog the 11.3 Solid Sample Preparation—Total nebulizer, a portion of the extract solution Recoverable Analytes may be filtered for their removal prior to 11.3.1 For the determination of total re- analysis. However, care should be exercised coverable analytes in solid samples, mix the to avoid potential contamination from fil- sample thoroughly and transfer a portion tration.) The sample extract is now ready for (>20 g) to tared weighing dish, weigh the analysis. Because the effects of various mat- sample and record the wet weight (WW). (For rices on the stability of diluted samples can- samples with <35% moisture a 20 g portion is not be characterized, all analyses should be sufficient. For samples with moisture >35% a performed as soon as possible after the com- larger aliquot 50–100 g is required.) Dry the pleted preparation. sample to a constant weight at 60 °C and record the dry weight (DW) for calculation of 11.4 Sample Analysis percent solids (Section 12.6). (The sample is dried at 60 °C to prevent the loss of mercury 11.4.1 Prior to daily calibration of the in- and other possible volatile metallic com- strument inspect the sample introduction pounds, to facilitate sieving, and to ready system including the nebulizer, torch, injec- the sample for grinding.) tor tube and uptake tubing for salt deposits, 11.3.2 To achieve homogeneity, sieve the dirt and debris that would restrict solution dried sample using a 5-mesh polypropylene flow and affect instrument performance. sieve and grind in a mortar and pestle. (The Clean the system when needed or on a daily sieve, mortar and pestle should be cleaned basis. between samples.) From the dried, ground 11.4.2 Configure the instrument system to material weigh accurately a representative the selected power and operating conditions ± 1.0 0.01 g aliquot (W) of the sample and as determined in Sections 10.1 and 10.2. transfer to a 250 mL Phillips beaker for acid 11.4.3 The instrument must be allowed to extraction (Sections 1.6, 1.7, 1.8, and 1.9). become thermally stable before calibration 11.3.3 To the beaker add 4 mL of (1 + 1) and analyses. This usually requires at least HNO3 and 10 mL of (1 + 4) HCl. Cover the lip of the beaker with a watch glass. Place the 30 to 60 minutes of operation. After instru- beaker on a hot plate for reflux extraction of ment warmup, complete any required optical the analytes. The hot plate should be located profiling or alignment particular to the in- in a fume hood and previously adjusted to strument.

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11.4.4 For initial and daily operation cali- luted with reagent water that has been acidi- brate the instrument according to the in- fied in the same manner as calibration blank strument manufacturer’s recommended pro- and reanalyzed (see Section 11.4.8). Also, for cedures, using mixed calibration standard so- the interelement spectral interference cor- lutions (Section 7.9) and the calibration rection routines to remain valid during sam- blank (Section 7.10.1). A peristaltic pump ple analysis, the interferant concentration must be used to introduce all solutions to must not exceed its LDR. If the interferant the nebulizer. To allow equilibrium to be LDR is exceeded, sample dilution with acidi- reached in the plasma, aspirate all solutions fied reagent water and reanalysis is required. for 30 seconds after reaching the plasma be- In these circumstances analyte detection fore beginning integration of the background limits are raised and determination by an- corrected signal to accumulate data. When other approved test procedure that is either possible, use the average value of replicate more sensitive and/or interference free is integration periods of the signal to be cor- recommended. related to the analyte concentration. Flush 11.4.8 When it is necessary to assess an op- the system with the rinse blank (Section erative matrix interference (e.g., signal re- 7.10.4) for a minimum of 60 seconds (Section duction due to high dissolved solids), the 4.4) between each standard. The calibration tests described in Section 9.5 are rec- line should consist of a minimum of a cali- ommended. bration blank and a high standard. Rep- licates of the blank and highest standard 11.4.9 Report data as directed in Section provide an optimal distribution of calibra- 12.0. tion standards to minimize the confidence 11.5 If the method of standard additions band for a straight-line calibration in a re- (MSA) is used, standards are added at one or sponse region with uniform variance.20 more levels to portions of a prepared sample. 11.4.5 After completion of the initial re- This technique 21 compensates for enhance- quirements of this method (Sections 10.3 and ment or depression of an analyte signal by a 10.4), samples should be analyzed in the same matrix. It will not correct for additive inter- operational manner used in the calibration ferences such as contamination, interele- routine with the rinse blank also being used ment interferences, or baseline shifts. This between all sample solutions, LFBs, LFMs, technique is valid in the linear range when and check solutions (Section 7.10.4). the interference effect is constant over the 11.4.6 During the analysis of samples, the range, the added analyte responds the same laboratory must comply with the required as the endogenous analyte, and the signal is quality control described in Sections 9.3 and corrected for additive interferences. The 9.4. Only for the determination of dissolved simplest version of this technique is the sin- analytes or the ‘‘direct analysis’’ of drinking gle-addition method. This procedure calls for water with turbidity of <1 NTU is the sample two identical aliquots of the sample solution digestion step of the LRB, LFB, and LFM to be taken. To the first aliquot, a small vol- not required. ume of standard is added; while to the second 11.4.7 Determined sample analyte con- aliquot, a volume of acid blank is added centrations that are 90% or more of the equal to the standard addition. The sample upper limit of the analyte LDR must be di- concentration is calculated by the following:

