SEPA United States Environmental Protection Agency Analytical Methods Approved for Compliance Monitoring under the Revised Total Coliform Rule Analysis for the following contaminants shall be conducted in accordance with the methods in the following table, or their equivalent as determined by EPA. The methods and monitoring requirements for these contaminants are specified in 40 CFR 141.852(a)(5). Additional methods are listed in Appendix A to Subpart C of Part 141. The CFR is the legal reference for approved methods and takes precedent over this table. The table should accurately reflect the analytical methods information published in 40 CFR 141. §141.852 - Analytical Methods and Laboratory Certification (a) Analytical methodology (1) The standard sample volume required for analysis, regardless of analytical method used, is 100 mL. (2) Systems need only determine the presence or absence of total coliforms and E. coli; a determination of density is not required. (3) The time from sample collection to initiation of test medium incubation may not exceed 30 hours. Systems are encouraged but not required to hold samples below 10°C during transit. (4) If water having residual chlorine (measured as free, combined, or total chlorine) is to be analyzed, sufficient sodium thiosulfate (Na2S203) must be added to the sample bottle before sterilization to neutralize any residual chlorine in the water sample. Dechlorination procedures are addressed in Section 9060A.2 of Standard Methods for the Examination of Water and Wastewater (20th and 21st editions). (5) Systems must conduct total coliform and E.coli analyses in accordance with one of the analytical methods in the table at §141.852 or one of the alternative methods listed in Appendix A to Subpart C of Part 141. The procedures must be done in accordance with the documents listed at paragraph (c) of §141.852 or one of the alternative methods listed in Appendix A to Subpart C of Part 141. For Standard Methods Online, the year in which each method was approved by the Standard Methods Committee is designated by the last two digits following the hyphen in the method number. The methods listed are the only online versions of the method that maybe used. For vendor methods, the date of the method listed in paragraph (c) of §141.852 or Appendix A to Subpart C of Part 141 is the date/version of the approved method. The methods listed are the only versions that may be used for compliance with this rule. Laboratories should be careful to use only the approved versions of the methods as product packaging inserts may not be the same as the approved versions of the methods. (b) Laboratory certification- systems must have all compliance samples required under this subpart analyzed by a laboratory certified by EPA or a primacy state to analyze drinking water samples. The laboratory used by the system must be certified for each method (and associated contaminant(s)) used for compliance monitoring analyses under this rule. (c) Incorporate by reference - The standards required in §141.852 (a) are incorporated by reference into the regulation. Contaminant Office of Water (MS - 4606 M) EPA 815-B-19-007 April 2019 ------- Page 2 of 13 Total Coliforms - lactose fermentation methods: Method Organization Reference Title/Source Date Notes 9221 B.l, B.2 Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 Lactose broth, as commercially available, may be used in lieu of lauryl tryptose broth, if the system conducts at least 25 parallel tests between lactose broth and lauryl tryptose broth using the water normally tested, and if the findings from this comparison demonstrate that the false-positive rate and the false-negative rate for total coliforms, using lactose broth, is less than 10 percent 9221 B.l, B.2 Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 Lactose broth, as commercially available, may be used in lieu of lauryl tryptose broth, if the system conducts at least 25 parallel tests between lactose broth and lauryl tryptose broth using the water normally tested, and if the findings from this comparison demonstrate that the false-positive rate and the false-negative rate for total coliforms, using lactose broth, is less than 10 percent 9221 B.l, B.2 Standard Methods Standard Methods for the Examination of Water and Wastewater, 22nd Edition Standard Methods 2012 Lactose broth, as commercially available, may be used in lieu of lauryl tryptose broth, if the system conducts at least 25 parallel tests between lactose broth and lauryl tryptose broth using the water normally tested, and if the findings from this comparison demonstrate that the false-positive rate and the false-negative rate for total coliforms, using lactose broth, is less than 10 percent 9221 B.l, B.2, B.3, B.4 Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 Lactose broth, as commercially available, may be used in lieu of lauryl tryptose broth, if the system conducts at least 25 parallel tests between lactose broth and lauryl tryptose broth using the water normally tested, and if the findings from this comparison demonstrate that the false-positive rate and the false-negative rate for total coliforms, using lactose broth, is less than 10 percent 9221 B.l, B.2-99 Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. Standard Methods 1999 Lactose broth, as commercially available, may be used in lieu of lauryl tryptose broth, if the system conducts at least 25 parallel tests between lactose broth and lauryl tryptose broth using the water normally tested, and if the findings from this comparison demonstrate that the false-positive rate and the false-negative rate for total coliforms, using lactose broth, is less than 10 percent. 9221 B.l, B.2-06 Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the 2006 Lactose broth, as commercially available, may be used in lieu of lauryl tryptose broth, if the system conducts at least 25 parallel tests between lactose broth and lauryl tryptose broth using the ------- Page 3 of 13 Method Organization Reference Title/Source Date Notes regulations or in Appendix A to Subpart C of Part 141 are approved. Standard Methods water normally tested, and if the findings from this comparison demonstrate that the false-positive rate and the false-negative rate for total coliforms, using lactose broth, is less than 10 percent 9221 D.l, D.2 Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 A multiple tube enumerative format, as described in Standard Methods for the Examination of Water and Wastewater 9221, is approved for this method for use in presence-absence determination under this regulation. 