&EPA
g^,protec,ior Analytical Methods Approved for Compliance Monitoring under the Long
Term 2 Enhanced Surface Water Treatment 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 for Cryptosporidium are listed at 40 CFR 141.704, the methods for enumeration of E. coli in source
water are listed in Table 1H at 40 CFR 136.3(a) and the methods for turbidity are listed at 40 CFR 141.74. Additional approved methods
are listed in Appendix A to Subpart C of Part 141.
The CFR is the legal reference for approved methods and takes precedence over this table. The table should accurately reflect the
analytical methods information published in 40 CFR 141. If discrepancies are found, please notify the Safe Drinking Water Hotline (800-
426-4791) so that EPA can correct the table.
Contaminant
Cryptosporidium: Systems must analyze at least a 10 L sample or a packed pellet volume of at least 2 mL. Systems unable to process a 10 L sample must
analyze as much sample volume as can be filtered by two filters approved by EPA for the methods listed, up to a packed pellet volume of
at least 2 mL.
Method
Organization
Reference Title
Date
EPA Publication
Number
1622
EPA
Cryptosporidium in Water by Filtration/IMS/FA
December
2005
EPA-815-R-05-001
1623
EPA
Cryptosporidium and Giardia in Water by
Filtration/IMS/FA
December
2005
EPA-815-R-05-002
1623.1
EPA
Cryptosporidium and Giardia in Water by
Filtration/IMS/FA
January 2012
EPA-816-R-12-001
Office of Water (MS - 4606 M)
EPA 821-F-17-001
February 2017

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Page 2 of 12
Contaminant
Escherichia coli: The time from sample collection to initiation of analysis may not exceed 30 hours. The State may approve on a case-by-case basis the
holding of an E.coli sample for up to 48 hours between sample collection and initiation of analysis if the State determines that analyzing
an E.coli sample within 30 hours is not feasible. E. coli samples held between 30 to 48 hours must be analyzed by the Colilert reagent
version of Standard Method 9223B as listed in § 136.3 (a) Table 1H of this title.
Systems must maintain samples between 0°C and 10°C during storage and transit to the laboratory.
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 organism density of the water sample.
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.
Method
Organization
Reference Title
Date
Notes
9221B.2 F-
2006
Standard
Methods
Online
Online version. Approval
year is designated by the last
4 digits. Only online versions
cited in the regulations or in
Appendix A to Subpart C of
Part 141 are approved.
2006
Samples shall be enumerated by the multiple-tube or multiple-well procedure.
Using multiple-tube procedures, employ an appropriate tube and dilution
configuration of the sample as needed and report the Most Probable Number
(MPN).
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.
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 + 3 h of incubation shall be submitted to 9221 F-
2006. Commercially available EC-MUG medium or EC medium supplemented
in the laboratory with 50 ng/mL of MUG may be used.
9223 B-2004
Colilert®
Standard
Methods
Online
Online version. Approval
year is designated by the last
4 digits. Only online versions
cited in the regulations or in
Appendix A to Subpart C of
Part 141 are approved.
2004
Multiple tube or multiple well
These tests are collectively known as defined substrate tests, where, for
example, a substrate is used to detect the enzyme p-glucuronidase produced
by E. coli
Descriptions of the Colilert®, Colilert-18®, and Quanti-Tray® may be obtained
from IDEXX Laboratories Inc.

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Page 3 of 12
Method
Organization
Reference Title
Date
Notes
9223 B-2004
Colilert-18®
Standard
Methods
Online
Online version. Approval
year is designated by the last
4 digits. Only online versions
cited in the regulations or in
Appendix A to Subpart C of
Part 141 are approved.
2004
Multiple tube or multiple well
These tests are collectively known as defined substrate tests, where, for
example, a substrate is used to detect the enzyme p-glucuronidase produced
by E. coli
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.
Descriptions of the Colilert®, Colilert-18®, and Quanti-Tray® may be obtained
from IDEXX Laboratories Inc.
991.15
Colilert®
AOAC
International
Official Methods of Analysis
of AOAC International, 16th
Edition, Volume 1, Chapter 17
1995
Multiple tube or multiple well
These tests are collectively known as defined substrate tests, where, for
example, a substrate is used to detect the enzyme p-glucuronidase produced
by E. coli
Descriptions of the Colilert®, Colilert-18®, and Quanti-Tray® may be obtained
from IDEXX Laboratories Inc.
991.15
Colilert-18®
AOAC
International
Official Methods of Analysis
of AOAC International, 16th
Edition, Volume 1, Chapter 17
1995
Multiple tube or multiple well
These tests are collectively known as defined substrate tests, where, for
example, a substrate is used to detect the enzyme p-glucuronidase produced
by E. coli
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.
Descriptions of the Colilert®, Colilert-18®, and Quanti-Tray® may be obtained
from IDEXX Laboratories Inc.

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Page 4 of 12
Method
Organization
Reference Title
Date
Notes
1103.1
EPA
EPA 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.
2010
A 0.45-nm 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.
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.
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 organism
density of the water sample.
When the MF method has not been used previously to test waters with high
turbidity, large numbers of nonconform 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.
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.