where: one or more elements (not in the samples C = Concentration of the standard solution and verified not to cause an uncorrected (mg/L) interelement spectral interference) at the S1 = Signal for fortified aliquot same concentration (which is sufficient for S2 = Signal for unfortified aliquot optimum precision) to the prepared samples V1 = Volume of the standard addition (L) (blanks and standards) that are affected the V2 = Volume of the sample aliquot (L) used same as the analytes by the sample matrix. for MSA Use the ratio of analyte signal to the inter- nal standard signal for calibration and quan- For more than one fortified portion of the titation. prepared sample, linear regression analysis can be applied using a computer or calcu- 12.0 Data Analysis and Calculations lator program to obtain the concentration of the sample solution. An alternative to using 12.1 Sample data should be reported in the method of standard additions is use of units of mg/L for aqueous samples and mg/kg the internal standard technique by adding dry weight for solid samples.

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12.2 For dissolved aqueous analytes (Sec- in processing or additional dilutions required tion 11.1) report the data generated directly to complete the analysis. from the instrument with allowance for sam- 12.4 For analytes with MDLs <0.01 mg/L, ple dilution. Do not report analyte con- round the data values to the thousandth centrations below the IDL. place and report analyte concentrations up 12.3 For total recoverable aqueous to three significant figures. For analytes analytes (Section 11.2), multiply solution with MDLs <0.01 mg/L round the data values analyte concentrations by the dilution fac- to the 100th place and report analyte con- tor 0.5, when 100 mL aliquot is used to centrations up to three significant figures. produce the 50 mL final solution, and report Extract concentrations for solids data should data as instructed in Section 12.4. If a dif- ferent aliquot volume other than 100 mL is be rounded in a similar manner before cal- used for sample preparation, adjust the dilu- culations in Section 12.5 are performed. tion factor accordingly. Also, account for 12.5 For total recoverable analytes in any additional dilution of the prepared sam- solid samples (Section 11.3), round the solu- ple solution needed to complete the deter- tion analyte concentrations (mg/L) as in- mination of analytes exceeding 90% or more structed in Section 12.4. Report the data up of the LDR upper limit. Do not report data to three significant figures as mg/kg dry- below the determined analyte MDL con- weight basis unless specified otherwise by centration or below an adjusted detection the program or data user. Calculate the con- limit reflecting smaller sample aliquots used centration using the equation below:

where: Do not report analyte data below the esti- C = Concentration in extract (mg/L) mated solids MDL or an adjusted MDL be- V = Volume of extract (L, 100 mL = 0.1L) cause of additional dilutions required to D = Dilution factor (undiluted = 1) complete the analysis. W = Weight of sample aliquot extracted (g × 12.6 To report percent solids in solid sam- 0.001 = kg) ples (Section 11.3) calculate as follows:

where: used to avoid boron and silica contamination DW = Sample weight (g) dried at 60 ßC from glassware with the final dilution to 50 WW = Sample weight (g) before drying mL completed in polypropylene centrifuged tubes. The listed MDLs for solids are esti- Note: If the data user, program or labora- mates and were calculated from the aqueous tory requires that the reported percent sol- ids be determined by drying at 105 °C, repeat MDL determinations. the procedure given in Section 11.3 using a 13.2 Data obtained from single laboratory separate portion (>20 g) of the sample and method testing are summarized in Table 6 dry to constant weight at 103–105 °C. for five types of water samples consisting of drinking water, surface water, ground water, 12.7 The QC data obtained during the analyses provide an indication of the quality and two wastewater effluents. The data pre- of the sample data and should be provided sented cover all analytes except cerium and with the sample results. titanium. Samples were prepared using the procedure described in Section 11.2. For each 13.0 Method Performance matrix, five replicate aliquots were prepared, analyzed and the average of the five deter- 13.1 Listed in Table 4 are typical single minations used to define the sample back- laboratory total recoverable MDLs deter- mined for the recommended wavelengths ground concentration of each analyte. In ad- using simultaneous ICP–AES and the oper- dition, two pairs of duplicates were fortified ating conditions given in Table 5. The MDLs at different concentration levels. For each were determined in reagent blank matrix method analyte, the sample background con- (best case situation). PTFE beakers were centration, mean percent recovery, standard