9221 D.l, D.2 Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 A multiple tube enumerative format, as described in Standard Methods for the Examination of Water and Wastewater 9221, is approved for this method for use in presence-absence determination under this regulation. 9221 D.l, D.2, D.3 Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 A multiple tube enumerative format, as described in Standard Methods for the Examination of Water and Wastewater 9221, is approved for this method for use in presence-absence determination under this regulation. 9221 D.l, D.2-99 Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. Standard Methods 1999 A multiple tube enumerative format, as described in Standard Methods for the Examination of Water and Wastewater 9221, is approved for this method for use in presence-absence determination under this regulation. Total coliforms - membrane filtration methods: Method Organization Reference Title/Source Date Notes 9222 B, C Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. 9222 B, C Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. 9222 B, C Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. ------- Page 4 of 13 Method Organization Reference Title/Source Date Notes 9222 B, C-97 Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. Standard Methods 1997 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. 1604 EPA Method 1604: Total Coliforms and Escherichia coli in Water by Membrane Filtration Using a Simultaneous Detection Technique (Ml Medium), September 2002 EPA Method 1604 2002 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. m-ColiBlue24®Test Hach Company Membrane Filtration Method m- ColiBlue24® Broth, Revision 2, August 17,1999 m-ColiBlue 24 Test 1999 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. 9222 J m-ColiBlue24®Test Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. Chromocult EMD Millipore Chromocult® Coliform Agar Presence/Absence Membrane Filter Test Method for Detection and Identification of Coliform Bacteria and Escherichia coli for Finished Waters, November 2000, Version 1.0 Chromocult 2000 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. Total coliforms - enzyme substrate methods: Method Organization Reference Title/Source Date Notes 9223 B Colilert® Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. ------- Page 5 of 13 Method Organization Reference Title/Source Date Notes 9223 B Colilert® Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert® Standard Methods Standard Methods for the Examination of Water and Wastewater, 22nd Edition Standard Methods 2012 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert® Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B-97 Colilert® Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. 1997 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B-04 Colilert® Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. 2004 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colisure® Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. Colisure results may be read after an incubation time of 24 hours. 9223 B Colisure® Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. Colisure results may be read after an incubation time of 24 hours. 9223 B Colisure® Standard Methods Standard Methods for the Examination of Water and Wastewater, 22nd Edition Standard Methods 2012 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. ------- Page 6 of 13 Method Organization Reference Title/Source Date Notes Colisure results may be read after an incubation time of 24 hours. 9223 B Colisure® Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. Colisure results may be read after an incubation time of 24 hours. 9223 B-97 Colisure® Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. 1997 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. Colisure results may be read after an incubation time of 24 hours. 9223 B-04 Colisure® Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. 2004 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. Colisure results may be read after an incubation time of 24 hours. 9223 B Colilert-18® Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert-18® Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert-18® Standard Methods Standard Methods for the Examination of Water and Wastewater, 22nd Edition Standard Methods 2012 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert-18® Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B-04 Colilert-18® Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to 2004 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. ------- Page 7 of 13 Method Organization Reference Title/Source Date Notes Subpart C of Part 141 are approved. E*Colite® Charm Sciences Charm E*Colite™ Presence/Absence Test for Detection and Identification of Coliform Bacteria and Escherichia coli in Drinking Water, January 9,1998 Charm E*Colite 1998 Readycult® EMD Millipore Readycult® Coliforms 100 Presence/Absence Test for Detection and Identification of Coliform Bacteria and Escherichia coli in Finished Waters, January 2007, Version 1.1 Readvcult 2007 Modified Colitag® CPI International, Inc. Modified Colitag™ Test Method for the Simultaneous Detection of E. coli and other Total Coliforms in water (ATP D05- 0035), August 28, 2009 Modified Colitag 2009 Tecta EC/TC, v. 1.0 Veolia Water Solutions and Technologies Tecta™ EC/TC medium and the Tecta™ Instrument: A Presence/Absence Method for the Simultaneous Detection of Total Coliforms and Escherichia coli (E. coli) in Drinking Water, May 2014, Version 1.0 Tecta, Version 1.0 2014 Tecta EC/TC, v. 2.0 Pathogen Detection Systems, Inc. Tecta™ EC/TC medium and the Tecta™ Instrument: A Presence/Absence Method for the Simultaneous Detection of Total Coliforms and Escherichia coli (E. coli) in Drinking Water, March 2017, Version 2.0 Tecta, Version 2.0 2017 ------- Page 8 of 13 Contaminant Escherichia coli - Procedure (following Lactose Fermentation