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Page 5 of 12
Method
Organization
Reference Title
Date
Notes
9222 B-
2006/9222 G-
2006
Standard
Methods
Online
Online version. Approval
year is designated by the last
4 digits. Only online versions
cited in the regulations or in
Appendix A to Subpart C of
Part 141 are approved.
2006
A 0.45-nm 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.
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.
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 organism
density of the water sample.
When the MF method has not been used previously to test waters with high
turbidity, large numbers of nonconform 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.
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.
Subject total coliform positive samples determined by 9222B-2006 or other
membrane filter procedure to 9222G-2006 using NA-MUG medium.

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Page 6 of 12
Method
Organization
Reference Title
Date
Notes
9222 D/9222
G
Standard
Methods
Standard Methods for the
Examination of Water and
Wastewater, 20th edition
1998
A 0.45-nm 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.
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.
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 organism
density of the water sample.
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.
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.
Subject total coliform positive samples determined by 9222B-2006 or other
membrane filter procedure to 9222G-2006 using NA-MUG medium.

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Page 7 of 12
Method
Organization
Reference Title
Date
Notes
9213 D-2007
Standard
Methods
Online
Online version. Approval
year is designated by the last
4 digits. Only online versions
cited in the regulations or in
Appendix A to Subpart C of
Part 141 are approved.
2007
A 0.45-nm 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.
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.
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 organism
density of the water sample.
When the MF method has not been used previously to test waters with high
turbidity, large numbers of nonconform 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.
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.

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Page 8 of 12
Method
Organization
Reference Title
Date
Notes
D5392-93
ASTM
International
Annual Book of ASTM
Standards - Water and
Environmental Technology.
Section 11.02.
1996
A 0.45-nm 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.
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.
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 organism
density of the water sample.
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.
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.

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Page 9 of 12
Method
Organization
Reference Title
Date
Notes
D5392-93
ASTM
International
Annual Book of ASTM
Standards - Water and
Environmental Technology.
Section 11.02.
1999
A 0.45-nm 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.
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.
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 organism
density of the water sample.
When the MF method has not been used previously to test waters with high
turbidity, large numbers of nonconform 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.
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.

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Page 10 of 12
Method
Organization
Reference Title
Date
Notes




A 0.45-nm 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.




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.
D5392-93
ASTM
International
Annual Book of ASTM
Standards - Water and
Environmental Technology.
Section 11.02.
2000
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 organism
density of the water sample.
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.
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.
1603
EPA
EPA 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.
2014

1604
EPA
EPA Method 1604: Total
Coliforms and Escherichia
coli (E.coli) in Water by
Membrane Filtration by
Using a Simultaneous
Detection Technique (Ml
Medium), EPA 821-R-02-024,
September 2002.
2002
Preparation and use of Ml agar with a standard membrane filter procedure is
set forth in the article, Brenner et al. 1993. New Medium for the Simultaneous
Detection of Total Coliform and Escherichia coli in Water. Appl. Environ.
Microbiol. 59: 3534-3544

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Page 11 of 12
Method
Organization
Reference Title
Date
Notes
mColiBlue-
24®
Hach Company


A description of the mColiBlue24® test may be obtained from Hach Company.
Water Quality Parameters
Turbidity: §141.704(c) Systems must use methods for turbidity measurement approved in 141.74 (a]
Method
Organization
Reference Title
Date
Notes
2130 B
Standard Methods
Standard Methods for the
Examination of Water and
Wastewater, 18th Edition
1992
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stabilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
2130 B
Standard Methods
Standard Methods for the
Examination of Water and
Wastewater, 19th Edition
1995
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
2130 B
Standard Methods
Standard Methods for the
Examination of Water and
Wastewater, 20th Edition
1998
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
2130 B
Standard Methods
Standard Methods for the
Examination of Water and
Wastewater, 21st Edition
2005
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
2130 B
Standard Methods
Standard Methods for the
Examination of Water and
Wastewater, 22nd Edition
2012
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
180.1
EPA
Methods for the
Determination of Inorganic
Substances in Environmental
Samples,
EPA/600/R-93/100,
August 1993
1993
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
Method 2
Great Lakes Instruments
Great Lakes Instruments
Method 2, Turbidity,
November 2, 1992
1992
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
1).

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Page 12 of 12
Method
Organization
Reference Title
Date
Notes
10133
Hach
Hach FilterTrak Method
10133
Determination of Turbidity by
Laser Nephelometry
January 2000
Revision 2.0
2000
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stabilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
M5271
Leek Mitchell
Mitchell Method M5271,
Revision 1.1,
Determination of Turbidity by
Laser Nephelometry,
March 5, 2009
2009
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
M5331
Leek Mitchell
Mitchell Method M5331,
Revision 1.1,
Determination of Turbidity by
LED Nephelometry,
March 5, 2009
2009
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
AMI Turbiwell
Swan Analytische Instrumente
AG
Continuous Measurement of
Turbity Using ASWAN AMI
Turbiwell Turbidimeter,
August 2009
2009
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
AQ4500
Thermo Scientific
Orion Method AQ.4500,
Revision 1.0, Determination
of Turbidity by LED
Nephelometry,
May 8, 2009
2009
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
M5331, Rev.
1.2
Leek Mitchell
Mitchell Method M5331,
Revision 1.2,
Determination of Turbidity by
LED or Laser Nephelometry,
February 2016
2016
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin
10258
Hach Company
Hach Method 10258,
Determination of Turbidity by
360° Nephelometry,
January 2016
2016
Styrene divinyl benzene beads (e.g. AMCO-AEPA-1 or
equivalent) and stablilized formazin (e.g. Hach StablCal™
or equivalent) are acceptable substitutes for formazin

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