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deviation of the percent recovery, and rel- agement for Waste Reduction’’, available ative percent difference between the dupli- from the American Chemical Society’s De- cate fortified samples are listed in Table 6. partment of Government Relations and The variance of the five replicate sample Science Policy, 1155 16th Street NW., Wash- background determinations is included in ington, DC 20036, (202) 872–4477. the calculated standard deviation of the per- cent recovery when the analyte concentra- 15.0 Waste Management tion in the sample was greater than the 15.1 The Environmental Protection Agen- MDL. The tap and well waters were proc- cy requires that laboratory waste manage- essed in Teflon and quartz beakers and di- ment practices be conducted consistent with luted in polypropylene centrifuged tubes. all applicable rules and regulations. The The nonuse of borosilicate glassware is re- Agency urges laboratories to protect the air, flected in the precision and recovery data for water, and land by minimizing and control- boron and silica in those two sample types. ling all releases from hoods and bench oper- 13.3 Data obtained from single laboratory ations, complying with the letter and spirit method testing are summarized in Table 7 of any sewer discharge permits and regula- for three solid samples consisting of EPA 884 tions, and by complying with all solid and Hazardous Soil, SRM 1645 River Sediment, hazardous waste regulations, particularly and EPA 286 Sludge. Samples the hazardous waste identification rules and were prepared using the procedure described land disposal restrictions. For further infor- in Section 11.3. For each method analyte, the mation on waste management consult ‘‘The sample background concentration, mean per- Waste Management Manual for Laboratory cent recovery of the fortified additions, the Personnel’’, available from the American standard deviation of the percent recovery, Chemical Society at the address listed in the and relative percent difference between du- Section 14.2. plicate additions were determined as de- scribed in Section 13.2. Data presented are 16.0 References for all analytes except cerium, silica, and ti- tanium. Limited comparative data to other 1. U.S. Environmental Protection Agency. methods and SRM materials are presented in Inductively Coupled Plasma—Atomic Reference 23 of Section 16.0. Emission Spectrometric Method for 13.4 Performance data for aqueous solu- Trace Element Analysis of Water and tions independent of sample preparation Wastes—Method 200.7, Dec. 1982. EPA–600/ from a multilaboratory study are provided in 4–79–020, revised March 1983. Table 8.22 2. U.S. Environmental Protection Agency. 13.5 Listed in Table 9 are regression equa- Inductively Coupled Plasma Atomic tions for precision and bias for 25 analytes Emission Spectroscopy Method 6010, SW– abstracted from EPA Method Study 27, a 846 Test Methods for Evaluating Solid multilaboratory validation study of Method Waste, 3rd Edition, 1986. 200.7.1 These equations were developed from 3. U.S. Environmental Protection Agency. data received from 12 laboratories using the Method 200.7: Determination of Metals total recoverable sample preparation proce- and Trace Elements in Water and Wastes dure on reagent water, drinking water, sur- by Inductively Coupled Plasma—Atomic face water and three industrial effluents. For Emission Spectrometry, revision 3.3, a complete review and description of the EPA 600 4–91/010, June 1991. study, see Reference 16 of Section 16.0. 4. U.S. Environmental Protection Agency. Inductively Coupled Plasma—Atomic 14.0 Pollution Prevention Emission Spectrometry Method for the 14.1 Pollution prevention encompasses Analysis of Waters and Solids, EMMC, any technique that reduces or eliminates the July 1992. quantity or toxicity of waste at the point of 5. Fassel, V.A. et al. Simultaneous Deter- generation. Numerous opportunities for pol- mination of Wear Metals in Lubricating lution prevention exist in laboratory oper- Oils by Inductively-Coupled Plasma ation. The EPA has established a preferred Atomic Emission Spectrometry. Anal. hierarchy of environmental management Chem. 48:516–519, 1976. techniques that places pollution prevention 6. Merryfield, R.N. and R.C. Loyd. Simulta- as the management option of first choice. neous Determination of Metals in Oil by Whenever feasible, laboratory personnel Inductively Coupled Plasma Emission should use pollution prevention techniques Spectrometry. Anal. Chem. 51:1965–1968, to address their waste generation (e.g., Sec- 1979. tion 7.8). When wastes cannot be feasibly re- 7. Winge, R.K. et al. Inductively Coupled duced at the source, the Agency recommends Plasma—Atomic Emission Spectroscopy: recycling as the next best option. An Atlas of Spectral Information, Phys- 14.2 For information about pollution pre- ical Science Data 20. Elsevier Science vention that may be applicable to labora- Publishing, New York, New York, 1985. tories and research institutions, consult 8. Boumans, P.W.J.M. Line Coincidence Ta- ‘‘Less is Better: Laboratory Chemical Man- bles for Inductively Coupled Plasma

