&EPA
Off ice of Water
www.epa.gov
United States
Environmental Protection
Agency
Protocol for Review and Validation of
Alternate Test Procedures for
Regulated Organic and Inorganic
Analytes in Wastewater Under EPA's
Alternate Test Procedure Program
February 2016
-------
U.S. Environmental Protection Agency
Office of Water
Engineering and Analysis Division
1200 Pennsylvania Avenue, NW(4303T)
Washington, DC 20460
EPA 821-B-16-002
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Foreword
This document ("protocol") provides guidance on how the U.S. Environment Protection Agency (EPA)
will evaluate certain test procedures under its National Alternate Test Procedure program for inclusion as
an approved 40 Code of Federal Regulations (C.F.R.) Part 136 method. The protocol applies to alternate
test procedures (ATP) for measuring an organic or inorganic analyte for which there is already at least
one existing Part 136 method to measure the analyte. The protocol outlines in substantial detail the kind
of information and evidentiary showing EPA would expect is necessary to demonstrate the suitability of a
method for approval and inclusion in Part 136. The protocol also includes guidance regarding obtaining
approval of methods for measurement of method-defined analytes or parameters (MDPs) for which there
is already at least one existing Part 136 method. This protocol applies to modifications of an EPA-
approved method or a procedure that uses the same determinative technique and measures the same
analyte(s) of interest as an approved method.
The protocol provides guidance for validation, submission, and EPA review of ATP applications under
EPA's National ATP Program submitted for modifications of an EPA-approved method or a procedure
that uses the same determinative technique and measures the same analyte(s) of interest as an approved
method. Methods that use a different determinative technique to measure the same analyte(s) of interest
or methods that measure a different form or species of an analyte or parameter than the approved method
are considered new methods. The requirements for EPA approval of new methods are detailed in a
separate protocol. The protocol provides supplementary information for complying with the ATP
requirements at 40 C.F.R. § §136.4 and 136.5.
This protocol supersedes the 1999 version of the Protocol for EPA Approval of Alternate Test Procedures
for Organic and Inorganic Analytes in Wastewater. With respect to ATP applications for methods that
measure MDPs, this guidance recommends side-by-side comparison studies to validate that there are no
systematic differences in performance between the ATP and the EPA-approved methods. This protocol
continues the recommended current practices for ATP applications involving other types of methods for
measurement of organic and inorganic analytes (i.e., applicants should conduct validation studies in the
recommended number of laboratories depending upon the type of approval being sought to demonstrate
acceptable method performance by meeting or exceeding the quality control (QC) acceptance criteria
associated with EPA-approved reference methods for the corresponding combination of analyte(s) and
determinative technique).
Under EPA's ATP program, in certain circumstances, a method developer may apply for approval for the
use of an ATP to test for a specific regulated constituent. The recommended procedures described herein
will likely expedite the approval of these methods for organic and inorganic analytes, encourage the
development of innovative technologies, and enhance the overall utility of the EPA-approved methods for
compliance monitoring under the National Pollution Discharge Elimination System (NPDES) permit
program.
Disclaimer
This guidance generally describes the approval process for EPA's program for establishing test
procedures for organic and inorganic analytes that are used in Clean Water Act programs and codified at
40 C.F.R. Part 136. It describes EPA's conclusions about the types of data and information EPA will
need in order to evaluate whether to approve any particular ATP for such analytes. It includes a model
application form for use when requesting EPA approval for ATPs for such analytes. Although the
guidance provides additional explanation of EPA's requirements, it does not alter or substitute for any of
the regulations at 40 C.F.R. Part 136. The guidance, including the model application form, is not a rule
and is not legally enforceable. It does not confer legal rights or impose legal obligations on any federal,
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
state agency or any member of the public. It does not create any rights, substantive or procedural,
enforceable at law by a party to litigation with EPA or the United States. In the event there is an apparent
conflict between the guidance and any statute or regulation, the guidance is not controlling. EPA has
made every effort to ensure the accuracy of information in the guidance, but the requirements for EPA
approval of test procedures for use in its CWA programs are determined by the relevant statutes,
regulations or other legally binding requirements.
This protocol represents EPA's "best thinking" about the information that is useful in making the
determination of whether or not to approve use of any ATP for organic and inorganic analytes. This
guidance document reflects EPA views about what data and information sound scientific practice would
require for approval of an ATP for such analytes. Where the guidance uses the word "should," or in some
cases "must," this is only intended to apprise the applicant of the kind of information that, in EPA's view,
will demonstrate the adequacy of a given method for use under the CWA and thus its suitability for EPA
approval. Applicants may provide other data or information for use in EPA's determination and remain
free to deviate from the recommendations EPA has provided here. EPA will make the decision to
approve or disapprove any ATP for such analytes based on the record before it, and that decision is
subject to challenge and judicial review.
40 C.F.R. §§ 136.4 and 136.5 establish the procedures and regulatory requirements for applying for and
for EPA approval of alternate test procedures for nationwide use and for limited use. The regulations
require submission of an application that, among other things, provides comparability data for the
performance of the alternate test procedure as compared to the performance of the approved Part 136
method for which it is a proposed alternative. (40 C.F.R. § 136.4(a)(4) and 40 C.F.R. § 136,5(a)(5)).
This guidance explains in more detail the information that EPA expects will be necessary for EPA to
determine comparability or justify using the alternate test procedures instead of the approved Part 136
method for organic and inorganic analytes.
EPA may decide to revise the guidance without public notice. The public may offer suggestions to EPA
for clarifications at any time.
Neither the United States Government nor any of its employees, contractors, or their employees make any
warranty, expressed or implied, or assumes any legal liability or responsibility for any third party's use of
apparatus, product, or process discussed in this document, or represents that its use by such party would
not infringe on privately owned rights. Mention of company or trade names or commercial products in
this protocol does not constitute endorsement or recommendation for use.
Questions or comments regarding this document or the ATP program should be directed to:
Lem Walker
Clean Water Act ATP Coordinator
U.S. Environmental Protection Agency
Office of Science and Technology
Engineering and Analysis Division (EAD)
1200 Pennsylvania Avenue, NW
Mail Code - 4303T
Washington, DC 20460
Fax: (202) 566-1053
walker.lemuel@epa.gov
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
TABLE OF CONTENTS
1.0 INTRODUCTION 1
1.1 Background and Objectives 1
1.2 Tiered System for Validation of Alternate Test Procedures 2
1.3 Scope of Alternate Test Procedures 3
1.3.1 EPA-approved Reference Methods 3
1.3.2 Modifications to Front-end Techniques 4
1.3.3 Adding New Target Analytes 4
1.3.4 Method-defined Analytes 5
2.0 OVERVIEW OF THE ATP APPROVAL PROCESS 6
3.0 APPLICATION 7
3.1 Submission Addresses 7
3.2 Application Information 8
3.2.1 Justification for the ATP 9
3.2.2 EPA Method Format 10
3.2.3 Method Comparison Table 10
3.2.4 Validation Study Report 10
3.2.5 Method Information and Documentation 10
4.0 METHOD VALIDATION 12
4.1 Introduction 12
4.2 Summary of Validation Study Designs 12
4.2.1 Tier 1 Validation Studies for Wastewater 14
4.2.2 Tier 2 Validation Studies for Wastewater 14
4.2.3 Tier 3 Validation Studies for Wastewater 15
4.3 Detailed Procedures for Conducting Validation Studies 15
4.3.1 Method Compilation 15
4.3.2 Method Detection Limit Study 16
4.3.3 Calibration 16
4.3.4 Initial Precision and Recovery 16
4.3.5 Field Sample Collection and Analyses 16
4.3.6 Ongoing Precision and Recovery 18
4.3.7 Calibration Verification 18
4.3.8 Contamination Level in Blanks 18
4.3.9 Surrogate or Labeled Compound Recovery 19
4.3.10 Absolute and Relative Retention Time 19
4.3.11 New Analytes 19
4.3.12 Proficiency Testing Results 19
5.0 EPA REVIEW AND APPROVAL 20
5.1 EPA Review of Applications 20
5.2 Approval Recommendation 21
5.3 Rulemaking Process 21
6.0 REFERENCES 22
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
APPENDICES
Appendix A ATP Application Form A-l
Appendix B Data Collection Certification B-l
Appendix C Headquarters and Regional ATP Contacts C-l
Appendix D Standard EPA Method Format D-l
Appendix E Validation Study Plan and Report E-l
Appendix F ATP Data Reporting Form F-l
Appendix G Quality Control Acceptance Criteria G-l
Appendix H Requirements For Method-Defined Parameters (MDPs) H-l
Appendix I Checklist for Methods to be Considered by EPA for use in Compliance Monitoring
Programs under the Clean Water Act 1-1
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
1.0 INTRODUCTION
1.1 Background and Objectives
In accordance with section 304(h) of the Clean Water Act (CWA), the U.S. Environmental Protection
Agency (EPA) promulgates guidelines establishing test procedures (analytical methods) for the analysis
of pollutants. EPA regulations require the use of these methods where measurements of waste
constituents are required in applications for National Pollutant Discharge Elimination System (NPDES)
permits or for reports required under NPDES permits. 40 C.F.R. § 136.1. EPA has codified these
approved test procedures in the Code of Federal Regulations (C.F.R.) at 40 C.F.R. Part 136. For the
purposes of this protocol, these test procedures are referred to as "EPA-approved" methods, regardless of
whether they were developed by EPA, a voluntary consensus standards body (VCSB) such as ASTM
International, or another government entity such as the U.S. Geological Survey (USGS).
EPA's regulations at 40 C.F.R. §§ 136.4, and 136.5 also establish procedures for EPA to review and
approve the use of an alternate test procedure (ATP) in place of an EPA-approved method. These
regulations govern the Agency's Alternate Test Procedure (ATP) program for CWA methods1. Section
136.4 describes the process for obtaining approval for nationwide use of an ATP. Section 136.4(a) first
requires a written application for review of an ATP for nationwide use. Required elements of that
application include, among other things, a detailed description of the proposed ATP and studies
confirming the general applicability of the ATP for analysis of the pollutant or parameter for which
approval is requested. The applicant must also provide comparability data for the performance of the
ATP as compared to the existing approved method. Section 136.4(a)(4). The National Coordinator of the
ATP program reviews the application and notifies the applicant of its suitability for use in CWA
programs (Section 136.4(c)). If approval is recommended, EPA will propose to amend Part 136 to include
the ATP and following public comment make a final decision on approving the ATP. In the event that the
National Coordinator recommends against approval, the Coordinator will specify what additional
information might lead to a recommendation for approval. These requirements are the basis for EPA's
CWA ATP program administered by the Office of Water, Office of Science and Technology, Engineering
and Analysis Division (EAD). Section 136.5 describes the process for obtaining approval for limited use
of an ATP. Section 136.5 first requires a written application for review of an ATP for limited use to be
submitted to the director of the State agency having responsibility for issuance of NPDES permits in
cases where the request for use of an ATP concerns use in a State with an NPDES permit program
approved pursuant to Section 403 for the Clean Water Act. In cases where the request is made in a State
that has not been grated authority to administer the NPDES permit program or in cases where the Sate is
the applicant, the request is submitted directly to the Regional ATP Coordinator who has the final
authority to approve or reject applications for use of an ATP. Limited use approval may be restricted to
use by a single facility on one or more discharges. In cases where the National ATP Coordinator has
approved an applicant's request for nationwide use of an ATP, an applicant may request limited use
approval of the method under §136.5. In these instances, limited use approval maybe extended all
dischargers or facilities (and their associated laboratories) specified in the approval for the Region at the
discretion of the Regional ATP Coordinator. The Regional ATP Coordinator will forward a copy of every
approval and rejection notification to the National Alternate Test Procedure Coordinator.
In addition, as specified at 40 C.F.R. § 136.6, EPA allows users to make certain modifications to an
approved method to address matrix interferences without the extensive review and approval process
specified for an alternate test procedure at 40 C.F.R. §§ 136.4 and 136.5. Acceptable reasons for an
1 EPA also promulgates analytical methods under the Safe Drinking Water Act (SDWA) and has a similar ATP
program. This protocol only addresses the CWA ATP program and does not apply to the SDWA ATP program.
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
analyst to modify a method include analytical practices that lower detection limits, improve precision,
reduce interferences, lower laboratory costs, and promote environmental stewardship by reducing
generation of laboratory wastes. Acceptable modifications may use existing or emerging analytical
technologies that achieve these ends provided that they do not depart substantially from the underlying
chemical principles in methods currently approved in 40 C.F.R. Part 136. The flexibility to modify
methods without the need for approval as an ATP and the associated requirements that must be met
before such modified methods may be used for CWA compliance monitoring are described in more detail
at 40 C.F.R. § 136.6.
An ATP is a modification of an approved method or a procedure that uses the same determinative
technique and measures the same analyte(s) of interest as the approved method. An ATP also may
involve adding new analyte(s) of interest required in a specific permit to the target analyte list of an
approved reference method. The ATP program provides laboratory professionals with the opportunity to
enhance compliance monitoring and encourages use of innovative technologies. Approval for an ATP
may be sought when the alternate procedure reduces analytical costs, overcomes matrix interference
problems, improves laboratory productivity, or reduces the amount of hazardous materials used and/or
produced. The applicant is responsible for validating its proposed alternate test procedure.
This protocol sets out EPA's views about what information and data will support approval of an ATP for
organic and inorganic analytes under the ATP program for use in NPDES Compliance monitoring. As
such, it provides a detailed explanation of the kinds of information and studies that generally will support
a finding of a method's comparability to an existing approved method and thus its appropriateness for
approval as an ATP for such analytes. This version of the ATP protocol describes validation processes
for modifying methods that measure MDPs. Details regarding these MDP validation procedures are
found in Appendix H of this document.
The use of a different determinative technique to measure the same analyte(s) of interest or a method that
measures a different form or species of analyte or parameter than the approved method is considered a
new method. EPA has established a different set of requirements for validation, submission and approval
of new methods that are detailed in a separate protocol (USEPA 2015).
Note: Methods developed by voluntary consensus standard bodies (VCSBs) and other federal agencies
are not processed for approval under the ATP Program. Instead EPA has developed a separate
path to approval for these keeping with the National Technology Transfer and Advancement Act
(NTTAA). EPA considers VCSB methods and those from other agencies in regulatory actions
when periodically updating the list of approved methods at 40 C.F.R. Part 136. EPA's "Checklist
for Methods to be Considered by EPA for Use in Compliance Monitoring Programs under the
Clean Water Act" (Appendix I) provides a list of items and information EPA considers in
evaluating all new, updated, and ATP methods for use in wastewater compliance monitoring for
approval.
1.2 Tiered System for Validation of Alternate Test Procedures
EPA recognizes that a formal interlaboratory method validation may not be necessary to demonstrate
suitability for approval for all situations and may be prohibitively costly to implement, especially for
small laboratories and regulated entities. Therefore, the protocol describes a three-tiered, cost-effective
approach to method validation that would tailor the validation study to reflect the intended use of the
method. EPA has specified approved methods that contain (or are supplemented with) QC acceptance
criteria (Appendix G) for most combinations of analyte and determinative technique. When considering
how to demonstrate that its ATP for organic and inorganic analytes is able to meet or exceed the QC
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
acceptance criteria of the EPA-approved reference method (see Section 1.3.1) for the applicable
combination of analyte and determinative technique, an applicant should review the tiers below and
decide what the most appropriate tier for the applicant's ATP is based on its intended use. An applicant is
required to demonstrate that its ATP is able to meet or exceed the QC acceptance criteria of the EPA-
approved reference method (see Section 1.3.1) for the applicable combination of analyte and
determinative technique. The three method validation tiers are listed below.
Tier 1: These types of ATP should be validated for use in one or more matrix type(s). EPA approval of a
Tier 1 ATP would generally require successful single-laboratory testing in the matrix type(s) of
interest. Tier 1 ATPs are reviewed by the State issuing the NPDES permit where the State is not
the requesting party, and forwarded to EPA Regional staff, along with a recommendation for or
against approval. Where the State is the requesting party, applications for Tier 1 ATPs are sent
directly to the EPA Regional staff
Tier 2: ATPs for use by all laboratories for nationwide use for only one matrix type. The application for
approval should generally demonstrate successful testing of the ATP in a three-laboratory
validation study. Tier 2 ATPs will be reviewed by the National ATP staff at EPA Headquarters
and if positively reviewed, will be recommended for approval. These methods are then proposed
for promulgation in the C.F.R.
Tier 3: ATPs for use by all laboratories (nationwide use) for all matrix types. The application for
approval should generally demonstrate successful testing of the ATP in a nine-laboratory
validation study. Tier 3 ATPs are reviewed by the National ATP staff at EPA Headquarters and
if positively reviewed are recommended for approval. These methods are then proposed for
promulgation in the C.F.R.
Note: Matrix type, in the context of these tiers, is defined as a sample medium (e.g., air, soil, water,
sludge) with common characteristics across a given industrial subcategory. For example, C-stage
effluents from chlorine bleach mills, effluent from the continuous casting subcategory of the iron
and steel industrial category, publicly owned treatment works (POTW) sludge, and in-process
streams in the Atlantic and Gulf Coast Hand-shucked Oyster Processing subcategory are each a
matrix type. (A list of industrial categories with existing effluent guidelines can be found
at: http://water.epa. gov/scitech/wastetech/guide/industry.cfm).
1.3 Scope of Alternate Test Procedures
This protocol for validation, submission, and approval of an ATP offers flexibility to modify approved
methods, provided that a laboratory demonstrates and documents that the modified method produces
results equal or superior to those produced by the EPA-approved reference method for the applicable
combination of analyte and determinative technique.
1.3.1 EPA-approved Reference Methods
EPA has approved one or more reference methods that contain (or are supplemented with) standardized
QC procedures and QC acceptance criteria for each combination of regulated analyte and determinative
technique. Appendix G of this document contains the QC acceptance criteria for the approved inorganic
methods. The approved organic methods include the QC acceptance criteria within the text of the method
itself.