Methods): Method Organization Reference Title/Source Date Notes 9221 F.l Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 9221 F.l Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 9221 F.l Standard Methods Standard Methods for the Examination of Water and Wastewater, 22nd Edition Standard Methods 2012 9221 F.l Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 9221 F.l-06 Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. Standard Methods 2006 Contaminant Escherichia coli - Partition methods: Method Organization Reference Title/Source Date Notes 9222 G.lc(2) Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 9222 G.lc(2) Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 ------- Page 9 of 13 Method Organization Reference Title/Source Date Notes 9222 H Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 9222 G.lc(l) Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 9222 G.lc(l) Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 9222 1 Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 Contaminant Escherichia coli - membrane filtration methods: Method Organization Reference Title/Source Date Notes 1604 EPA Method 1604: Total Coliforms and Escherichia coli in Water by Membrane Filtration Using a Simultaneous Detection Technique (Ml Medium), September 2002 EPA Method 1604 2002 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. m- ColiBlue24®Test Hach Company Membrane Filtration Method m- ColiBlue24® Broth, Revision 2, August 17,1999 m-ColiBlue 24 Test 1999 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. 9222 J Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. ------- Page 10 of 13 Method Organization Reference Title/Source Date Notes Chromocult EMD Millipore Chromocult® Coliform Agar Presence/Absence Membrane Filter Test Method for Detection and Identification of Coliform Bacteria and Escherichia coli for Finished Waters, November 2000, Version 1.0 Chromocult 2000 All filtration series must begin with membrane filtration equipment that has been sterilized by autoclaving. Contaminant Escherichia coli - enzyme substrate methods: Method Organization Reference Title/Source Date Notes 9223 B Colilert® Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert® Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert® Standard Methods Standard Methods for the Examination of Water and Wastewater, 22nd Edition Standard Methods 2012 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert® Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert® B-97 Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. 1997 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. ------- Page 11 of 13 Method Organization Reference Title/Source Date Notes 9223 B Colilert® B-04 Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. 2004 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colisure® Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. Colisure results may be read after an incubation time of 24 hours. 9223 B Colisure® Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. Colisure results may be read after an incubation time of 24 hours. 9223 B Colisure® Standard Methods Standard Methods for the Examination of Water and Wastewater, 22nd Edition Standard Methods 2012 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. Colisure results may be read after an incubation time of 24 hours. 9223 B Colisure® Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. Colisure results may be read after an incubation time of 24 hours. 9223 B-97 Colisure® Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. 1997 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. Colisure results may be read after an incubation time of 24 hours. 9223 B Colisure® B-04 Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. 2004 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. Colisure results may be read after an incubation time of 24 hours. ------- Page 12 of 13 Method Organization Reference Title/Source Date Notes 9223 B Colilert-18® Standard Methods Standard Methods for the Examination of Water and Wastewater, 20th Edition Standard Methods 1998 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert-18® Standard Methods Standard Methods for the Examination of Water and Wastewater, 21st Edition Standard Methods 2005 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert-18® Standard Methods Standard Methods for the Examination of Water and Wastewater, 22nd Edition Standard Methods 2012 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B Colilert-18® Standard Methods Standard Methods for the Examination of Water and Wastewater, 23rd Edition Standard Methods 2017 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. 9223 B-04 Colilert-18® Standard Methods Online Online version. Approval year is designated by the last 2 digits. Only online versions cited in the regulations or in Appendix A to Subpart C of Part 141 are approved. 2004 Multiple-tube and multi-well enumerative formats for this method are approved for use in presence-absence determination under this regulation. E*Colite® Charm Sciences Charm E*Colite™ Presence/Absence Test for Detection and Identification of Coliform Bacteria and Escherichia coli in Drinking Water, January 9,1998 Charm E*Colite 1998 Readycult® EMD Millipore Readycult® Coliforms 100 Presence/Absence Test for Detection and Identification of Coliform Bacteria and Escherichia coli in Finished Waters, January 2007, Version 1.1 Readvcult 2007 ------- Page 13 of 13 Method Organization Reference Title/Source Date Notes Modified Colitag® CPI International, Inc. Modified Colitag™ Test Method for the Simultaneous Detection off. coli and other Total Coliforms in water (ATP D05- 0035), August 28, 2009 Modified Colitag 2009 Tecta EC/TC, v. 1.0 Veolia Water Solutions and Technologies Tecta™ EC/TC medium and the Tecta™ Instrument: A Presence/Absence Method for the Simultaneous Detection of Total Coliforms and Escherichia coli (E. coli) in Drinking Water, May 2014, Version 1.0 Tecta, Version 1.0 2014 Tecta EC/TC, v. 2.0 Pathogen Detection Systems, Inc. Tecta™ EC/TC medium and the Tecta™ Instrument: A Presence/Absence Method for the Simultaneous Detection of Total Coliforms and Escherichia coli (E. coli) in Drinking Water, March 2017, Version 2.0 Tecta, Version 2.0 2017 ------- |