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Atomic Emission Spectrometry, 2nd edi- Technical Information Service (NTIS) as tion. Pergamon Press, Oxford, United PB 85–248–656. Kingdom, 1984. 17. Botto, R.I. Quality Assurance in Oper- 9. Carcinogens—Working With Carcinogens, ating a Multielement ICP Emission Spec- Department of Health, Education, and trometer. Spectrochim. Acta, 39B(1):95– Welfare, Public Health Service, Center 113, 1984. for Disease Control, National Institute 18. Wallace, G.F., Some Factors Affecting for Occupational Safety and Health, Pub- the Performance of an ICP Sample Intro- lication No. 77–206, Aug. 1977. Available duction System. Atomic Spectroscopy, from the National Technical Information Vol. 4, p. 188–192, 1983. Service (NTIS) as PB–277256. 19. Koirtyohann, S.R. et al. Nomenclature 10. OSHA Safety and Health Standards, Gen- System for the Low-Power Argon Induc- eral Industry, (29 CFR 1910), Occupa- tively Coupled Plasma, Anal. Chem. tional Safety and Health Administration, 52:1965, 1980. OSHA 2206, (Revised, January 1976). 20. Deming, S.N. and S.L. Morgan. Experi- 11. Safety in Academic Chemistry Labora- mental Design for Quality and Produc- tories, American Chemical Society Pub- tivity in Research, Development, and lication, Committee on Chemical Safety, Manufacturing, Part III, pp. 119–123. 3rd Edition, 1979. Short course publication by Statistical 12. Proposed OSHA Safety and Health Stand- Designs, 9941 Rowlett, Suite 6, Houston, ards, Laboratories, Occupational Safety TX 77075, 1989. 21. Winefordner, J.D., Trace Analysis: and Health Administration, FEDERAL Spectroscopic Methods for Elements, REGISTER, July 24, 1986. Chemical Analysis, Vol. 46, pp. 41–42. 13. Rohrbough, W.G. et al. Reagent Chemi- 22. Jones, C.L. et al. An Interlaboratory cals, American Chemical Society Speci- Study of Inductively Coupled Plasma fications, 7th edition. American Chem- Atomic Emission Spectroscopy Method ical Society, Washington, DC, 1986. 6010 and Digestion Method 3050. EPA–600/ 14. American Society for Testing and Mate- 4–87–032, U.S. Environmental Protection rials. Standard Specification for Reagent Agency, Las Vegas, Nevada, 1987. Water, D1193–77. Annual Book of ASTM 23. Martin, T.D., E.R. Martin and SE. Long. Standards, Vol. 11.01. Philadelphia, PA, Method 200.2: Sample Preparation Proce- 1991. dure for Spectrochemical Analyses of Total 15. Code of Federal Regulations 40, Ch. 1, Pt. Recoverable Elements, EMSL ORD, 136 Appendix B. USEPA, 1989. 16. Maxfield, R. and B. Mindak. EPA Method Study 27, Method 200.7 Trace Metals by 17.0 Tables, Diagrams, Flowcharts, and ICP, Nov. 1983. Available from National Validation Data

TABLE 1—WAVELENGTHS, ESTIMATED INSTRUMENT DETECTION LIMITS, AND RECOMMENDED CALIBRATION

Estimated Wavelengtha Calibratec Analyte (nm) detection to (mg/L) limitb (μg/L)

Aluminum ...... 308.215 45 10 Antimony ...... 206.833 32 5 Arsenic ...... 193.759 53 10 Barium ...... 493.409 2.3 1 Beryllium ...... 313.042 0.27 1 Boron ...... 249.678 5.7 1 Cadmium ...... 226.502 3.4 2 Calcium ...... 315.887 30 10 Cerium ...... 413.765 48 2 Chromium ...... 205.552 6.1 5 Cobalt ...... 228.616 7.0 2 Copper ...... 324.754 5.4 2 Iron ...... 259.940 6.2 10 Lead ...... 220.353 42 10 Lithium ...... 670.784 d 3.7 5 Magnesium ...... 279.079 30 10 Manganese ...... 257.610 1.4 2 Mercury ...... 194.227 2.5 2 Molybdenum ...... 203.844 12 10 Nickel ...... 231.604 15 2 Phosphorus ...... 214.914 76 10 Potassium ...... 766.491 e 700 20 Selenium ...... 196.090 75 5 d Silica (SiO2) ...... 251.611 26 (SiO2) 10 Silver ...... 328.068 7.0 0.5

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TABLE 1—WAVELENGTHS, ESTIMATED INSTRUMENT DETECTION LIMITS, AND RECOMMENDED CALIBRATION—Continued

Estimated Wavelengtha Calibratec Analyte (nm) detection to (mg/L) limitb (μg/L)