The QC acceptance criteria associated with the EPA-approved reference methods are the performance
criteria against which ATPs are evaluated. Method performance is deemed to be acceptable when results
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
produced by an ATP meet or exceed the QC acceptance criteria associated with the corresponding EPA-
approved reference method. Using these established QC acceptance criteria as the method performance
measure allows EPA to implement the ATP program more efficiently.
1.3.2 Modifications to Front-end Techniques
A front-end technique is any technique in the analytical process that precedes the determinative
technique. Front-end techniques include all procedures, equipment, solvents, etc., that are used in the
preparation and cleanup of a sample prior to analyte detection and measurement. Laboratories may
modify any and all front-end techniques for non-MDPs provided that the modification:
• Is not explicitly prohibited in the corresponding approved method, and
Can be demonstrated to produce results equal or superior to results produced by the approved method.
This flexibility includes the ability to modify the chemistry of the front-end of the method, for example,
changing the extraction solvent and substituting liquid-liquid for solid-liquid extraction. ATP approval is
not required if changes to the front-end techniques are within the allowed flexibility of 40 C.F.R. § 136.6.
Note: Changes to the front-end chemistry or extraction solvent may affect the stability of the analyte(s)
of interest, potentially leading to analyte transformation or degradation. Depending on the nature
of the front-end change, the developer of a modified method may need to demonstrate that
analyte stability is not adversely affected in either the original sample or in the sample extract for
at least the duration of the established holding time(s) in the reference methods.
The developer of a modified method always has the option of asking EPA or another regulatory authority
for a technical opinion on the acceptability of the validation data that supports the method modification.
1.3.3 A dding New Target A nalytes
EPA will permit method developers to modify the scope of an approved method by adding additional
analytes if required in a specific permit. This allowance is in response to public comment on previous
rules (59 FR 62456, December 5, 1994; 58 FR 65622, December 15, 1993). Method developers seek this
approval when they want to adapt an existing method to obtain occurrence data for a new analyte. EPA
believes these requests have merit when there is a potential for new regulatory requirements and when
technological advances make the measurement of additional analytes feasible (e.g., adding lead to the
scope of EPA Method 200.7). Under this ATP protocol, developers can obtain approval for adding
analytes to an approved method if the following conditions are met:
(1) It has been demonstrated that the added analyte does not interfere with determination of the analytes
of concern in the approved method.
(2) QC acceptance criteria are developed and used for determination of the added analyte; see Protocol
for EPA Approval of New Methods for Organic and Inorganic Analytes in Wastewater and Drinking
Water.
(3) The reason for adding the analyte is not to avoid the sample preservation or sample (or extract)
holding time conditions that are already required for that analyte in another approved method. (This
criterion precludes the addition of analytes to an approved method with less rigid sample collection or
holding time criteria.)
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
1.3.4 Method-defined Analytes
As specified at 40 C.F.R. § 136.6, the term "method-defined analyte " means an analyte (or parameter)
that is defined solely by the method used to determine the analyte (generically referred to in this
document as a method-defined parameter or MDP). Such an analyte may be a physical parameter, a
parameter that is not a specific chemical, or a parameter that may be comprised of a number of
substances. Examples include, but are not limited to:
• Acidity,
• Alkalinity,
Biological oxygen demand (BOD),
• Chemical oxygen demand (COD),
• Color,
Oil and grease,
• pH (hydrogen ion),
Conductivity (specific conductance),
• Temperature,
• Total dissolved solids (TDS),
Total organic carbon (TOC),
• Total suspended solids (TSS),
• Total phenolics, and
Turbidity.
Modifications to methods that measure MDPs have the potential to change what is being measured.
Therefore, any modifications to those methods beyond that specifically allowed in the approved methods
require EPA review and approval as alternate test procedures by the appropriate approval authority (see
Table 1).
In order to more clearly distinguish the ATP requirements for MDPs from those for the more traditional
type of analytes, the discussion data and information that, in EPA's view, will generally demonstrate the
suitability of the ATP for measurement of MDPs has been placed in Appendix H of this document.
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
2.0 OVERVIEW OF THE ATP APPROVAL PROCESS
The process for obtaining approval of an ATP for organic and inorganic analytes is summarized in Figure
1. Depending on the tier, ATPs may be reviewed by (1) the State authority that issues the NPDES permit,
and/or by EPA Regional staff, or (2) EPA Headquarters staff. The relevant authority will review the
application, including the justification for the ATP provided by the applicant and determine whether an
ATP is necessary (e.g., the approved method may already allow the modification proposed or the
modification falls within the flexibility allowed at 40 C.F.R. § 136.6, so ATP approval is not needed or
warranted). Where the State is not the requesting party, the State will review Tier 1 ATP applications and
forward these to EPA Regional staff with a recommendation for or against approval. Where the State is
the requesting party, the EPA Regional staff will review the Tier 1 ATP applications. If, after initial
review, EPA Headquarters accepts a Tier 2 or Tier 3 application, the applicant should move forward with
preparing a method development and validation study plan in consultation with National ATP staff.
/
/
1 <"
en
|l
LJ
Apply through
\ Regional ATP
\j coordinator or
A Director of State
C y Agency, as
appropriate
Acronyms:
I^^^^fp 1 MUK - Method Updat
application 1 OW = Office of Water
CD
f>
^
CD
11
•-
TO
Iz
WW = Wastewater
V
K
e Rule
strator
\
/
V
"CD
c
o
s
\
\
Regional
review
( Forward to OW for review
i OW recommendation
OW review and
decision
1
RA provides letter of
approval or disapproval
to the applicant, OW,
and the appropriate
State Agency
Figure 1. Flowchart summarizing the ATP application process for methods under the CWA Program
In order to expedite the approval process, the applicant should submit to EPA its plan for developing the
necessary data to demonstrate the suitability of ATP for approval. For Tier 2 and Tier 3 ATPs, once the
applicant has received EPA's view concerning its study plan, the applicant should move forward with the
study and submit the study report to the ATP staff. If the validation study results confirm that the
applicant's method is sufficiently rugged and provides data of comparable quality, EPA will generally
notify the applicant that it intends to pursue approval via the rulemaking process. If this is not the case,
ATP staff may identify additional information or data required. If the laboratory studies fail to
satisfactorily verify the comparability of the applicant's method, the applicant should address the
problems encountered and follow-up with further laboratory validation studies.
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
3.0 APPLICATION
This section describes the information that EPA would expect in an application for approval of an ATP
for organic and inorganic analytes to demonstrate its appropriateness for approval and provides
information on the approval authorities for the three tiered approach described in Section 1.2 of this
protocol. This section also describes how to treat any proprietary information submitted with an
application.
Note: Where the State is not the requesting party, Tier 1, Limited Use ATPs are subject to State
authority review prior to EPA Regional approval. State authorities may have additional
requirements and/or authority-specific application forms that are beyond the scope of this
protocol. Therefore, applicants for Tier 1 ATPs should consult such authorities regarding Tier 1
ATP requirements.
Applications may be submitted by email, in hardcopy, or on electronic media by U.S. mail or other
carrier. Hard copy applications and supporting documentation should be submitted in triplicate.
Applicants are advised to consult the recipient before submitting large files via email.
3.1 Submission Addresses
A summary of where to submit ATP applications and the approval authorities for each tier is provided in
Table 1.
Table 1: Submission of Alternate Test Procedure Applications
Tier
Tierl
Tier 2
TierS
Level of Use
Limited Use for
Wastewater2
Nationwide Use
in a Single
Wastewater
Matrix Type
Nationwide Use
in All
Wastewater
Matrix Types
Typical Applicant
EPA Regional laboratories, States,
commercial laboratories, individual
dischargers, or permittees in States
that do not have authority to
implement the NPDES permit program
Commercial laboratories, individual
dischargers, or permittees in States
that have authority to implement the
NPDES permit program
All applicants
All applicants
Submit
Application
to1
EPA Regional
ATP
Coordinator3
Director of
State Agency
issuing the
NPDES
permit4
National ATP
Coordinator,
EPA
Headquarters
National ATP
Coordinator,
EPA
Headquarters
Approval
Authority
EPA Regional
ATP
Coordinator
(as designated
by the EPA
Regional
Administrator)
EPA
Administrator
EPA
Administrator
1 See Appendix C for EPA addresses.
2 Per 40 C.F.R. § 136.4(c)(5): "Whenever the National Coordinator has approved an applicant's request for nationwide use of an
alternate test procedure, any person may request an approval of the method for limited use under §136.5 from the EPA
Region." In these instances, limited use approval maybe extended all dischargers or facilities (and their associated laboratories)
specified in the approval for the Region (limited use approval under § 136.5) at the discretion of the Regional ATP
Coordinator.
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
3 The Regional ATP Coordinator may choose to request assistance with the Tier 1 (limited use) applications from the National
ATP Coordinator for an approval recommendation.
4 Per 40 C.F.R. § 136.5, in States with authority to issue NPDES permits, the State agency has primary responsibility for
reviewing Tier 1 ATP applications. The State agency will forward the application to the Regional ATP Coordinator with a
recommendation for or against approval. Where the State is the applicant for the ATP, the application goes directly to the
Regional ATP Coordinator.
On receipt of the application, the ATP Coordinator will assign an identification number to the application.
The applicant should use the identification number in all future communications about the application.
3.2 Application Information
A copy of a model ATP application form is included in Appendix A. The information requested on the
ATP application form includes the following:
Name and address of the applicant,
• Application submission date,
• Method number and title of the proposed ATP,
Citation (i.e., number) of the EPA-approved method that was modified to develop the proposed ATP,
• Approved reference method (see Section 1.3.1) that contains the QC acceptance criteria that will be
used for comparison,
Analytes(s) for which the ATP is proposed,
• Level of use desired (i.e., limited use or nationwide use),
Tier at which the proposed ATP will be validated, and
Applicant's NPDES permit number, issuing agency, type of permit, and the discharge serial number
(if applicable).
In addition the applicant should provide the following items:
The proposed ATP prepared in standard EPA method format,
A table that gives a side-by-side comparison of the proposed ATP and the EPA-approved method that
was modified,
The method validation study report, including supporting data,
• For nationwide applications that will undergo rulemaking, method development information and
documentation that EPA can use in preparing the preamble and docket for the proposed rule, and
For limited use applications, applicants should identify the NPDES permit numbers for all discharges
for which the applicant is seeking approval to apply the alternate test procedure (if applicable).
Note 1: Not all of these documents would need to be submitted with the initial application. The
applicant should submit a validation study plan for EPA review and comment before
proceeding with ATP validation. Recommended study plan elements are described in
Appendix E of this protocol.
Note 2: As stated in Section 1.3, the information that should demonstrate the suitability for approval of
ATPs that measure MDPs may be found in Appendix H of this document.
If an applicant is unsure whether or not a modification is allowed within the method-specified flexibility,
the applicant may request that EPA determine the necessity for a full ATP validation. The minimum
information required for EPA to make this determination or begin reviewing an application is the
completed application form, the proposed method in standard EPA format, and the method comparison
table. From this information, EPA can determine whether a full ATP validation is required or whether
the proposed modification is within the allowed flexibility of 40 C.F.R. § 136.6.
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
The elements that should be provided for an application at each tier are presented in Table 2. For Tier 2
and 3 applications, the National ATP Coordinator at EPA Headquarters will not process an application
until the Coordinator determines that the applicant has submitted adequate information to evaluate the
application. As noted at the beginning of Section 3.0, Tier 1 applicants should consult the relevant State
authority issuing the NPDES permit to determine if there are also State requirements for those
applications.
Table 2. Recommended Application Elements
Tier
Level of Use
Application Elements
TieM
Limited Use
Completed application form submitted to the EPA Regional ATP
Coordinator or the Director of State Agency issuing the NPDES permit
Justification for the ATP
Method in EPA format
Validation Study Plan1
Method comparison table
Validation study report
Tier 2
TierS1
Nationwide
Use
Completed application form submitted to National ATP Coordinator,
EPA Headquarters
Justification for the ATP
Method in EPA format
Validation Study Plan1
Method comparison table
Validation study report
Method information and documentation
1 The applicant should submit a validation study plan with the initial application for a Tier 2 or 3 ATP for EPA review and
comment before proceeding with the study.
3.2.1 Justification for the ATP
Because EPA review and evaluation of proposed ATPs can entail considerable effort, EPA strives to
minimize the submission of unnecessary method modifications or modifications that are already allowed
in approved methods. Therefore, the entity that proposes an ATP should provide a brief justification for
why the ATP is being proposed. Examples of useful justifications include, but are not limited to:
• The ATP successfully overcomes some or all of the interferences associated with the approved
method
The ATP reduces the amount of hazardous wastes generated by the laboratory
• The cost of or time required for analyses is reduced, or
The quality of the data is improved.
The Agency acknowledges that there may be some trade-offs between meeting QC acceptance criteria
and encouraging use of potentially beneficial alternate methods. For example, a proposed ATP may be
far more rapid and less expensive to perform, but have slightly lower precision than the currently
approved methods for a given analyte. Depending on the chemical being measured, ATP staff may
consider the ATP application because the alternate method could allow more frequent monitoring with no
added cost. More frequent monitoring may result in enhanced information quality for that chemical. The
Agency may consider relaxing certain QC acceptance criteria for a given ATP, depending on the analyte
and the benefits likely to be realized.
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
It is highly recommended that the method developer consult with ATP staff concerning their
proposed candidate method and its justification prior to extensive method development. Candidate
methods that are insufficiently justified will not be considered further.
3.2.2 EPA Method Format
In accordance with the standard EPA format originally developed by EPA's Environmental Monitoring
Management Council in 1996 (Reference 4), methods should contain 17 specific topical sections in a
designated order. These 17 sections are listed in Appendix D. Any additional numbered sections should
be inserted starting with Section 11.0, Procedure, as appropriate for a particular method. For detailed
information on the EPA format for proposed methods, refer to Guidelines and Format for Methods to Be
Proposed at 40 C.F.R. Part 136 or Part 141 (Guidelines and Format document), EPA-821-B-96-003.
3.2.3 Method Comparison Table
Applicants should perform an in-depth comparison between their proposed ATP and the corresponding
EPA-approved method and document the comparison in a two-column table. The table should include the
number and title of each method, the latest revision date of the proposed ATP, and a detailed discussion
of each of the 17 topics required by the standard EPA method format. Each topic should be discussed on
a separate row. The applicant should highlight any differences between the proposed ATP and the
approved method. If the proposed ATP is an automation of a previously approved manual method, any
differences in kinetics and interferences should be presented and a comparison of the final ratios of the
concentrations of the reactants in the proposed and approved methods included.
3.2.4 Validation Study Report
The applicant should conduct a validation study of the ATP that meets the validation study design
described in Section 4.2 of this protocol. Once the validation study is complete, the applicant should
prepare a comprehensive report on the validation study and submit a copy of that report with the ATP
application. The validation study report should include the following elements, which are described
further in Appendix E:
Background
Study Design and Objectives
• Study Implementation
Data Reporting and Validation
• Results
• Data Analysis/Discussion
Conclusions
• Appendix A - The Method
• Appendix B - Validation Study Plan
Appendix C - Supporting Data (Raw Data and Example Calculations)
3.2.5 Method Information and Documentation
For Tier 2 and 3 applications, a successful ATP will be approved by the EPA Administrator through
rulemaking. In these cases, in order to expedite the approval process, the applicant should provide
information and documentation that will aid EPA in preparing the preamble and docket for publication of
a proposed rule in the Federal Register. Specifically, it will be useful for the applicant to submit
information that:
10 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
• Defines the purpose and intended use of the method.
States what the method is based upon, noting any relationship of the method to other existing
analytical methods and indicating whether the method is associated with a sampling method.
• Identifies the matrix type(s) for which the method has been found satisfactory.
Describes method limitations and indicates any means of recognizing cases where the method may
not be applicable to the specific matrix types.
• Outlines the basic steps involved in sample and data analysis.
Lists options within the method, if applicable.
• Describes and discusses the validation study in a study report that includes study design and
objectives, study limitations, study management, technical approach, data reporting and validation,
results, data analysis discussion (including, for MDPs, development of QC acceptance criteria), and
conclusions.
Copies of all relevant supporting documents used in developing the ATP (including any other studies
conducted during method development and validation), for EPA's possible inclusion in the rule
docket.
Previous method rules that may serve as examples of the type of information and the appropriate level of
detail necessary include: 49 FR 43234, October 26, 1984; 56 FR 5090, February 7, 1991; 60 FR 53988,
October 18, 1995; and 61 FR 1730, January 23, 1996.
3.3 Proprietary Information in Applications
All information provided to the Federal government is subject to Freedom of Information Act
requirements. Therefore, any information submitted with the proposed ATP application that the applicant
considers proprietary must be marked as "business confidential." EPA staff will handle such information
according to the regulations in subparts A and B of 40 C.F.R. Part 2.
In accordance with 40 C.F.R. § 2.203, a business that submits information to EPA may assert a business
confidentiality claim covering the information by placing on (or attaching to) the information at the time
it is submitted to EPA, a cover sheet, stamped or typed legend, or other suitable form of notice employing
language such as trade secret, proprietary, or company confidential.
Note: Confidential Business Information (CBI) must be submitted as hard copy and must not be
emailed.
Confidential claims to portions of otherwise non-confidential documents should be clearly identified by
the business, and may be submitted separately to facilitate identification and handling by EPA. If the
business desires confidential treatment only until a certain date, or until the occurrence of a certain event,
the notice should state this. However, applicants are advised that any methods to be proposed in the
Federal Register cannot involve claims of confidential business information.
If a claim of business confidentiality is not made at the time of submission, EPA will make such efforts as
are administratively practicable to associate a late claim with copies of previously submitted information
in EPA files. However, EPA cannot ensure that such efforts will be effective in light of the possibility of
prior disclosure or widespread prior dissemination of the information.