Sodium ...... 588.995 29 10 Strontium ...... 421.552 0.77 1 Thallium ...... 190.864 40 5 Tin ...... 189.980 25 4 Titanium ...... 334.941 3.8 10 Vanadium ...... 292.402 7.5 2 Zinc ...... 213.856 1.8 5 a The wavelengths listed are recommended because of their sensitivity and overall acceptability. Other wavelengths may be substituted if they can provide the needed sensitivity and are treated with the same corrective techniques for spectral inter- ference (see Section 4.1). b These estimated 3-sigma instrumental detection limits 16 are provided only as a guide to instrumental limits. The method de- tection limits are sample dependent and may vary as the sample matrix varies. Detection limits for solids can be estimated by di- viding these values by the grams extracted per liter, which depends upon the extraction procedure. Divide solution detection lim- its by 10 for 1 g extracted to 100 mL for solid detection limits. c Suggested concentration for instrument calibration.2 Other calibration limits in the linear ranges may be used. d Calculated from 2-sigma data.5 e Highly dependent on operating conditions and plasma position.

TABLE 2—ON-LINE METHOD INTERELEMENT TABLE 3—MIXED STANDARD SOLUTIONS SPECTRAL INTERFERANCES ARISING FROM Solu- INTERFERANTS AT THE 100 MG/L LEVEL tion Analytes

Wave- I ...... Ag, As, B, Ba, Ca, Cd, Cu, Mn, Sb, and Se Analyte length Interferant* II ...... K, Li, Mo, Na, Sr, and Ti (nm) III ..... Co, P, V, and Ce IV .... Al, Cr, Hg, SiO , Sn, and Zn Ag ...... 328.068 Ce, Ti, Mn 2 V ..... Be, Fe, Mg, Ni, Pb, and Tl Al ...... 308.215 V, Mo, Ce, Mn As ...... 193.759 V, Al, Co, Fe, Ni B ...... 249.678 None TABLE 4—TOTAL RECOVERABLE METHOD Ba ...... 493.409 None Be ...... 313.042 V, Ce DETECTION LIMITS (MDL) Ca ...... 315.887 Co, Mo, Ce Cd ...... 226.502 Ni, Ti, Fe, Ce MDLs (2) Analyte (1) Solids, mg/kg Ce ...... 413.765 None Aqueous, mg/L Co ...... 228.616 Ti, Ba, Cd, Ni, Cr, Mo, Ce Cr ...... 205.552 Be, Mo, Ni Ag ...... 0.002 0.3 Cu ...... 324.754 Mo, Ti Al ...... 0.02 3 Fe ...... 259.940 None As ...... 0.008 2 Hg ...... 194.227 V, Mo B ...... 0.003 — K ...... 766.491 None Ba ...... 0.001 0.2 Li ...... 670.784 None Be ...... 0.0003 0.1 Mg ...... 279.079 Ce Ca ...... 0.01 2 Mn ...... 257.610 Ce Cd ...... 0.001 0.2 Mo ...... 203.844 Ce Ce ...... 0.02 3 Na ...... 588.995 None Co ...... 0.002 0.4 Ni ...... 231.604 Co, Tl Cr ...... 0.004 0.8 P ...... 214.914 Cu, Mo Cu ...... 0.003 0.5 Pb ...... 220.353 Co, Al, Ce, Cu, Ni, Ti, Fe Fe ...... *0.03 6 Sb ...... 206.833 Cr, Mo, Sn, Ti, Ce, Fe Se ...... 196.099 Fe Hg ...... 0.007 2 K ...... 0.3 60 SiO2 ...... 251.611 None Sn ...... 189.980 Mo, Ti, Fe, Mn, Si Li ...... 0.001 0.2 Sr ...... 421.552 None Mg ...... 0.02 3 Tl ...... 190.864 Ti, Mo, Co, Ce, Al, V, Mn Mn ...... 0.001 0.2 Ti ...... 334.941 None Mo ...... 0.004 1 V ...... 292.402 Mo, Ti, Cr, Fe, Ce Na ...... 0.03 6 Zn ...... 213.856 Ni, Cu, Fe Ni ...... 0.005 1 P ...... 0.06 12 * These on-line interferences from method analytes and tita- nium only were observed using an instrument with 0.035 nm Pb ...... 0.01 2 resolution (see Section 4.1.2). Interferant ranked by mag- Sb ...... 0.008 2 nitude of intensity with the most severe interferant listed first Se ...... 0.02 5 in the row. SiO2 ...... 0.02 — Sn ...... 0.007 2 Sr ...... 0.0003 0.1 Tl ...... 0.001 0.2 Ti ...... 0.02 3 V ...... 0.003 1

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TABLE 4—TOTAL RECOVERABLE METHOD TABLE 5—INDUCTIVELY COUPLED PLASMA DETECTION LIMITS (MDL)—Continued INSTRUMENT OPERATING CONDITIONS

MDLs Analyte Solids, mg/kg(2) Incident rf power ...... 1100 watts Aqueous, mg/L(1) Reflected rf power ...... <5 watts Zn ...... 0.002 0.3 Viewing height above work coil ...... 15 mm Injector tube orifice i.d...... 1 mm (1) MDL concentrations are computed for original matrix with allowance for 2x sample preconcentration during preparation. Argon supply ...... liquid argon Samples were processed in PTFE and diluted in 50-mL plas- Argon pressure ...... 40 psi tic centrifuge tubes. Coolant argon flow rate ...... 19 L/min. (2) Estimated, calculated from aqueous MDL determinations. Aerosol carrier argon flow rate ...... 620 mL/min. — Boron not reported because of glassware contamination. Silica not determined in solid samples. Auxiliary (plasma) argon flow rate .. 300 mL/min. * Elevated value due to fume-hood contamination. Sample uptake rate controlled to ... 1.2 mL/min.