11 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
4.0 METHOD VALIDATION
4.1 Introduction
ATP validation is the process by which an applicant demonstrates that the modified method accurately
measures the concentration of an analyte in an environmental sample and can meet or exceed the QC
acceptance criteria in the EPA-approved reference method or other EPA-specified document. The
validation recommendations described below were developed to reflect the level of intended use of the
ATP. This is accomplished through a three-tiered approach, as shown in Table 3.
Table 3. Tiered Validation Strategy
Tier
TieM
Tier 2
TierS
Laboratory Use
Limited use1
All Laboratories (Nationwide use)
All Laboratories (Nationwide use)
Applicable to ...
One or more matrix types from one or more industries.
Approved in Regions for use within the Region.2
One matrix type J within one industrial subcategory
All matrix typesj from all industrial subcategories
1 Whenever the National Coordinator has approved an applicant's request for nationwide use of an alternate test procedure, any
person may request an approval of the method for limited use under §136.5 from the EPA Region (40 C.F.R. § 136.4(c)(5)). In
these instances, limited use approval may be extended to all dischargers or facilities (and their associated laboratories) specified
in the approval for the Region at the discretion of the Regional ATP Coordinator (40 C.F.R. § 136.5(d)).
2 See 40 C.F.R. §136.5
3 Section 4.2 provides more information on the matrix types applicable to each tier.
Please contact the appropriate Regional ATP Coordinator for specific method validation
recommendations applicable to Tier 1 ATPs. Methods intended for multi-laboratory use in a given
industrial subcategory (Tier 2), or for multi-laboratory use for all industrial subcategories (Tier 3), should
be validated through interlaboratory testing as described in the Section 4.2.
4.2 Summary of Validation Study Designs
Approval of ATPs will require the applicant to show that the ATP performs comparably to an existing
part 136 method. That is the applicant should validate that the ATP is capable of yielding reliable data for
compliance monitoring purposes. For most ATPs, applicants are should demonstrate acceptable method
performance by meeting or exceeding the QC acceptance criteria associated with the EPA-approved
reference methods for different combinations of regulated analyte and determinative technique.
Appendix G to this protocol contains the QC acceptance criteria for inorganic methods. The QC
acceptance criteria for organic methods generally are contained in the text of the methods. For organic
methods that do not contain QC acceptance criteria, applicants should consult with EPA to determine how
best to proceed.
Note: The exception to the summary requirements above is for ATPs that measure MDPs. Validation
requirements for ATPs that measure MDPs are provided in Appendix H of this document.
All validation study results should be documented in accordance with the validation study designs
outlined below. Table 4 and Sections 4.2.1 - 4.2.3 below summarize the validation study designs for
non-MDP wastewater ATPs at each of the three tier levels.
All ATPs must be approved by the proper approval authority before they can be used or reported for
compliance monitoring.
12 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Note: The validation requirements specified in this document for Tier 1 (limited use) ATPs are intended
to serve as guidance for the Regions regarding the minimum validation that would be required if a
recommendation for or against approval is requested from the National ATP Coordinator. The
Regions or States may impose more stringent validation requirements at their discretion.
Table 4. Summary of Recommended Validation Approaches for Non-MDP Wastewater Alternate
Test Procedures
d)
Method Application
Tier 1 - Single-lab
First matrix type
Each additional matrix
type (8 max.)
Tier 2 - Multi-lab, single
matrix type
Tier 3 - Multi-lab, all matrix
types
Number of
Labs
1
1
3
9
Matrix
types
1
1-8
1
9
Number of Analyses
Back-
ground
Analysis
1
1-8
3
9
I PR-
Reagent
Water121
4
Q<6)
12
36
PT
Sample'31
1
0
3
9
MS/
MSD'4'
2
2(7)
(16 max)
6<7>
18<7)
MDL'5'
7
0(6)
21
63
Total
15
3
(24 max)
45
135
Notes:
(1) Numbers of analyses in this table do not include additional QC tests such as calibration, blanks, etc. Nine is the
maximum number of matrix types (or facilities) to validate a modified wastewater method at Tier 1 or Tier 3.
(2) Initial precision and recovery (IPR) reagent water analyses are used to validate a method modification. The
number of IPR analyses is four times the number of laboratories used to validate a method modification because
each laboratory performs a four-replicate IPR test.
(3) The proficiency testing (PT) sample should be obtained from a third party vendor and should be analyzed by
each laboratory participating in the study. If sewage sludge or ocean water are matrices of interest, PT samples
for those matrices are required as well.
(4) The matrix spike/matrix spike duplicate (MS/MSD) test would demonstrate that the EPA-approved method
MS/MSD QC acceptance criteria have been met.
(5) A method detection limit (MDL) test would be performed in each laboratory, using the alternate test procedure.
40 C.F.R. Part 136 Appendix B requires a minimum of seven analyses per laboratory to determine an MDL. If
Appendix B is modified at a later date, then the validation study must conform to the most recent MDL
requirements. Each lab involved in validation of a wastewater method modification must demonstrate that the
modified method would achieve the detection limits specified in the EPA-approved reference method.
(6) The MDL and reagent water IPR tests do not have to be repeated after the first matrix type is validated.
(7) The MS/MSD analyses would demonstrate that MS/MSD recovery and precision criteria associated with the
EPA-approved reference method have been met. The number of MS/MSD analyses is two times the number of
matrix types tested (i.e., one MS/MSD pair per laboratory).
13
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
4.2.1 Tier 1 Validation Studies for Wastewater
Any person may request the Regional Alternate Test Procedure (ATP) Coordinator to approve the use of
an alternate test procedure in the Region. The primary intent of Tier 1 is to allow use of a modified
method by a single laboratory. Tier 1 is expected to be used by commercial laboratories, dischargers, and
state and municipal laboratories repetitively testing samples from the same site(s) on a routine basis. Tier
1 can be applied to one or more matrix types. Additional Information regarding the application and
validation requirements for and approval of limited use ATPs may be found at 40 C.F.R. § 136.5. Please
contact the appropriate Regional ATP Coordinator for additional information regarding specific method
validation study designs for these types of ATPs. See Appendix C for a list of Regional ATP
Coordinators.
Tier 1 - Single Matrix Type
Tier 1 - Single Matrix Type validation studies are performed in a single laboratory on a single matrix type
plus analysis of a proficiency testing (PT) sample (see Section 4.3.12). Results of the validation study
and the method modification are applicable in the laboratory that validated the ATP for this matrix type,
and the results may not be used by another laboratory or for another matrix type.
Tier 1 - Multiple Matrix Types
If a laboratory intends to apply the method to fewer than nine matrix types, the laboratory should validate
the method on each matrix type. Results of the validation study and the method modification are
applicable in the laboratory that validated the ATP for these matrix types; the results may not be used by
another laboratory or for another matrix type. The maximum number of matrix types to which the ATP
should be applied to demonstrate that it will likely be successful for all other matrix types is nine. The
specific tests to be conducted on the first matrix type and for each additional matrix type are shown in
Table 4.
Matrices that must be tested for a multiple matrix type validation of a wastewater ATP for use in all
matrix types are given in Table 5.
4.2.2 Tier 2 Validation Studies for Wastewater
The primary intent of Tier 2 is to allow all regulated entities and laboratories to apply an ATP to a single
sample matrix type from a single industry. EPA has determined that Tier 2 will encourage the
development and application of techniques that overcome matrix interference problems specific to
effluents of certain industrial subcategories, lower detection limits, improve the reliability of results,
lower the costs of measurements, and/or improve overall laboratory productivity when analyzing samples
from a given industry.
Tier 2 validation studies are performed in a minimum of three laboratories. Samples of the same matrix
type (e.g., final effluent, extraction-stage effluent) are collected from one or more facilities in the same
industrial subcategory. In contrast to Tier 1, once an ATP has been validated under Tier 2, the results can
be used by other laboratories as long as it is applied to samples from the validated matrix type within the
industrial subcategory, and as long as the other laboratories meet or exceed all of the method's QC
acceptance criteria. If the ATP is to be applied to another matrix type, the modification should be
validated separately on that matrix type.
14 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
4.2.3 Tier 3 Validation Studies for Wastewater
The primary intent of Tier 3 is to allow nationwide use of an ATP by all regulated entities and
laboratories for all matrix types. Tier 3 validation studies are performed in a minimum of nine
laboratories, each with a different matrix type, for a total of nine samples. Suggested sample matrix types
that should be used in the validation study are given in Table 5.
Table 5. Matrix Types Recommended for Multiple Matrix Type Validation Studies
1.
2.
3.
4.
5.
Effluent from a POTW
ASTM D 5905 - 98 (Reapproved 2013), Standard Specification for Substitute Wastewater
Sewage sludge, if sludge will be in the permit
ASTM D 1141 - 98 (Reapproved 2013), Standard Specification for Substitute Ocean Water,
ocean water will be in the permit
Untreated and treated wastewaters up to a total of nine matrix types (see
http://water.epa.gov/scitech/wastetech/guide/industry.cfm
for a list of industrial categories with existing effluent guidelines)
if
At least one of the above wastewater matrix types should have at least one of the following characteristics:
Total suspended solids (TSS) greater than 40 mg/L
Total dissolved solids (TDS) greater than 100 mg/L
Oil and grease greater than 20 mg/L
NaCI greater than 120 mg/L
CaCO3 greater than 140 mg/L
4.3 Detailed Procedures for Conducting Validation Studies
When validating ATPs, laboratories must adhere to the standardized QC operations and criteria detailed
in the EPA-approved reference method (or other EPA-specified document) and incorporate these
operations and criteria into the ATP. QC acceptance criteria for most inorganic analyte-method
combinations can be found at Appendix G of this document. QC acceptance criteria for other classes of
analytes (e.g., pesticides) are often published in the reference method or in other EPA documents.
Laboratories should use both a reference matrix (usually reagent water) and field samples for the
validation study. For multi-lab validation studies (e.g., Tiers 2 and 3), the applicant is responsible for
ensuring that each laboratory in the study fulfills the validation study design specifications detailed in
Sections 4.3.2 to 4.3.11 and provides all of the data that support the ATP application. However, it is
important that the validation study accurately reflect the ruggedness of the ATP and any limitations
regarding clarity of the ATP procedures. Therefore, a vendor or other applicant to should not directly
assist laboratories participating in the validation study with implementation of the ATP methodology or
equipment during the course of the study (e.g., the vendor or applicant may provide training and advice to
participant laboratories regarding the equipment or methodology prior to the start of the study, but the
study samples are to be analyzed by the study participants under "routine" conditions). Direct
participation by the vendor or applicant will compromise the results of the study. The applicant also is
responsible for the technical and statistical evaluation of the validation study results in order to produce
the validation study report.
4.3.1 Method Compilation
Prior to conducting a validation study, the applicant responsible for modifying the method should detail
the full method in accordance with EPA's Guidelines and Format for Methods to Be Proposed at 40
C.F.R. Part 136 or Part 141 (Guidelines and Format document), EPA-821-B-96-003. The documented
15 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
method should be distributed to each laboratory participating in the validation study to ensure each
laboratory is validating the same set of procedures.
4.3.2 Method Detection Limit Study
Each laboratory participating in the Tier 1, 2, or 3 validation study must perform a method detection limit
(MDL) study in accordance with the procedure given at 40 C.F.R. Part 136, Appendix B while using the
procedures specified in the modified method. The final results for each MDL study aliquot must be
provided by each laboratory in the validation study, along with the details of the spiking levels and MDL
calculations, and each laboratory should keep the raw data that supports those MDL study results on file
and available for review.
In order to successfully validate the ATP, each laboratory participating in the validation study must
demonstrate that it can achieve an MDL that is less than or equal to the minimum level (ML) of the EPA-
approved reference method, or less than 1/10 the regulatory compliance limit, whichever is greater. For
approved methods that do not explicitly include ML values or other quantitation levels, consult Appendix
G of this document for default ML targets.
The allowance for an MDL higher than that of the approved reference method, but that supports a
regulatory compliance limit, recognizes that a method modification that overcomes interferences may not
achieve an MDL that is as low as the MDL achieved by the reference method (or other EPA-specified
document), but is potentially more valuable in allowing determination of the analyte(s) of interest at the
regulatory compliance limit in a complex sample matrix.
4.3.3 Calibration
Each laboratory participating in the validation study must perform a calibration in accordance with the
procedures specified in the ATP. Each participating laboratory must demonstrate that it can meet or
exceed the calibration criterion and achieve an ML or other quantitation level that is specified in the EPA-
approved reference method (or other EPA-specified document), or in the applicable regulations.
4.3.4 Initial Precision and Recovery
Each laboratory participating in the study must obviously perform initial precision and recovery (IPR)
analyses using only the procedures specified in the method. The IPR test is performed by analyzing four
replicates of reagent water spiked with the analytes of interest. This IPR test should be performed for
both the ATP and the corresponding approved method.
In order to successfully validate the ATP, each participating laboratory must demonstrate that it can meet
or exceed the IPR precision and recovery criteria given for the EPA-approved reference method (or other
EPA-specified document) using both the ATP and the corresponding approved method.
4.3.5 Field Sample Collection and Analyses
After laboratories participating in the Tier 1, 2, or 3 validation study have successfully completed the IPR
analyses, the method modification should be validated on the matrix type(s) chosen for the validation
study. The numbers of analyses required are described below.
Samples of each matrix type should be properly collected in sufficient quantity to support the validation
study. The volume required will vary by tier, and by the volume required in the analytical method or
ATP. Because the composition of many treated effluents may vary over time, composite sampling
16 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
equipment may be used to minimize that temporal variability. When a regulation or a reference method
specifies collecting grab samples for compliance monitoring, it still may be feasible to use composite
sampling equipment to collect a bulk effluent sample for use in a validation study. Alternatively, multiple
grab samples may be collected and combined to create a bulk sample of sufficient quantity to support the
validation study.
Note: The validation study plan should describe the sample collection procedures that will be employed
and the homogenization procedures that will be used to produce replicate aliquots of the bulk
sample for distribution and/or testing by the study participants.
All field samples should be analyzed by the laboratory as received from the study coordinator to
determine the background concentration of the target analyte prior to preparation of the MS and MSB
aliquots. This will ensure that the MS and MSB aliquots are fortified at an appropriate concentration.
That is, the MS/MSB pair shall be fortified with the target analyte a concentration equal to the regulatory
limit, if the ATP is for use to demonstrate compliance with a specific permit, or at one to five times the
background concentration of the sample, whichever is higher.
Note: Analyzing the field samples before preparing the MS/MSB aliquots may contradict the specific
requirements in some reference methods that stipulate that the MS/MSB aliquots be prepared and
analyzed in the same batch as the field samples. However, for the purposes of validating an ATP,
it is essential that the MS/MSB aliquots generate meaningful data about the performance of the
ATP in the matrix of interest.
4.3.5.1 Tier 1 - Single Matrix Type Validation Studies
In a Tier 1 - Single Matrix Type study performed to validate an ATP, the laboratory should determine the
background concentration of an unspiked sample prior to analyzing an MS/MSB pair for the matrix type
being tested, for a total of three field sample analyses (i.e., background, MS, and MSB). The laboratory
performing the validation study must demonstrate that it can meet or exceed the MS/MSB precision and
recovery QC acceptance criteria given for the EPA-approved reference method (or other EPA-specified
document). In all, Tier 1- single matrix type validation studies for ATPs will require at minimum,
analysis of seven MBL samples, four IPR reagent water samples, one PT sample and three field samples
(one background, one MS and one MSB), for a total of 15 analyses.
4.3.5.2 Tier 1 - Multiple Matrix Type Validation Studies
In Tier 1 - Multiple Matrix Type studies performed to validate ATPs, the laboratory should determine the
background concentration and analyze an MS/MSB pair for each matrix type being tested, up to a total of
nine matrix types. Since three field sample analyses are required for each matrix type (one background,
one MS, and one MSB), and between two and nine matrix types may be tested, a Tier 1- Multiple Matrix
Type validation study will require analysis of 6 - 21 field samples. The laboratory performing the study
should demonstrate that it can meet or exceed the MS/MSB precision and recovery QC acceptance
criteria given for the EPA-approved reference method (or other EPA-specified document) for each matrix
type being tested, all, Tier 1- multiple matrix type validation studies for ATPs will require at minimum,
analysis of seven MBL samples, four IPR reagent water samples, one PT sample and between 6 and 24
field sample analyses (one background, one MS, and one MSB for each additional matrix type to a
maximum of eight additional matrix types). A Tier 1- multiple matrix type validation study will require a
minimum of between 20 and 39 total analyses since between two and eight additional matrix types may
be tested.
17 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
4.3.5.3 Tier 2 Single Matrix Type Validation Studies
In a Tier 2 validation study, each of the three laboratories will determine the background concentration
and analyze an MS/MSD pair for the field sample received. Because there are three laboratories, each of
which performs three field sample analyses (one background, one MS, and one MSB), Tier 2 validation
studies will require analysis of 9 field samples in total. Each laboratory participating in the study should
demonstrate that it can meet or exceed the MS/MSD precision and recovery QC acceptance criteria given
for the EPA-approved reference method (or other EPA-specified document). Since there are three
laboratories, each of which performs analysis of seven MDL samples, four IPR reagent water samples,
one PT sample and three field samples (one background, one MS, and one MSB), Tier 2 validation
studies will require a minimum of 45 total analyses.
4.3.5.4 Tier3 Validation Studies
In a Tier 3 validation study, each of the nine laboratories participating in the study will determine the
background concentration and analyze an MS/MSD pair for the field sample received. Because there are
a total of nine laboratories, each performing three field sample analyses (one background, one MS, and
one MSB), a Tier 3 validation study will require analysis of 27 field samples in total. Each laboratory
participating in the study should demonstrate that it can meet or exceed the MS/MSD precision and
recovery QC acceptance criteria given for the EPA-approved reference method (or other EPA-specified
document). Since there are nine laboratories, each of which performs analysis of seven MDL samples,
four IPR reagent water samples, one PT sample and three field samples (one background, one MS, and
one MSD), a Tier 3 validation study will require a minimum of 135 total analyses.
4.3.6 Ongoing Precision and Recovery
Each batch of samples that includes field samples, but not the IPR samples, must include an OPR sample.
(As noted above, field samples are analyzed after each laboratory participating in the study has
successfully completed the IPR analyses.) In order to successfully validate the ATP, each participating
laboratory must demonstrate it can meet or exceed the OPR recovery criteria given in the EPA-approved
reference method or other EPA-specified document.
4.3.7 Calibration Verification
The field samples discussed in Section 4.3.5 should be analyzed in a separate batch from the initial
calibration sequence, so that calibration verification is performed. In order to successfully validate the
ATP, each laboratory participating in a Tier 1, 2, or 3 validation study should verify calibration as
described in the method. In order to successfully validate the ATP, each participating laboratory also
should demonstrate it can meet or exceed the acceptance criteria given for the EPA-approved reference
method (or other EPA-specified document) for calibration verification.
4.3.8 Method Blanks
Each laboratory that participates in a Tier 1, 2, or 3 validation study should prepare and analyze at least
one method blank with the sample batch containing the matrix samples. The actual number of blank
samples analyzed by each laboratory must meet or exceed the frequency specified in the method. In order
to successfully validate the ATP, each participating laboratory should demonstrate it can meet or exceed
the QC acceptance criteria for blanks that are specified in the reference method or other EPA-specified
document.
18 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
4.3.9 Surrogate or Labeled Compound Recovery
For methods that use surrogates or labeled compounds, each laboratory participating in the Tier 1, 2, or 3
validation study should spike all field and QC samples with the surrogates/labeled compounds at the
concentrations specified in the method. In order to successfully validate the ATP, each participating
laboratory must demonstrate it can meet or exceed the surrogate or labeled compound recovery criteria
specified in the EPA-approved reference method (or other EPA-specified document).
4.3.10 Absolute and Relative Retention Time
Each laboratory participating in a Tier 1, 2, or 3 validation study of a chromatographic method should
determine the absolute and/or relative retention times of the analytes of interest where required by the
method. To successfully validate the ATP, each participating laboratory should demonstrate that it can
meet or exceed the absolute and relative retention time criteria that are specified in the EPA-approved
reference method (or other EPA-specified document) if applicable.
4.3.11 New Analytes
As described in Section 1.3.3, EPA will allow the addition of new analytes to approved methods as
method modifications under this protocol when required by a specific permit. Laboratories will be
required to demonstrate acceptable method performance in accordance with the requirements summarized
above for other Tier 1, 2, and 3 ATPs. In addition, laboratories are required either to develop QC
acceptance criteria for the added analyte or demonstrate that the existing QC acceptance criteria can be
met for the added analyte; see Protocol for EPA Approval of New Methods for Organic and Inorganic
Analytes in Wastewater.
4.3.12 Proficiency Testing Results
Each laboratory participating in a Tier 1, 2, or 3 validation study should include analysis of a proficiency
testing (PT) sample obtained from an approved vendor. An example list of approved vendors can be
found at: http://www.nelac-institute.org/ptproviders.php (other lists may exist as well). This PT sample
will be analyzed in addition to each of the matrix types required to be analyzed as part of the validation
study and will be analyzed as it is received from the vendor. The same PT sample or samples obtained
from the same vendor with the same lot number or preparation batch number will be analyzed by all
laboratories participating in the validation study.
The concentrations of the target analytes in the PT sample should be relevant to any regulatory limits
associated with the matrix type(s) of interest. PT vendors that prepare samples for periodic Discharge
Monitoring Report Quality Assurance (DMRQA) studies may be able to provide assistance with selection
of concentrations for the PT samples.
The study coordinator will be responsible for obtaining the PT sample from the vendor, along with the
certificate of analysis that specifies the certified value and acceptance limits for reporting results. The
study coordinator will also be responsible for distributing the sample to the laboratories that will be
performing the analyses for the validation study (or in the case of a Tier 1 study to the analyst responsible
for performing the analyses) without providing them with the certificate of analysis (e.g., "blind" as to the
expected results). The study coordinator is also responsible for informing each laboratory participating in
the validation study (or in the case of a Tier 1 study, the analyst responsible for performing the analyses)
that the sample is to be analyzed only once just as it is received and is not to be diluted or fortified for
analysis as an MS/MSD pair. In addition, the study coordinator should include a copy the certificate of
analysis as an addendum to the validation study report.
19 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
5.0 EPA REVIEW AND APPROVAL
5.1 EPA's Office of Water Review of ATP Applications
All requests for approval of ATPs must undergo review and approval by the approval organization listed
in Table 1 of Section 3.1. Limited-use ATPs (Tier 1) will be approved by the EPA Regional ATP
Coordinator. ATP applications for nationwide use (Tiers 2 and 3) will be approved through rulemaking.
ATPs prepared under this protocol should demonstrate an improvement when compared to the EPA-
approved reference method that offers one or more of the following advantages: better method sensitivity
or selectivity, lower analytical costs, fewer matrix interference problems, improvement in laboratory
productivity, or reduction in the amount of hazardous materials used and/or produced in the laboratory.
EPA's Office of Water (OW) will review all Tier 2 and Tier 3 nationwide use ATPs and will review
limited-use (Tier 1) applications if requested by the EPA Regional Office or state agency. OW may be
assisted in its technical review by contractor personnel. When a formal ATP application is received, it
will be checked for completeness. If the documentation is incomplete, OW will contact the applicant and
request missing documentation before proceeding with its review.
At a minimum, an application should include a completed ATP application form, the test procedure in
EPA standard format, and the method comparison table, before OW will review the package. If these
elements are present, OW will assess the application to determine if the modification falls within the
flexibility provided at 40 C.F.R. § 136.6. If the modification falls within the flexibility provided at 40
C.F.R. § 136.6 the application will be returned to the applicant with no further action. If the modification
does not fall within the flexibility provided, then a full ATP validation is required.
Once all elements of the ATP application are present, including the validation study report and supporting
data, OW will begin its internal review of the ATP for scientific merit, consistency, and appropriateness.
The internal review may involve multiple programs and workgroups. Should any problems or questions
arise during the review, OW or its technical support contractor will communicate with the applicant to
resolve outstanding issues. Depending on the circumstances, OW may return the application to the
applicant for revision. OW review of ATP applications will involve the three steps briefly described
below.
The first step of OW's technical review will evaluate the description of the alternate method and method
comparison table, and assess the ATP's applicability for approval at 40 C.F.R. 136. If the alternate
method is not applicable to 40 C.F.R. 136 and/or the method description or method comparison table are
not acceptable, OW will notify the applicant and describe the basis for rejection of the application. If this
information is acceptable, the evaluation will proceed.
In the second step of OW's review, the performance of the ATP is compared to the performance of the
corresponding EPA-approved method. At a minimum, results produced using the ATP must meet the QC
acceptance criteria of the corresponding reference method (for methods addressing non-method-defined
parameters) or demonstrate that there are no systematic differences in performance between the ATP and
the corresponding EPA-approved method (for methods addressing method-defined parameters). If
method performance is acceptable, the review will continue.
As the third and final step, OW will perform a detailed audit of the alternate method test data. The
evaluation of test data in applications can be accomplished more quickly if machine-readable files of test
data (and analysis software where different from EPA software) are provided with the application. Data
files should be in a PC-compatible format, suitable for input directly into statistical analysis software.
20 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Note: Although EPA will review the data from the validation study and conduct its own statistical test
on the study results, the applicant is responsible for the technical and statistical evaluation of the
validation study results prior to submitting the study report.
5.2 Approval Recommendation
EPA will complete its review and notify the applicant of its approval recommendation as expeditiously as
practicable after receipt of an application containing the information necessary for EPA's evaluation. For
limited-use applications (Tier 1), the Regional ATP Coordinator will notify the applicant and the
appropriate State agency of approval or rejection of the use of the alternate test procedure. The EPA
Region will issue the formal approval for use of the Tier 1 ATP. The approval may be restricted to use
only with respect to a specific discharge or facility (and its laboratory) or, at the discretion of the Regional
ATP Coordinator, to all dischargers or facilities (and their associated laboratories) specified in the
approval for the Region.
For all nationwide use ATP applications for use in Clean Water Act programs (Tiers 2 or 3), OW will
notify the applicant of EPA's recommendation, and if the ATP is recommended for approval, will initiate
the rulemaking process through which the ATP is formally approved by the EPA Administrator.
5.3 Rulemaking Process
EPA periodically updates the lists of analytical methods approved for Clean Water Act compliance
monitoring at 40 C.F.R. 136 to provide increased flexibility to the regulated community and laboratories
in their selection of analytical methods for use in Clean Water Act programs. EPA also uses these
periodic "method update rules" (MURs) to formalize the approval status of nationwide ATPs which have
been positively reviewed. Using the method information provided with the ATP application to develop
the justification and record support, EPA will prepare the proposed rule for approval of wastewater
methods, compile the rule docket, pass the proposed rule through internal and/or external review at EPA,
and submit it to the Office of the Federal Register (OFR) for publication. Preparation, approval, and
publication of a proposed rule generally requires a minimum of nine months, but may take longer,
depending on the number of methods involved in the rulemaking effort. When published, the proposed
rule requests public comment and allows a specified comment period. At the end of the comment period,
EPA may forward any significant comments to the ATP applicant with a request that they provide
technical assistance to EPA in drafting responses to comments. All comments that have scientific or legal
merit, or raise substantive issues with the proposed rule, must be answered to complete the rulemaking
process.
EPA will review any technical responses provided by the applicant and complete the response-to-
comments document for the final rule. EPA will then prepare the final rule, compile the rule docket, and
submit the final rule to the OFR for publication. The final rule will state the date that the rule becomes
effective, typically 30 days after rule publication. As of this effective date, the method is approved by
EPA and will be included in the appropriate table(s) at 40 C.F.R. 136 in the next C.F.R. update. It
generally requires a minimum of fifteen months, but may take longer, after the proposed rule is published
to receive and respond to comments, prepare and process the final rule through internal EPA review, and
publish the final rule in the Federal Register.
21 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
6.0 REFERENCES
1. ASTM, 1994. Standard Practice for Determination of Precision and Bias of Applicable Methods of
Committee D-19 on Water. Designation D-2777-86 (Reapproved 1994). Annual Book of ASTM
Standards,. Vol. 11.04.
2. Youden, W.J. and E.H. Steiner, 1975. Statistical Manual of the AOAC. AOAC- International. 1111 N.
19th Street; Suite 210, Arlington, VA 22209.
3. Wernimont, G.T., 1985. Use of Statistics to Develop and Evaluate Analytical Methods. AOAC-
International.
4. USEPA, 1996. Guidelines and Format for Methods to Be Proposed at 40 C.F.R. Part 136 or Part
141 (Guidelines and Format document). U.S. Environmental Protection Agency. Office of Water,
Engineering and Analysis Division. Washington, DC EPA-821-B-96-003.
5. USEPA, 1999. Protocol for EPA Approval of New Methods for Organic and Inorganic Analytes in
Wastewater and Drinking Water. U.S. Environmental Protection Agency. Office of Water,
Engineering and Analysis Division. Washington, DC EPA 821-B-98-003.
6. USEPA, 1999. Protocol for EPA Approval of Alternate Test Procedures for Organic and Inorganic
Analytes in Wastewater and Drinking Water. U.S. Environmental Protection Agency. Office of
Water, Engineering and Analysis Division. Washington, DC EPA 821-B-98-002.
7. USEPA, 2016. Protocol for Review and Validation of New Methods for Regulated Organic and
Inorganic Analytes in Wastewater Under EPA's Alternate Test Procedure Program. U.S.
Environmental Protection Agency. Office of Water, Engineering and Analysis Division. Washington,
DCEPA821-B-16-001.
22 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
APPENDIX A - Sample ATP APPLICATION FORM
EPA Office of Water
Alternate Test Procedure Application Form for Chemical Analytes
Applicant Name and Address:
Date Application Submitted:
Alternate Test Procedure:
(Method number & title)
Alternate to Approved Method:
EPA-Approved Reference
Method used for Comparison:
Analyte(s):
Type (WW, DW, or WW/DW):
Level of Use:
(Limited Use or Nationwide Use)
EPA Use Only
ATP Case No.
Is this a Method-Defined
Parameter (Yes/No)?
Validation Tier: (1 , 2 or 3)
FOR LIMITED-USE APPLICATIONS ONLY:
ID number of existing or
pending permit:
Issuing agency:
Type of permit:
Discharge serial number:
ATTACHMENTS: Each item below includes a reference to the section of the ATP protocol that describes
the material in detail
Justification for ATP (Sec. 3.2.1)
Alternate Test Procedure (Method in standard EPA format) (Sec. 3.2.2)
Method Comparison Table (Sec. 3.2.3)
Validation Study Plan (Appendix E)
Validation Study Report (Sec. 3.2.4)
Method Information anc
1 Documentation for Preamble and Docket (Sec. 3.2.5)
Other
Submit Application and Attachments in Triplicate
A-l
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
APPENDIX B - DATA COLLECTION CERTIFICATION
It is the expectation of the ATP program that all data will be collected as outlined in the validation study
plan. If a data set needs to be recollected (e.g., QC failure, instrument failure, matrix effects etc.) this
should be clearly be documented in the final report and the initial data along with the recollected data
should be submitted. It is not permissible to collect multiple data sets and submit the "best one".
Occasionally, blind samples (performance evaluation samples) will be distributed by the ATP program to
assess method performance. Successful analysis of these samples will be required as part of the candidate
method approval process. Laboratory fraud is a serious issue and applicants must attest on the application
that the data collection was performed as outlined in the validation study plan.
The applicant hereby certifies that the data included with this application was collected as outlined
in the validation study plan.
Applicant (print name)
Applicant (signature) (Date)
Questions, comments or applications should be directed to:
Lem Walker
Clean Water Act ATP Coordinator
U.S. Environmental Protection Agency
Office of Science and Technology
Engineering and Analysis Division (EAD)
1200 Pennsylvania Avenue, NW
Mail Code - 4303T
Washington, DC 20460
Fax: (202) 566-1053
walker.lemuel@epa.gov
B-l February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
APPENDIX C - HEADQUARTERS AND REGIONAL ATP CONTACTS
Headquarters
Lem Walker
Clean Water Act ATP Coordinator
USEPA
Office of Science and Technology
Engineering and Analysis Division (EAD)
1200 Pennsylvania Avenue, NW
Mail Code - 4303T
Washington, DC 20460
walker.lemuel@epa.gov
Region 1
Ann Jefferies
QA Chemist
USEPA Region 1
11 Technology Drive
North Chelmsford, MA 01863-2431
jefferies.ann@epa.gov
Region 2
Donna Ringel
USEPA Region 2
Raritan Depot
2890 Woodbridge Avenue (MS-220)
Building 10
Edison, NJ 08837-3679
ringel.donna@epa.gov
Region 3
Terry Simpson
Regional Quality Assurance Manager
USEPA Region 3
Environmental Science Center
701 Mapes Road
Fort Meade, MD 20755-5350
simpson.terry@epa.gov
Region 4
Ray Terhune
Chemist/ATP Coordinator
USEPA Region 4
Room: SESD
980 College Station Road
Athens, GA 30605-2720
terhune.ray@epa.gov
Region 5
Kenneth Gunter
USEPA Region 5
77 W. Jackson Blvd.
WC-15J
Chicago, IL 60604-3507
gunter.kenneth@epa.gov
Region 6
David Stockton
USEPA Region 6 Laboratory
Houston Branch
10625 Fallstone Road (6MD)
Houston, TX 77099
stockton.david@epa.gov
Region 7
Isabel Velasquez
USEPA Region 7
Science and Technology Center
300 Minnesota Avenue
Kansas City, KS 66101
velasquez.isabel@epa.gov
Region 8
Jeff Pritt
Regional Quality Assurance Officer
USEPA Region 8
1595 Wynkoop Street (8TMS-Q)
Denver, CO 80202-1129
pritt.jeff@epa.gov
Region 9
Roseanne Sakamoto
USEPA Region 9
75 Hawthorne Street,
MTS-3
San Francisco, CA 94105
sakamoto.roseanne@epa.gov
Region 10
Donald M. Brown
Office of Environmental Assessment
USEPA Region 10
1200 6th Avenue, Suite 900, OEA-40
Seattle, Washington 98101
brown.donaldm@epa.gov
Note: The names and addresses in this list are current as of the date of this document, and are subject to change.
Please consult with the individual EPA Regional Office for the current ATP contact.
C-l
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
APPENDIX D - STANDARD EPA METHOD FORMAT
The following is a listing of the 17 elements of the standard EPA method format. Applicants should
consult the Guidelines and Format document (USEPA, 1996, Reference 4 in Section 6 of the main body
of this document) for a detailed description of the required content for each section and other formatting
guidelines and conventions.
1.0 Scope and application
This section outlines the purpose, range, limitations, and intended use of the method, and identifies
target analytes.
2.0 Summary of Method
This section provides an overview of the method procedure and quality assurance.
3.0 Definitions
This section includes definitions of terms, acronyms, and abbreviations used in the method. If
preferred, definitions may be provided in a glossary at the end of the method or manual. In this
case, the definitions section should still appear in the method, with a notation that definitions are
provided in a glossary at the end of the method. Refer to the specific section number of the
glossary.
4.0 Interferences
This section identifies known or potential interferences that may occur during use of the method,
and describes ways to reduce or eliminate interferences.
5.0 Safety
This section describes special precautions needed to ensure personnel safety during the
performance of the method. Procedures described here should be limited to those which are above
and beyond good laboratory practices. The section should contain information regarding specific
toxicity of analytes or reagents.
6.0 Equipment and Supplies
This section lists and describes all non-consumable supplies and equipment needed to perform the
method.
7.0 Reagents and Standards
This section lists and describes all reagents and standards required to perform the method, and
provides preparation instructions and/or suggested suppliers as appropriate.