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mg/L High spike ATRICES M QUEOUS A S (R) RPD ATA IN D R (%) Average recovery Tap Water Pond Water ECOVERY R mg/L Low spike RECISION AND mg/L conc. 6—P Sample ABLE T Analyte ...... 20.0 96 1.1 2.3 104 3.3 3.4 6.5 5.0 2 Ag ...... Al ...... As ...... B0.05 95 0.7 2.1 0.2 ...... <0.002 96 0.0 Ba ...... 101 0.7 2.0 0.05 105 3.0 3.1 108 1.4 3.7 0.2 98 8.8 1.7 0.2 Be0.05 0.185 ...... <0.008 Ca ...... Cd 0.023 0.1 98 0.2 0.0 0.4 ...... 100 0.0 0.1 102 1.6 2.2 0.2 98 0.2 0.5 0.01 99 0.0 0.05 98 0.4 0.8 Co <0.0003 0.042 ...... Cr ...... 103 2.0 0.9 105 3.5 9.5 0.1 20.0 Cu0.01 98 0.0 101 8.8 1.7 ...... <0.001 35.2 5.0 100 0.0 0.2 Fe0.02 99 0.5 1.5 ...... <0.002 102 0.0 110 0.0 0.1 Hg0.01 ...... 101 1.2 3.5 <0.004 103 1.8 4.9 0.2 K0.02 ...... <0.003 Li ...... 100 0.4 1.0 105 0.3 0.5 103 0.7 1.9 0.2 106 1.0 1.8 0.4 Mg0.05 0.008 0.1 ...... <0.007 Mn ...... Mo107 0.7 1.7 110 1.9 4.4 109 1.4 2.3 20...... 103 6.9 3.8 0.2 1.98 5.0 0.02 Na 0.006 ...... 100 0.7 1.1 100 0.0 0.1 20.0 Ni0.01 99 0.0 ...... 104 2.2 1.5 <0.001 8.08 5.0 P ...... <0.004 0.02 Pb ...... 95 104 1.1 2.9 108 1.8 4.7 0.2 3.5 10.5 Sb106 1.0 1.6 0.02 20.0 ...... 0.2 10.3 5.0 <0.005 108 99 3.0 2.0 Se 0.5 ...... 1.4 SiO 0.045 13.1 0.1 102 9.4 0.05 100 0.2 0.5 0.4 104 95 0.7 2.1 0.2 3.2 <0.01 Sn0.05 102 0.7 2.0 1.3 99 0.7 2.0 0.2 ...... <0.008 Sr ...... <0.02 0.1 Tl 87 1.1 3.5 0.4 ...... 99 0.8 2.3 101 1.8 5.0 103 2.1 5.8 0.2 V0.05 ...... <0.007 Zn ...... 105 0.8 1.0 102 3.3 2.1 0.4 0.181 0.1 101 0.1 0.3 10.9 0.4 101 0.7 2.0 0.2 101 3.9 0.05 99 0.2 0.5 <0.02 0.1 <0.003 101 3.7 9.0 0.2 0.05 98 0.9 2.5 Ag 0.005 ...... Al ...... As ...... B0.05 92 0.0 0.2 ...... <0.002 94 0.0 Ba ...... 100 2.9 3.7 10.0 5.0 0.8 102 0.0 0.2 Be 0.819 0.2 0.05 98 1.4 4.1 88 ...... <0.008 Ca ...... 103 2.0 0.0 111 8.9 6.9 0.4 Cd 0.034 0.1 ...... 0.05 0.01 96 0.9 0.0 0.2 95 0.4 1.1 0.2 Co 0.029 97 0.3 0.5 <0.0003 95 0.0 ...... Cr ...... 107 0.0 0.1 0.01 97 0.0 53.9 5.0 <0.001 100 2.7 7.5 0.2 0.02 97 0.7 2.1 * <0.002

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High + spike ATRICES M OLID S S (R) RPD ATA IN D (%) ECOVERY Average recovery R R ¥ ¥ EPA Hazardous Soil #884 mg/kg Low + spike RECISION AND 7—P conc. mg/kg Sample ABLE T Analyte ...... 100 2.2 3.0 20.0 6.83 5.0 99 6.8 1.7 2 S (R) Standard deviation of percent recovery. RPD Relative percent difference between duplicate spike determinations.