8.0 Sample Collection, Preservation, and Storage
This section provides requirements and instructions for collecting, preserving, and storing samples.
D-l February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
9.0 Quality Control
This section cites the procedures and analyses required to fully document the quality of data
generated by the method. The required components of the laboratory's quality assurance (QA)
program and specific quality control (QC) analyses are described in this section. For each QC
analysis, the complete analytical procedure, the frequency of required analyses, and interpretation
of results are specified.
10.0 Calibration and Standardization
This section describes the method/instrument calibration and standardization process, and required
calibration verification. Corrective actions are described for cases when performance specifications
are not met.
11.0 Procedure
This section describes the sample processing and instrumental analysis steps of the method, and
provides detailed instructions to analysts.
12.0 Data Analysis and Calculations
This section provides instructions for analyzing data, and equations and definitions of constants
used to calculate final sample analysis results.
13.0 Method Performance
This section provides method performance criteria for the method, including precision/bias
statements regarding detection limits and source/limitations of data produced using the method.
14.0 Pollution Prevention
This section describes aspects of the method that minimize or prevent pollution known to be or
potentially attributable to the method.
15.0 Waste Management
This section describes minimization and proper disposal of waste and samples.
16.0 References
This section lists references for source documents and publications that contain ancillary
information. Note: Each method should be a free-standing document, providing all information
necessary for the method user to perform the method may be found. References within a method
should be restricted to associated or source material. Procedural steps or instructions should not be
referenced as being found elsewhere, but should be included in total within the method.
77.0 Tables, Diagrams, Flowcharts, and Validation Data
This section contains all method tables and figures (diagrams and flowcharts), and may contain
validation data referenced in the body of the method.
D-2 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
APPENDIX E -Validation Study Plan and Study Report
1.1 Development of a Validation Study Plan
Prior to conducting Tier 1, 2, or 3 validation studies, the ATP applicant (e.g., the organization responsible
for conducting the study) should prepare and submit a detailed study plan. As noted earlier, for ATPs that
measure method-defined parameters, a detailed validation study plan should be submitted and agreed
upon prior to conducting the study (see Appendix H). For Tier 1 ATP validation studies involving
analytes which are not method-defined, development of a validation study plan is not required, though it
is recommended.
The validation study plan should contain the elements described in Sections 1.1.1 through 1.1.6.
1.1.1 Background
The Background section of the validation study plan should:
Identify the ATP method as a modification of an approved method
• Identify intended use of the ATP method (Tier 1, Tier 2 or Tier 3)
• Include a summary of the ATP method
• Cite the organization and method number (given in 40 C.F.R. Parts 136 or 405 - 503) for the
approved method (e.g., EPA Method 353.1)
• Describe the reasons for and extent of the modification, the logic behind the technical approach to the
modification, and the result of the modification
• Identify the matrices, matrix types, and/or media to which the ATP method is believed to be
applicable
List the analytes measured by the ATP method including corresponding CAS Registry numbers (if
applicable)
• Indicate whether any, some, or all known metabolites, decomposition products, or known commercial
formulations containing the analyte are included in the measurement. For example, a method
designed to measure acid herbicides should include the ability to measure the acids and salts of these
analytes; a total metals method should measure total metals.
1.1.2 Objectives
The Objectives section of the validation study plan should describe overall objectives and data quality
objectives of the study.
1.1.3 Study Management
The Study Management section of the validation study plan should:
• Identify the organization responsible for managing the study
Identify laboratories, facilities, and other organizations that will participate in the study
• Delineate the study schedule
E-l February 2106
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
1.1.4 Technical Approach
The Technical Approach section of the validation study plan should:
Indicate at which tier the study will be performed
• Describe the approach that will be followed by each organization involved in the study
• Describe how sample matrices and participating laboratories will be selected
Explain how samples will be collected and distributed
• Specify the numbers and types of analyses to be performed by the participating laboratories
Describe how analyses are to be performed
1.1.5 Data Reporting and Evaluation
This section of the validation study plan should explain the procedures that will be followed for reporting
and validating study data, and should address statistical analysis of study results.
1.1.6 Limitations
The Limitations section of the validation study plan should explain any limiting factors related to the
scope of the study.
1.2 Validation Study Report
Applicants responsible for developing ATPs at Tiers 1, 2, or 3 should document the results of the
validation study in a formal validation study report that contains the elements described in this section and
presents these elements in the same order described in this section. In all cases, a copy of all required
validation data should be maintained at the laboratory or other organization responsible for developing the
ATP.
The information and supporting data required in the validation study report should be sufficient to enable
EPA to support a claim of acceptable performance of a method modification. If data are collected by a
contract laboratory, the organization responsible for using the method (e.g., permittee, POTW, or other
regulated entity) is responsible for ensuring that all method-specified requirements are met by the contract
laboratory and that the validation study report contains all required data.
Like the validation study plan, the validation study report contains background information and describes
the study design. In addition, the validation study report details the process and results of the study,
provides an analysis and discussion of the results, and presents study conclusions. If a validation study
plan was prepared, it should be appended to and referenced in the validation study report. The validation
study report should identify and discuss any deviations from the study plan that were made in
implementing the study.
The validation study report should contain a signed Data Collection Certification form (see Appendix B
of this document) and the elements described in Sections 1.2.1 through 1.2.10 below.
1.2.1 Background
The Background section of the validation study report should describe the method modification that was
validated and identify the organization responsible for developing the ATP. The background section of
the validation study report should:
E-2 February 2106
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
• Include a method summary
Cite the organization and method number and title for the ATP
• Cite the method number (given in 40 C.F.R. Part 136) of the approved method that is being modified
• Cite the method number (given in 40 C.F.R. Part 136) of the EPA-approved reference method that is
being used to demonstrate acceptable ATP performance
• Describe the reasons for and extent of the modification, the logic behind the technical approach to the
modification, and the result of the modification
Identify the matrices, matrix types, and/or media to which the modified method is intended to apply
• List the analytes measured by the modified method including corresponding CAS Registry numbers
(Alternatively, this information may be provided on the data reporting forms in the Supporting Data
appendix to the validation study report.)
• Indicate whether any, some, or all known metabolites, decomposition products, or known commercial
formulations containing the analyte are included in the measurement. (For example, a method
designed to measure acid herbicides should include the ability to measure the acids and salts of these
analytes.)
State the purpose of the study.
1.2.2 Study Design and Objectives
The Study Design and Objectives section of the validation study report should describe the study design,
and identify overall objectives and data quality objectives of the study. Any study limitations should be
identified. The validation study plan may be appended to the validation study report to provide the
description of the study design. If no validation study plan was prepared, the study design should be
described in this section (see Section 4.3, Detailed Procedures for Conducting Validation Studies, in the
main body of this document for required elements of the study design).
1.2.3 Study Implementation
The Study Implementation section of the validation study report should describe the methodology and
approach undertaken in the study. This section should:
• Identify the organization that was responsible for managing the study
Identify the laboratories, facilities, and other organizations that participated in the study; describe how
those participants were selected; and explain the role of each organization involved in the study
• Indicate at which Tier level the study was performed
Delineate the study schedule that was followed
• Describe how sample matrices were chosen, including a statement of compliance with Tier specific
validation study specifications for matrix type selection
Explain how samples were collected and distributed
• Specify the numbers and types of analyses performed by the participating laboratories
• Describe how analyses were performed
Identify any problems encountered or deviations from the study plan and their resolution/impact on
study performance and/or results
1.2.4 Data Reporting and Validation
This section of the validation study report should describe the procedures that were used to report and
validate study data. While EPA does not require the use of a standard format for analytical data
submission, a validation study data reporting form may be found in Appendix F of this document.
E-3 February 2106
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
1.2.5 Results
This section of the validation study report presents the study results. Raw data and example calculations
are required as part of the results and shall be included in an appendix to the validation study report (see
Section 1.2.10 below).
1.2.6 Data Analysis/Discussion
This section of the validation study report should provide a statistical analysis and discussion of the study
results. The discussion should address any discrepancies between the results and the QC acceptance
criteria of the EPA-approved reference method.
1.2.7 Conclusions
The Conclusions section of the validation study report should describe the conclusions drawn from the
study based on the data analysis discussion. The Conclusions section should contain a statement(s)
regarding achievement of the study objective(s).
1.2.8 Appendix A - The Method Compilation
A written version of the modified method prepared in accordance with EPA's Guidelines and Format
document, should be appended to the validation study report (see Reference 4 in Section 6 of the main
body of this document).
1.2.9 Appendix B - Validation Study Plan
If a validation study plan was prepared, it should be appended to the validation study report.
1.2.10 Appendix C - Supporting Data
The validation study report should be accompanied by raw data and example calculations that support the
results presented in the report.
1.2.10.1 Raw Data
The Results section of the validation study report should be supported by an appendix containing all raw
data that will allow an independent reviewer to verify each determination and calculation performed by
the laboratory. This verification consists of tracing the instrument output (peak height, area, or other
signal intensity) to the final result reported. Raw data are method-specific and may include any of the
following:
Sample numbers or other identifiers used by the both the ATP applicant and the laboratory(ies) that
participated in the study
Sample preparation (extraction/digestion) dates
Analysis dates and times
• Sequence of analyses or run logs
Sample volume
Extract volume prior to each cleanup step
• Extract volume after each cleanup step
Final extract volume prior to injection
• Digestion volume
E-4 February 2106
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
• Titration volume
Percent solids or percent moisture
• Dilution data, differentiating between dilution of a sample and dilution of an extract or digestate
• Instrument(s) and operating conditions
GC and/or GC/MS operating conditions, including detailed information on
o Columns used for determination and confirmation (column length and diameter, stationary phase,
solid support, film thickness, etc.)
o Analysis conditions (temperature programs, flow rates, etc.)
o Detectors (type, operating conditions, etc.)
Chromatograms, ion current profiles, bar graph spectra, library search results
Quantitation reports, data system outputs, and other data to link the raw data to the results reported.
(Where these data are edited manually, explanations of why manual intervention was necessary
should be included)
Direct instrument readouts; i.e., strip charts, printer tapes, etc., and other data to support the final
results
Laboratory bench sheets and copies of all pertinent logbook pages for all sample preparation and
cleanup steps, and for all other parts of the determination
Raw data are required for all samples, calibrations, verifications, blanks, matrix spikes and duplicates, and
other QC analyses required by the EPA-approved reference method. Data should be organized so that an
analytical chemist can clearly understand how the analyses were performed. The names, titles, addresses,
and telephone numbers of the analysts who performed the analyses and of the quality assurance officer
who will verify the analyses should be provided. For instruments involving data systems (e.g., GC/MS),
raw data should be made available in appropriate electronic formats upon request.
1.2.10.2 Example Calculations
The validation study report should provide example calculations that will allow the data reviewer to
determine how the laboratory used the raw data to arrive at the final results. Useful examples include
both detected compounds and undetected compounds. If the laboratory or the method employs a
standardized reporting level for undetected compounds, this should be made clear in the example, as
should adjustments for sample volume, dry weight (solids only), etc.
E-5 February 2106
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
APPENDIX F - Sample DATA REPORTING FORM
This appendix provides an example data reporting form. The form illustrates those aspects of data
reporting which are expected, regardless of the specific format used; specifically, data should be
presented in a clear and logical format, and should be labeled clearly.
In addition to using an appropriate data reporting format, submitting the data in an appropriate electronic
format can be very helpful in expediting the review of an ATP. Data files should be in PC-compatible
format, suitable for input directly into statistical analysis software.
F-l February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Sample ATP Data Reporting Form1
I ATP Method
Title*
Revision
Date
_/_/_
Include Method Number and Revision Number
Please record all data and quality control (QC) performance results (for comparison against QC
acceptance criteria) from your validation study using this data form. If you have additional data, please
attach it to this form in a tabular format, being sure to label all columns and rows clearly.
For Tier 1 Studies (Single Laboratory Use): Complete 1_ form for each matrix type
For Tier 2 (Nationwide Use; Single Matrix) or Tier 3 (Nationwide Use; Multiple Matrices): Complete 1 form
for each participant laboratory.
Units of Concentration:
Linear Calibration Data
Units of Response: Number of Points:.
Analyte Cone.
Response
RF/CF/RR*
*Response Factor/Calibration Factor/Relative Response
Method Detection Limit (MDL) Data
Spiking Concentration used for MDL Study (include units):
MDL Data
I
I
Initial Precision Recovery (IPR) Data
Spiking Concentration used for IPR Study (include units):
IPR Data
Matrix Spike / Matrix Spike Duplicate (MS/MSD) Data
Spiking Concentration used for MS/MSD Study (include units):
MS Concentration
MSD Concentration
Background Concentration
ATP QC Performance Results
Calibration | Spike | IPR Recovery and Precision
Points I! Lin | Cone | Low | High
Precision
OPR Data Precision | MS/MSD Recovery and RPD | MDL/ML
Low
High 1 Low | High | RPD | MDL
ML
1 For multi-analyte methods, present additional Data and QC acceptance criteria for each analyte in a tabular format, making sure to include
proper labels, and attach to this form.
F-2
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
APPENDIX G - QUALITY CONTROL ACCEPTANCE CRITERIA
Table G1 - Standardized QC and QC Acceptance Criteria for Methods in 40 C.F.R. Part 136, Table 1B
No
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
Analyte- Detector
Aluminum- Flame
" - Furnace
11 -ICP
Ammonia - distill
" - Nessler
11 -Titr
11 -ISE
" -Phenate
Antimony - Flame
Antimony - Furnace
Antimony - ICP
Arsenic
11 - Hydride
" - Furnace
11 - ICP
11 - Color (SDDC)
Barium - Flame
" - Furnace
11 - ICP
Beryllium - Flame
" - Furnace
11 - ICP
Boron - Color
11 - ICP
Bromide
Cadmium - Flame
Cadmium - Furnace
Cadmium - ICP
Calcium - Flame
Calcium - ICP
Calcium - Titr
Chloride - Titr/Hg
Chloride - Auto
Chlorine - Ampere
Chlorine - lodo
Chlorine- Back titr
Chlorine -DPD-FAS
Chlorine - Spectra
Chromium VI - AA
Chromium - Flame
Chromium - Furnace
Chromium - ICP
Cobalt - Flame
Cobalt - Furnace
Cobalt - ICP
Copper -Flame
Copper -Furnace
Copper -ICP
Cyanide - Spectra
Reference
Method
202.1
202.2
200.7
350.2
350.2
350.3
350.1
204.1
204.2
200.7
206.3
206.2
200.7
206.4
208.1
208.2
200.7
210.1
210.2
200.7
212.3
200.7
320.1
213.1
213.2
200.7
215.1
200.7
215.2
325.3
325.1
330.1
330.3
330.2
330.4
330.5
218.4
218.1
218.2
200.7
219.1
219.2
200.7
220.1
220.2
200.7
335.2
Spike
cone.
500 |jg/L
500 |jg/L
500|jg/L
1 mg/L
1 mg/L
1 mg/L
1 mg/L
1 mg/L
200 |jg/L
200 |jg/L
100|jg/L
100|jg/L
100|jg/L
40 |jg/L
1 mg/L
1 mg/L
1 mg/L
100|jg/L
50 |jg/L
100|jg/L
240 |jg/L
1 mg/L
2.8 mg/L
100|jg/L
100|jg/L
100|jg/L
200 |jg/L
10 mg/L
10 mg/L
100 mg/L
100 mg/L
1 mg/L
1 mg/L
1 mg/L
1 mg/L
1 mg/L
100|jg/L
100|jg/L
100|jg/L
100|jg/L
500 |jg/L
100|jg/L
100|jg/L
100|jg/L
100|jg/L
100|jg/L
250 |jg/L
Calibra-
tion
points
3
5
3
3
3
3
1
1
5
3
3
3
3
3
3
5
3
3
5
3
5
3
3
3
3
3
3
3
3
3
3
3
5
3
3
3
3
3
3
3
3
3
3
3
5
3
3
Lin-
earity
10%
25%
10%
10%
10%
10%
___
___
25%
10%
10%
10%
10%
10%
10%
25%
10%
10%
25%
10%
25%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
25%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
25%
10%
10%
Specification
IPR
% Recovery and Precision
Low
81
71
81
81
73
79
87
77
70
71
71
82
73
72
97
82
90
85
79
79
54
76
70
88
84
84
82
86
84
92
93
79
78
68
79
82
84
67
83
84
85
85
86
90
86
86
65
High
117
127
121
121
129
127
115
117
118
121
127
118
129
128
101
122
110
109
119
119
146
126
122
110
114
118
120
120
124
108
109
115
116
124
119
120
112
123
117
118
113
113
116
110
112
116
129
SD
18
28
20
20
28
24
14
20
24
25
28
18
28
28
2.0
20
10
12
20
20
46
25
26
11
15
17
19
17
20
7.6
8.2
18
19
28
20
19
14
28
17
17
14
14
15
10
13
15
32
OPR
% Recovery
Low
79
68
79
79
70
77
86
75
68
68
68
80
70
69
97
80
89
84
77
77
49
74
67
87
83
83
80
84
82
92
82
77
76
65
77
80
83
64
82
82
84
83
84
89
84
84
62
High
119
130
123
123
132
129
116
119
120
124
130
120
132
131
101
124
111
110
121
121
151
128
125
111
115
119
122
122
126
108
110
117
118
127
121
122
113
126
118
119
114
115
118
111
114
118
132
MS/MSD
% Recovery
Low
79
68
79
79
70
77
86
75
68
68
68
80
70
69
97
80
89
84
77
77
49
74
67
87
83
83
80
84
82
92
82
77
76
65
77
80
83
64
82
82
84
83
84
89
84
84
62
High
119
130
123
123
132
129
116
119
120
124
130
120
132
131
101
124
111
110
121
121
151
128
125
111
115
119
122
122
126
108
110
117
118
127
121
122
113
126
118
119
114
115
118
111
114
118
132
RPD
20
31
22
22
31
26
15
22
26
28
31
20
31
31
2.2
22
11
13
22
22
51
27
29
12
16
18
21
19
22
8.4
9.0
20
21
31
22
21
15
31
18
18
15
16
17
11
15
17
35
ML
15|jg/L
20 |jg/L
50 |jg/L
50 |jg/L
1 .0 mg/L
30 |jg/L
10|jg/L
1 .0 mg/L
20 |jg/L
20 |jg/L
2.0 |jg/L
5.0 |jg/L
20 |jg/L
10|jg/L
1 .0 mg/L
10|jg/L
2M9/L
50 |jg/L
1.0|jg/L
1.0|jg/L
100|jg/L
10|jg/L
2 mg/L
50 |jg/L
0.5 |jg/L
2M9/L
200 |jg/L
20 |jg/L
2 mg/L
___
1 mg/L
___
0.1 mg/L
___
0.1 mg/L
0.2 mg/L
10|jg/L
15|jg/L
5|jg/L
10|jg/L
500 |jg/L
5M9/L
5M9/L
100|jg/L
5M9/L
10|jg/L
60 |jg/L
G-l
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Table G1 - Standardized QC and QC Acceptance Criteria for Methods in 40 C.F.R. Part 136, Table 1B
No
18.
19.
20.
21.
22.
23.
24.
25.
26.
27.
28.
29.
30.
31.
32.
33.
34.
35.
36.
37.
Analyte- Detector
Fluoride - Elec/man
Fluoride -SPADNS
Fluoride - Auto
Hardness - Color/auto
Hardness -Titr/EDTA
pH - Electrode
Iron - Flame
Iron - Furnace
Iron-ICP
TKN - Digest
TKN - Titr
TKN - Nessler
TKN - Electrode
TKN - Phenate
TKN - Block/color
Lead - Flame
Lead - Furnace
Lead-ICP
Magnesium - Flame
Magnesium - ICP
Manganese - Flame
Manganese - Furnace
Manganese - ICP
Mercury - CV/Man
Mercury - CV/Auto
Molybdenum - Flame
Molybdenum - ICP
Nickel - Flame
Nickel - Furnace
Nickel - ICP
Nitrate
N02-N03-Cd/Man
N02-N03-Cd/Auto
N02-N03-Cd/Hydra
o-Phosphate - Auto
o-Phosphate - Man
DO - Winkler
DO - Electrode
Phenol - Color/Man
Phenol - Color/Auto
Phosphorus - Asc/Man
Phosphorus - Asc/Man
Phosphorus - Asc/Auto
Phosphorus - Block
Potassium - Flame
Potassium - ICP
Selenium - Furnace
Selenium - ICP
Silica - Color/Man
Silica -ICP
Reference
Method
340.2
340.1
340.3
130.1
130.2
150.1
236.1
236.2
200.7
351.3
351.3
351.3
351.3
351.1
351.2
239.1
239.2
200.7
242.1
200.7
243.1
243.2
200.7
245.1
245.2
246.1
200.7
249.1
249.2
200.7
352.1
353.3
353.2
353.1
365.1
365.2
360.2
360.1
420.1
420.2
365.2
365.3
365.1
365.4
258.1
200.7
270.2
200.7
370.1
200.7
Spike
cone.
1 mg/L
1 mg/L
1 mg/L
100 mg/L
100 mg/L
N/A
500 |jg/L
100|jg/L
500 |jg/L
2 mg/L
5 mg/L
5 mg/L
5 mg/L
5 mg/L
5 mg/L
300 |jg/L
100|jg/L
300 |jg/L
2 mg/L
2 mg/L
100|jg/L
100|jg/L
100|jg/L
4|jg/L
4|jg/L
300 |jg/L
100|jg/L
100|jg/L
100|jg/L
100|jg/L
1 mg/L
1 mg/L
1 mg/L
1 mg/L
300 |jg/L
300 |jg/L
1 mg/L
1 mg/L
500 |jg/L
500 |jg/L
1 mg/L
1 mg/L
1 mg/L
1 mg/L
10 mg/L
10 mg/L
100|jg/L
300 |jg/L
5 mg/L
1 mg/L
Calibra-
tion
points
3
3
3
3
3
2
3
5
3
5
3
5
5
5
3
3
3
3
3
3
3
3
3
5
3
3
3
3
3
3
5
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
5
3
5
Lin-
earity
10%
10%
10%
10%
10%
___
10%
25%
10%
25%
10%
25%
25%
25%
10%
10%
10%
10%
10%
10%
25%
10%
10%
25%
10%
10%
10%
10%
10%
10%
25%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
25%
10%
25%
Specification
IPR
% Recovery and Precision
Low
85
79
87
93
93
87
80
88
49
82
78
69
78
79
87
84
84
83
84
86
83
86
84
77
67
80
83
84
82
77
79
88
88
86
89
98
98
59
41
82
79
81
80
84
82
77
80
64
-82
High
115
127
117
109
107
113
124
116
153
118
122
129
122
119
113
116
118
115
120
112
113
114
126
121
131
118
117
116
120
125
119
110
110
112
113
102
102
123
121
112
115
111
112
116
120
117
120
120
190
SD
15
24
15
8.4
7.2
2.2
13
22
14
52
18
22
30
22
20
13
16
17
16
18
13
15
14
26
22
32
19
17
16
19
24
20
11
11
13
12
2.0
2.0
32
40
15
18
15
16
16
19
20
20
28
136
OPR
% Recovery
Low
84
77
85
92
92
86
78
86
44
80
76
66
76
77
86
82
82
81
82
85
81
84
71
75
64
78
81
83
80
75
77
87
87
84
87
98
98
56
37
81
77
80
79
82
80
75
78
61
-96
High
116
129
119
110
108
114
126
118
158
120
124
132
124
121
114
118
120
117
122
113
115
116
129
123
134
120
119
117
122
127
121
111
111
114
115
102
102
126
125
113
117
112
113
118
122
119
122
123
204
MS/MSD
% Recovery
Low
84
77
85
92
92
86
78
86
44
80
76
66
76
77
86
82
82
81
82
85
81
84
71
75
64
78
81
83
80
75
77
87
87
84
87
98
98
56
37
81
77
80
79
82
80
75
78
61
-96
High
116
129
119
110
108
114
126
118
158
120
124
132
124
121
114
118
120
117
122
113
115
116
129
123
134
120
119
117
122
127
121
111
111
114
115
102
102
126
125
113
117
112
113
118
122
119
122
123
204
RPD
16
26
17
9.2
7.9
2.4
14
24
16
57
20
24
33
24
22
14
18
19
18
20
14
17
16
29
24
35
21
19
17
21
26
22
12
12
15
14
2.2
2.2
35
44
16
20
16
17
18
21
22
22
31
150
ML
100|jg/L
100|jg/L
50 |jg/L
10 mg/L
30 mg/L
N/A
300 |jg/L
5M9/L
100|jg/L
50 |jg/L
50 |jg/L
50 |jg/L
50 |jg/L
50 |jg/L
100|jg/L
40 |jg/L
5|jg/L
20 |jg/L
20 |jg/L
50 |jg/L
100|jg/L
1M9/L
2M9/L
o.2Mg/L
0.2 |jg/L
300 |jg/L
10|jg/L
0.2 |jg/L
5|jg/L
20 |jg/L
0.1 mg/L
10|jg/L
50 |jg/L
10|jg/L
10|jg/L
10|jg/L
50 |jg/L
50 |jg/L
5M9/L
2Mg/L
10|jg/L
10|jg/L
10|jg/L
10|jg/L
100|jg/L
1 mg/L
5M9/L
50 |jg/L
2 mg/L
50 |jg/L
G-2
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Table G1 - Standardized QC and QC Acceptance Criteria for Methods in 40 C.F.R. Part 136, Table 1B
No
38.
39.
40.
41.
42.
43.
44.
45.
46.
Analyte- Detector
Silver - Flame
Silver - Furnace
Silver -ICP
Sodium - Flame
Sodium - ICP
Sulfate - Color/Auto
Sulfate - Grav
Sulfate - Turbid
Surfactants
Thallium - Flame
Thallium - Furnace
Thallium - ICP
Tin -Flame
Titanium - Flame
Vanadium - Flame
Vanadium - Furnace
Vanadium - ICP
Zinc- Flame
Zinc - Furnace
Zinc -ICP
Reference
Method
272.1
272.2
200.7
273.1
200.7
375.1
375.3
375.4
425.1
279.1
279.2
200.7
282.1
283.1
286.1
286.2
200.7
289.1
289.2
200.7
Spike
cone.
100|jg/L
100|jg/L
100|jg/L
30 |jg/L
10mg/L
50 mg/L
50 mg/L
50 mg/L
3 mg/L
100|jg/L
100|jg/L
100|jg/L
10 mg/L
2 mg/L
2 mg/L
200 |jg/L
200 |jg/L
100|jg/L
100|jg/L
100|jg/L
Calibra-
tion
points
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
Lin-
earity
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
10%
Specification
IPR
% Recovery and Precision
Low
88
83
83
90
86
83
85
83
83
85
81
73
83
85
81
82
87
87
81
83
High
112
115
117
116
122
115
113
115
119
115
115
127
109
115
121
118
113
113
119
121
SD
12
16
17
13
18
16
14
16
18
15
17
27
13
15
20
18
13
13
19
19
OPR
% Recovery
Low
86
82
82
88
85
82
83
81
81
83
80
70
32
84
79
80
86
85
79
81
High
114
116
118
118
123
116
115
117
121
117
116
130
110
116
123
120
114
115
121
123
MS/MSD
% Recovery
Low
86
82
82
88
85
82
83
81
81
83
80
70
32
84
79
80
86
85
79
81
High
114
116
118
118
123
116
115
117
121
117
116
130
110
116
123
120
114
115
121
123
RPD
14
17
18
15
19
17
16
18
20
17
18
30
14
16
22
20
14
15
21
21
ML
100|jg/L
1M9/L
5|jg/L
30 |jg/L
100|jg/L
10 mg/L
10|jg/L
1 mg/L
25 |jg/L
600 |jg/L
5M9/L
50 |jg/L
10 mg/L
2 mg/L
2 mg/L
10|jg/L
10|jg/L
50 |jg/L
o.2Mg/L
5M9/L
Legend for acronyms and abbreviations in Table G1:
Reference Method: QC acceptance criteria are for modifications to the reference method specified in Table IB.
Spike cone. The concentration at which the QC acceptance criteria were determined.
Calibration points: The number of points required for calibration
Linearity: The relative standard deviation (RSD) of the calibration factor or response factor below which an averaged calibration factor or
response factor may be used in place of a calibration curve. For an averaged response or calibration factor above this number, a
calibration curve must be used. For reference methods that allow the use of a correlation coefficient (r) to judge linearity (e.g., more
recent versions of Method 200.7), the same r value may be used in place of the RSD value listed in this table.
% Recovery: The amount of analyte recovered expressed as a percent (applies to recovery entries for the IPR, OPR, and MS/MSD)
IPR and OPR The lower and upper QC acceptance criteria for % recovery in the initial precision and recovery (IPR) test or the ongoing precision
recovery (low/high) and recovery (OPR) test. For the IPR, these limits apply individually to the recovery in each aliquot, not to the mean recovery of all
four aliquots.
SD: The standard deviation (SD) of the four % recoveries in the IPR test.
MS/MSD recovery The lower and upper QC acceptance criteria for % recovery of the matrix spike and matrix spike duplicate
(low/high):
RPD: The upper limit on the QC acceptance criterion for precision expressed as the relative percent difference (RPD) for the MS/MSD
test. RPD = 100% x [*MS - MSD* / V2(MS + MSD)]
ML value: The minimum level (ML) is the concentration in a sample that is equivalent to the concentration of the lowest calibration point, taking
into account all method-specified sample processing weights and volumes.
G-3
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Table G2 - Standardized QC, QC Acceptance Criteria, and Performance Data for Methods for Method-defined Analytes
in 40 C.F.R. Part 136, Table IB1
Analyte - Detector
Acidity - endpoint
Alkalinity - endpoint
BOD 5 - lodometric
COD-
Spectrophotometric
Color -
Spectrophotometric
Hydrogen ion -
Electrometric
Oil and grease-HEM
- Gravimetry
TOO - Persulfate-UV
Oxidation
Total solids -
Gravimetry
Total dissolved solids
- Gravimetry
Total suspended
solids - Gravimetry
Temperature -
Thermometer
Reference
Method
SM2310B
SM 2320B
SM5210B
EPA 410.4
NCASI 253
SM4500-H+B
EPA1664A
SM5310C
SM 2540B
SM 2540C
SM 2540D
SM 2550B
Spike
cone
20 mg/L
120mg/L
300mg/L
50 mg/L
100CU
7.3 pH
40 mg/L
10 mg/L
293 mg/L
24 mg/L
Calibration
#Pt
3
6
2
Linearity
R2 >0.991
Calibration
Verification
Low
90
High
110
Note 2
Precision and Recovery
Initial (IPR)
Recovery (%)
Low
90
80
83
High
110
120
101
Preci-
sion
RSD
10
11
Ongoing (OPR)
Recovery (%)
Low
56
75
78
High
76
125
114
Matrix Spike/Matrix
Spike Duplicate
(MS/MSD)
Recovery (%)
Low
90
78
High
110
114
Preci-
sion
RPD
18
Detection or
Quantitation
Limit
ML
LDL
2 mg/L
Range
3 mg/L
MDC
10 CU
0.1 pH
5 mg/L
0.1 °C
Method
Performance
Rec (%)
100
93
66
93
93
93
RSD
9
4.2
15.4
14
SD0.26
PH
8.7
7
SD6.0
7.2
10
Note 1. Some QC acceptance criteria may not be appropriate for some analytes in this table.
Note 2 Within +10% of Class S weight at 2 mg and with +0.5% at 1000 mg
G-4
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
APPENDIX H - METHOD-DEFINED PARAMETERS (MDPs)
This appendix provides the recommended validation requirements associated with ATPs for a method-
defined parameter (MDP). As noted throughout the main document, these details are provided in this
appendix to distinguish the validation requirements for ATPs for MDPs more clearly from the validation
requirements of ATPs for the more traditional analytes.
1.1 Definition of a Method-defined Analyte or Parameter
As defined at 40 C.F.R. § 136.6 and noted in Section 1.3.4 in the main body of this document, the term
"method-defined analyte " means an analyte (or parameter) that is defined solely by the method used to
determine the analyte (generically referred to in this document as an MDP). Such an analyte may be a
physical parameter, a parameter that is not a specific chemical, or a parameter that may be comprised of a
number of substances. Examples include, but are not limited to:
• Acidity,
• Alkalinity,
Biological oxygen demand (BOD),
• Chemical oxygen demand (COD),
• Color,
Oil and grease,
• pH (hydrogen ion),
Conductivity (specific conductance),
• Temperature,
• Total dissolved solids (TDS),
Total organic carbon (TOC),
• Total suspended solids (TSS),
• Total phenolics, and
Turbidity.
ATPs that measure MDPs have the potential to change what is being measured. Therefore, all ATPs that
measure MDPs require EPA approval prior to use in NPDES compliance monitoring. Furthermore, the
three-tiered validation approach to ATPs described in the main body of this document for non-MDPs
should not be used in the case of ATPs for MDPs. Rather, all ATPs for MDPs should be validated and
reviewed using the process described in this appendix.
1.2 Approaches to Validation of ATPs for MDPs
EPA would not expect to be able to approve any applications for ATPs that failed to establish the
suitability of the method to measure the MDP through side-by-side comparison studies. These are
necessary to ensure there are no systematic differences in method performance, and that the comparison
data may be evaluated to ensure that any differences in what is being measured are not masked by
between-sample variability.
1.2.1 Tier 1: Side-by-side Comparison for Use in a Single Laboratory
For ATPs that measure MDPs that are intended for limited use in a single laboratory (Tier 1), the
laboratory must perform and document side-by-side comparison of the ATP and the EPA approved
reference method. This study should include analysis of a minimum of three replicate samples collected
on any seven days over a minimum 30-day period using each method. This will require analysis of a total
of 42 field samples (21 by the ATP and 21 by the EPA approved reference method for a single matrix
H-l February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
study). If the laboratory wishes to use the ATP for analysis of more than one matrix type a similar model
should be used for each additional matrix type up to a maximum of nine matrix types. If the laboratory
wishes to use the ATP for analysis of any matrix type, the study design should be similar to the Phase I
single laboratory study comparison study described in Section 1.2.3.1 for Tier 3 ATPs.
If all six results for a given day associated with any sample are less than the minimum level (< ML) of the
reference method, these results should not be used in the comparison because it is necessary to have
actual measured values to test equivalency. In the event that a test result less than the ML is obtained,
samples should be collected on an additional day (i.e., the number of tests should be increased to provide
a minimum of seven paired triplicate results for the comparison).
1.2.2 Tier II: Side-by-side Comparison for Nationwide Use in a Single Matrix Type
Similarly in the case of ATPs that measure MDPs that are intended for nationwide use in a single matrix
type (Tier 2), in order to establish its suitability for use, the applicant should provide side-by-side
comparison studies that are conducted in two phases: a single-laboratory phase, and a multi-laboratory
phase. In the single-laboratory phase, comparability would be established by performing a statistical
comparison of the results obtained from the analysis of minimum of three replicate samples of the
appropriate matrix type collected on any seven days over a minimum 30-day period by both the ATP and
the approved reference method. The single laboratory comparison study should also include analysis of a
proficiency testing sample obtained from an approved vendor and analyzed in triplicate using both the
ATP and the approved reference method. If the ATP single-laboratory data are determined to be
generally comparable to those from the approved reference method, then a second phase will be
conducted to further demonstrate comparability and to generate method performance data across multiple
laboratories and applicable quality control (QC) acceptance criteria.
Given the nature of the side-by-side testing, a carefully prepared validation study plan is an essential
component of the validation and approval process for ATPs that measure MDPs. The applicant may
prepare separate study plans for the two phases of the process, or where practical, a single plan may be
developed that supports both phases.