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TABLE 8—ICP–AES INSTRUMENTAL PRECISION AND ACCURACY FOR AQUEOUS SOLUTIONS A

c Mean conc. b Accurace Element (mg/L) N RSD (%) (% of Nominal)

Al ...... 14.8 8 6.3 100 Sb ...... 15.1 8 7.7 102 As ...... 14.7 7 6.4 99 Ba ...... 3.66 7 3.1 99 Be ...... 3.78 8 5.8 102 Cd ...... 3.61 8 7.0 97 Ca ...... 15.0 8 7.4 101 Cr ...... 3.75 8 8.2 101 Co ...... 3.52 8 5.9 95 Cu ...... 3.58 8 5.6 97 Fe ...... 14.8 8 5.9 100 Pb ...... 14.4 7 5.9 97 Mg ...... 14.1 8 6.5 96 Mn ...... 3.70 8 4.3 100 Mo ...... 3.70 8 6.9 100 Ni ...... 3.70 7 5.7 100 K ...... 14.1 8 6.6 95 Se ...... 15.3 8 7.5 104 Na ...... 14.0 8 4.2 95 Tl ...... 15.1 7 8.5 102 V ...... 3.51 8 6.6 95 Zn ...... 3.57 8 8.3 96 a These performance values are independent of sample preparation because the labs analyzed portions of the same solutions using sequential or simultaneous instruments. b N = Number of measurements for mean and relative standard deviation (RSD). c Accuracy is expressed as a percentage of the nominal value for each analyte in the acidified, multi-element solutions.

TABLE 9—MULTILABORATORY ICP PRECISION AND ACCURACY DATA*

Concentration Total recoverable digestion Analyte μg/L μ/L

Aluminum ...... 69–4792 X = 0.9380 (C) + 22.1 SR = 0.0481 (X) + 18.8 Antimony ...... 77–1406 0.8908 (C) + 0.9 SR = 0.0682 (X) + 2.5 Arsenic ...... 69–1887 X = 1.0175 (C) + 3.9 SR = 0.0643 (X) + 10.3 Barium ...... 9–377 X = 0.8.80 (C) + 1.68 SR = 0.0826 (X) + 3.54 Beryllium ...... 3–1906 X = 1.0177 (C) ¥ 0.55 SR = 0.0445 (X) ¥ 0.10 Boron ...... 19–5189 X = 0.9676 (C) + 18.7 SR = 0.0743 (X) + 21.1 Cadmium ...... 9–1943 X = 1.0137 (C) ¥ 0.65 SR = 0.0332 (X) + 0.90 Calcium ...... 17–47170 X = 0.9658 (C) + 0.8 SR = 0.0327 (X) + 10.1 Chromium ...... 13–1406 X = 1.0049 (C) ¥ 1.2 SR = 0.0571 (X) + 1.0 Cobalt ...... 17–2340 X = 0.9278 (C) + 1.5 SR = 0.0407 (X) + 0.4 Copper ...... 8–1887 X = 0.9647 (C) ¥ 3.64 SR = 0.0406 (X) + 0.96 Iron ...... 13–9359 X = 0.9830 (C) + 5.7 SR = 0.0790 (X) + 11.5 Lead ...... 42–4717 X = 1.0056 (C) + 4.1 SR = 0.0448 (X) + 3.5 Magnesium ...... 34–13868 X = 0.9879 (C) + 2.2 SR = 0.0268 (X) + 8.1 Manganese ...... 4–1887 X = 0.9725 (C) + 0.07 SR = 0.0400 (X) + 0.82 Molybdenum ...... 17–1830 X = 0.9707 (C) ¥ 2.3 SR = 0.0529 (X) + 2.1 Nickel ...... 17–47170 X = 0.9869 (C) + 1.5 SR = 0.0393 (X) + 2.2 Potassium ...... 347–14151 X = 0.9355 (C) ¥ 183.1 SR = 0.0329 (X) + 60.9 Selenium ...... 69–1415 X = 0.9737 (C) ¥ 1.0 SR = 0.0443 (X) + 6.6 Silicon ...... 189–9434 X = 0.9737 (C) ¥ 22.6

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TABLE 9—MULTILABORATORY ICP PRECISION AND ACCURACY DATA*—Continued

Concentration Total recoverable digestion Analyte μg/L μ/L

SR = 0.2133 (X) + 22.6 Silver ...... 8–189 X = 0.3987 (C) + 8.25 SR = 0.1836 (X) ¥ 0.27 Sodium ...... 35–47170 X = 1.0526 (C) + 26.7 SR = 0.0884 (X) + 50.5 Thallium ...... 79–1434 X = 0.9238 (C) + 5.5 SR = 0.0106 (X) + 48.0 Vanadium ...... 13–4698 X = 0.9551 (C) + 0.4 SR = 0.0472 (X) + 0.5 Zinc ...... 7–7076 X = 0.9500 (C) + 1.82 SR = 0.0153 (X) + 7.78 *—Regression equations abstracted from Reference 16. X = Mean Recovery, μg/L. C = True Value for the Concentration, μg/L. SR = Single-analyst Standard Deviation, μg/L.

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Net Emision Intensity Counts (X103 ) 32r------~

30

28

26

24

22

20

18

16

14

12~--~----~----L---~----~----~--~ 476 526 575 626 676 726 776 826 Nebulizer Argon Flow Rate - ml/min Figure 1

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[77 FR 29813, May 18, 2012] centrates containing various levels of this element were added to reagent water, surface APPENDIX D TO PART 136—PRECISION water, drinking water and three effluents. AND RECOVERY STATEMENTS FOR These samples were digested by the total di- METHODS FOR MEASURING METALS gestion procedure, 4.1.3 in this manual. Re- sults for the reagent water are given below. Two selected methods from ‘‘Methods for Results for other water types and study de- Chemical Analysis of Water and Wastes,’’ tails are found in ‘‘EPA Method Study 31, EPA–600/4–79–020 (1979) have been subjected Trace Metals by Atomic Absorption (Fur- to interlaboratory method validation stud- nace Techniques),’’ National Technical Infor- ies. The two selected methods are for Thal- mation Service, 5285 Port Royal Road, lium and Zinc. The following precision and Springfield, VA 22161 Order No. PB 86–121 704/ recovery statements are presented in this ap- AS, by Copeland, F.R. and Maney, J.P., Jan- pendix and incorporated into Part 136: uary 1986. For a concentration range of 0.51–189 μg/L Method 279.2 X = 1.6710(C) + 1.485 For Thallium, Method 279.2 (Atomic Ab- S = 0.6740(X) ¥ 0.342 sorption, Furnace Technique) replace the SR = 0.3895(X)¥ 0.384 Precision and Accuracy Section statement Where: with the following: C = True Value for the Concentration, μg/L Precision and Accuracy X = Mean Recovery, μg/L μ An interlaboratory study on metal anal- S = Multi-laboratory Standard Deviation, g/ yses by this method was conducted by the L μ Quality Assurance Branch (QAB) of the Envi- SR = Single-analyst Standard Deviation, g/ ronmental Monitoring Systems Laboratory— L Cincinnati (EMSL–CI). Synthetic con- [77 FR 29833, May 18, 2012] centrates containing various levels of this element were added to reagent water, surface water, drinking water and three effluents. PART 140—MARINE SANITATION These samples were digested by the total di- DEVICE STANDARD gestion procedure, 4.1.3 in this manual. Re- sults for the reagent water are given below. Sec. Results for other water types and study de- 140.1 Definitions. tails are found in ‘‘EPA Method Study 31, 140.2 Scope of standard. Trace Metals by Atomic Absorption (Fur- 140.3 Standard. nace Techniques),’’ National Technical Infor- 140.4 Complete prohibition. mation Service, 5285 Port Royal Road, 140.5 Analytical procedures. Springfield, VA 22161 Order No. PB 86–121 704/ AS, by Copeland, F.R. and Maney, J.P., Jan- AUTHORITY: 33 U.S.C. 1322, as amended. uary 1986. SOURCE: 41 FR 4453, Jan. 29, 1976, unless For a concentration range of 10.00–252 μg/L otherwise noted. X = 0.8781(C) ¥ 0.715 S = 0.1112(X) + 0.669 § 140.1 Definitions. SR = 0.1005(X) + 0.241 For the purpose of these standards Where: the following definitions shall apply: C = True Value for the Concentration, μg/L (a) Sewage means human body wastes μ X = Mean Recovery, g/L and the wastes from toilets and other S = Multi-laboratory Standard Deviation, μg/ L receptacles intended to receive or re- SR = Single-analyst Standard Deviation, μg/ tain body wastes; L (b) Discharge includes, but is not lim- ited to, any spilling, leaking, pumping, Method 289.2 pouring, emitting, emptying, or dump- For Zinc, Method 289.2 (Atomic Absorp- ing; tion, Furnace Technique) replace the Preci- (c) Marine sanitation device includes sion and Accuracy Section statement with any equipment for installation onboard the following: a vessel and which is designed to re- Precision and Accuracy ceive, retain, treat, or discharge sew- age and any process to treat such sew- An interlaboratory study on metal anal- yses by this method was conducted by the age; Quality Assurance Branch (QAB) of the Envi- (d) Vessel includes every description ronmental Monitoring Systems Laboratory— of watercraft or other artificial con- Cincinnati (EMSL–CI). Synthetic con- trivance used, or capable of being used,

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