1.2.2.1 Phase I: Side-by-side Comparison in a Single Laboratory
In Phase I of the comparison study, a minimum of three replicate samples of the appropriate matrix type
collected on any seven days over a minimum 30-day period will be analyzed in a single laboratory by
both the ATP and the approved reference method, and should be used to assess whether there is a
statistically significant difference between the results produced by the ATP and the results produced by
the approved corresponding reference method.
The design of the side-by-side comparison is left up to the ATP applicant. However, a detailed validation
study plan should be prepared by the applicant and submitted to EPA for review and comment, and the
plan agreed upon by all parties prior to conducting the comparison study. This will ensure that the plan
provides the demonstration necessary for EPA to evaluate the ATP MDP's suitability. Although EPA
may be consulted for additional guidance during the development of the study plan, it is the applicant's
responsibility to write the study plan and submit it to EPA for review. The minimum elements to provide
the showing necessary for EPA's evaluation for the design of the Phase I study are provided below and
summarized in Section 1.2.2.3, Table H-l of this appendix.
• Number/Types of Real-World Sample Types: A minimum of three replicate samples of the appropriate
matrix type types should be collected on any seven days over a minimum 30-day period and analyzed
by each method. If preparation of multiple spike levels is feasible for the method-defined parameter,
then use of multiple spike levels is recommended, but a minimum of seven samples per spike level is
H-2 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
expected unless the applicant explains why they are unnecessary. However, in most cases, seven
samples are the minimum number needed to capture the expected variability. If spiking is not
feasible, a range of samples should be targeted that would be expected to yield background
concentrations that vary by at least one order of magnitude.
• Laboratories: The Phase I Comparison Study should be performed in a single laboratory to minimize
the sources of variability. This laboratory should have familiarity with both the approved method and
the ATP to ensure that any differences in performance are not the result of inexperience with one or
both methods. However, it is important that the validation study accurately reflect the ruggedness of
the ATP and any limitations regarding clarity of the ATP procedures. Therefore, the laboratory
should not be affiliated with the ATP applicant.
Replication: The recommended number of replicates to be analyzed per method and sample within
the side-by-side study is three.
To ensure the laboratory can perform both methods acceptably, the laboratory must meet all QC analysis
criteria specified in the approved method using both, the approved method and ATP, prior to the
statistical comparisons of the method data. Moreover, the specific statistical tests that will be used to
compare the results of the ATP with those from the reference method must be described in the study
plan. See Section 1.3 of this appendix for a discussion of the relevant statistical considerations.
If a statistical assessment indicates that Phase I study results produced by the ATP are comparable to
those produced by the designated reference method based on the statistical test described in the validation
study plan, then the ATP will be deemed to be sufficiently comparable to proceed to Phase II.
1.2.2.2 Phase II: Interlaboratory Study
In Phase II of the comparison study, results of the analyses of synthetic and real-world samples in three
laboratories will be used to demonstrate method equivalence and establish method performance and inter-
laboratory QC acceptance criteria for the alternate test procedure. The study design and specific QC tests
for the Phase II study will generally follow the guidelines presented for Tier 2 validation as described in
Section 4.3 of this document, and acceptance criteria will be developed as described in Appendix G of the
"Protocol for Validation and Review of New Methods for Regulated Organic and Inorganic Analytes in
Wastewater". However, not all QC tests will be applicable to all method-defined parameters. For
example, matrix spike samples are not applicable to methods that measure method-defined analytes such
as pH or temperature.
As with Phase I, the specific statistical tests that will be used to compare the results of the ATP with those
from the reference method must be described in the study plan. See Section 1.3 of this appendix for a
discussion of the relevant statistical considerations.
Despite careful planning, situations may arise in which the results from one of the three laboratories in the
study may not represent the performance of the ATP or the other laboratories. Applicants may wish to
plan for such a contingency in the Phase II study plan by utilizing more than three laboratories, or by
documenting relevant corrective action procedures that all laboratories in the study will use prior to
repeating study analyses.
Outlier testing is not recommended for either the single-lab or multi-laboratory phases of the study.
However, if the applicant has reason to believe that some of the results from the validation study truly do
not represent the performance of the method, then they should contact EPA to discuss whether and how
an outlier test could be applied.
H-3 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
It is important that Phase 2 accurately reflect the ruggedness of the ATP and any limitations regarding
clarity of the ATP procedures. Therefore, a vendor or other applicant should not directly assist
laboratories participating in Phase 2 of the study with implementation of the ATP methodology or
equipment during the course of the study (e.g., the vendor or applicant may provide training and advice to
participant laboratories regarding the equipment or methodology prior to the start of the study, but the
study samples are to be analyzed by the study participants under "routine" conditions). Direct
participation by the vendor or applicant will compromise the results of the study.
1.2.2.3 Analyses Recommended for Both Phases of a Tier 2 Validation Study of an A TP
foraMDP
The following table summarizes the recommended minimum numbers of analyses involved in both
phases of the validation study for an ATP involving an MDP
Table H-1 Summary of Validation Recommendations for Tier 2 MDP ATPs1
Study
Phase
Phase I
Phase II
Procedure
ATP
Reference Method
ATP
Reference Method
Number of
Labs
1
1
3
3
Matrix
Samples2
7
7
3
3
Number of Analyses Required
Replicates
per Matrix
Sample3
3
3
3
3
IPRin
Reagent
Water4
4
4
12
12
PT
Sample
1
1
3
3
MS/MSD5
14
14
18
18
MD
|_(6)
7
7
21
21
Total
51
51
63
63
Numbers of analyses in this table do not include additional QC tests such as calibration, blanks, etc.
In Phase I, the matrix samples are collected on any seven days over a minimum 30-day period and analyzed using
each method. In Phase II, the matrix samples are collected on any three days over a minimum 30-day period and
analyzed using each method.
Each laboratory analyzes each matrix sample in triplicate, regardless of the phase.
The IPR analyses only apply to MDPs where the reference method also includes the IPR test.
Each laboratory analyzes one MS/MSD pair for each matrix sample.
The current MDL procedure requires 7 samples, validation studies will comply with most updated MDL study
requirements published in Appendix B of 40 C.F.R. Part 136.
1.2.3 Tier 3: Side-by-side Comparison for Nationwide Use in Any Matrix Type
ATPs that measure MDPs that are intended for nationwide use in all matrix types (Tier 3), shall require
side-by-side comparison studies that will be conducted in two phases: a single-laboratory phase, and a
multi-laboratory phase. In the single-laboratory phase, comparability will be established by performing a
statistical comparison of the results obtained from the analysis of various sample types by both the ATP
and the approved reference method, including the analysis of a proficiency testing sample obtained from
an approved vendor. If the ATP single-laboratory data are determined to be generally comparable to
those from the approved reference method, then a second phase will be conducted to further demonstrate
comparability and to generate method performance data across multiple laboratories and applicable
quality control (QC) acceptance criteria.
Given the nature of the side-by-side testing, a carefully prepared validation study plan is an essential
component of the validation and approval process for ATPs that measure MDPs. The applicant may
prepare separate study plans for the two phases of the process, or where practical, a single plan may be
developed that supports both phases.
H-4
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
1.2.3.1 Phase I: Side-by-side Comparison in a Single Laboratory
In Phase I of the comparison study, a wide variety of synthetic and real-world samples agreed upon (by
EPA and the applicant) prior to analysis will be analyzed in a single laboratory, and will be used to assess
whether there is a statistically significant difference between the results produced by the ATP and the
results produced by the approved corresponding reference method.
The design of the side-by-side comparison is left up to the ATP applicant. However, a detailed validation
study plan must be prepared by the applicant and submitted to EPA for review and comment, and the
plan must be agreed upon by all parties prior to conducting the comparison study. Although EPA may be
consulted for additional guidance during the development of the study plan, it is the applicant's
responsibility to write the study plan and submit it to EPA for review. The minimum requirements
regarding the design of the Phase I study are provided below and summarized in Section 1.2.3.3, Table H-
2 of this appendix.
• Number/Types of Real-World Sample Types: A minimum of nine real-world sample types must be
collected from a variety of sources and analyzed by each method. To better identify any sample-
specific differences between the ATP and the approved reference method, analyses should be
performed across a wide range of sample types (a list of industrial categories with existing effluent
guidelines can be found at: http://water.epa.gov/scitech/wastetech/guide/industry.cfm). If preparation
of multiple spike levels is feasible for the method-defined parameter, then use of multiple spike levels
is recommended, but a minimum of nine sample types per spike level are required. If spiking is not
feasible, a range of sample types should be targeted that would be expected to yield background
concentrations that vary by at least one order of magnitude.
• Laboratories: The Phase I Comparison Study should be performed in a single laboratory to minimize
the sources of variability. This laboratory should have familiarity with both the approved method and
the ATP to ensure that any differences in performance are not the result of inexperience with one or
both methods. However, it is important that the validation study accurately reflect the ruggedness of
the ATP and any limitations regarding clarity of the ATP procedures. Therefore, the laboratory
should not be affiliated with the ATP applicant.
Replication: The recommended number of replicates to be analyzed per method and sample within
the side-by-side study is three.
To ensure the laboratory can perform both methods acceptably, the laboratory must meet all QC analysis
criteria specified in the approved method using both, the approved method and ATP, prior to the
statistical comparisons of the method data. Moreover, the specific statistical tests that will be used to
compare the results of the ATP with those from the reference method must be described in the study
plan. See Section 1.3 of this appendix for a discussion of the relevant statistical considerations.
If a statistical assessment indicates that Phase I study results produced by the ATP are comparable to
those produced by the designated reference method based on the statistical test described in the validation
study plan, then the ATP will be deemed to be sufficiently comparable to proceed to Phase II.
1.2.3.2 Phase II: Interlaboratory Study
In Phase II of the comparison study, results of the analyses of synthetic and real-world samples in nine
laboratories will be used to demonstrate method equivalence and establish method performance and inter-
laboratory QC acceptance criteria for the alternate test procedure. The study design and specific QC tests
for the Phase II study will generally follow the guidelines presented for Tier 3 validation as described in
Section 4.3 of this document, and acceptance criteria will be developed as described in Appendix G of the
H-5 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
"Protocol for Validation and Review of New Methods for Regulated Organic and Inorganic Analytes in
Wastewater". However, not all QC tests will be applicable to all method-defined parameters. For
example, matrix spike samples are not applicable to methods that measure method-defined analytes such
as pH or temperature.
As with Phase I, the specific statistical tests that will be used to compare the results of the ATP with those
from the reference method must be described in the study plan. See Section 1.3 of this appendix for a
discussion of the relevant statistical considerations.
Despite careful planning, situations may arise in which the results from one of the nine laboratories in the
study may not represent the performance of the ATP or the other laboratories. Applicants may wish to
plan for such a contingency in the Phase II study plan by utilizing more than nine laboratories, or by
documenting relevant corrective action procedures that all laboratories in the study will use prior to
repeating study analyses.
Outlier testing is not recommended for either the single-lab or multi-laboratory phases of the study.
However, if the applicant has reason to believe that some of the results from the validation study truly do
not represent the performance of the method, then they should contact EPA to discuss whether and how
an outlier test could be applied.
It is important that Phase 2 accurately reflect the ruggedness of the ATP and any limitations regarding
clarity of the ATP procedures. Therefore, it is not permissible for a vendor or other applicant to directly
assist laboratories participating in Phase 2 of the study with implementation of the ATP methodology or
equipment during the course of the study (e.g., the vendor or applicant may provide training and advice to
participant laboratories regarding the equipment or methodology prior to the start of the study, but the
study samples are to be analyzed by the study participants under "routine" conditions).
1.2.3.3 Analyses Required for Both Phases of a Tier 3 Validation Study of an A TP for a
MDP
The following table summarizes the recommended minimum numbers of analyses for both phases of the
validation study for an ATP involving an MDP.
Table H-2 Summary of Validation Recommendations for Tier 3 MDP ATPs1
Study
Phase
Phase 1
Phase II
Procedure
ATP
Reference Method
ATP
Reference Method
Number of
Labs
1
1
9
9
Matrix
types
9
9
9
9
Number of Analyses Required
Replicates
per Matrix
Type2
3
3
3
3
IPRin
Reagent
Water3
4
4
36
36
PT
Sample
1
1
9
9
MS/MSD4
18
18
18
18
MDL'51
7
7
63
63
Total
57
57
153
153
Numbers of analyses in this table do not include additional QC tests such as calibration, blanks, etc. Nine is the
maximum number of matrix types that is should be used to validate a modified wastewater method at Tier 1 or
TierS.
In Phase I the laboratory analyzes each of the nine matrix types in triplicate by each method. In Phase II each
laboratory is assigned one of the nine matrix types and analyzes it in triplicate by each method.
The IPR analyses only apply to MDPs where the reference method also includes the IPR test.
In Phase I, the laboratory should analyze one MS/MSD pair for each of the nine matrix types by each method. In
Phase II, each laboratory should analyze one MS/MSD pair for the assigned matrix type by each method.
The current MDL procedure requires 7 samples, validation studies will comply with most updated MDL study
requirements published in Appendix B of 40 C.F.R. Part 136.
H-6
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
1.3 Statistical Considerations in Evaluating for MDPs
Demonstrating comparability of the results for an ATP for a MDP presents a number of challenges for
both the applicant and EPA. By their very nature, the results for method-defined parameters are a direct
function of the sum of all of the steps in the method used to generate them. Thus, an ATP that achieves
"better" results for an MDP is not an appropriate goal, and common statistical tests such as the Student's
t-test of mean results, the F-test of variances, or an analysis of variance (ANOVA) are not useful for
MDPs.
For the purposes of evaluating ATPs for MDPs, EPA employs the Root Mean Square Deviation (RMSD).
The RMSD measures variations in the ATP results both above and below the results from the reference
method. For example, the average results for the ATP across all samples may be close to those obtained
with the reference method, yet the variability of the ATP data may be quite high (results are accurate on
average but are imprecise), or the differences between the methods vary widely from sample to sample.
The RMSD computes the squared deviation of the results from the ATP from the results of the reference
method on the same sample, and sums those squared deviations across all the samples in the validation
study to provide an overall measure of agreement between the two sets of results (ATP and reference
method). A generalized formula for the RMSD applicable to an ATP evaluation is shown below:
RMSD =
n
where: XRMj = The "jth" sample mean from the reference method
XATPJ = The "jth" sample mean from the ATP, and
n = The total number of replicates per sample and method
The calculated RMSD is then compared to the upper limit RMSDmax, determined using the formula
below:
F**
2MSE.
'"(0.95;J,2*J*(n-l))
where: J= the total number of samples, and
n = the total number of replicates per sample and method, and
MSE = the mean-squared error, as calculated below:
1 J 2
MSE = —
O * T Z_
Z J i=
where: Sjk = the standard deviation of the replicates for sample j and method k (i.e., where the
approved method is method 1 and the ATP is method 2), and
J = the total number of samples
Due to the natural variation in the MDP across samples, it is recommended that all results from both
methods be log-transformed prior to calculating the RMSD and RMSDmax
H-7 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Using the RMSD, the goal is to demonstrate whether or not there is a statistically significant difference
between the performance characteristics of the ATP and the reference method2. By its derivation, the
RMSD sums the deviations in both directions (i.e., ATP results above the reference method results and
those below), rather than looking at the simple "inequality" of the two sets of results. If no statistically
significant difference is observed with the RMSD, then the results for the ATP may be judged acceptable.
Another advantage of the RMSD relative to other common statistical tests is that using the other tests will
generate a large number of statistical outcomes that would not produce a clear picture of the overall
performance of an ATP relative to the reference method. For example, for the Phase I study of an ATP
application for nationwide use, 27 analyses are required (e.g., 9 separate sample matrices, analyzed in
triplicate, in a single laboratory). Using t-tests and F-tests to compare the results across even nine
samples could well result in a mix of outcomes across all the samples (i.e., 5 samples with statistically
significant differences and 4 without such differences). Such a mix of outcomes for the ATP would be
difficult, if not impossible, to interpret in the context of comparability with the reference method.
1.4 Other Recommendations for ATPs for MDPs
Despite the more rigorous side-by-side testing warranted for ATPs for MDPs, all other aspects of the ATP
development and approval process still apply. For example:
• The applicant should comply with the application in Section 3.2 of this document, use of the
application form in Appendix A of this document, and inclusion of the Data Collection Certification
form in Appendix B of this document with their validation study report
• The applicant should follow the procedures for proprietary information in Section 3.3 of this
document.
• Requirements in Section 4.3 for documenting the ATP in EPA format, providing MDL data and the
routine QC operation data described in Sections 4.3.3 to 4.3.10 of this document continue to apply.
EPA review, approval, and rulemaking framework described in Section 5 of this document continue
to apply
Note: As noted in Section 1.3 of this appendix, demonstrating comparability of the results for an ATP
for a MDP presents a number of challenges. However, even if the use of the RMSD demonstrates
there are not any statistically significant differences between the performance characteristics of
the ATP and the reference method, EPA may choose not to consider ATPs for MDPs that alter
the fundamental chemistry of the overall analytical process, including the determinative technique
used for measurement of the MDP.
Given the nature of MDPs, all ATP applications for MDPs must be submitted to the National ATP
Coordinator at EPA Headquarters. EPA Regional and State authorities may not approve ATPs for MDPs.
Note: As for all other ATP applications, the applicant is responsible for the technical and statistical
evaluation of the validation study results and preparation of the study report.
2 The significance test used in the RMSD is equivalent to an F-test of significant difference that tests the compound null
hypothesis that the mean log concentration is equal between the two methods, for each sample in the study.
H-8 February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
APPENDIX I - Checklist for Methods To Be Considered by EPA for Use in Compliance Monitoring
Programs under the Clean Water Act
EPA uses the following checklist to evaluate requests for consideration of Alternate Test Procedures (ATPs), new methods, or modified1 methods
for use in Clean Water Act (CWA) compliance monitoring programs. The checklist addresses minimum submission requirements (documented in
40 C.F.R. § § 136.6 and 136.7 and in the ATP Protocols), as well as other laws, regulations, and policies that EPA staff must consider when
evaluating method submissions. Although the checklist is for internal use by EPA, applicants are encouraged to review the checklist to better
understand the Agency's process for reviewing and considering applicant submissions. Two attachments are provided at the end of the checklist to
assist EPA users and applicants in understanding the requirements. In addition, the checklist contains references to applicable laws, regulations,
and policies that are not explicitly covered at 40 C.F.R. § § 136.6 and 136.7 and in the ATP Protocols.
Reviewer #1, Name and Organization:
Reviewer #2, Name and Organization:
Reviewer # 1 Initials and Date:
Reviewer # 2 Initials and Date:
Use the check boxes to identify if following items were submitted or indicated. The Not Applicable (N/A) box may not be used to answer a question if it is
blacked out.
YES NO N/A
D D •
D D •
ITEM OR QUESTION TO BE ADDRESSED
la. Is this a completed ATP or a new method that has been reviewed by EPA? If yes, indicate type
below.
ATP New Method
Ib. If no, is this a request for EPA to consider approval of a method from a Voluntary Consensus
Standards Body (VCSB) or other Government Agency (or their designated representative)?
VCSB Method Other Government Agency Method
COMMENTS/NOTES
1 For the purposes of this checklist, the terms modified method, revised method, and updated method are synonymous and are intended to mean any method
changes, updates, or revisions that are being submitted to EPA for review and approval at 40 C.F.R. Part 136 to support CWA programs. Due to the increased
flexibility allowed for method modifications under 40 C.F.R. 136.6, most method changes, updates, or revisions submitted to EPA will consist of ATP
applications for procedures involving method-defined analytes, procedures that involve changes to the chemistry of the method, determinative techniques, or
applications for consideration of updated versions of previously approved methods.
1-1
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Use the check boxes to identify if following items were submitted or indicated. The Not Applicable (N/A) box may not be used to answer a question if it is
blacked out.
YES NO N/A
D D •
D D •
D D •
D D •
D D •
D D D
D D D
D D D
ITEM OR QUESTION TO BE ADDRESSED
2. Is a justification provided for consideration of the ATP, new method, or VCSB or other Government
Agency method for use in CWA compliance monitoring programs?
This may include advantages over approved method(s) or may state that the method is a revised or updated
version of an already approved method.
3. Is a copy of the method written in standard EPA format included? (See the Guidelines and Format
document at http://epa. sov/waterscience/methods/suide/40cfr. html.) Alternatively, method(s) may be
written in another organization's format but must address and reference the topics specified below
in Attachment A and Attachment B.
EPA Format Other Format that Addresses topics below and Attachments A and B
4. Does the method include all appropriate quality control (QC) elements or are they included as part
of a compendium and referenced in the method? (see 40 C.F.R. § 136. 7, reprinted as Attachment B to
this checklist, for a list of required QC elements)
Included in method Included in compendium and referenced in method
5. Does the method specify acceptance criteria for required QC tests equal to or better than the
method currently approved at 40 C.F.R Part 136?
6a. Does the method include a unique method number and date/revision date?
6b. For methods submitted by a VCSB or another Government Agency, does the method contain a
revision date or date of approval?
Enter N/A if the application is not for a VCSB or other Government Agency method.
1. Is a copy of the approved reference method (with red-line strikeouts and additions) enclosed if the
application is for a modified method or a revised version of an approved method?
This applies to method modifications/revisions. Enter N/A if the submission is for a new method
application.
8. Would utilization of the method be practical and comply with existing law and be compatible with
agency and departmental missions, authorities, priorities and budget resources? [stipulated by the
National Technology Transfer Advancement Act, 15 U.S.C. §3701 et seq. (1996)]
This applies to VCSB applications. Enter N/A if the application is not for a VCSB method.
COMMENTS/NOTES
1-2
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Use the check boxes to identify if following items were submitted or indicated. The Not Applicable (N/A) box may not be used to answer a question if it is
blacked out.
YES NO N/A
ITEM OR QUESTION TO BE ADDRESSED
COMMENTS/NOTES
9. If the method is from a VCSB or other Government Agency is the method in its final form and has it
been approved/published by that VCSB or Government Agency?
This applies to VCSB and other Government Agency methods only. Enter N/A for all other types of
applications.
Questions lOa through lOc address method validation study plans. These requirements only apply to new
method applications, applications for methods involving method-defined parameters, and other ATP
applications that go beyond the modifications explicitly allowed at 40 C.F.R. § 136.6.
Enter N/A to questions 1 Oa through 10 c if the application is for an update to a previously approved method
and the revisions do not affect the chemistry of the method, determinative technique or QC acceptance criteria.
lOa. Was EPA consulted or did EPA participate in the development of the original study plan for
validation of the method?
lOb. If EPA was consulted or participated in the development of the original study plan for validation of
the method, does the application include written documentation of EPA's participation (e.g., copies
of correspondence and records of any verbal communications with EPA staff by phone or in meetings)?
lOc. If EPA was consulted or participated in the development of the original study plan for validation of
the method, were all EPA recommendations incorporated into the study plan?
If yes, this must be documented in writing. If no, the submission should include a written explanation
regarding EPA recommendations that were not adopted.
11. Is a copy of the validation study plan with validation study report and reference to the
organization's study data requirements provided?
Enter N/A if the application is for an update to a previously approved method and the revisions do not
affect the chemistry of the method, determinative technique or QC acceptance criteria (including the
Method Detection Limit).
A "yes" answer is required for consideration of new method applications, applications for methods
involving method-defined parameters, and other ATP applications that go beyond the modifications
explicitly allowed at 40 C.F.R. § 136.6.
1-3
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Use the check boxes to identify if following items were submitted or indicated. The Not Applicable (N/A) box may not be used to answer a question if it is
blacked out.
YES NO N/A
ITEM OR QUESTION TO BE ADDRESSED
COMMENTS/NOTES
Questions 12a through 12j address method validation study reports and supporting documentation. These
requirements only apply to new method applications, applications for methods involving method-defined
parameters, and other ATP applications that go beyond the modifications explicitly allowed at 40 C.F.R.
§ 136.6. A yes answer is required for such applications.
Enter N/A to questions 12a through 12j if the application is for an update to a previously approved method and
the revisions do not affect the chemistry of the method, determinative technique or QC acceptance criteria.
12a. Are supporting data documenting the Method Detection Limit (MDL) was determined as a part of
the method validation study provided?
Note: EPA requires that all methods approved at 40 C.F.R. 136, including ATPs, be supported by an
MDL determined as specified at 40 C.F.R. 136, Appendix B. This includes VCSB and other Government
Agency methods, even if those organizations normally use other approaches for defining and determining
detection limits.
12b. Does the method validation include real world samples? (see list of effluent guidelines promulgated by
EPA, sorted by industry category, http://water.epa.gov/scitech/wastetech/guide/industry.cfm)
12c. Was the method validated to demonstrate compliance with existing analyte concentration ranges,
sample collection, preservation, preparation and holding time requirements of the approved
method?
(Data demonstrating compliance should be included in the submission)
12d. Are quantitation range and limits supporting data provided?
A quantitation range corresponds to the range of analyte concentration (or other quantity) characterized
for measurement accuracy (trueness and precision) during method validation, (see Validation and Peer
Review of U.S. Environmental Protection Agency Chemical Methods of
Analysis, http://www.epa.gov/fem/pdfs/chemmethod_validity_guide.pdf)
12e. Are supporting data that address instrument calibration provided?
The performance characteristic is sometimes referred to as "instrument linearity." (see Validation and
Peer Review of U.S. Environmental Protection Agency Chemical Methods of
Analysis, http://www.epa.sov/fem/pdfs/chemmethod validity suide.pdf
1-4
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
12f. Are supporting data that address bias/trueness provided?
Trueness is a performance characteristic that addresses sources of known systematic error and bias is a
measure of trueness. (see Validation and Peer Review of U.S. Environmental Protection Agency
Chemical Methods of Analysis, http://www.epa.gov/fem/pdfs/chemmethod_validity_guide.pdf
12g. Are supporting data that address precision (repeatability and reproducibility) provided?
Precision is a performance characteristic that reflects sources of random error in a measurement process.
Methods designed for demonstrating compliance with regulatory requirements should be evaluated for
both repeatability (within lab) and reproducibility (among labs), (see Validation and Peer Review of
U.S. Environmental Protection Agency Chemical Methods of
Analysis, http://www. epa. gov/fem/pdfs/chemmethod_validity_guide.pdf)
12h. Are data demonstrating method selectivity provided?
Selectivity is a performance characteristic that demonstrates the ability of the method to yield useful data
for the analytes, analytes levels and matrices defined within the scope of the method. Selectivity is
demonstrated by providing information that substantiates the identity of the analyte in presence of
expected matrix constituents, (see Validation and Peer Review of U.S. Environmental Protection Agency
Chemical Methods of Analysis, http://www. epa. gov/fem/pdfs/chemmethod_validity_guide.pdf)
12i. Are data demonstrating method ruggedness provided?
Ruggedness refers to the capacity of analytical method to remain unaffected by small variations in
operating conditions or environmental conditions. The changes should reflect expected, reasonable
variations that are likely to be encountered in different labs, (see Validation and Peer Review of U.S.
Environmental Protection Agency Chemical Methods of
Analysis, http://www. epa. gov/fem/pdfs/chemmethod_validity_guide.pdf)
12\. Are interlaboratory study/studies as defined in the ATP and New Method Protocols documents
provided?
Interlaboratory studies determine whether an analytical method can be transferred for use in other
laboratories and used for regulatory testing. Data from the interlaboratory study should be reported in
tabular form and the raw data should be maintained and available for review. If appropriate, there should
be a discussion describing the details of, and rationale for, any changes made to the method resulting
from the interlaboratory study, (see Validation and Peer Review of U.S. Environmental Protection
Agency Chemical Methods of Analysis, http://www.epa.gov/fem/pdfs/chemmethod^validity_guide.pdf)
1-5
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
ODD
D D D
D D D
D D D
D D D
D D D
Questions 13a through 13g address applications for methods involving method-defined parameters. A yes
answer is required for such applications.
Enter N/A to questions 13a through 13g if the application does not involve method-defined parameters.
13a. Is the request for one or more well-defined analytes that are NOT a 40 C.F.R § 136.6 Method-
Defined Parameter. The following is a list of some Method-Defined Parameters - Acidity,
Alkalinity, BODS, COD, Color, Oil & Grease, Total Solids, Total Dissolved Solids, Total Organic
Carbon, Total Suspended Solids, Total Phenols, Temperature, or pH? Other parameters may be
added at EPA's discretion.
13b. If the request is for a 40 C.F.R § 136.6 Method-Defined Parameter, does the application include 1)
comparative raw data resulting from side-by-side split sample or grab sample analyses performed
in triplicate using both the new method and the approved method in a minimum of 9 distinct real
world matrix types and 2) data from all required QC analyses performed using each method?
13c. If the request is for a 40 C.F.R § 136.6 Method-Defined Parameter, is the chemistry or
determinative step the same as the approved method?
13d. If the request is for a 40 C.F.R § 136.6 Method-Defined Parameter (MDP) AND the chemistry or
determinative step is different than the approved method, are the chemistry and determinative step
used to identify and measure the MDP well explained and clearly defined as well as any potential
interferences or difficulties with the method?
13e. Are data provided from a routinely run, freshly prepared method calibration curve that was used
to quantify the analyte(s) in the samples analyzed as part of the validation study, including
verification of the calibration curve using independent second source, quality certified, traceable
standards?
13f. If the request is for a 40 C.F.R § 136.6 Method-Defined Parameter, do the data submitted
demonstrate comparable performance of the new method to the approved method?
Note: Comparable performance is determined by comparing the achievement of statistical RMSD
comparability between the new method and the approved method from analyses of samples from a
minimum of 9 distinct real world matrix types (split or grab - collected and analyzed at the same time),
performed in triplicate AND by comparison of the QC acceptance criteria of the two methods.
13g. Will the new method and the approved method measure the same forms and species of analyte?
1-6
February 2016
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Attachment A
Topics to be Covered in Written Method Submission
Scope and Application - This section of the method should clearly state the analyte(s) determined
and the types of matrices to which the method is applicable. This section also may list the
detection limit of the method and the range of concentrations over which the method is
applicable.
Summary - This section briefly states the sample preparation (if any) and the underlying
chemistry and determinative technique used in measurement of the target analyte(s). It also may
list the method detection limit and the range of concentrations over which the method is
applicable.
Definitions - This section should define the terms and abbreviations that are used in the method.
The section should include definitions for abbreviations, especially those that relate to quality
control, for example: LRB - Laboratory Reagent Blank, LFB - Laboratory Fortified Blank,
LFM - Laboratory Fortified Matrix, MS and MSD - Matrix Spike and Matrix Spike Duplicate,
MDL - Method Detection Limit, and QCS - Quality Control Sample.
Interferences - This section should identify common interferences, and where applicable, list
ways to eliminate, reduce, or overcome them. Of particular note are interferences that may lead
to loss or under reporting of target analyte(s).
Safety - This section should adequately address any safety concerns associated with the
performance of the method (e.g., toxicity, carcinogenic reagents, or explosion risks).
Equipment and Supplies - This section should list all equipment (apparatus) and supplies to
perform the procedures of the method.
Reagents and Standards - This section of the method should clearly list all reagents and standards
needed to perform the analysis. It also may detail both preparation and storage of stock standard
solutions from neat materials and preparation and storage of working standard solutions.
Sample Collection, Preservation and Storage - This section should list the proper types of sample
containers, preservation techniques and holding times per the requirements of 40 C.F.R. § 136.3,
Table II.
Quality Control - This section should list the minimum QC requirements and acceptance criteria
for each of the QC tests applicable to the method (see 40 C.F.R. § 136.7 for a listing of QC
elements that are required where applicable).
Calibration and Standardization - This section of the method should list the procedures for
calibration of the instrument and the type of calibration used (i.e., linear, 2n order). It should
specify a sufficient number of standards used to establish linearity or to clearly define any non-
linear portion of the curve. This section also may specify procedures for periodic verification of
calibration standards and specify acceptance criteria listed for calibration verification.
Checklist for Methods to be Considered by EPA for Use in CWA Compliance Monitoring Programs A-l
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Procedure - This section should contain all of the critical steps required to perform the analysis
of samples. If sample preparation steps such as distillation, digestion, or pH adjustment are
required prior to analysis these steps should also be specified or referenced.
Data Analysis and Reporting - This section should explain how to calculate and report sample
results. A statement indicating that only results that fall between the lowest and highest
calibration standards should be reported unless the result is flagged as an estimated value. In
addition, a statement should be included that samples with results exceeding the highest
calibration standard should be diluted and re-analyzed.
Method Performance - This section should present any data or other information that
demonstrate or indicate the expected performance characteristics of the method.
Pollution Prevention - This section should contain information on minimizing or preventing
pollution known to be potentially attributable to use of the method.
Waste Management - This section should contain information on the minimization and proper
disposal of any hazardous wastes known to be generated by use of the method?
References - This section should cite proper references and sources used in the development of
the method. References should be restricted to associated or source material.
Tables, Diagrams, and Validation Data - This section of the method should contain all method
tables and figures (diagrams and flowcharts). If performance data are included here, they should
support the MDL, method range and QC acceptance criteria listed in the method.
Checklist for Methods to be Considered by EPA for Use in CWA Compliance Monitoring Programs A-2
-------
ATP Protocol for Regulated Organic and Inorganic Analytes in Waste-water
Attachment B
40 C.F.R. § 136.7 Quality Assurance and Quality Control
The permittee/laboratory shall use suitable QA/QC procedures when conducting compliance
analyses with any Part 136 chemical method or an alternative method specified by the permitting
authority. These QA/QC procedures are generally included in the analytical method or may be
part of the methods compendium for approved Part 136 methods from a consensus organization.
For example, Standard Methods contain QA/QC procedures in the Part 1000 section of the
Standard Methods Compendium. The permittee/laboratory shall follow these QA/QC
procedures, as described in the method or methods compendium. If the method lacks QA/QC
procedures, the permittee/laboratory has the following options to comply with the QA/QC
requirements:
(a) Refer to and follow the QA/QC published in the "comparable" EPA method for that
parameter that has such QA/QC procedures;
(b) Refer to the appropriate QA/QC section(s) of an approved Part 136 method from a consensus
organization compendium;
(c)(l) Incorporate the following twelve quality control elements, where applicable, into the
laboratory's documented standard operating procedure (SOP) for performing compliance
analyses when using an approved Part 136 method when the method lacks such QA/QC
procedures. One or more of the twelve QC elements may not apply to a given method and may
be omitted if a written rationale is provided indicating why the element(s) is/are inappropriate for
a specific method.
(i) Demonstration of Capability (DOC);
(ii) Method Detection Limit (MDL);
(iii) Laboratory reagent blank (LRB), also referred to as method blank (MB);
(iv) Laboratory fortified blank (LFB), also referred to as a spiked blank, or laboratory control
sample (LCS);
(v) Matrix spike (MS) and matrix spike duplicate (MSD), or laboratory fortified matrix (LFM)
and LFM duplicate, may be used for suspected matrix interference problems to assess
precision;
(vi) Internal standards (for GC/MS analyses), surrogate standards (for organic analysis) or
tracers (for radiochemistry);
(vii) Calibration (initial and continuing), also referred to as initial calibration verification (ICV)
and continuing calibration verification (CCV);
(viii) Control charts (or other trend analyses of quality control results);
(ix) Corrective action (root cause analysis);
(x) QC acceptance criteria;
(xi) Definitions of preparation and analytical batches that may drive QC frequencies; and
(xii) Minimum frequency for conducting all QC elements.
(2) These twelve quality control elements must be clearly documented in the written standard
operating procedures (SOP) for each analytical method not containing QA
Checklist for Methods to be Considered by EPA for Use in CWA Compliance Monitoring Programs B-l
------- |